WO2021243372A1

WO2021243372A1 - Closed-loop antimicrobial photo-plasmapheresis

Info

Publication number: WO2021243372A1
Application number: PCT/US2021/070633
Authority: WO
Inventors: Reka SZIGETI; Richard KELLERMAYER
Original assignee: Baylor College of Medicine
Current assignee: Baylor College of Medicine
Priority date: 2020-05-28
Filing date: 2021-05-28
Publication date: 2021-12-02
Anticipated expiration: 2022-11-28

Abstract

The present disclosure concerns systems and methods related to the treatment, prevention, delay of onset, or reduction of severity of a microbial infection in an individual, including closed-loop plasmapheresis and extracorporeal light irradiation of the plasma of the individual. The systems and methods further relate to irradiation of the plasma of the individual with UV and/or visible light, optionally in the presence of a photosensitizer. The systems and methods further relate to light irradiation inactivation of microbial particles in the plasma taken from the individual. The systems and methods further relate to the optional filtration of microbial particles from UV light-treated plasma. The systems and methods further relate to reinfusion of the UV light-treated, optionally filtered, plasma back into the individual. The systems and methods further relate to treating, preventing, delaying onset of, or reducing in severity a viral infection, particularly a viral infection caused by a Coronaviridae family virus, at least including SARS-CoV-2. The systems and methods are further configured to reduce viral load and/or lymphocyte reinfection rate in the individual.

Description

CLOSED-LOOP ANTIMICROBIAL PHOTO-PLASMAPHERESIS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of priority to United States Provisional Application No. 63/031,015 filed May 28, 2020, which is incorporated by reference herein in its entirety.

TECHNICAL FIELD

[0002] The present disclosure is directed at least to the fields of virology, bacteriology, biochemistry, cell biology, chemistry, molecular biology, immunology, infectious disease, and medicine.

BACKGROUND

[0003] The severity of the COVID-19 (SARS-CoV-2) pandemic is undisputable. Every means possible should be considered to save lives and speed the resolution of this worldwide crisis. Ultraviolet (UV) light irradiation is a well-established method for viral inactivation in transfusion medicine (Chin et ah, 1997) but has not been exploited to treat viral infections. SARS-CoV-2 is a medium-sized, enveloped, positive-sense, single- stranded RNA vims of the Coronaviridae family. The vims is the pathogen of the third largest severe respiratory syndrome outbreak caused by Coronavimses (CoV) (1. SARS emerged in late 2002 and disappeared by 2004; 2. MERS, emerged in 2012 and remains in circulation in camels).

[0004] Such viruses are highly sensitive to UV light radiation either by itself (UVC) or in the presence of photosensitizers such as riboflavin (UVB) or amotosalen (UVA), and even to visible light in the presence of photosensitizers such as methylene blue. (Eickmann et ah, 2018; Chang et ah, 2020) The pathogenesis of COVID-19 was recently proposed by Lin, et ah, 2020. Their hypothesis outlines three stages of the disease. The first phase (I.) is viremia, when the vims disseminates in the body reaching extrapulmonary organs; the second stage (II.) is acute (pneumonia), during which the viral load reaches its peak in association with progressive lymphopenia, and if the immune system cannot control the vims, the disease progresses to the third severe phase (III.) with further decline in lymphocyte count. The third, severe stage is characterized by persistent viremia and multi organ failure. [0005] This pathomechanism is very similar to the previously described multi organ infection model for SARS-CoV-1 (SARS). (Gu et ah, 2005) COVID-19 resembles SARS in many ways, including genetic homology, disease dynamics, and transmission route. Both viruses use similar mechanisms (key molecule being angiotensin converting enzyme 2 [ACE2]) to enter cells. The incubation time and how the disease progresses are also very similar, along with risk factors for developing severe disease. (Wilder-Smith et ah, 2020) As for SARS, a multi organ infection model theory was developed by Gu et ah, 2005, based on autopsy findings of 8 confirmed cases. In their model, the key step for SARS evolution is the infection of lymphocytes by SARS, which facilitates its widespread dissemination. Observations from Li et ah, 2003, supports this model, and showed active viral replication in lymphocytes. As a result, extrapulmonary organ damage occurs, including the lymphoid system, which is reflected by atrophic lymphoid organs and by declining lymphocyte counts. (Gu et ah, 2005) It is the weakened immune response that can contribute to the progression of the pneumonia and potentially extrapulmonary manifestations. This model emphasized that the extent of the immune system damage, manifesting as lