RU2795769C1 - Antipneumococcal hyperimmune globulin for the treatment and prevention of pneumococcal infection - Google Patents

Abstract

FIELD: pharmaceutical industry.

SUBSTANCE: group of inventions relates to immunoglobulin, a method for its production and methods for treating an infection caused by Streptococcus pneumonia. A method of producing an immunoglobulin containing an opsonophagocytic antibody titer of at least 1:256 for 50% or more of Streptococcus pneumonia serotypes selected from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F, the said method comprising the following steps: immunizing healthy adult human plasma donors aged 18 to 60 years with a primary immunization with a polyvalent S. pneumoniae conjugate vaccine followed by a booster immunization with a polyvalent polysaccharide S. pneumoniae vaccine; collecting plasma from the said plasma donors after the said booster immunization; pooling of plasma from vaccinated donors to obtain pooled plasma containing an opsonophagocytic antibody titer of at least 1:256 for 50% or more of Streptococcus pneumonia serotypes selected from serotypes 1, 3, 4, 5, 6B, 7F, 9V , 14, 18C, 19A, 19F and 23F; obtaining immunoglobulin from plasma pooled in accordance with the previous stage. Immunoglobulin obtained by the method described above. A method for treating S. pneumonia infection in a subject, comprising administering a therapeutically effective amount of an immunoglobulin to the said subject. A method of performing immunotherapy for a subject, comprising administering a therapeutically effective amount of an immunoglobulin to the said subject.

EFFECT: above method allows to create a new composition of immunoglobulin obtained from the plasma of donors vaccinated with one or more pneumococcal vaccines with an increased titer of specific anti-pneumococcal antibodies, and the resulting composition has an increased opsonic activity compared to the control sample.

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Description

FIELD OF TECHNOLOGY

This invention relates to compositions and methods for the treatment of infection caused by or associated with Streptococcus pneumonia. In particular, the invention provides human hyperimmune globulin and compositions thereof for the prevention or treatment of pneumococcal infection. The invention relates to methods for producing immune globulins containing high titers of opsonic anti-pneumococcal antibodies, compositions containing the same, and methods for using these compositions in the prevention and treatment of pneumococcal infections (for example, infections of the upper respiratory tract (for example, sinusitis, otitis media, pharyngitis and etc. (for example, in an immunologically "compromised" subject))).

BACKGROUND OF THE INVENTION

There are many components of the immune system, all of which work together to fight off foreign invasive pathogens. While most people have an intact immune system that serves the purpose of protecting them from the wide range of infectious organisms that commonly infect humans, including viruses, bacteria, and fungi, many people's immune systems are compromised or compromised.

Primary immunodeficiencies (PID) are a group of more than 200 genetically inherited disorders characterized by deficiency of certain components of the innate or adaptive immune system, with the clinical result of increased susceptibility to infection. For example, a defect in the humoral immune system that impairs the body's ability to produce antibodies makes a person susceptible to many infections. To be considered a primary immunodeficiency, the cause of the immune deficiency must not be secondary in nature (eg, caused by another disease, drug, or exposure to environmental toxins). Most primary immunodeficiencies are genetic disorders and are diagnosed in children, although less severe forms may not be recognized until adulthood. About 1 in 500 people are born with a primary immunodeficiency.

Intravenous administration of immune globulin has been shown to restore the ability of immunocompromised people to defend themselves against infection. Because immune globulin is pooled from many donors, antibody titers for the many infectious organisms against which protection must be provided vary widely and may or may not be sufficient to meet immune requirements in the event of infection in immunocompromised individuals.

Most commercially available immunoglobulins are derived from human plasma collected, processed and distributed for sale by the blood and plasma product industry. The first purified human immunoglobulin G (IgG) preparation used clinically was serum immune globulin, developed in the 1940s (Cohn, E.J., et al., J. Am Chem. Soc, 68:459-475 (1946)) and Oncely, J.L. et al., J. Am Chem Soc. 71:541-550 (1949). Gammaglobulin obtained in this manner shows a high molecular weight distribution as analyzed by high resolution size exclusion chromatography.

Standard immune globulin (IVIG) has been shown to have many variations in opsonic activity against a widely distributed commensal organism ubiquitous on human skin, S. epidermidis (L. A. Clark and C. S. F. Easmon, J. Clin. Pathol 39:856 (1986)). For example, in the Clark and Easmon study, one-third of IVIG batches tested had weak opsonic activity with complement, and only two out of fourteen were opsonic without complement. Thus, although batches of IVIG were made from large pools of donor plasma, the proper opsonic S. epidermidis antibody was not uniformly present.

IVIG is generally successful in preventing severe lower respiratory tract infections in immunocompromised patients. However, although immunocompromised patients receiving IVIG appear to have acceptable levels of total immunoglobulin, as well as sufficient levels of anti-S. pneumoniae antibodies to prevent serious S. pneumoniae bacterial infections, there is a significant percentage patients who develop disabling upper respiratory tract infections and non-inflammatory infections that reduce quality of life, increase the use of antibiotics that are ineffective, and increase medical costs (see, for example, Favre et al., Allergy 2005 60 :385-390).

SUMMARY OF THE INVENTION

This invention relates to compositions and methods for the treatment of infection caused by Streptococcus pneumonia. In particular, the invention provides human immune globulin and compositions thereof for the prevention or treatment of pneumococcal infection. The invention provides methods for producing immune globulins containing high titers of anti-pneumococcal antibodies with opsonic activity (for example, to a variety of serotypes of S. pneumonia), compositions based on them, and methods for using the compositions for the prevention and treatment of pneumococcal infection. Additionally, the invention provides methods for preventing or treating pneumococcal infections (e.g., upper respiratory tract infections (e.g., pharyngitis, otitis media, sinusitis, etc.)) in immunocompromised subjects (e.g., by administering immune globulin compositions of the invention (e.g., containing a high titer of opsonic anti-pneumococcal antibodies) to immunocompromised subjects who were not adequately protected by regular infusions of standard IVIG (IVIG)).

In one embodiment of the invention, the object of the invention is to provide a novel immune globulin composition obtained from plasma donors according to the methods of the invention (e.g., human plasma donors (e.g., healthy human plasma donors) who have been vaccinated with one or more pneumococcal vaccines in accordance with methods described here) that contains an increased titer of specific anti-pneumococcal antibodies, the administration of the antibodies has opsonic activity compared to the control sample.

The invention is not limited to the kind of control sample used. For example, in one embodiment of the invention, the control sample is an immune globulin obtained from the plasma of healthy human plasma donor(s) prior to vaccination (e.g., such that titer levels of specific opsonic anti-pneumococcal antibodies in human donor plasma prior to immunization are used in as a baseline for measuring the increase in specific opsonic anti-pneumococcal antibody titers after immunization). In another embodiment, the control sample is an immune globulin obtained from a mixture of plasma samples obtained from random human plasma donors (e.g., 100, 300, 500, 1000 or more random human plasma donors) who have not been vaccinated with pneumococcal vaccine ( for example, such that titer levels of specific opsonic anti-pneumococcal antibodies in unvaccinated human plasma donors are used as the basis for measuring the increase in specific opsonic anti-pneumococcal antibody titers present in human plasma donors vaccinated according to the methods of the invention). Those skilled in the art will appreciate that other controls may be used. The invention is not limited to a method or assay used to measure the opsonic (eg, opsonophagocytic) titer of an antibody present. In fact, various assays can be used, including but not limited to those described herein (eg, in Example 1).

In one embodiment, the object of the invention is to provide a novel immune globulin composition containing a high titer of opsonic anti-pneumococcal antibodies. Another object of the invention is to provide a new composition of immune globulin for intravenous administration containing a high titer of opsonic anti-pneumococcal antibodies (for example, "anti-pneumococcal hyperimmune globulin") and methods for its preparation and use (for example, for the prevention and/or treatment of pneumococcal infection (for example, in immunocompromised patients). The term "high titer" in this context means the presence of opsonic pneumococcal antibody in an amount that is 2 or more times, for example, 3, 5, 7, 10, 15, 20 or more times higher than in the control sample.

Accordingly, a novel immune globulin formulation (e.g., anti-pneumococcal hyperimmune globulin) containing a high titer of opsonic anti-pneumococcal antibodies of the invention differs from standard/conventional immune globulin preparations (e.g., standard, conventional IVIG) and differs from other IVIG preparations (IVIG ) (e.g., other IVIG hyperimmune drugs) in that it contains a high titer of human opsonic anti-pneumococcal antibodies that are functional and react with a wide range of S. pneumoniae serotypes, enhance phagocytosis, and cause elimination of S. pneumoniae (e.g., in vitro and/or in vivo (eg, the antibodies are opsonophagocytic)), regardless of the total amount of binding anti-pneumococcal antibodies (eg, as measured by enzyme immunoassay, ELISA) that are present in the composition.

This is unexpected as the data obtained and disclosed herein indicate that there was no systematic or predictable correlation between total anti-pneumococcal IgG antibody titer and opsonic anti-pneumococcal antibody titer present in plasma or immune globulin obtained from vaccinated donors. Due to the unexpected finding of a large discrepancy between the overall titer level of binding anti-pneumococcal IgG antibodies and the titer of opsonic anti-pneumococcal antibodies present in plasma or plasma-derived immune globulin from the plasma of vaccinated donors described in this description, it is another object of the invention to provide a composition of plasma and/ or an immune globulin (eg, anti-pneumococcal hyperimmune globulin) containing significantly increased titers of functional opsonic anti-pneumococcal antibodies (eg, regardless of the total titer of binding anti-pneumococcal antibodies (eg, by pooling plasma and/or immune globulin derived from the plasma of vaccinated donors, or by combining the plasma of vaccinated donors with immune globulin derived from unvaccinated donors who may not have a high titer of opsonic anti-pneumococcal antibodies, but when combined with donor plasma and/or immune globulin with a significantly elevated titer of opsonic anti-pneumococcal antibodies, it does not dilute the total opsonic titer to levels that do not provide protection)) compared with the control sample. The invention includes serum/plasma as well as immune globulin (eg, hyperimmune globulin) derived from substances containing significantly elevated titers of broad-spectrum functional and opsonic anti-pneumococcal antibodies. In one embodiment, the invention provides plasma/serum and/or immune globulin obtained from substances containing an increased titer of specific opsonic anti-pneumococcal antibodies, which is at least 2 times, 3 times, 4 times, 5 times, 7 times, 10 or more times higher for at least about 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more) S. pneumonia 1 serotypes, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F) compared with titers of specific opsonic anti-pneumococcal antibodies against the same serotypes present in a control sample (immune globulin obtained from a mixture of plasma samples obtained from random human plasma donors (eg, 100, 300, 500, 1000 or more random human plasma donors who have not been vaccinated with pneumococcal vaccine).

In another embodiment, the invention provides a composition of hyperimmune globulin (for example, IVIG) containing a titer of broad-spectrum opsonic antibodies specific for 70% or more of S. pneumonia serotypes selected from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23 that is 2 or more times (e.g., 3-25 or more times) higher than the titer of opsonic antibodies specific for the same S. pneumoniae serotypes present in a control (e.g., immune globulin obtained from a mixture of plasma samples obtained from random human plasma donors (e.g., 100, 300, 500, 1000 or more random human plasma donors) that have not been vaccinated with pneumococcal vaccine In another embodiment, plasma/serum and /or an immune globulin derived from it containing a titer of specific opsonic anti-pneumococcal antibodies ranging from 1:64 to 1:8192 (for example, for at least 50% or more (for example, 60%, 65%, 70%, 75%, 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F).

Furthermore, it is an object of the invention to provide a novel immune globulin (e.g., anti-pneumococcal hyperimmune globulin) composition containing a high titer of opsonic anti-pneumococcal antibodies that enhances immunity in vivo and/or prophylactically and therapeutically suppresses an infection (e.g., an upper respiratory tract infection), in immunocompromised patients in whom an adequate response is not achieved with regular infusions of conventional IVIG (IVIG).

Another preferred object of the present invention is that, although standard pools of immunoglobulin from normal donors (e.g., used to produce commercially available standard/conventional IVIG (IVIG)) do not contain systematic, reproducible and fully protective levels of opsonic antibody to S. pneumonia , a hyperimmune globulin composition (e.g., anti-pneumococcal hyperimmune globulin) containing a high titer of opsonic anti-pneumococcal antibodies of the invention, when administered intravenously, immediately provides specific functional (e.g., not simply binding) antibodies that promote phagocytosis and cause elimination of S. pneumonia by phagocytes. The invention is not limited to the serotype or number of S. pneumoniae serotypes for which functional opsonic antibodies present in hyperimmune globulin (eg, IVIG) promote phagocytosis and/or cause elimination. In fact, another object of the invention is that the composition (for example, the composition of hyperimmune plasma and/or hyperimmune globulin (for example, IVIG))) according to the invention contains broad-spectrum opsonic antibodies against at least 7 out of 12, 8 of 12, 9 of 12, 10 of 12, 11 of 12, or all 12 of 12 of the following S. pneumonia serotypes: 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. Thus, the invention provides compositions that contain an increased titer of specific opsonic anti-pneumococcal antibodies that is at least 2-fold, 3-fold, 4-fold, 5-fold, 7-fold, 10 or more times higher than the specific opsonic anti-pneumococcal antibody titer. present in a control sample (e.g., for at least about 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more)) pneumococcal serotypes selected from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F), and methods for their preparation. In one embodiment, the invention provides an immune globulin (e.g., hyperimmune globulin (e.g., IVIG))) containing a broad-spectrum opsonic antibody titer specific for 70% or more of S. pneumoniae serotypes selected from serotypes 4, 6B , 9V, 14, 18C, 19F, and 23, which is 2 or more times (e.g., 3-25 times or more) higher than the titer of opsonic antibodies specific for S. pneumonia serotypes present in the control sample ( for example, immune globulin obtained from a mixture of plasma samples obtained from random human plasma donors (for example, 100, 300, 500, 1000 or more random human plasma donors) who have not been vaccinated with pneumococcal vaccine, or immune globulin obtained from a healthy pre-vaccination human plasma donor(s) (e.g., such that serotype-specific opsonic antibody titer levels in human plasma donors prior to immunization are used as a baseline to measure the rise in serotype-specific opsonic antibody titers after immunization)) . In another embodiment, the invention provides an immune globulin (e.g., hyperimmune globulin (e.g., IVIG)), which contains a high titer of broad-spectrum opsonic antibodies, at least against 50%, at least against 55%, at least at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, 95%, 98% or more of each serotype present in a vaccine or multiple vaccines used to immunize one or more plasma donors from which the immune globulin composition is derived.

Another advantage and purpose of the invention is to provide a hyperimmune globulin (e.g. IVIG) composition containing broad spectrum opsonic antibodies specific for S. pneumonia for a patient (e.g. an immunocompromised patient (e.g. a PIDD patient) ), in the treatment or prevention of pneumococcal infection in a patient (for example, by suppressing the growth of S. pneumonia and / or clearance of S. pneumonia from the patient's blood).Another advantage and purpose of the invention is to provide a composition of hyperimmune globulin (for example, IVIG) of the invention, containing broad-spectrum opsonic antibodies specific for S. pneumonia, to a patient (for example, an immunocompromised patient (for example, a patient with PID (PIDD)) in the treatment of pneumococcal infection in a patient (for example, by improving or increasing the clearance of S. pneumonia from A further advantage and object of the invention is to provide a hyperimmune globulin (e.g., IVIG) composition of the invention, comprising a broad spectrum opsonic antibody specific for S. pneumoniae, to a patient (e.g., an immunocompromised patient (e.g., a patient with PID (PIDD)) to prevent upper respiratory tract infections in a patient that cannot be prevented with conventional IVIG therapy. The invention is not limited to the type of upper respiratory tract infection being prevented and/or treated and may include, but is not limited to, rhinosinusitis (sinusitis), otitis media, pharyngitis, epiglottitis, laryngotracheitis, and laryngotracheobronchitis. Similarly, the compositions and methods of their use (e.g., administration) are used to prevent and/or treat the signs or symptoms of an upper respiratory tract infection, including, but not limited to, cough, sneezing, nasal discharge, nasal congestion, runny nose, fever, sore or sore throat and nasal breathing.

In one embodiment, the invention provides compositions and methods for preparing a composition containing pooled plasma samples (e.g., plasma from multiple donors (e.g., donors who have been vaccinated with one or more pneumococcal vaccines)) that contain high titers of opsonic anti-pneumococcal antibodies.

Thus, it is an object of the invention to provide methods for preparing compositions (eg blood, plasma and/or immune globulin (eg hyperimmune globulin) compositions) containing a high titer of opsonic anti-pneumococcal antibodies. In one embodiment of the invention, one or more healthy adults (eg, people who do not have known diseases) are administered a pneumococcal immunogen, a recombinant pneumococcal protein, or a combination thereof. In some embodiments, the S. pneumonia immunogen is an S. pneumoniae cell membrane sugar (eg, a polysaccharide). In some embodiments, the S. pneumonia immunogen is an S. pneumonia vaccine comprising one or more S. pneumonia proteins (eg, recombinant or isolated proteins). In some embodiments, the S. pneumonia immunogen is a conjugate vaccine (eg, conjugated with a carrier and/or adjuvant (eg, a protein or other carrier molecule)). In some embodiments, the S. pneumonia immunogen is a non-conjugated vaccine. In some embodiments, the conjugate vaccine or non-conjugate vaccine contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 , 21, 22, 23 or more different immunogens (eg, from an equal number of different S. pneumoniae serotypes). In some embodiments, one or more different serotypes of S. pneumonia include, but are not limited to, serotypes 1, 2, 3, 4, 5, 6A, 6B, 7A, 7B, 1C, 7D, 7E, 7F, 8, 9A-9V, 12, 14, 18C, 19A-19F, 23A-23F, and 25. In some embodiments, one or more different S. pneumonia serotypes are selected from any of more than 90 different known S. pneumonia serotypes. In some embodiments of the invention, one or more different serotypes of S. pneumonia are newly identified.

In a further embodiment of the invention, one or more healthy individuals (eg, individuals with no known disease) are administered a pneumococcal immunogen, a recombinant pneumococcal protein, or a combination present in a commercial pneumococcal vaccine. The invention is not limited to the type of commercial pneumococcal vaccine. In fact, any pneumococcal vaccine known in the art may be used, including, but not limited to, pneumococcal conjugate vaccine (PCV13 or PREVNAR13, Wyeth Pharmaceuticals, Collegeville, PA), SYNFLORIX, and/or polysaccharide pneumococcal vaccine (PPSV23 or PNEUMOVAX23, Merck Sharp & Dohme Corp., North Wales, PA). In one embodiment of the invention, one or more healthy individuals receive a first or primary vaccination with a first pneumococcal vaccine and a subsequent booster vaccination with a first pneumococcal vaccine or a second, different pneumococcal vaccine. For example, in one embodiment of the invention, one or more healthy individuals receive a first or primary vaccination/immunization with a first pneumococcal vaccine (e.g., PREVNAR) and then receive a booster vaccination/immunization (e.g., after 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 or more weeks after primary vaccination/immunization) with a second pneumococcal vaccine (eg PNEUMOVAX23). At a time point following sequential vaccination (eg, 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 or more weeks after sequential vaccination), serum/plasma is collected from vaccinated healthy human plasma donors. Plasma from vaccinated donors can be pooled (with each other and/or with plasma from unvaccinated donors), followed by isolation and/or purification of immune globulin from it (for example, to obtain anti-pneumococcal hyperimmune globulin according to the invention). Methods for collecting plasma as well as extracting immune globulin are well known to one of ordinary skill in the art.

In one embodiment, this invention relates to a method for producing hyperimmune globulin with a high titer of opsonophagocytic antibodies against Streptococcus pneumoniae (e.g., a titer that is at least 2 times, 3 times, 4 times, 5 times, 7 times, 10 or more times higher for at least about 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5 , 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F) compared with the titer of specific opsonophagocytic anti-pneumococcal antibodies against the same serotypes that are present in the control sample; or a titer specific for 70% or more of S. pneumonia serotypes selected from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23 that is 2 or more times (e.g., 3-25 or more times) higher than the titer of opsonophagocytic antibodies specific for the same S. pneumoniae serotypes present in the control sample; or a titer in the range of 1:64 to 1:8192 (e.g., at least 1:256 (e.g., at least 50% or more (e.g., 60%, 65%, 70%, 75% , 80%, 85%, or more)) for S. pneumoniae serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) as determined by the opsonophagocytic elimination assay described in described herein), the method comprising the steps of immunizing healthy adult human plasma donors aged 18 to 60 years by primary immunization with a polyvalent S. pneumonia vaccine and subsequent booster immunization with a polyvalent S. pneumonia vaccine that is the same or may be different. from the primary vaccine; plasma collection from plasma donors after booster immunization; pooling plasma of vaccinated donors in order to obtain pooled plasma containing a high titer of opsonophagocytic antibodies against S. pneumonia; and obtaining immune globulin from the pooled plasma. In a further embodiment of the invention, the method comprises converting the immune globulin thus obtained into a form suitable for intravenous injection. The immune globulin may be provided in solution and/or the pH and ionic strength of the solution may be adjusted to make it suitable for intravenous injection. The invention is not limited to the number of people vaccinated in accordance with the methods described here. For example, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400 or more healthy adult human plasma donors may be vaccinated and plasma collected from the donors. In one embodiment, pooled plasma is obtained from a plasma pool of 1000 or more different vaccinated healthy adult human plasma donors. In one embodiment, the pooled plasma contains an opsonophagocytic antibody titer that is at least 2-fold, 3-fold, 4-fold, 5-fold, 7-fold, 10-fold or more for, at least 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) compared with specific opsonophagocytic anti-pneumococcal antibody titers against the same serotypes present in a control sample (eg, immune globulin obtained from the plasma of 1000 or more random unvaccinated human plasma donors). In another embodiment, the pooled plasma contains an opsonophagocytic antibody titer specific for 70% or more of S. pneumoniae serotypes selected from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23, i. 2 or more times (e.g., 3-25 times or more) higher than the titer of opsonophagocytic antibodies specific for the same S. pneumoniae serotypes present in a control sample (e.g., immune globulin obtained from pooled plasma from 1000 or more random unvaccinated human plasma donors). In another embodiment, the pooled plasma contains an opsonophagocytic antibody titer in the range of 1:64 to 1:8192 (e.g., at least 50% or more (e.g., 60%, 65%, 70%, 75% , 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) as determined by the opsonophagocytic elimination assay described herein or known in the art.

In addition, the invention provides an immune globulin (eg, hyperimmune globulin) obtained by the method described above. The immune globulin thus obtained (eg, hyperimmune globulin) can be used in various ways. For example, the immune globulin can be used in a method of treating an S. pneumoniae infection in a subject (eg, comprising administering to the subject a therapeutically effective amount of immune globulin). Additionally, the immune globulin can be used in a method of administering immunotherapy to a subject (eg, comprising administering to the subject a therapeutically effective amount of immune globulin).

Additionally, in one embodiment, the invention provides a method for producing a hyperimmune globulin having enhanced opsonophagocytic bactericidal activity against at least seven serotypes of Streptococcus pneumoniae for the prevention or treatment of S. pneumoniae infection, the method comprising the steps of immunizing healthy adult human plasma donors by a primary immunization with a polyvalent S. pneumoniae conjugate vaccine followed by a booster immunization with a polyvalent S. pneumoniae polysaccharide vaccine that is different from the primary vaccine; collection and pooling of plasma from immunized plasma donors; and obtaining immune globulin from pooled plasma, wherein the immune globulin contains an opsonophagocytic antibody titer specific for each of the at least seven S. pneumonia serotypes that is two or more times higher (e.g., 3-25 or more times higher) than the titer of opsonophagocytic antibodies specific for each of at least seven S. pneumoniae serotypes present in a control sample (e.g., immune globulin obtained from pooled plasma of 1000 or more random unvaccinated human plasma donors ). In addition, the invention relates to hyperimmune globulin obtained in accordance with the above method. The hyperimmune globulin thus obtained can be used in various ways. For example, hyperimmune globulin can be used in a method of treating an S. pneumoniae infection in a subject, comprising administering to the subject a therapeutically effective amount of hyperimmune globulin. Additionally, hyperimmune globulin can be used in a method of administering immunotherapy to a subject, comprising administering to the subject a therapeutically effective amount of hyperimmune globulin.

In some embodiments, this invention relates to a method for producing compositions (e.g., compositions of blood, plasma and/or immune globulin (e.g., hyperimmune globulin)) containing a high titer of opsonic anti-pneumococcal antibodies, including plasma screening (e.g., pools of plasma or immunoglobulin , immunoglobulin, or immunoglobulin preparations) for total anti-pneumococcal binding antibodies using an in vitro antigen binding assay (e.g., ELISA) together with an opsonophagocytic elimination assay (e.g., an opsonophagocytic elimination assay as described herein or known in the art). ). In some embodiments, this invention relates to a method for obtaining compositions (for example, blood, plasma and/or immune globulin compositions) containing a high titer of opsonic, anti-pneumococcal antibodies, and the method includes plasma screening (for example, plasma or immunoglobulin pools, immunoglobulin or immunoglobulin preparations) for the total titer of opsonic anti-pneumococcal antibodies using an opsonophagocytic bactericidal assay (for example, the assay described in Example 1). In another embodiment, the invention provides a method for producing compositions (for example, blood, plasma and/or immune globulin compositions) containing a high titer of opsonic, anti-pneumococcal antibodies, including screening of plasma (for example, plasma or immunoglobulin pools, immunoglobulin or immunoglobulin preparations) not total anti-pneumococcal binding antibodies using an in vitro antigen-binding assay (eg, ELISA), followed by plasma screening (eg, plasma or immunoglobulin pools, immunoglobulin, or immunoglobulin preparations) for total opsonic anti-pneumococcal antibody titer using an opsonophagocytic bactericidal analysis (for example, the analysis described in Example 1). In yet another embodiment, protective efficacy can be documented in vivo by assaying the protective activity of compositions (e.g., blood, plasma, and/or immune globulin compositions) containing a high titer of opsonic anti-pneumococcal antibodies using an animal model of S. pneumoniae infection (e.g., to determine morbidity, mortality and/or clearance of bacteria). The invention is not limited to a method or assay used to measure the opsonophagocytic titer of antibodies present. In fact, various assays may be used, including, but not limited to, those described herein (eg, in Example 1).

In one embodiment, the invention provides a method for preparing a composition comprising obtaining plasma samples of donors (e.g., 50, 100, 300, 500, 1000 or more donors (e.g., healthy adult human plasma donors)) vaccinated with one or more pneumococcal vaccines, and pooling plasma samples (eg, with each other and/or plasma from unvaccinated donors) to form a pooled plasma composition. In a further embodiment of the invention, immune globulin is obtained from pooled plasma samples (eg, immune globulin is fractionated, purified and/or isolated from plasma by any method known in the art, including those described herein). In a further embodiment of the invention, the immune globulin of the invention is converted into a form suitable for intravenous injection (eg, providing the immune globulin in solution, adjusting the pH, adjusting the ionic strength, etc.). As described herein, in one embodiment, the pooled plasma composition of the invention contains an opsonophagocytic antibody titer that is at least 2-fold, 3-fold, 4-fold, 5-fold, 7-fold, 10 or more times higher for at least about 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) compared with specific opsonic anti-pneumococcal antibody titers against the same serotypes that are present in a control sample (e.g., immune globulin obtained from pooled plasma 1000 or more random unvaccinated human plasma donors). In another embodiment, the pooled plasma composition contains an opsonophagocytic antibody titer specific for 70% or more of S. pneumoniae serotypes selected from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23, which is 2 or more times (e.g., 3-25 times or more) higher than the titer of opsonic antibodies specific for the same S. pneumoniae serotypes present in a control sample (e.g., immune globulin obtained from pooled plasma of 1000 or more number of random unvaccinated human plasma donors). In yet another embodiment, the pooled plasma composition contains an opsonophagocytic antibody titer in the range of 1:64 to 1:8192 (e.g., at least 50% or more (e.g., 60%, 65%, 70%, 75 %, 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) as determined by the opsonophagocytic elimination assay described herein.

In one embodiment of the invention, plasma samples (eg, from vaccinated and/or unvaccinated donors) are screened to confirm the absence of bloodborne pathogens (eg, before or after pooling). In some embodiments of the invention, plasma samples and/or antibodies are obtained from donor subjects in the form of donated or acquired biological material (eg, blood or plasma). In some embodiments of the invention, plasma and/or antibody samples (eg, blood, plasma, isolated antibodies, etc.) are obtained from a commercial source. In some embodiments, a plasma sample and/or antibodies, donated blood, or donated plasma is screened for the presence of pathogens and purified or discarded if certain pathogens are present. In some embodiments of the invention, the screening is carried out before the pooling of the donor sample with other donor samples. In other embodiments of the invention, screening is performed after the pooling of samples. Antibodies, blood and/or plasma can be obtained from any suitable subjects. In some embodiments, antibodies, blood, and/or plasma are obtained from a subject who has recently (e.g., within 1 year, within 6 months, within 2 months, within 1 month, within 2 weeks, within 1 week) , within 3 days, within 2 days, within 1 day) has been vaccinated or exposed to one or more pneumococcal vaccines.

In some embodiments of the invention, samples of blood, plasma and/or immune globulin are identified (for example, in accordance with the methods described herein) as containing a high titer of opsonic anti-pneumococcal antibodies, combined (for example, pooled) to obtain a composition according to the invention ( for example, hyperimmune globulin) containing a high titer of opsonic anti-pneumococcal antibodies.

Any suitable method for obtaining plasma, antibody samples, pooled plasma and/or immunoglobulin compositions thereof is within the scope of this invention. In addition, any suitable method for obtaining, producing, purifying, fractionating, enriching, etc. antibody samples and/or plasma pools is within the scope of this invention. Typical methods and procedures for obtaining antibody samples and obtaining plasma pools are given, for example, in US patent No. 4174388; US patent No. 4346073; US patent No. 4482483; US patent No. 4587121; US Pat. No. 4,617,379; US patent No. 4659563; US patent No. 4665159; US patent No. 4717564; US Pat. No. 4,717,766; US patent No. 4801450; US patent No. 4863730; US Pat. No. 5,505,945; US patent No. 5582827; US patent No. 6692739; US patent No. 6966200; US patent No. 6984492; US patent No. 7045131; US patent No. 7488486; US patent No. 7597891; US patent No. 6372216; US Patent Application No. 2003/0118591; US Patent Application No. 2003/0133929; US Patent Application No. 2005/0053605; US Patent Application No. 2005/0287146; US Patent Application No. 2006/0110407; US Patent Application No. 2006/0198848; US Patent Application No. 2006/0222651; US Patent Application No. 2007/0037170; US Patent Application No. 2007/0249550; US Patent Application No. 2009/0232798; US Patent Application No. 2009/0269359; US Patent Application No. 2010/0040601; US Patent Application No. 2011/0059085; and US Patent Application No. 2012/0121578; which are incorporated herein in their entirety by reference. In embodiments of the present invention, any suitable combination of methods, methods, or compositions from the aforementioned references may be used.

The isolated immune globulin of the invention may be from an IgG fraction or isotype, but the isolated immune globulin is not limited to any particular fraction or isotype and may be IgG, IgM, IgA, IgD, IgE, or any combination thereof. It is further preferred that the isolated immune globulin be pure or antigenically human immune globulin.

In one embodiment of the invention, a composition containing a high titer of opsonic pneumococcal antibodies of the invention is a sterile solution with a pH of about 6.0-7.8 (e.g., 5.0-6.0, 6.0, 6.1, 6 .2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4 , 7.5, 7.6, 7.7, 7.8 or higher). In another embodiment of the invention, a composition containing a high titer of opsonic anti-pneumococcal antibodies of the invention is prepared in accordance with US FDA standards for an immune globulin preparation (see, for example, 37 CFR, §640.100, 640.101, 640.102, 640.103, 640.104, April 1, 2013 G.). In one embodiment of the invention, a composition containing a high titer of opsonic anti-pneumococcal antibodies of the invention (e.g., the hyperimmune globulin described herein, in particular in the Examples) contains at least a minimum level of antibody titers against Corynebacterium diphtheria, measles virus, and poliomyelitis virus recommended by the FDA (eg see 37 CFR §640.104) for the treatment of patients with immunodeficiency disease.

In one embodiment of the invention, the pooled plasma composition does not contain detectable levels (e.g., detectable using any method known in the art (e.g., recommended by the U.S. Food and Drug Administration)) human immunodeficiency virus (HIV). ) 1 (HIV-1), HIV-2, Treponema pallidum, Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, Plasmodium knowlesi, hepatitis B virus (HBV), hepatitis C virus (HCV), prions, West Nile virus, parvovirus, Typanosoma cruzi, SARS coronavirus and/or vaccinia virus. In one embodiment, each individual plasma sample used in the method or composition of the invention is only collected from FDA-approved blood services and tested with serological tests (e.g., FDA-approved serological tests) for human immunodeficiency virus (HIV) 1 (HIV-1). ), HIV-2, Treponema pallidum, Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, Plasmodium knowlesi, hepatitis B virus (HBV), hepatitis C virus (HCV), prions, West Nile virus, parvovirus, Typanosoma cruzi, coronavirus SARS and/or vaccinia virus. In another embodiment, an individual plasma sample and/or pooled plasma composition of the invention is tested for the presence of HIV-1, HIV-2, HBV, HCV, or another infectious agent (e.g., pathogen) using nucleic acid testing (NAT )) and are used in the method or composition of the invention only if the absence of pathogens is confirmed.

The invention is not limited to the type of subject (eg, mammal, non-human primate, human, etc.) administered or treated with the composition of the invention (eg, pooled plasma samples and/or immunotherapeutic composition containing them). For example, the invention is not limited to the type of patient receiving compositions (eg blood, plasma and/or immune globulin (eg hyperimmune globulin) compositions) that contain a high titer of opsonic, anti-pneumococcal antibodies. The aim of the invention is to provide and/or administer the composition of the invention to the immunocompromised patients described herein.

In one embodiment, the pooled plasma comprises plasma samples obtained from 1000-3000 or more (e.g., more than 1000, 1250, 1500, 1750, 2000, 2500, 3000, 3500, 4000 or more) human subjects. In one embodiment, the composition comprising the pooled plasma samples further comprises a pharmaceutically acceptable carrier (eg, natural and/or non-natural carriers). In one embodiment, the pooled plasma composition is used to produce immune globulin (eg, for intravenous administration to a subject). In one embodiment of the invention, the pooled plasma and/or immune globulin composition provides a therapeutic benefit to the subject to whom the composition is administered that is not achievable by administering a mixture of plasma samples obtained from 1000 or more random human subjects and/or immunoglobulin derived from them. (eg, prevents or treats an upper respiratory tract infection in a subject that is not prevented or treated by conventional IVIG). The invention is not limited to the type of therapeutic benefit. In fact, various therapeutic benefits can be achieved, including those described herein.

In one embodiment, the pooled plasma and/or the immunoglobulin prepared therefrom of the invention reduces the incidence of infection (eg, upper respiratory tract infection) in a subject to whom the composition is administered. In another embodiment of the invention pooled plasma and/or immunoglobulin of the invention prepared from it reduces the number of days during which a subject administered with pooled plasma and/or immunoglobulin of the invention needs to receive antibiotics (for example, to treat an infection ( e.g. upper respiratory tract infection). In yet another embodiment, the pooled plasma and/or the immunoglobulin prepared therefrom of the invention increases the minimum level of circulating opsonic anti-pneumococcal antibodies in a subject (e.g., increases the titer of S. pneumoniae-specific opsonic antibodies (e.g., thereby conferring protective specific anti-pneumococcal antibody levels between scheduled administration dates for pooled plasma and/or immune globulin derived from it of the invention (eg, not maintained in a subject receiving conventional IVIG))).

Additional objects, features and advantages of the present invention will become apparent from the following detailed description. However, it is to be understood that the detailed description and specific examples indicating preferred embodiments of the invention are provided for purposes of illustration only, as various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art based on this detailed description.

BRIEF DESCRIPTION OF GRAPHICS

On FIG. 1 shows titers of binding, serotype-specific anti-pneumococcal IgG antibodies in individual sera from vaccinated donors taken at the following time points: immediately before primary vaccination with PREVNAR (Pre (Pre)), immediately before booster vaccination with PNEUMOVAX23 at week 4 (week 4), at 8 weeks, 10 weeks, 12 weeks and 16 weeks. Mean titers of binding, serotype-specific anti-pneumococcal IgG antibodies are given for pre-vaccination and at 4, 8, 10, and 12 weeks, and for pre-vaccination and 12 weeks pooled samples. In addition, the multiplicity of increase in titers of binding, serotype-specific anti-pneumococcal IgG antibodies before vaccination and at week 12 is given.

Fig. 2 illustrates the titer of serotype-specific opsonic (opsonophagocytic) elimination (OPE (OPK)) pneumococci in individual serum samples of vaccinated donors taken at the following time points: immediately before the primary vaccination with PREVNAR (Pre), immediately before the secondary vaccination with PNEUMOVAX23 at week 4 (4 week), at 8 weeks, at 10 weeks, at 12 weeks and at 16 weeks specific for different pneumococcal serotypes. Mean anti-pneumococcal serotype-specific opsonic titers are given before vaccination and at 4, 8, 10 and 12 weeks, and for pooled samples before vaccination and at 12 weeks. In addition, the multiplicity of the increase in antipneumococcal serotype-specific titers of OFE in the period from the time before vaccination to the 12th week is given.

Fig. 3 illustrates the titer of binding, serotype-specific anti-pneumococcal IgG antibodies in pooled donated sera from 40 subjects, which was obtained at the following time points: immediately prior to PREVNAR primary vaccination (Pre (Pre)), week 8, week 10, 12 week, 16 weeks and 5 months.

Fig. 4 illustrates the increase in fold concentrations of binding, serotype-specific anti-pneumococcal IgG antibodies in pooled donor serum obtained at the following time points: immediately before PREVNAR primary vaccination (Pre (Pre)), week 8, week 10, week 12 and at 16 weeks and 5 months (* = baseline).

Fig. 5 illustrates pneumococcal serotype-specific opsonic elimination (OPK) titers in pooled donor sera from 40 subjects obtained at the following time points: week, at 12 weeks and at 16 weeks and at 5 months.

Fig. 6 illustrates titers of serotype-specific anti-pneumococcal IgG binding antibodies in purified immune globulin (IG(IG)) from pooled donated sera from 40 individuals that were obtained at the indicated time points.

Fig. 7 illustrates pneumococcal serotype-specific OFE (OPK) titers in purified IgG from pooled donor sera from 40 individuals that were obtained at the indicated time points.

Fig. 8 illustrates a comparison of the titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in pooled human sera from vaccinated donors compared to the titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in several different traditional commercially available IVIG ( IVIG). Each column represents a single unique S. pneumonia serotype. Each row is a unique pattern. Samples A-I are different batches of conventional, commercially available IVIG. "IVIG (IVIG) average" is the average titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in a pooled sample containing equal amounts of each conventional, commercially available IVIG (IVIG). "Prevaccination Donor Serum" represents the titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in the pooled pre-vaccination human donor serum described in Example 1. "Vaccinated Donor Serum" represents the titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in pooled human donor sera for twelve weeks after the vaccination described in Example 1. "Titer increase compared to commercial IVIG (IVIG)" is the titer increase fold of serotype-specific OFE (OPK) functional/opsonic antibodies present in pooled human donor serum twelve weeks after the vaccination described in Example 1 compared to the mean titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in a pooled sample containing equal the amount of each of the conventional commercially available IVIG (IVIG).

Fig. 9A-9L illustrate the titer of serotype-specific opsonic antibodies against S. pneumonia present in nine different IVIG commercial lots (Samples A-I) compared to the titer of serotype-specific opsonic S. pneumonia antibodies present in the hyperimmune globulin of the invention containing high titer of opsonic anti-pneumococcal antibodies to each serotype.

Fig. 10 illustrates the percentage of primary immunocompromised subjects whose residual anti-S. pneumoniae binding antibody level falls below protective levels (1.2 mg/mL) following conventional IVIG (IVIG) infusion.

DEFINITIONS

As used herein, the term "subject" refers to any human or animal (eg, non-human primate, rodent, feline, canine, large ruminant, porcine, equine, etc.).

As used herein, the term "sample" is used in its broadest sense to include materials derived from any source and may be used, for example, to refer to materials derived from a biological source, such as derived from animals (including humans), and additionally includes any liquids, solids and tissues. In specific embodiments of this invention, the biological samples include blood and blood products such as plasma, serum, and the like. However, these examples should not be construed as limiting the types of samples that find use in this invention.

As used herein, the term "antibody" refers to an immunoglobulin molecule, which typically consists of two identical pairs of polypeptide chains, each pair containing one "light" (L) chain and one "heavy" (H) chain. Human light chains are classified as kappa and lambda light chains. Heavy chains are classified as mu, delta, gamma, alpha or upsilon and define the isotype of an antibody as IgM, IgD, IgG, IgA and IgE, respectively. In the light and heavy chains, the variable and constant regions are joined by a "J" region containing about 12 or more amino acids, with the heavy chain further comprising a "D" region containing about 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (abbreviated herein as HCVR or V _H ) and a heavy chain constant region. The heavy chain constant region consists of three domains: CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated as LCVR or V _L ) and a light chain constant region. The light chain constant region consists of a single CL domain. Antibody constant regions can mediate immunoglobulin binding to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (C1q) of the classical complement system. The V _H and V _L regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) that alternate with more conserved regions called framework regions (FRs). Each V _H and V _L consists of three CDRs and four FRs in the following order from amino to carboxy: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of each heavy/light chain pair (V _H and V _L ), respectively, form an antibody binding site. The term "antibody" includes an antibody that is part of an antibody polymer (polymeric form of an antibody), such as dimers, trimers, or higher order polymers of monomeric antibodies. In addition, it includes an antibody that is associated or attached to or otherwise physically or functionally connected to a fragment that is not an antibody. In addition, the term "antibody" is not limited to any particular method for producing an antibody. For example, it includes but is not limited to recombinant antibodies, synthetic antibodies, monoclonal antibodies, polyclonal antibodies, bispecific antibodies, and polyspecific antibodies.

As used herein, the term "antibody derivative" or "derivative" of an antibody refers to a molecule that is capable of binding to the same antigen as the antibody from which it is derived and contains an amino acid sequence that is the same or similar to the antibody. associated with an additional molecular entity. The amino acid sequence of an antibody that is contained in an antibody derivative may be a full length antibody, or may be any portion or portions of a full length antibody. The additional molecular entity may be a chemical or biological molecule. Examples of additional molecular entities include chemical groups, amino acids, peptides, proteins (such as enzymes, antibodies), and chemical compounds. The additional molecular entity may be useful from any point of view, for example, for use as a detection agent, label, marker, pharmaceutical or therapeutic agent. The amino acid sequence of an antibody may be attached to or linked to an additional entity through chemical bonding, genetic synthesis, non-covalent association, or otherwise. In addition, the term "antibody derivative" includes chimeric antibodies, humanized antibodies, and molecules that are derived from modifications to the amino acid sequences of an antibody, such as conservative substitutions, additions, and insertions of amino acids.

As used herein, the terms "antigen" and "immunogen" are used interchangeably to refer to any substance that is capable of inducing an adaptive immune response. The antigen may be a whole cell (eg, a bacterial cell), a virus, a fungus, or an antigenic portion or component thereof. Examples of antigens include, but are not limited to, microbial pathogens, bacteria, viruses, proteins, glycoproteins, lipoproteins, peptides, glycopeptides, lipopeptides, toxoids, carbohydrates, tumor-specific antigens, and antigenic portions or components thereof.

As used herein, the term "antigen-binding fragment" of an antibody refers to one or more portions of a full length antibody that retain the ability to bind to the same antigen that the antibody binds to.

As used herein, the terms "immunoglobulin", "immune globulin", "immunoglobulin molecule", and "IU" include (1) antibodies, (2) antibody antigen-binding fragments, and (3) antibody derivatives, each as defined herein. As described herein, immune globulin can be obtained from (eg, fractionated from, isolated from, purified from, concentrated from, etc.) pooled plasma compositions (eg, for administration to a subject). As used herein, the term "intravenous immune globulin (IVIG)" refers to conventional immunoglobulin derived from human plasma from large amounts (e.g., 250, 500, 1000 or more) random donors (e.g., HIZENTRA, immune subcutaneous globulin, CSL Behring), while the terms "anti-pneumococcal immune globulin" or "anti-pneumococcal IVIG (IVIG)" or "anti-pneumococcal hyperimmune globulin" (for example, as described in this description and in the Examples) refers to immune globulin obtained from plasma donors (e.g., human plasma donors) according to the methods of the invention (e.g., large numbers (e.g., 250, 500, 1000 or more) of human plasma donors (e.g., healthy human plasma donors) that have been vaccinated with one or more anti- pneumococcal vaccines in accordance with the methods described in this description), which contains an increased titer of specific opsonic anti-pneumococcal antibodies compared to the control sample.

The terms "opsonophagocytic antibody" and "opsonic antibody" are used interchangeably herein to refer to antibodies that function to actively cause bacteria or other foreign bodies (e.g., cells or cell products) to be susceptible to phagocytes (e.g., opsonic antibodies coat bacteria, causing bacterial cells to become susceptible to phagocytosis as a result of the interaction between opsonic antibodies coating bacterial cells and receptors on the surface of phagocytes). For example, an opsonic pneumococcal antibody is an antibody that binds to and coats the surface of S. pneumoniae, causing the bacterium to be susceptible to phagocytosis/elimination by phagocytes.

The terms "opsonophagocytic antibody titer" and "opsonic antibody titer" are used interchangeably herein to refer to a titer of functionally active opsonophagocytic antibodies (eg, that correlate with protection against infection and/or disease). For example, an opsonic anti-pneumococcal antibody titer (e.g., as determined by the OFE assay (OPK) described herein) is the titer of S. pneumoniae-specific opsonophagocytic antibodies in a sample (e.g., which is independent of and different from the total antibody count, able to bind to S. pneumonia, present in the sample (for example, according to the analysis of ELISA (ELISA)).

As used herein, the term "hyperimmune globulin" refers to an immunoglobulin obtained from the plasma of donors with a high antibody titer (eg, opsonic antibody) against a particular organism (eg, S. pneumonia). For example, as used herein, "anti-pneumococcal hyperimmune globulin" is an immunoglobulin containing an elevated titer of opsonic anti-pneumococcal antibodies, independent of (eg, independent of, and distinct from) total titer or amount of antibodies capable of binding to S. pneumonia.

As described herein, an elevated titer of specific opsonic pneumococcal antibodies is at least 2-fold, 3-fold, 4-fold, 5-fold, 7-fold, 10-fold or greater for at least about 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F) compared with specific opsonic anti-pneumococcal antibody titers specific for the same serotypes present in the control sample; or specific for 70% or more of S. pneumonia serotypes selected from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23, being 2 or more times higher (e.g., 3-25 or more times) than the titer of opsonic antibodies specific for the same S. pneumoniae serotypes that are present in the control sample; or one that is in the range from 1:64 to 1:8192 (for example, at least 50% or more (for example, 60%, 65%, 70%, 75%, 80%, 85% or more ) S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) according to the opsonophagocytic elimination assay described herein; or one that is at least 2-fold, 3-fold, 4-fold, 5 or more times higher than the titer of specific opsonic anti-pneumococcal antibodies that are present in the control sample, for example, for at least 55 % or more pneumococcal serotypes selected from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F (i.e., at least 2, 3, 4, A 5-fold or greater increase in titers of opsonic anti-pneumococcal antibodies specific for 7/12, 8/12, 9/12, 10/12, 11/12, or 12/12 pneumococcal serotypes compared to a control). The invention is not limited to the kind of control sample used. For example, in one embodiment of the invention, the control sample is an immune globulin obtained from the plasma of healthy human plasma donor(s) prior to vaccination (e.g., such that titer levels of specific opsonic anti-pneumococcal antibodies in human donor plasma prior to immunization are used in as a baseline for measuring the increase in specific opsonic anti-pneumococcal antibody titers after immunization). In another embodiment of the invention, the control sample is an immune globulin obtained from a mixture of plasma samples obtained from 1000 or more random human plasma donors who have not been vaccinated with pneumococcal vaccine (for example, so that titer levels of specific opsonic anti-pneumococcal antibodies in unvaccinated human plasma donors were used as the basis for measuring the increase in titers of specific opsonic pneumococcal antibodies present in human plasma donors vaccinated according to the methods of the invention). Those skilled in the art will appreciate that other controls may be used. In addition, an elevated specific opsonic anti-pneumococcal antibody titer may be a titer that is used to measure bacterial clearance and/or that is used as an accurate surrogate for predicting the effectiveness of an immunoglobulin preparation in protecting against S. pneumoniae infections.

As used herein, the expression "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and do not typically cause an unacceptable allergic or similar adverse reaction when administered to a human.

PNEUMOVAX (Merck Sharp & Dohme Corp., North Wales, RA) refers to a pneumococcal polysaccharide vaccine consisting of purified pneumococcal capsular polysaccharide preparations. PPSV23 contains a polysaccharide antigen from the following 23 pneumococcal serotypes: 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, PA, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20 , 22F, 23F and 33F. It contains 25 micrograms of each antigen per dose and contains 0.25% phenol as a preservative.

PREVNAR-13 (Wyeth Pharmaceuticals, Collegeville, PA) is a pneumococcal conjugate vaccine that contains a purified capsular polysaccharide from 13 Streptococcus pneumoniae serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 19A, 19F , 18C and 23F) conjugated to a non-toxic variant of the diphtheria toxin known as CRM 197. A 0.5 milliliter (mL) dose of PCV13 contains approximately 2.2 micrograms (μg) of each of the 12 serotypes of polysaccharide and approximately 4.4 μg of serotype 6B polysaccharide ; the total concentration of CRM 197 is approximately 34 mcg; it contains 0.02% polysorbate 80 (P80), 0.125 milligrams (mg) aluminum as aluminum phosphate (AlPO4) adjuvant, and 5 ml succinate buffer. The vaccine does not contain thimerosal as a preservative.

As used herein, the term "antibody sample" refers to a composition containing antibodies (e.g., liquid (e.g., plasma, blood, purified antibodies, blood or plasma fractions, blood or plasma components, etc.)) obtained from or provided by a donor (eg, a natural source) or derived from a synthetic, recombinant, other in vitro or commercial source. An antibody sample may contain an elevated titer of a particular antibody or set of antibodies based on pathogenic/antigenic exposures (eg, natural exposure or vaccination) to the donor, or antibodies engineered to be produced in a synthetic, recombinant, or in vitro context. In this case, an antibody sample with an increased titer of antibody X is referred to as a "sample with an elevated level of antibody X". For example, an antibody sample with an elevated titer of antibodies against S. pneumonia is called an "antibody sample with an increased titer against S. pneumonia".

As used herein, the term "isolated antibody" or "isolated binding molecule" refers to an antibody or binding molecule that has been identified and separated from at least one contaminant with which it is normally associated in the source. Examples of isolated antibodies include: an antibody that: (1) is not associated with one or more of its naturally associated components that accompany it in its natural state; (2) essentially free of other proteins from the source of origin; or (3) expressed recombinantly, in vitro or without cells, or synthesized and the medium in which it was produced is removed.

As used herein, the terms "pooled plasma", "pooled plasma samples", and "pooled plasma composition" refer to a mixture of two or more plasma samples and/or a composition (eg, immune globulin) derived from it. An elevated titer of a particular antibody or set of antibodies in pooled plasma reflects elevated titers of the antibody samples that make up the pooled plasma. For example, plasma samples can be obtained from subjects who have been vaccinated (e.g., with a pneumococcal vaccine) and therefore have a high antibody titer (e.g., high total binding antibody titer and/or high opsonic antibody titer) to a pathogen (S. pneumonia) compared to the antibody level(s) found in the general population. By pooling plasma samples, a pooled plasma composition (eg, containing an increased titer of antibodies specific to a particular pathogen) is obtained. In this case, the pooled plasma with elevated antibody X titer (eg, where "X" is a microbial pathogen) is referred to as "antibody pool with increased X". For example, pooled plasma with an elevated titer of S. pneumoniae binding antibodies is referred to as an "anti-S. pneumoniae antibody pool." Pooled plasma compositions can be used to produce immune globulin (eg, which is then administered to a subject) by methods known in the art (eg, fractionation, purification, isolation, etc.). The invention contemplates that both the pooled plasma compositions and the immune globulin derived therefrom may be administered to a subject to provide prophylactic and/or therapeutic benefits to the subject. Accordingly, the term pooled plasma composition may refer to immune globulin obtained from pooled plasma/pooled plasma samples.

As used herein, the term "antibody-loaded pool" refers to pooled plasma loaded with or pooled with antibodies or other immunoglobulin produced synthetically, recombinantly, or by other means in vitro.

As used herein, the term "purified" or "purify" means the result of any process that removes a portion of a contaminant from a component of interest, such as a protein (eg, antibody) or nucleic acid. Thus, the percentage of purified component in the sample is increased.

As used herein, the term "immunotherapeutic agents" refers to a chemical or biological agent that can enhance the immune response (eg, specific or general) of a mammal. As used herein, the term "donor" refers to an entity that provides a biological sample (eg, blood, plasma, etc.). The donor/donor specimen may be screened for the presence or absence of specific pathogens (e.g., using the U.S. Food and Drug Administration (FDA) Blood Product Safety Standards Evaluation Guidelines for Evaluating Blood Product Safety Standards ( for example, issued by the FDA Blood Products Advisory Committee.) For example, a donor/donor specimen may be screened according to FDA guidelines to ensure that one or more blood-borne pathogens (eg, Human Immunodeficiency Virus (HIV) 1 (HIV- 1), HIV-2, Treponema pallidum (syphilis), Plasmodium falciparum, P. malariae, P. ovale, P. vivax or P. knowlesi (malaria), hepatitis B virus (HBV), hepatitis C virus HCV), prions ( Creutzfeldt-Jakob disease), West Nile virus, parvovirus, Typanosoma cruzi, SARS coronavirus (SARS), vaccinia virus, or other pathogen that is routinely screened for or recommended by a regulatory body such as the FDA).

As used herein, the term "immunostimulatory amount" refers to that amount of a vaccine (eg, pneumococcal vaccine) that is capable of stimulating an immune response. The immune response includes a set of biological effects that cause the body to produce immunoglobulins or antibodies in response to a foreign object. Accordingly, the immune response refers to the activation of B cells, in vivo or in culture, by stimulation of Ig receptor molecules on the cell surface of a B cell. Measurement of the immune response is within the normal purview of those skilled in the art, and includes the determination of antibody levels using the methods described in the Tijssen series, Laboratory Techniques in Biochemistry and Molecular Biology: Practice and Theory of Enzyme Immunoassays (Burdon & van Knippenberg eds., 3rd ed., 1985) Elsevier, New York; and Antibodies: A Laboratory Manual (Harlow & Lane eds., 1988), Cold Spring Harbor Laboratory Press; as well as techniques such as countercurrent immunoelectrophoresis (PTEF (GIEP)), radioimmunoassay, radioimmunoprecipitation, enzyme-linked immunosorbent assay (ELISA), dot blot assays, and sandwich assays, see U.S. Patent Nos. 4,376,110 and 4,486,530, all of which are incorporated by reference. In addition, measuring the immune response includes detecting or identifying B cell activation events that may precede antibody production or signal an increase in antibody production. Such measurements include B cell proliferation assays, phosphorylation assays, intracytoplasmic free calcium concentration assays, and other methods for determining B cell activation known in the art. Typical analyzes are presented in Mongini et al., J. Immunol. 159:3782-91 (1997); Frade, et al., BBRC 188:833-842 (1992); Tsokos et al., J. Immunol. 144:1640-1645 (1990); Delcayre et al., BBRC 159:1213-1220 (1989); and Nemerow et al., J. Immunol. 135:3068-73 (1985), each of which is incorporated by reference. In preferred embodiments of the invention, the practice of the invention includes promoting, enhancing or stimulating an immune response. These actions refer to the formation of an immune response that did not previously exist; to optimize or enhance the desired immune response; to the formation or increase of a secondary response, characterized by an increase in isotype switching, memory responses, or both; to provide a statistically increased immunoprotective action against the pathogen; to obtain an equivalent or more pronounced humoral immune response or other measure of activation of B-cells based on a reduced or threshold dose of antigen; generating an increased humoral immune response or other indicator of B cell activation in response to an equivalent dose of antigen; or lower the affinity threshold for B cell activation in vivo or in vitro. Preferably, an immunostimulatory amount refers to that amount of a vaccine that is capable of stimulating an immune response in a subject (e.g., a donor) from whom plasma, serum, or other blood component is being collected, for use in the compositions and methods of the invention (e.g., in therapeutic and/or prophylactic treatment of pneumococcal infection in a subject receiving treatment with the compositions and methods described herein)).

The terms "buffer" or "buffering agents" refer to materials that, when added to a solution, cause the solution to resist changes in pH.

The terms "reducing agent" and "electron donor" refer to a material that donates electrons to a second material in order to reduce the oxidation state of one or more atoms of the second material.

The term "monovalent salt" refers to any salt in which the metal (eg Na, K, or Li) has a net charge of 1+ in solution (i.e., one more proton than electrons).

The term "divalent salt" refers to any salt in which the metal (eg Mg, Ca or Sr) has a net 2+ charge in solution.

The terms "chelator" or "chelating agent" refers to any material containing more than one atom with a single pair of electrons that is available for bonding with a metal ion.

The term "solution" refers to an aqueous or non-aqueous mixture.

As used herein, the term "adjuvant" refers to any substance that can stimulate an immune response. Some adjuvants can cause the activation of an immune cell (for example, an adjuvant can cause an immune cell to produce and secrete a cytokine). Examples of adjuvants that can induce immune cell activation include, but are not limited to, saponins purified from the bark of the Q. saponaria tree, such as QS21 (a glycolipid that elutes at peak 21 on HPLC fractionation; Aquila Biopharmaceuticals, Inc. , Worcester, Massachusetts); poly(di(carboxylatophenoxy)phosphazene (PCPP (PCPP) polymer, Virus Research Institute, USA), lipopolysaccharide derivatives such as monophosphoryl lipid A (MPL, Ribi ImmunoChem Research, Inc., Hamilton, Montana), muramyl dipeptide (MDP, Ribi ) and threonyl-muramyl dipeptide (t-MDP, Ribi), OM-174 (glucosamine disaccharide related to lipid A, OM Pharma SA, Meuren, Switzerland), cholera toxin (TX(CT)) and Leishmania elongation factor (purified Leishmania protein, Corixa Corporation, Seattle, WA.) Conventional adjuvants are well known in the art and include, for example, aluminum phosphate or hydroxide salts ("alum"). In some embodiments, the compositions of this invention are administered with one or more adjuvants ( for example, to cause a bias in the immune response towards a Th1 and/or Th2 type response.) In some embodiments, the adjuvants described in US 2005158329, US 2009010964, US 2004047882 or US 6262029 are used (each of which is included in this application in its entirety via link).

As used herein, the term "an amount effective to induce an immune response" (e.g., pneumococcal vaccine) refers to the dose level required (e.g., when administered to a subject) to stimulate, generate, and/or induce an immune response in the subject. An effective amount may be administered in one or more administrations (eg, by the same or different route), applications, or doses, without being limited to a particular formulation or route of administration.

As used herein, the term "under conditions under which said subject generates an immune response" refers to any qualitative or quantitative induction, generation and/or stimulation of an immune response (e.g., innate or acquired).

As used in this description, the term "immune response" refers to the response of the subject's immune system. For example, immune responses include, but are not limited to, a detectable change (e.g., increase) in Toll-like receptor (TLR) activation, expression and/or secretion of a lymphokine (e.g., cytokine (e.g., Th1 or Th2 type) or chemokine), activation macrophages, dendritic cell activation, T cell activation (eg, CD4+ or CD8+ T cells), natural killer cell activation, PK (>1K) cells, and/or B cell activation (eg, antibody generation and/or secretion ). Additional examples of immune responses include binding of an immunogen (e.g., an antigen (e.g., an immunogenic polypeptide)) to a major histocompatibility complex molecule, OKH (MHC), and inducing a response in the form of a cytotoxic T lymphocyte ("CTL") that induces a B cell response (e.g., , antibody production) and/or a T-helper lymphocyte response and/or a delayed-type hypersensitivity (DTH) response against the antigen from which the immunogenic polypeptide is derived, expansion (e.g., cell population growth) of cells of the immune system ( e.g., T cells, B cells (e.g., any stage of development (e.g., plasma cells), and increased antigen processing and presentation by antigen-presenting cells. The immune response may be directed to immunogens that the subject's immune system considers foreign (e.g., non-self antigens from microorganisms (e.g., pathogens) or self-antigens recognized as foreign.) Thus, it should be understood that, as used herein, "immune response" refers to any type of immune response, including, but not limited to, innate immune response ( e.g., activation of the Toll receptor signaling cascade), cell-mediated immune responses (e.g., responses mediated by T cells (e.g., antigen-specific T cells) and non-specific cells of the immune system), and humoral immune responses (e.g., e.g., B-mediated responses). -cells (for example, through the generation and secretion of antibodies into plasma, lymph and/or tissue fluids). The term "immune response" includes all aspects of the ability of a subject's immune system to respond to antigens and/or immunogens (e.g., both primary response to an immunogen (e.g., pathogen) and acquired (e.g., memory) that result from an adaptive immune response).

As used herein, the term "pharmaceutically acceptable carrier" means any of the standard pharmaceutical carriers, including, but not limited to, phosphate-buffered saline, water, and various types of wetting agents (e.g., sodium lauryl sulfate), any and all solvents , dispersion media, coatings, sodium lauryl sulfate, isotonic and absorbent agents, disintegrants (eg potato starch or sodium starch glycolate), polyethylene glycol, other natural and non-natural carriers, and the like. In addition, the compositions may contain stabilizers and preservatives. Examples of carriers, stabilizers and adjuvants are described and known in the art (see, for example, Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, Pa. (1975), incorporated herein by reference) .

As used herein, the term "pharmaceutically acceptable salt" refers to any salt (eg, obtained by reaction with an acid or base) of a composition of this invention that is physiologically tolerated by the target subject. "Salts" of the compositions of this invention can be prepared with inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, chloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic, formic , benzoic, malonic, sulfonic, naphthalene-2-sulfonic, benzenesulfonic acid, and the like. Other acids, such as oxalic acid, which are not in themselves pharmaceutically acceptable, may be used in the preparation of salts useful as intermediates for preparing the compositions of the invention and their pharmaceutically acceptable acid addition salts. Examples of bases include, but are not limited to, alkali metal hydroxides (eg sodium), alkaline earth metal hydroxides (eg magnesium), ammonia, and compounds of the formula NW ₄ ⁺ where W is C _1-4 alkyl, and the like.

Examples of salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentane propionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptano at, hexanoate , chloride, bromide, iodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, etc. .P. Other examples of salts include anions of the compounds of this invention in combination with a suitable cation such as Na ⁺ , NH4 ₄ ⁺ and NW ₄ ⁺ (where W is a C _1-4 alkyl group) and the like. For therapeutic use, pharmaceutically acceptable salts of the compounds of this invention are included. However, salts of acids and bases that are not pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

For therapeutic use, pharmaceutically acceptable salts of the compounds of this invention are included. However, salts of acids and bases that are not pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

DETAILED DESCRIPTION OF THE INVENTION

Streptococcus pneumoniae is the leading cause of death and morbidity worldwide. Its main virulence factor is the capsular polysaccharide. Older age and antibody immunodeficiency (eg, as a result of primary immunodeficiency described herein) are major risk factors for pneumococcal infection. The World Health Organization (WHO) estimates that 1.6 million people die each year from pneumococcal disease. Although most of the more than 90 known capsular serotypes can cause serious illness, only a limited number cause most cases of invasive pneumococcal disease (IPD). Immunity to S. pneumonia is mediated by bacterial phagocytosis in the presence of complement and a serotype-specific opsonic antibody. Evaluation of serological responses to various pneumococcal vaccines in large clinical trials has provided the parameters of correlation with protection needed for the release of new vaccines. To predict IPD disease protection (IPD), WHO recommends a reference threshold of 0.2 to 0.35 µg/mL for pneumococcal conjugate vaccines (all serotypes), and for polysaccharide vaccines, the current guideline specifies 1.3 µg/mL. A more accurate surrogate marker that is indicative of protection, and which has been adopted by the FDA as being correlated with clinical efficacy, is opsonic antibody titer, and has been found to be accurate enough to replace the need for vaccine trials. For example, opsonic titers in a vaccinated individual of 1:8 are generally considered to be correlated with protection against various S. pneumoniae serotypes. If a vaccine induces an opsonic titer of 1:8 against a particular serotype, then the vaccine can be considered clinically effective even in the absence of a clinical trial demonstrating a reduction in infections in vaccinated people compared to unvaccinated people.

Primary immunodeficiencies (PID) are a group of more than 200 genetically inherited disorders characterized by deficiency of certain components of the innate or adaptive immune system, with the clinical result of increased susceptibility to infection. The most common immunodeficiency is antibody deficiency. The main clinical features of patients with antibody deficiency (also called hypogammaglobulinemia) are recurrent respiratory tract infections, with Streptococcus pneumoniae being the most commonly isolated bacterium (see, e.g., Fried and Bonilla, Clin Microbiol Rev., 2009; 22: 396-414; Busse et al., J Allergy Clin Immunol, 2002;109:1001-1004).

Over the past few decades, long-term use of polyvalent intravenous human immunoglobulin (IVIG, also referred to herein as "conventional IVIG") has been the main therapeutic option to reduce the severity and frequency of infections in immunocompromised individuals (e.g., antibody-deficient) (See e.g. Salehzadeh et al., J Microbiol Immunol Infect 2010, 43 (1): 11-17; Favre et al., Allergy 2005 60: 385-390). Infusion of IVIG (IVIG) results in a rapid peak in immunoglobulin G (IgG), followed by a decrease in IgG. The level of IgG immediately before the next infusion (ie, the residual level) is monitored to assess the adequacy of a particular infusion regimen. In general, PID patients treated with IVIG are infused every three or four weeks. Although IVIG infusion every three or four weeks has generally been successful in preventing serious lower respiratory tract infections in PID patients, there is a significant percentage of PID patients who continue to develop upper respiratory tract infections. , with associated morbidity and disease, during treatment. This is despite the fact that most immunocompromised patients receiving IVIG appear to have acceptable levels of total immunoglobulin as well as acceptable residual levels of anti-S. pneumonia IgG (see, e.g., Favre et al., Allergy 2005 60: 385-390). In addition, antibiotics have been used, but they are often ineffective.

Thus, the administration of exogenous pooled human IVIG (IVIG) has been a significant therapeutic option in clinical medicine for immunocompromised (eg, antibody-deficient) patients and appears to prevent serious lower respiratory tract infections. However, and in complete contrast, conventional IVIG therapy has failed to prevent or treat a significant incidence of less life-threatening upper respiratory tract infections (e.g. due to S. pneumonia) that occur in PID patients. (See, for example, Favre et al., Allergy 2005 60: 385-390, Simao-Gurge et al., Allergo Irnmunopathol 2017, 45: 55-62). In other words, while total IgG concentrations achieved by regular IVIG infusions prevent serious lower respiratory tract infections in PID patients, there remains a significant and significant incidence of less clinically serious upper respiratory tract infections. in patients with PID caused by S. pneumonia, which in turn leads to significant medical complications, reduced quality of life and indiscriminate use of broad-spectrum antibiotics in patients in an attempt to control infections.

This document discloses compositions and methods for treating an infection caused by Streptococcus pneumonia. In particular, the invention provides human hyperimmune globulin and compositions thereof for the prevention or treatment of pneumococcal infection. The invention relates to methods for producing hyperimmune globulins containing high titers of opsonic anti-pneumococcal antibodies, compositions based on them, and methods for using the compositions for the prevention and treatment of pneumococcal infection. The invention further provides methods for preventing or treating pneumococcal infections (e.g., upper respiratory tract infections (e.g., bronchitis, otitis media, sinusitis, etc.)) in immunocompromised subjects by administering hyperimmune globulin compositions of the invention (e.g., containing a high opsonic anti-pneumococcal antibody titer) in immunocompromised subjects.

As detailed herein (e.g., in the Examples), the invention provides a novel hyperimmune globulin formulation containing a high titer of opsonic anti-pneumococcal antibodies that is surprisingly and significantly different from conventional immune globulin preparations as well as other IVIG preparations (e.g., other hyperimmune drugs IVIG (IVIG)). In particular, as described in Examples 2-4, hyperimmune globulin prepared according to the methods of the invention was surprisingly found to contain an elevated titer of opsonic anti-pneumococcal antibodies that are functional, have a broad spectrum of activity against a variety of S. pneumoniae serotypes, enhance phagocytosis and cause elimination of S. pneumonia in vitro (eg, the antibodies are opsonophagocytic), regardless of the total amount of binding anti-pneumococcal antibodies (eg measured by ELISA) that are present in the composition. In other words, experiments performed during the development of embodiments surprisingly revealed that total IgG-binding antibody against S. pneumoniae capsular polysaccharide does not correlate with and is not predictive of the amount of functional opsonic antibody present in immune globulin derived from vaccinated host serum. (see, for example, Examples 2-4). Thus, low levels of binding antibody may be associated with high levels of protective opsonic antibodies and vice versa. Thus, measuring total antibody levels in immunoglobulin preparations alone does not predict or correlate with the protective efficacy of that preparation. Thus, in one embodiment, the invention provides for the identification and characterization of plasma and/or immune globulin compositions containing the desired functional opsonic antibody titer rather than one in which only the total amount of IgG is known (e.g., due to the detection of a lack of correlation between total titer anti-pneumococcal IgG and titer of opsonic anti-pneumococcal antibodies present in plasma or plasma-derived immune globulin from vaccinated donors).

In addition, a marked variability in serotype-specific opsonic titers was found in different commercial batches of conventional IVIG (see, e.g., Figs. 9A-9L, each of samples A-I represents a commercial IVIG) and that elevated titer to one serotype did not predict or correlate with elevated titer to any other serotype. This indicated that the individual increased response in any individual IVIG does not reflect an overall enhanced immune response against S. pneumonia, but rather a sporadic enhanced response(s) to very specific serotypes (see for example, Malgorzata et al Clin Diagn Lab Immunol 2004, 11 (6): 1158-1164). In contrast, the opsonic antibody titers observed for immune globulin obtained from immunized donors according to the invention were noted to be enhanced for all serotypes without exception (see, for example, Example 5, Fig. 8 and see Fig. 9A-9L, sample HIG - "hyperimmune globulin" according to the invention). Depending on the serotype, there was an increase in the titer of opsonic anti-pneumococcal antibodies in immune globulin from immunized donors from 3-256 times compared with various commercial batches of immune globulin (see, for example, Example 5 and Fig. 8, commercial/conventional IVIG samples (IVIG ) A-I compared with hyperimmune donor sera). Thus, in some embodiments of the invention, compositions and methods for preparing compositions of the invention (e.g., blood, plasma, and/or immune globulin compositions) containing an increased titer of opsonic anti-pneumococcal antibodies provide a heterogeneous composition containing titers of opsonic anti-pneumococcal antibodies specific for a plurality of pneumococcal serotypes (eg, 9, 10, 11, 12, 13, 14, 15, or more serotypes) and/or significantly increased (eg, 3-256 times higher) opsonic antibody titers compared to conventional commercial immune globulin.

Thus, due to the identification of an inconsistency between the total titer of binding anti-pneumococcal IgG antibodies and the titer of opsonic anti-pneumococcal antibodies present in plasma or plasma-derived immune globulin from the plasma of vaccinated donors described in this description, as another object of the invention, a composition of plasma and / or an immune globulin (e.g., anti-pneumococcal hyperimmune globulin) containing the desired increased titer of functional, opsonic anti-pneumococcal antibodies (e.g., at least 1:64 to about 1:8192 (e.g., regardless of the total titer of binding anti-pneumococcal antibodies (e.g., by combining plasma and/or immune globulin collected from the plasma of vaccinated donors and/or immune globulin identified as containing a high titer of opsonic anti-pneumococcal antibodies with each other and/or with vaccinated donor plasma and/or immune globulin, which may not contain high titer of opsonic anti-pneumococcal antibodies, but when combined with high-titer opsonic anti-pneumococcal antibody donor plasma and/or immune globulin, do not dilute the total opsonic titer to an undesirable level compared to the control sample))).

This description further describes the discovery that standard/conventional immunoglobulin pools from normal donors (e.g., used to produce commercially available standard/conventional IVIG (IVIG)) do not contain reliable and consistently high levels of opsonic antibodies for multiple serotypes of S. pneumonia, whereas an immune globulin composition (e.g., anti-pneumococcal hyperimmune globulin) containing a high titer of the broad-spectrum opsonic anti-pneumococcal antibodies of the invention against multiple serotypes, when administered intravenously, immediately provides functional, specific antibodies that promote phagocytosis and cause elimination of multiple S. pneumoniae serotypes by phagocytes. The invention is not limited to the serotype or number of S. pneumoniae serotypes for which functional, opsonic antibodies present in the hyperimmune globulin (eg, IVIG) of the invention promote opsonophagocytosis and/or elimination. In fact, the invention provides a composition (for example, a composition of hyperimmune plasma and/or hyperimmune globulin (for example, IVIG))) according to the invention, which contains broad-spectrum opsonic antibodies against at least 9 out of 12, 10 out of 12, 11 out of 12 or all 12 out of 12 of the following S. pneumonia serotypes: 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F (see, for example, Examples 2-5). Thus, the invention provides compositions and methods for their preparation, which contain an increased titer of specific opsonic anti-pneumococcal antibodies, which is at least 2 times, 3 times, 4 times, 5 times, 7 times, 10 or more times higher than the specific opsonic anti-pneumococcal antibody titer present in a control sample (e.g., at least 75% of pneumococcal serotypes selected from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A , 19F and 23F). In another embodiment, the invention provides a composition of hyperimmune globulin (for example, IVIG (IVIG)), which contains a high titer of broad-spectrum opsonic antibodies, at least against 70%, at least 75%, at least 80%, at least 85%, at least 90%, 95%, 98% or more for each of the serotypes present in the vaccine or multiple vaccines used to immunize one or more plasma donors from which immune globulin is derived.

Another advantage and object of the invention is to provide a hyperimmune globulin (e.g. IVIG) composition containing broad spectrum opsonic antibodies specific for S. pneumonia to a patient (e.g. an immunocompromised patient (e.g. a PID patient (PIDD)) , for the treatment (e.g., therapeutic and/or prophylactic) of a pneumococcal infection in a patient (e.g., by inhibiting the growth of S. pneumoniae and/or the clearance of S. pneumoniae from the patient's blood). (IVIG)), containing a broad-spectrum opsonic antibody specific for S. pneumonia, to a patient (e.g., an immunocompromised patient (e.g., a patient with PID (PID)D)) for the purpose of treating a pneumococcal infection in a patient (e.g., by improving or enhancing the clearance of S. pneumonia from the patient's blood).Additionally, the invention relates to a hyperimmune globulin composition (for example, IVIG) containing broad-spectrum opsonic antibodies specific for S. pneumonia, for a patient (for example, an immunocompromised patient ( for example, a patient with PID (PID-D)), to prevent an upper respiratory tract infection in a patient that cannot be prevented with conventional IVIG therapy. The invention is not limited to the type of upper respiratory tract infection being prevented and/or treated and may include, but is not limited to, rhinosinusitis (sinusitis), otitis media, pharyngitis, epiglottitis, laryngotracheitis, and laryngotracheobronchitis. Similarly, the compositions and methods of their use (e.g., administration) are used to prevent and/or treat signs or symptoms of an upper respiratory tract infection, including, but not limited to, cough, sneezing, nasal discharge, nasal congestion, runny nose, fever, sore or sore throat and nasal breathing.

The invention is not limited to the type of streptococcal infection being treated (eg prophylactically and/or therapeutically). In fact, any streptococcal infection caused by the streptococcal group of bacteria can be treated. There are over 90 different strains of Streptococcus pneumonia (S. pneumonia) bacteria (known as serotypes), some of which cause more serious infection than others. Symptoms of pneumococcal infection can vary depending on the type of infection. Common symptoms include: high temperature (fever) 38C (100, 4F), aching and sharp pains and / or headache. Pneumococcal infections usually fall into one of two categories: non-invasive pneumococcal infections - these occur outside of major organs or blood and tend to be less severe; and invasive pneumococcal infections, they occur within a major organ or bloodstream and tend to be more serious. Compositions of the invention and methods of using them find use in the treatment (eg, therapeutic and/or prophylactic) of both non-invasive and invasive pneumococcal infections.

In one embodiment, the invention provides compositions and methods for preparing a composition containing pooled plasma samples (e.g., plasma from multiple donors (e.g., donors who have been vaccinated with one or more pneumococcal vaccines)) that contain high titers of opsonic anti-pneumococcal antibodies. Thus, it is an object of the invention to provide methods for preparing compositions (eg, blood, plasma and/or immune globulin compositions) containing a high titer of opsonic anti-pneumococcal antibodies. In one embodiment of the invention, one or more healthy adults (eg, people with no known disease) are administered a pneumococcal immunogen, a recombinant pneumococcal protein, or a combination thereof. In some embodiments, the S. pneumonia immunogen is an S. pneumoniae cell membrane sugar (eg, a polysaccharide). In some embodiments, the S. pneumonia immunogen is a conjugate vaccine (e.g., conjugated with a carrier and/or an adjuvant (e.g., protein or other carrier molecule). In some embodiments, the S. pneumonia immunogen is a non-conjugated vaccine. In some embodiments of the invention, the conjugate vaccine or non-conjugated vaccine contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, or more different immunogens (e.g., from the same number of different S. pneumoniae serotypes) In some embodiments, one or more different S. pneumoniae serotypes include, but are not limited to, serotypes 1, 2, 3 , 4, 5, 6A, 6B, 7A, 7B, 7C, 7D, 7E, 7F, 8, 9A-9V, 12, 14, 18C, 19A-19F, 23A-23F, and 25. In some embodiments, one or more different S. pneumonia serotypes are selected from any of the more than 90 identified S. pneumonia serotypes. In some embodiments, one or more different serotypes of S. pneumoniae have been recently identified.

One or more healthy individuals (eg, individuals without any known disease) may be administered pneumococcal immunogen, recombinant pneumococcal protein, or a combination thereof, as found in a commercial pneumococcal vaccine. The invention is not limited to the type of commercial pneumococcal vaccine. In fact, any pneumococcal vaccine known in the art may be used, including, but not limited to, pneumococcal conjugate vaccine (PCV13 or PREVNAR13, Wyeth Pharmaceuticals, Collegeville, PA), SYNFLORIX, and/or pneumococcal polysaccharide vaccine (PPSV23 or PNEUMOVAX23, Merck Sharp & Dohme Corp., North Wales, PA). In one embodiment of the invention, one or more healthy individuals receive a first or primary vaccination with a first pneumococcal vaccine and a subsequent booster vaccination with a first pneumococcal vaccine or another pneumococcal vaccine. For example, in one embodiment, one or more healthy individuals receive a first or primary vaccination/immunization with a first pneumococcal vaccine (e.g., PREVNAR) and then receive a booster vaccination/immunization (e.g., after 2 weeks, 4 weeks, 6 weeks , 8 weeks, 10 weeks, 12 or more weeks after primary vaccination/immunization) with a second pneumococcal vaccine (eg, PNEUMOVAX23). At a time point following sequential vaccination (eg, 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 or more weeks after sequential vaccination), serum/plasma is collected from vaccinated healthy human plasma donors. Plasma from vaccinated donors can be pooled (with each other and/or with plasma from unvaccinated donors) followed by collection of immune globulin. Methods for collecting plasma as well as collecting immune globulin are well known to those skilled in the art.

In one embodiment, the present invention provides a method for producing hyperimmune globulin with a high titer of an opsonophagocytic antibody to Streptococcus pneumoniae (e.g., a titer that is at least 2-fold, 3-fold, 4-fold, 5-fold, 7 to 10, or more times for at least about 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85%, or more) of S. pneumonia serotypes 1, 3, 4, 5 , 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F) higher than the titers of specific opsonic anti-pneumococcal antibodies present in the control sample against the same serotypes; or titer specific for 70% or more of S. pneumonia serotypes selected from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23 by 2 or more times (e.g., 3-25 or more times) higher than the titer of opsonophagocytic antibodies present in the control sample, specific for the same serotypes of S. pneumonia; or a titer in the range of 1:64 to 1:8192 (e.g., for at least 50% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more) pneumoniae serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) according to the opsophagocytic elimination assay described herein), wherein the method includes the steps of immunizing healthy adult donors plasma of a person aged 18-60 years with a primary polyvalent vaccine against S. pneumonia, followed by immunization with a booster polyvalent vaccine against S. pneumonia, which is different from the primary vaccine; plasma collection from plasma donors after booster immunization; pooling the plasma of vaccinated donors in order to obtain pooled plasma containing a high titer of opsonophagocytic antibodies against S. pneumonia; and obtaining immune globulin from the pooled plasma. In a further embodiment of the invention, the method includes converting the resulting immune globulin into a form suitable for intravenous injection. The immune globulin may be provided in solution and/or the pH and ionic strength of the solution may be adjusted to make it suitable for intravenous injection. The invention is not limited to the number of people vaccinated in accordance with the methods described in this description. For example, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400 or more healthy adult human plasma donors may be vaccinated and plasma collected from the donors. In one embodiment, pooled plasma is obtained from a plasma pool of 1000 or more different healthy vaccinated adult human plasma donors. In one embodiment, the pooled plasma contains an opsonophagocytic antibody titer that is at least 2-fold, 3-fold, 4-fold, 5-fold, 7-fold, 10-fold or more for, at least 55% or more (e.g., 60%, 65%, 70%, 75%, 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) compared to the titers of specific opsonic anti-pneumococcal antibodies present in a control against the same serotypes (e.g., plasma or plasma-derived immune globulin pooled for 1000 or more random unvaccinated plasma donors). person). In another embodiment, the pooled plasma contains an opsonophagocytic antibody titer specific for 70% or more of S. pneumoniae serotypes selected from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23, which is 2 or more times (e.g., 3-25 times or more) higher than the titer of opsonophagocytic antibodies present in a control sample specific for the same S. pneumoniae serotypes (e.g., plasma or plasma-derived immune globulin pooled for 1000 or more random unvaccinated human plasma donors). In yet another embodiment, the pooled plasma contains an opsonophagocytic antibody titer in the range of 1:64 to 1:8192 (e.g., at least 50% or more (e.g., 60%, 65%, 70%, 75% , 80%, 85% or more) of S. pneumonia serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) as determined by the opsonophagocytic elimination assay described herein. In addition, the invention provides hyperimmune globulin obtained in accordance with the above method. The hyperimmune globulin thus obtained can be used in various ways. For example, hyperimmune globulin can be used in a method of treating an S. pneumoniae infection in a subject (eg, comprising administering to the subject a therapeutically effective amount of hyperimmune globulin). Additionally, hyperimmune globulin may be used in a method of administering immunotherapy to a subject (eg, comprising administering to the subject a therapeutically effective amount of hyperimmune globulin).

In one embodiment, the invention further provides a method for producing an immune globulin having enhanced opsonophagocytic bactericidal activity against at least seven serotypes of Streptococcus pneumoniae (e.g., serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F), for the prevention or treatment of S. pneumonia infection, comprising the steps of immunizing healthy adult human plasma donors with primary immunization with a polyvalent S. pneumoniae conjugate vaccine, followed by immunization with a polyvalent S. pneumoniae polysaccharide vaccine other than the primary vaccine; collection and pooling of plasma from immunized plasma donors; and obtaining immune globulin from the pooled plasma, wherein the immune globulin contains a titer of opsonophagocytic antibodies specific for each of at least seven S. pneumonia serotypes, which is two or more times higher than the titer of opsonophagocytic antibodies present in the control sample. antibodies specific for each of the at least seven S. pneumoniae serotypes (eg, immune globulin obtained from plasma pooled from 1000 or more random unvaccinated human plasma donors). In addition, the invention provides an immune globulin obtained by the method described above. The immune globulin thus obtained can be used in various ways. For example, immune globulin can be used in a method of treating an S. pneumoniae infection in a subject, comprising administering to the subject a therapeutically effective amount of immune globulin. Additionally, the immune globulin can be used in a method of administering immunotherapy to a subject, comprising administering to the subject a therapeutically effective amount of immune globulin.

Experiments conducted during the development of embodiments of the invention revealed that in a significant number of patients treated with IVIG (IVIG), residual concentrations of antibodies against S. pneumonia decreased below the level that is considered protective. For example, as shown in FIG. 10, it was found that a significant percentage of patients with primary immunodeficiency disease treated with conventional IVIG (IVIG) have residual levels of total binding serotype-specific antibodies against S. pneumonia, reduced below the protective level (below 1.2 μg / ml).

All immune globulins fractionated and/or isolated from serum and used for intravenous infusion contain little, if any, measurable levels of IgA, which may explain the inability of immune globulin to protect at mucosal surfaces (eg, thereby leading to an increased incidence of infection of the upper airways in immunocompromised patients receiving conventional IVIG). Thus, although understanding of the mechanism is not required for the practice of this invention, and although this invention is not limited to any particular mechanism of action, in some embodiments of the invention, a hyperimmune globulin composition of the invention is provided, containing a high concentration of broad spectrum opsonic antibodies against S. pneumonia (e.g., as provided in hyperimmune globulin compositions of the invention) allows gradient diffusion of higher concentrations of functional, opsonic, broad-spectrum anti-pneumococcal IgG across mucosal surfaces, thereby providing mucosal protection and reducing the incidence of upper respiratory tract infection (e.g., compared to with conventional IVIG, which does not contain a high concentration of broad-spectrum opsonic antibodies against S. pneumonia and, therefore, gradient diffusion is significantly less or absent, resulting in a high incidence of upper respiratory tract infections in immunocompromised patients treated with conventional IVIG (IVIG))).

Thus, the invention provides a hyperimmune globulin obtained by any of the methods described in this description, which contains an increased titer of opsonic antibodies specific for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or more different S. pneumonia serotypes, including but not limited to serotypes 1, 2, 3, 4, 5 , 6A, 6B, 7A, 7B, 1C, 7D, 7E, 7F, 8, 9A-9V, 12, 14, 18C, 19A-19F, 23A-23F, and 25. Although an understanding of the mechanism is not required to practice this invention, and although the invention is not limited to any particular mechanism of action, pooling of vaccinated donor sera provides desired levels (e.g., therapeutically and/or prophylactically protective levels) of broad-spectrum opsonic IgG against all serotypes present in one or more vaccines that are used to vaccinate healthy adult human plasma donors (eg, despite any variation from serotype to serotype and/or pooling dilution). Accordingly, in some embodiments of the invention, any individual difference in the concentration of opsonic serotype-specific IgG can be normalized without compromising the protective levels of such antibodies by pooling sera. Purification of immune globulin from serum did not lead to a deficiency of any serotype-specific opsonic anti-pneumococcal IgG, nor its functional activity.

Preparation of purified immunoglobulin. In one embodiment of the invention, a purified immunoglobulin is obtained that contains a high titer of specific opsonic anti-pneumococcal antibodies. The term "high titer" in this context means the presence of an antibody in an amount that is 2 or more times (eg, up to 10-20 or more times higher than that found in a normal population of 1000 random samples).

The blood product can be obtained by (i) collecting and purifying immunoglobulin from a donor who has been vaccinated (e.g., according to the method of the invention), wherein the immunoglobulin contains high titers of specific opsonic anti-pneumococcal antibodies, or (ii) pooling donor immunoglobulin from multiple individuals (e.g., 100, 200, 200-500, 500-1000 or 1000 or more subjects) who have been vaccinated according to the method of the invention.

Plasmapheresis can be used to collect serum/plasma from a vaccinated donor. The term plasmapheresis describes a method in which blood is removed from an animal's body, separated into cellular and plasma components, then the cells are returned to the animal's body and the plasma is stored. High volume plasmapheresis requires that the extracted plasma be replaced with a suitable fluid, and if this is done it is often referred to as plasma exchange. Any components found in plasma can be removed by plasma exchange. Plasma thus recovered for commercial sale is available for use in the preferred embodiment of the present invention.

In some embodiments of the invention, it is preferable to identify and maintain a constant donor group by repeated sampling of small amounts of blood, for example, drawing blood from people once a month. The frequency of withdrawal can affect the amount that can be safely retrieved. In general, it is desirable to collect as much blood as possible over time without compromising the health of the donor. This may dictate taking small amounts at a high frequency, or as much as possible at a reduced frequency. Donor blood volume can be estimated using standard formulas available from the CDC. Methods for collecting blood and plasma are generally standard and well known to those skilled in the art. Either method can be used to achieve the desired results.

After plasma isolation, immune globulin (antibodies) can be purified from other cellular products. This can be achieved using various protein isolation techniques known to those skilled in the art of immunoglobulin purification, such as Cohn fractionation, ion exchange, affinity purification, etc. Means for obtaining and characterizing antibodies are well known in the art. For example, serum samples can be run through Protein A or Protein G Sepharose columns for IgG binding (depending on isotype). The bound antibodies can then be eluted with, for example, citrate buffer pH 5.0. The eluting fractions that contain Ab are dialyzed against isotonic buffer. Alternatively, the eluate is additionally passed through an anti-immunoglobulin sepharose column. The Ab can then be eluted with 3.5 M magnesium chloride. Ab thus purified can then be tested for binding activity, e.g. by isotype-specific ELISA and immunofluorescence staining of target cells, or for opsonic (opsophagocytic) activity (e.g., using one or more the described opsonic elimination assays described herein).

In one embodiment of the invention, if plasma samples are collected and mixed (for example, from a plurality of subjects (for example, 100, 200, 200-500, 500-1000, 1000 or more subjects) immunized in accordance with the methods described herein) , then the mixed plasma or the immune globulin derived from it (for example, isolated and/or fractionated) contains seroprotective titers of antibodies against measles, diphtheria and/or poliomyelitis (for example, contains titers of antibodies against measles, diphtheria and/or poliomyelitis, providing a subject to whom administering, with a mixed plasma or immunoglobulin-derived composition, serum levels of antibodies specific for measles, diphtheria and poliomyelitis to prevent or protect against infection by these pathogens). In another embodiment of the invention, if plasma samples from multiple vaccinated subjects are mixed, then the mixed plasma or immune globulin derived from it (for example, fractionated) contains seroprotective titers of antibodies against measles, diphtheria, poliomyelitis and/or tetanus (for example, contain antibody titers against measles, diphtheria, poliomyelitis and/or tetanus, providing the subject to whom the administration is carried out with a mixed composition of plasma or immune globulin derived from it, serum levels of antibodies specific for measles, diphtheria, poliomyelitis and/or tetanus, to prevent or protect against infection with these pathogens (e.g., consistent with FDA-recommended antibody titer levels (e.g., for treatment of an immunodeficiency disease and/or treatment or prevention of infection in an immunocompromised subject)). the plasma pool contains plasma samples obtained from 1000-3000 or more (eg, more than 1000, 1250, 1500, 1750, 2000, 2500, 3000, 3500, 4000 or more human subjects). In one embodiment, the pooled plasma is used to produce an immunoglobulin (eg, for intravenous administration to a subject). In one embodiment, pooled plasma and/or immunoglobulin provides a therapeutic benefit to a subject receiving pooled plasma and/or immunoglobulin that is not achievable by administering a mixture of plasma samples (or immune globulin derived from it) obtained from 1000 or more number of random people. The invention is not limited to the type of therapeutic benefit. Indeed, various therapeutic benefits can be achieved, including those described herein (eg, treatment of pneumococcal infection (eg, upper respiratory tract infection)). In one embodiment of the invention pooled plasma and/or immune globulin has improved opsonophagocytic anti-pneumococcal properties compared to a mixture of plasma samples obtained from 1000 or more random people or immune globulin derived from it. For example, in one embodiment, the pooled plasma has enhanced opsonophagocytic anti-pneumococcal properties against ten or more different pneumococcal serotypes (eg, 10, 10-20, 20-30 or more serotypes). In yet another embodiment of the invention, enhanced anti-pneumococcal opsonophagocytic properties reduce and/or prevent infection in a subject administered the composition for a period of time that is longer and is not achieved in a subject administered a mixture of plasma samples obtained from 1000 or more random (e.g. , unvaccinated) human subjects. In one embodiment, pooled plasma and/or immune globulin derived therefrom reduces the incidence of infection in a subject to whom the composition has been administered. In another embodiment of the invention pooled plasma and/or derived from it immune globulin reduces the required number of days of administration of antibiotics to a subject who was administered pooled plasma and/or immunoglobulin (for example, to treat an infection). In yet another embodiment, pooled plasma and/or plasma-derived immune globulin increases the residual level of circulating anti-pneumococcal opsonophagocytic antibodies in a subject (e.g., increases the titer of opsonophagocytic antibodies specific for ten or more pneumococcal serotypes (e.g., thereby providing protective levels of anti-pneumococcal opsonophagocytic antibodies between scheduled administration dates of pooled plasma and/or pooled immune globulin not maintained in a subject administered a mixture of plasma samples obtained from 1000 or more random individuals or pooled immune globulin) ). In one embodiment, the composition comprising the pooled plasma samples further comprises a pharmaceutically acceptable carrier (eg, any natural or non-natural carrier(s) known in the art). In one embodiment of the invention, the average fold increase in the titer of anti-pneumococcal opsonophagocytic antibodies is, in a subject who was administered immunoglobulin obtained from pooled plasma in accordance with the invention, is at least 2 times, 3 times, 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 or more times at a time point of at least 1 to 14 days after administration (e.g. , 14 days, 15 days, 16 days, 17 days, 18 days, 19 or more days) immune globulin. The invention is not limited to the amount of immune globulin administered to a subject. In one embodiment, the subject is administered 100-5000 mg/kg immunoglobulin once or daily for two or more days (eg, 2, 3, 4 or more consecutive days). In another embodiment of the invention, such doses are administered intermittently, for example, every week, every two weeks, every three weeks, every four weeks, etc. In one embodiment, the subject is administered 750-1500 mg/kg of immune globulin on the first day and 750-1500 mg/kg of immune globulin on day 2. In one embodiment, the subject is administered 1500 mg/kg immune globulin on the first day and 750 mg/kg immune globulin on day 2. In another embodiment, the subject is administered 750 mg/kg immune globulin on the first day and 750 mg/kg immune globulin on day 2. In one embodiment, the subject is given immune globulin on day one, given immune globulin on day 2 as needed, and then given immune globulin again every 21 days. In one embodiment, the subject is given immune globulin on day one, optionally given immune globulin on day 2, and then given immune globulin again every 28 days. In one embodiment, pooled plasma and/or immune globulin derived from it reduces the incidence of pneumococcal infection in a subject to whom the composition has been administered. In another embodiment, pooled plasma and/or plasma-derived immune globulin increases the residual level of circulating anti-pneumococcal opsonophagocytic antibodies and increases the residual level of circulating specific antibodies against measles, diphtheria, poliomyelitis and/or tetanus (for example, thereby increasing the level of titers opsonophagocytic anti-pneumococcal antibodies specific for pneumococcal serotypes and measles, diphtheria, poliomyelitis, and/or tetanus (e.g., thereby providing protective levels of anti-pneumococcal opsonophagocytic antibodies and specific antibodies against measles, diphtheria, poliomyelitis, and/or tetanus between scheduled introduction dates of the pooled plasma and/or plasma-derived immune globulin not maintained in a subject who was administered a mixture of plasma samples obtained from 1000 or more random people (for example, unvaccinated subjects) or immune globulin derived from it)).

In some embodiments of the invention, the plasma samples and/or antibodies contain donor and/or purchased samples of body fluids, such as individual samples of blood or blood components (eg, plasma). Such samples may be cleaned and/or screened for the presence of pathogens or other contaminants (eg, before or after pooling). Multiple donor antibody samples (eg, donated plasma samples or other antibody-containing samples) can be pooled to provide pooled plasma samples. In some embodiments, pooled antibody samples are purified, screened, and/or concentrated. In one embodiment of the invention, pooling of samples (eg, 1000 or more samples) is performed in a manner that uses the fewest possible number of samples (eg, generated by the compositions and methods described herein), but which still retains the desired standardized and elevated levels of anti-pneumococcal opsonophagocytic antibodies.

As described herein, in some embodiments, the plasma of subjects who have been administered immunogenic substances (eg, vaccines) is used to obtain elevated levels of anti-pneumococcal opsonophagocytic antibodies in the subject. The invention is not limited to the type of vaccine and/or antigen (eg, S. pneumoniae antigen) used to administer to a subject (eg, a donor) to induce the expression of specific antibodies. In some embodiments, the antigen is an S. pneumoniae antigen, or a fragment or component thereof. In some embodiments, the antigen is one or more polysaccharides (eg, unconjugated or conjugated to a carrier or protein). In some embodiments, the antigen (eg, S. pneumoniae antigen) is a vaccine containing components capable of inducing specific antibodies (eg, antibodies specific for several different S. pneumoniae serotypes). In some embodiments of the invention, the vaccine is a commercially available vaccine. The invention is not limited to a vaccine. In fact, a variety of vaccines (eg, S. pneumoniae vaccines) may be used, including, but not limited to, PREVNAR, SYNFLORIX, PNEUMOVAX, and others known in the art. Likewise, the invention is not limited to the type or method of administration/immunization. Indeed, you can use any method/type of immunization, including, but not limited to, the methods described in US publications No. US 2008026002, US 2007009542; US2002094338; US2005070876; US2002010428; US2009047353; US2008066739; and US 2002038111), each of which is incorporated into this description by reference in its entirety. Likewise, the invention is not limited to the composition of the vaccine (eg S. pneumonia vaccine). In fact, any drug can be used, including, but not limited to, those described in US 2002107265, which is hereby incorporated by reference in its entirety. In some embodiments of the invention, the vaccine is a polyvalent vaccine, which add additional antigens (see, for example, US 2007161088, US 2006121059, each of which is included in this description by reference in its entirety). In some embodiments, the antigens are purified prior to use in a vaccine (eg, conjugate vaccine) (see, for example, US 2008286838, which is incorporated herein by reference). Methods for cultivating microorganisms useful in the manufacture of pneumococcal conjugate vaccines are described in US 2010290996, which is incorporated herein by reference in its entirety. Alternatively, some embodiments of the invention use antigens (eg, S. pneumonia antigens) that are not conjugated to a carrier protein (see, for example, US 2009136547, which is hereby incorporated by reference in its entirety). In some embodiments of the invention, immunomodulators are used (see, for example, US 2004156857, US 5985264 and WO 11041691, each of which is incorporated into this description by reference in its entirety). In some embodiments of the invention, therapeutic antibodies are produced in the body of a donor who is administered an antigen (eg, S. pneumoniae antigen) and/or a vaccine in accordance with the methods described in WO 05070458; US2009191217; and WO 10094720, each of which is incorporated herein by reference in its entirety. In some embodiments, antigens (e.g., vaccines (e.g., conjugated or non-conjugated vaccines)) are used to generate antibodies (e.g., present in serum and/or plasma) that are useful against infectious pathogens (e.g., as described, for example, in US 2003099672, WO 0062802, each of which is incorporated into this description by reference in its entirety).

In some embodiments, a polysaccharide vaccine is used (e.g., containing multiple S. pneumoniae serotypes (e.g., containing purified polysaccharides from 1, 2, 3, 4 or more or all 23 of the following S. pneumoniae serotypes: 1, 2, 3, 4, 5, 6b, 7F, 8.9N, 9V, 10A, PA, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F, and 33F). of the invention, and the invention is not limited to any particular mechanism of action, in some embodiments of the invention, an antigen (e.g., S. pneumoniae antigen) or vaccine that stimulates B cells (e.g., plasma cells) to generate and secrete a specific (e.g., specific for S. pneumonia) immunoglobulin without the help of T cells finds use in the invention.

In some embodiments, a conjugate vaccine is used that contains capsular polysaccharides (eg, from a variety of S. pneumoniae serotypes) covalently linked to a carrier and/or adjuvant (eg, diphtheria toxoid CRM 197). Although it is not necessary to understand the mechanism for the practice of this invention, and this invention is not limited to any particular mechanism of action, in some embodiments of the invention, any antigen (for example, S. pneumonia antigen) or vaccine that stimulates B cells (for example, plasma cells) to generate and secrete a specific (e.g., S. pneumoniae IgM and/or IgG specific) immunoglobulin by interacting with specific type 2 helper T cells) and/or the production of memory B cells (e.g., memory B cells, specific for S. pneumonia) finds use in the invention.

Thus, in some embodiments, the present invention provides methods for stimulating high levels of opsonophagocytic anti-pneumococcal antibodies in a donor, which comprises administering to an animal, e.g., a human, a pharmaceutically acceptable composition comprising an immunologically effective amount of an antigenic composition (e.g., an S. pneumoniae antigen composition). The composition may contain partially or substantially purified antigens (e.g., S. pneumoniae antigens (e.g., polysaccharide, protein and/or peptide epitopes derived from natural or recombinant sources, which can be obtained naturally or chemically synthesized or alternatively , obtained in vitro from recombinant host cells expressing DNA segments that encode such epitopes)).

Methods for determining the effectiveness of immunization (for example, determining the level of antibody titers specific for S. pneumonia) are known in the art, and any known method can be used to evaluate the effectiveness of immunization (for example, the effectiveness of inducing opsonophagocytic anti-pneumococcal antibodies (for example, described in this description or an opsonophagocytic elimination assay known in the art)). In some embodiments of the invention, detection methods are used to evaluate the effectiveness of a vaccine (eg, pneumococcal conjugate vaccine), as described, for example, in US 2005260694; US 4308026; US 4185084; or US 2005208608, each of which is incorporated into this description by reference in its entirety.

In some embodiments, kits and methods are provided that identify samples and/or pools with specific antibody titers (eg, elevated opsonophagocytic anti-pneumococcal antibody titers). In one embodiment, a suitable amount of a detection reagent (eg, an antibody specific for an antibody, antigen, or other reagent known in the art) is immobilized on a solid support and labeled with a detectable agent. Antibodies can be immobilized on various solid substrates by known methods. Suitable solid support materials include materials containing a membrane or coating applied to or attached to sticks, balls, cups, flat packs, or other solid support. Other solid materials include cell culture plates, ELISA plates, tubes, and polymeric membranes. The antibodies can be labeled with a detectable agent such as a fluorochrome, radioactive label, biotin or another enzyme such as horseradish peroxidase, alkaline phosphatase and 2-galactosidase. If the detectable reagent is an enzyme, means for detecting the detectable reagent may be included in the kit. A suitable means for detecting a detectable agent uses an enzyme as the detectable agent and an enzyme substrate that changes color upon contact with the enzyme. In addition, the kit may include a tool for evaluating the product of the analysis, such as a color chart or a numerical reference chart. Some suitable methods for characterizing samples and pools are given in the references included in this description by reference. This invention is not limited to the method used to characterize samples and pools as containing increased titer. Additionally, assays can be used to determine the level of anti-pneumococcal opsonophagocytic antibodies in plasma, immune globulin and/or pools thereof (eg, as described in the Examples).

In some embodiments, compositions (eg, antibody samples, pooled plasma samples, immune globulins, etc.) are provided in which antibodies have been purified and/or isolated from one or more contaminants. Human immunoglobulins were first isolated on a large scale in the 1940s by F.J. Cohn. In some embodiments of the invention, the methods proposed by Cohn (Cohn et al., J. Am. Chem., 1946; 68: 459-475, incorporated herein by reference in its entirety), or modified methods according to Cohn, are used to obtain immune globulins. In some embodiments of the invention, various purification and isolation methods are used to obtain substantially unmodified, unaltered, undenatured and/or native immune globulin molecules in high purity. Typical techniques are given, for example, in US patent No. 4482483, included in this description by reference in its entirety. In some embodiments, the compositions (eg, antibody pools) contain >50% immunoglobulin (eg, >60%, >70%, >80%, >90%, >95%, >99%). Various methods can be used to prepare such compositions, including, for example, standard protein purification and isolation methods, as well as fractionation of biological fluids (eg, plasma). A description of the fractionation of antibodies for use in immunotherapy, is contained, for example, in US patent No. 4346073 and other references presented in this description, each of which is incorporated by reference in its entirety. In some embodiments of the invention, immunoglobulins are purified by fractional precipitation, ion exchange chromatography, size exclusion chromatography, ultrafiltration, affinity chromatography, or any suitable combination thereof (see, for example, US patent No. 7597891, US patent No. 4256631, US patent No. 4305870, Lullau et al ., J. Biol., 1996, 271: 16300-16309, Corthesy, Biochem, Soc, 1997, 25: 471-475 and Crottet et al., Biochem. J 1999, 341: 299-306, which are included in this description by reference in full).

The composition of the invention (e.g., pooled plasma and/or immune globulin derived from it) can be administered by any suitable route, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and, if desired, for topical treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intra-arterial, intraperitoneal or subcutaneous administration. In addition, the compositions of the invention may be administered by pulse infusion, especially at lower doses. Dosing may be by any suitable route, for example by injection, such as intravenous or subcutaneous injection, depending on whether the administration is acute or chronic.

The composition of the invention may be formulated, measured and/or administered in accordance with good medical practice. Factors to be considered in this context include the particular disorder being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, route of administration, schedule of administration, and other factors known to practitioners. Compositions of the invention need not, but may optionally, be formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of antibody present in the formulation, the type of disorder or treatment, and other factors discussed above. They are generally administered at the same doses and routes of administration as described herein, or from about 1% to about 99% of the doses described herein, or at any dose and by any route empirically/clinically determined to be appropriate.

For the prevention or treatment of a disease, the appropriate doses of the composition of the invention (when used as monotherapy or in combination with one or more other additional therapeutic agents) may depend on a number of factors, including the type of disease being treated, the type of antibody, patient size, surface area body, age, specific compound administered, sex, time and route of administration, general health, interactions with concomitantly administered other drugs, severity and course of disease, whether the antibody is administered for prophylactic or therapeutic purposes, prior therapy, and the patient's clinical history .

Exact dosages may be determined by the individual physician, taking into account the patient to be treated. Doses and administration are adjusted to provide sufficient levels of active fragment (eg plasma pool) or to maintain the desired effect. Additional factors that may be taken into account include the severity of the condition; age, body weight and gender of the patient; diet, time and frequency of administration, drug combination(s), response sensitivity and tolerability/response to therapy. Long acting pharmaceutical compositions may be administered every 3-4 days, every week, or every two weeks, four weeks, six weeks, eight or more weeks, depending on the half-life and clearance rate of the particular drug.

The composition according to the invention can be administered to the patient once or in several doses. Depending on the type and severity of the disease, from about 1 μg/kg to 5000 mg/kg (for example, from 0.5 mg/kg to 1500 mg/kg) of the composition of the invention may be a potential initial dose for administration to a patient, whether, for example, , once or in several doses or as a continuous infusion. As described herein, additional drugs or agents (eg, antimicrobials (eg, antibiotics, antivirals, antifungals, etc.), other immunoglobulin compositions (eg, conventional IVIG), anti-inflammatory and/or healing compounds, etc.) may be administered simultaneously with the pooled plasma composition of the invention. A typical daily dose of such an agent may be from about 1 to 100 mg/kg or more. With repeated administration over several days or longer, depending on the condition, treatment can generally be maintained until the desired suppression of disease symptoms occurs. One typical dose of a composition of the invention will be in the range of about 0.05 mg/kg to about 10 mg/kg. Thus, one or more doses of about 0.5 mg/kg, 2.0 mg/kg, 4.0 mg/kg, or 10 mg/kg (or any combination thereof) may be administered to a patient. Such doses may be administered intermittently, for example every week or every two to three weeks. The physician can easily control the therapeutic administration of the composition of the invention and can, in turn, determine whether higher or lower doses of the composition should be administered.

Compositions of the invention may be administered to a patient (eg, intravenously, orally, intramuscularly, subcutaneously, etc.) in a pharmaceutically acceptable carrier such as saline. Such methods are well known to those skilled in the art.

Accordingly, in some embodiments of the present invention, the composition of the invention may be administered to a patient alone or in combination with other drugs or in pharmaceutical compositions where it is mixed with excipient(s) or other pharmaceutically acceptable carriers. In one embodiment of this invention, the pharmaceutically acceptable carrier is pharmaceutically inert. Depending on the condition to be treated, the pharmaceutical compositions may be formulated and administered systemically or topically. Formulation and administration methods can be found in the latest edition of Remington's Pharmaceutical Sciences (Mack Publishing Co, Easton Pa.). Suitable methods may, for example, include oral or transmucosal administration; as well as parenteral administration, including intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal or intranasal administration.

For injection, the composition of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution or phosphate buffered saline. For introduction into tissues or cells in the preparation, penetrants are used that correspond to the specific barrier through which penetration must be ensured. Such penetrants are generally known in the art.

In other embodiments, the compositions of this invention (eg, pharmaceutical compositions) may be formulated using pharmaceutically acceptable carriers well known in the art at doses suitable for oral administration. Such carriers make it possible to formulate pharmaceutical compositions in the form of tablets, pills, capsules, liquids, gels, syrups, suspensions, suspensions, and the like. for oral or nasal administration by the patient to be treated.

Pharmaceutical compositions suitable for use in this invention include compositions in which the active ingredients are contained in an amount effective to achieve the intended purpose. For example, an effective amount of a composition of the invention may be an amount that results in growth inhibition and/or killing of bacteria in the subject's body. Determination of effective amounts is within the skill of the art, especially in light of the disclosure herein.

In addition to the active ingredients, the pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers containing excipients and additional substances which facilitate the formulation of the compositions of the invention into preparations which can be used in the pharmaceutical field.

Pharmaceutical compositions of this invention can be produced in a manner which is known per se (eg, by conventional mixing, dissolving, granulating, panning, levigating, emulsifying, encapsulating, trapping, or lyophilizing methods).

Pharmaceutical preparations for parenteral administration include aqueous solutions of the compositions in water-soluble form. In addition, suspensions of the compositions may be prepared as suitable oily injections. Suitable lipophilic solvents or carriers include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may further contain suitable stabilizers or agents that increase the solubility of the compositions to provide highly concentrated solutions.

Compositions of the invention formulated in a pharmaceutically acceptable carrier may be formulated, placed in an appropriate container, and labeled for the treatment of said condition. Conditions listed on the label may include the treatment or prevention of a viral or bacterial infection.

The pharmaceutical composition may be in the form of a salt and may be formed with many acids including, but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend to be more soluble in aqueous or other protic solvents than their corresponding free base forms. In other cases, the preferred formulation may be a lyophilized powder in 1-50 mM histidine, 0.1-2% sucrose, 2% mannitol, with a pH in the range of 4.5 to 5.5, which is combined with a buffer before use.

Compositions of the invention (eg, pneumococcal hyperimmune globulin) may be combined with additional agents (eg, antibodies, antibody fragments, antibody-like molecules, monoclonal antibodies, antimicrobials, other immune globulin formulations (eg, conventional IVIG or other proteins or small molecules) to enhance the immunotherapeutic and/or anti-inflammatory effect.Such additional agents can be obtained by recombinant, synthetic, in vitro, etc. The present invention is not limited to the types of additional agents with which antipneumococcal hyperimmune globulin is co-administered and/or combined In some embodiments, recombinant or synthetic antibodies (e.g., humanized monoclonal antibodies) or antibody fragments (e.g., directed to a specific pathogen or antigen) are co-administered and/or added. e.) against certain bacteria and viruses can be co-administered and/or added to compositions. In some embodiments of the invention, various therapeutic agents (eg, anti-inflammatory agents, chemotherapeutic agents), stabilizers, buffer solutions, etc. co-administered and/or added to anti-pneumococcal hyperimmune globulin, for example, to further enhance efficacy, stability, suitability for administration, duration of action, range of applications, etc. In one embodiment, antipneumococcal hyperimmune globulin is co-administered with conventional IVIG (e.g., to a patient with a primary immunodeficiency disease (e.g., to treat (e.g., prophylactic and/or therapeutic) an infection (e.g., caused by S. pneumonia, Corynebacterium diphtheria, measles virus and/or polio virus)).

The compositions may optionally contain carriers such as preservatives, wetting, emulsifying and dispersing agents. Prevention of exposure to microorganisms can be provided by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. In addition, it may be desirable to administer isotonic agents, for example sugars, sodium chloride, and the like. Prolonged absorption of immunoglobulins can be achieved by the use of agents that delay absorption, such as aluminum monostearate and gelatin.

In some embodiments, the anti-pneumococcal hyperimmune globulin of the invention is administered to a subject to provide therapeutic, prophylactic, prophylactic, and/or other benefits.

Diseases and conditions in which the anti-pneumococcal hyperimmune globulin of the invention should be administered for therapeutic or prophylactic purposes include, but are not limited to: common variable immunodeficiency, IgA deficiency, human immunodeficiency virus (HIV) infection, bacterial and viral infections such as respiratory tract infection influenza, respiratory syncytial virus infection, rhinovirus respiratory tract infection, adenovirus respiratory tract infection: protozoan infections such as giardiasis, yeast infections; chronic lymphocytic leukemia; multiple myeloma; macroglobulinemia; Chronical bronchitis; bronchiectasis; asthma; immunosuppression associated with bone marrow transplantation; immunosuppression associated with the administration of cyclophosphamide; immunosuppression associated with the administration of azathioprine; suppression of immunity associated with the introduction of methotrexate; immunosuppression associated with the introduction of chlorambucil; suppression of immunity associated with the introduction of agents based on mustard nitrogen; immunosuppression associated with the administration of 6-mercaptopurine; immunosuppression associated with the administration of thioguanine; severe combined immunodeficiency; deficiency of adenosine deaminase; deficiency of the main histocompatibility complex class I (empty leukocyte syndrome) and class II; deficiency of purine nucleoside phosphorylase; diGeorge syndrome; transient hypogammaglobulinemia of infancy; X-linked agammaglobulinemia; X-linked agammaglobulinemia with growth hormone deficiency; transcobalamin II deficiency; immunodeficiency with thymoma; immunodeficiency with a hereditary defective response to the Epstein-Barr virus; deficiency of immunoglobulin with elevated IgM; P chain deficiency; ataxia telangiectasia; immunodeficiency with partial albinism; consequences of selective IgA deficiency such as rheumatoid arthritis; juvenile rheumatoid arthritis; systemic lupus erythematosus; thyroiditis; pernicious anemia; dermatomyositis; Coombs-positive hemolytic anemia; idiopathic Addison's disease; cerebral vasculitis and idiopathic thrombocytopenic purpura.

In some embodiments, the compositions and methods of this invention provide anti-inflammatory benefits when administered to a subject. Pooled immunoglobulins have been shown to have anti-inflammatory effects when administered passively (see, for example, Nimmerjahn and Ravetch, Annu Rev. Immunol. 2008.26:513-33, Ramakrishna et al. Plos Pathogens. 2011.7:6: e 1002071, included in this description by reference in its entirety). In some embodiments, the anti-pneumococcal hyperimmune globulin of the invention has an enhanced anti-inflammatory effect (e.g., 10% improvement, 20% increase, 50% increase, 2-fold increase, 3-fold increase, 5-fold increase, 10-fold increase). or more times) compared to the anti-inflammatory effect of a mixture of plasma samples obtained from random people (for example, 1000 or more random people). Although an understanding of the mechanism is not essential to the practice of this invention, and although the invention is not limited to any particular mechanism, in one embodiment of the invention, the anti-pneumococcal hyperimmune globulin of the invention exhibits a significantly improved anti-inflammatory effect compared to conventional IVIG because the composition is combined The pool of plasma according to the invention contains plasma from at least 1000 donors (for example, compared with conventional hyperimmune globulin obtained from a limited number of donors (for example, in one embodiment of the invention, the greater the number of different plasma samples, the more pronounced the observed anti-inflammatory effect (for example, the more pronounced the histopathological effect (for example, a decrease in the death of epithelial cells)))).

In some embodiments of this invention, the compositions of the invention are administered separately, while in other embodiments of the invention, the compositions are preferably present in a pharmaceutical composition containing at least one active ingredient/agent as defined above, together with a solid support or, alternatively, together with one or more pharmaceutically acceptable carriers and optionally other therapeutic agents. Each carrier must be "acceptable" in the sense that it is compatible with the other ingredients of the formulation and harmless to the subject.

In one embodiment, the anti-pneumococcal hyperimmune globulin provided herein further comprises one or more biologically active agents. The invention is not limited to the type of biologically active agent/material. In fact, various biologically active agents/materials can be used, including, but not limited to, antibodies, antitoxin material, anti-inflammatory agent, anticancer agent, antimicrobial agent, therapeutic agent, antihistamine, cytokine, chemokine, vitamin, mineral, and the like. In one embodiment of the invention, the biologically active agent is an antitoxin agent. In one embodiment, the antitoxin agent is a monospecific, bispecific, or polyspecific antibody with specificity for a viral, bacterial, or fungal toxin. In another embodiment, the bacterial or fungal toxin is selected from botulinum neurotoxin, tetanus toxin, E. coli toxin, Clostridium difficile toxin, Vibrio RTX toxin, staphylococcal toxins, cyanobacteria toxin, and mycotoxins. In another embodiment of the invention, the immunotherapeutic composition further comprises an aliquot of one or more monoclonal antibodies with one or more specificities (for example, the immunogenic composition may be coated with one or more antibodies or biologically active substance (for example, a monoclonal antibody of any specificity, antitoxin agent and etc.)). The invention is not limited to the type of one or more antibodies added (eg, loading) to the immunogenic composition. In fact, any one or more antibodies (e.g., specific for a pathogen or pathogen product) can be used, including, but not limited to, standard antibodies, bispecific antibodies, polyspecific antibodies, and the like known in the art. (eg, specific for one or more antigens).

The invention is not limited to the kind of subject being treated with the compositions and methods of the invention. In fact, a variety of subjects can be treated in this manner, including, but not limited to, a subject at risk of developing an infection (e.g., upper respiratory tract or other type of infection (e.g., thereby reducing the risk of infection in a subject at increased risk of infection). )). In one embodiment, the subject being treated with the composition of the invention has or has been diagnosed with primary immunodeficiency disease (PIDD). In another embodiment, the subject is a patient with end-stage renal disease (ESRD); cancer patient receiving immunosuppressive therapy, AIDS patient, diabetic patient, neonate, transplant patient receiving immunosuppressive therapy patient, patient with PID (PIDD) and other immunodeficiencies, patient with impaired immune system function, patient with autoimmune disease, elderly person age in a nursing home-type facility, a patient on immunosuppressive therapy with an autoimmune disease, a transplant patient, an invasive surgery patient, a burn patient, or another patient in the emergency department.

EXPERIMENTAL PART

The following examples are provided for the purpose of demonstrating and further illustrating certain preferred embodiments of the invention and aspects of the present invention and should not be construed as limiting its scope.

Example 1

Generation of donors of hyperimmune plasma, hyperimmune serum and immune globulin derived from them

Experiments were conducted during the development of embodiments of the invention with the aim of generating hyperimmune plasma donors that have hyperimmune plasma/serum with a broad spectrum of action against pneumococcal serotypes. The following is a non-limiting example of such experiments.

Healthy people (men and women) aged 18 to 45 were selected for immunization. In particular, in order to participate in the study and function as a vaccinated human plasma donor, each subject was required to be free of any known pathological conditions.

Each study participant was given the following vaccination schedule: a primary vaccination/dose of PREVNAR (PCV13) (0.5 ml IM injection according to the manufacturer's instructions) on day 0, followed by a booster vaccination with PNEUMOVAX23 (0.5 ml IM injection according to the manufacturer's instructions). manufacturer) at 4 weeks. Blood was drawn from each participant (10 ml) at the following time points: immediately before the primary vaccination with PREVNAR, immediately before the secondary vaccination with PNEUMOVAX23 at 4 weeks, at 8 weeks, at 10 weeks, at 12 weeks and at 16 weeks. Serum samples were prepared from each blood sample, divided into aliquots and stored at -70°C ± 10°C. Serum samples were subjected to various studies (eg, described below).

Quantification of serotype-specific anti-pneumococcal antibodies against polysaccharide (PS (PS)) (IgG testing). Pneumococcal serotype (ST(ST))-specific IgG is quantified using previously described methods (see, for example, Lai et al., 2005 J bnmunolo Meth 296: 135-147). Briefly, spectrally distinct Luminex beads are conjugated with pneumococcal (Pnc) CT (ST)-specific capsular polysaccharide (PS (PS)), obtained from the American Type Culture Collection, AKTC (ATCC). Serum test samples are mixed with PS(P8)-conjugated beads and CT(ST)-specific IgG is quantified with 89SF human control serum. Serotype-specific concentration in serum samples was expressed as μg/ml.

Opsonophagocytic elimination assay (OPE (OPK)). Functional antibody concentrations (titer) in serum samples are tested using previously described methods (see, for example, Romero-Steiner et al., 1997 Clin Diagn Lab Immunol. Jul, 4 (4): 415-22.). Briefly, HL-60 cells (human promyelocytic leukocytes) differentiated into a polymorphonuclear (PMN) line are used as effector cells. Live serotype-specific strains of S. pneumoniae are used as targets, and baby rabbit complement (PELFREEZE, Inc, Rogers, Arkansas) is used as a source of complement for opsonophagocytic elimination. Reagents stored at 4±2° C. are brought to room temperature prior to analysis. To each well of a 96-well plate, add 10 µl of Hanks' balanced salt solution, HBSS (+) with gelatin, hereinafter referred to as assay buffer, except for row A. To wells A1-A3, add 20 µl of undiluted quality control serum. In wells A4-A12 add 20 µl of the diluted sample. Serum QC (QC) and unknown samples are serially diluted 1:2. Complement control wells and cell control wells at this point contain 10 µl of assay buffer.

The working bacterial suspension of S. pneumonia is diluted to a concentration of 8.0 x 105 colonies per 20 µl. 20 µl of the bacterial suspension is added to each well of the plate, including control wells. The plate is incubated for 30 minutes at room temperature on a plate shaker set at 200 rpm. 10 µl of frozen rabbit complement is added to each well, except for cell control. 10 µl of assay buffer is added to control cells. The plate is incubated for 15 minutes at room temperature on a plate shaker set at 200 rpm. 40 μl of differentiated PMN (PMN) HL60 is added to each well of the plate. The cell concentration is determined to add 100,000 cells per well. The plate is incubated for 30 minutes at room temperature on a plate shaker set at 200 rpm.

Using a multichannel pipette, 5 µl from each well in a row is simultaneously transferred to a 150 x 15 mm petri dish containing chocolate agar. The dish is tilted to allow the sample volume to flow down the agar plate in parallel rows. The plates are incubated overnight at 37°C and 5% CO2. The next day, the plates are photographed and the colony forming units (CFU) are counted. Bacterial elimination is calculated as percent kill per well (cfu per well/average cfu of complement control * 100). The reciprocal of the dilution at which >50% kill of the target bacterium occurs compared to the complement control is recorded as the OPE titer (OPK). Since the first dilution is 1:8, untitered serum samples are recorded as 1:4 for calculation purposes.

Example 2

Identification and characterization of the specific and general response to pneumococcal serotype in the form of functional antibodies in immunized individuals

An individual serum sample is collected from study participants at different time points (immediately prior to primary vaccination with PREVNAR, immediately prior to secondary vaccination with PNEUMOVAX23 at 4 weeks, at 8 weeks, at 10 weeks, at 12 weeks and at 16 weeks). Serum is analyzed for the concentration of IgG specific for pneumococcal (Pnc) serotype (CT (ST)) (see IgG test in Example 1 above). In addition to individual serum samples, pooled serum is obtained by pooling an equal amount of serum from each participant before and after week 12, and pooled serum samples are characterized to evaluate the effect of pooling on IgG test result. In addition, both individual and pooled serum samples are tested for functional antibody response using the opsophagocytic elimination (OPK) assay described in Example 1.

Concentrations/titer of serotype-specific anti-pneumococcal IgG. Serum samples from each individual (n=10) immunized with PREVNAR-PNEUMOVAX23 prime-boost were analyzed for ST-specific IgG Pnc. CT (ST)-specific IgG Pnc titers are shown in FIG. 1. Variations in IgG response in serum samples between donors and between STs have been reported. While 9/10 donors showed a reliable response to Pnc ST (ST) in the form of IgG, there are differences in ST (ST)-specific responses. Among donors, 7/10 had peak IgG concentrations within 4 weeks of PNEUMOVAX23 vaccination (booster response to PNEUMOVAX23 eight weeks after primary vaccination) and then a gradual decrease in levels that remained several times above baseline even at 12 weeks. Despite these variations, all donors demonstrated concentrations of ST-specific IgG Pnc above the protective level (>0.2 µg/mL) of immunity (see, e.g., Balmer et al., Clin Exp Immunol, 2003 Sep, 133 ( 3): 364-9). The pooled serum IgG concentration showed a 2.2-30 fold increase in IgG concentration at the end of the study period (week 12) compared to baseline (before). Among them, the highest increase in IgG response was recorded for the ST 1 serotype, followed by the ST23F and ST4 serotypes. Serotype ST19A shows the lowest fold increase in IgG concentration (by 2.2 times), followed by ST 3 (by 2.6 times) and ST14 (by 3.2 times).

Titer of functional/opsonic antibodies (OPK). On FIG. 2 shows data related to OPE titer (OPK) in donor serum samples specific for different STs (ST) Pnc. The baseline OFE titer (OPK) is indicated along with the IgG concentration. The multiplicity of increase in the titer of functional antibodies increased by 4-256 times (see Fig. 2). Surprisingly, it was found that the total amount of IgG (eg depicted in Fig. 1) does not correlate with the amount of total functional opsonic antibodies. For example, for the ST14 serotype, only a moderate increase in the total IgG titer (by 3.2 times, see Fig. 1) was registered. However, he recorded the largest increase in functional response (opsonic antibody titer) of 256 times compared to baseline.

Example 3

Quantification of total and functional serotype-specific anti-pneumococcal antibodies in pooled human serum

Serum samples from vaccinated human plasma donors (n=34) were obtained at one month intervals, starting 1 month before vaccination and ending 5 months after vaccination. At these 6 time points, serum samples were pooled and analyzed for the concentration of ST (ST)-specific IgG Pnc (IgG testing) and functional antibodies using OPE (OPK) described in Example 1 above. The concentrations of antibodies (serotype-specific IgG and OFE (OPK)) and the fold increase compared to the initial level before vaccination were determined.

Serum samples from vaccinated individuals (immunized with PREVNAR-PNEUMOVAX23 Prime Boost) are pooled every 1 month between 1 month before vaccination and up to 5 months after vaccination, and the pooled sera for each time point are analyzed for ST (ST) -specific IgG Pnc. The titer of ST-specific IgG Pnc in pooled donor sera obtained at each time point is depicted in FIG. 3. The average concentration of IgG in serum samples after immunization ranged from 4.4 to 37.59 µg/ml. The decrease in total IgG concentrations compared to the 12 week endpoint is possibly due to the gradual decline in IgG levels after the spike observed at 4 weeks after booster vaccination, resulting in a decrease in IgG concentrations over time. However, the fold increase in IgG concentration from baseline (see FIG. 4) showed a pattern similar to that shown in FIG. 1, with a higher response to ST1, ST23F and ST4.

OFE (OPK). OFE (OPK) titers in pooled donor serum specific for different STs (ST) Pnc are depicted in FIG. 5. The mean OFE titer (OPK) after immunization for ST1 was exceptionally high (36044), while all other STs (ST) showed a titer at least a logarithm lower. Although high OPE (OPK) titers were reported in individual serum samples tested at week 12 (see Fig. 2), the titers were not logarithm higher than for the other serotypes.

Example 4

Identification and characterization of generic and functional serotype-specific anti-pneumococcal antibodies in purified human IgG

Serum samples obtained from study participants at different time points (n=34) were pooled at each of the individual time points (before, 4, 8, 10, 12, 16 weeks and 5 months) and IgG was purified from pooled at a pool of samples using standard protein A immunosorbent columns. Purified IgG was analyzed for the concentration of ST (ST)-specific IgG Pnc (IgG testing) and functional antibodies using OFE (OPK) as described in Example 1. Antibody concentrations were characterized ( serotype-specific IgG) and functional antibody titer (OFE (OPK)). In addition, characterization of results was performed to understand the relationship between in vitro and ex vivo methods based on regression analysis of individual results.

Concentrations of serotype-specific pneumococcal IgG. Serum samples from individuals vaccinated using the PREVNAR-PNEUMOVAX23 prime boost schedule were pooled at one month intervals starting 1 month before vaccination and up to 5 months after, sera for each time point were pooled and IgG purified. Pooled serum samples were analyzed for ST (ST)-specific IgG Pnc. ST(ST)-specific IgG Pnc pooled serum is shown in FIG. 6. Mean IgG concentration in pooled serum after immunization ranged from 3.2 to 23.3 μg/ml, resulting in a decrease in IgG concentration over time. However, the increase in IgG concentration from baseline (see FIG. 4) showed a pattern similar to that depicted in FIG. 1, with a higher response for ST1, ST23F and ST4.

OFE (OPK): The titer of OFE (OPK) in pooled donor serum ST-specific for various Pncs is depicted in FIG. 7. The mean titer after immunization with OFE (OPK) ranged from 213 to 5826.7 with the highest titer for ST7F (5826.7) followed by ST23F (4778.7) and ST5 (4096).

Example 5

Comparison of OFE serotype (OPK) titer present in pooled human sera from vaccinated donors versus OFE serotype (OPK) titer present in a conventional, commercially available IVIG assay (IVIG)

OFE analysis (OPK). The OFE analysis (OPK) described in Example 1 above was used. Briefly, reagents stored at 4±2° C. are brought to room temperature prior to analysis. To each well of a 96-well plate, add 10 µl of HBSS (+) with gelatin, hereinafter referred to as assay buffer, except row A. To wells A1-A3, add 20 µl of undiluted serum for quality control. In wells A4-A12 add 20 µl of the diluted sample. Serum QC (QC) and unknown samples are serially diluted 1:2. Complement control wells and cell control wells at this point contain 10 µl of assay buffer.

The working bacterial suspension of S. pneumonia is diluted to a concentration of 8.0 x 105 colonies per 20 µl. 20 µl of the bacterial suspension is added to each well of the plate, including the control wells. The plate is incubated for 30 minutes at room temperature on a plate shaker set at 200 rpm. 10 µl of frozen rabbit complement is added to each well, except for cell control. 10 µl of assay buffer is added to control cells. The plate is incubated for 15 minutes at room temperature on a plate shaker set at 200 rpm. 40 μl of differentiated PMN (PMN) HL60 is added to each well of the plate. The cell concentration is determined to add 100,000 cells per well. The plate is incubated for 30 minutes at room temperature on a plate shaker set at 200 rpm.

Using a multichannel pipette, 5 µl from each well in a row is simultaneously transferred to a 150 x 15 mm petri dish containing chocolate agar. The dish is tilted to allow the sample volume to flow down the agar plate in parallel rows. The plates are incubated overnight at 37°C and 5% CO2. The next day, the plates are photographed and the colony forming units (CFU) are counted. Bacterial elimination is calculated as percent kill per well (cfu per well/average cfu of complement control * 100).

On FIG. 8 shows a comparison of the titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in pooled human sera from vaccinated donors compared to the titer of serotype-specific OFE (OPK) functional/opsonic antibodies present in nine random and different conventional , commercially available IVIG (IVIG). Each column represents a single, unique S. pneumoniae serotype. Each row represents a unique pattern. Samples A-I are conventional commercially available IVIG.

As shown in FIG. 8, marked variability in serotype-specific opsonic titers was found in IVIG commercial batches. In addition, an increased titer for one serotype was not found to predict or correlate with an increased titer for any of the other serotypes, suggesting that the individual increased response does not reflect an overall enhanced immune response to S. pneumonia, but rather a sporadic (e) enhanced ( f) response(s) to very specific serotypes. In marked contrast, opsonic antibody titers observed for immune globulin from immunized donors were noted to be elevated for all serotypes without exception. In addition, a 3-256-fold increase in opsonic anti-pneumococcal antibody titer in immune globulins was observed in immune globulin from immunized donors compared to commercial batches of immune globulin. Thus, in some embodiments, the compositions and methods of the invention for producing compositions (e.g., blood, plasma, and/or immune globulin compositions) containing an elevated titer of opsonic anti-pneumococcal antibodies provide a homogeneous composition containing titers of opsonic anti-pneumococcal antibodies specific for against all multiple pneumococcal serotypes (eg, 9, 10, 11, 12, 13, 14, 15 or more serotypes) and/or significantly elevated opsonic antibody titers compared to conventional commercial immune globulin (eg, it is 2, 3 , 4, 5, 6, 7, 8, 9, 10 or more times the opsonic antibody titer).

Claims (11)

1. A method for producing an immunoglobulin containing an opsonophagocytic antibody titer of at least 1:256 for 50% or more of Streptococcus pneumonia serotypes selected from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F, said method comprising the following steps: (a) immunizing healthy adult human plasma donors aged 18 to 60 years with a primary immunization with a polyvalent S. pneumonia conjugate vaccine followed by a booster immunization with a polyvalent polysaccharide vaccine against S. pneumonia ; (b) collecting plasma from said plasma donors after said booster immunization; (c) pooling plasma from vaccinated donors to obtain pooled plasma containing an opsonophagocytic antibody titer of at least 1:256 for 50% or more of Streptococcus pneumonia serotypes selected from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F; And (d) obtaining immunoglobulin from plasma pooled in accordance with stage (c). 2. The method of claim 1, further comprising step (e), in which the immunoglobulin obtained in accordance with step (d) is made suitable for administration by intravenous injection. 3. The method of claim 2, wherein the immunoglobulin is provided in solution, the pH and ionic strength of said solution being adjusted to make it suitable for intravenous injection. 4. The method of claim 1 wherein the pooled plasma is obtained by pooling 1000 or more different immunized human plasma donors into a plasma pool. 5. Immunoglobulin obtained by the method according to paragraph 1. 6. A method of treating an S. pneumonia infection in a subject, comprising administering to said subject a therapeutically effective amount of an immunoglobulin obtained by the method of claim 1. 7. A method of administering immunotherapy to a subject, comprising administering to said subject a therapeutically effective amount of an immunoglobulin obtained by the method of claim 1.

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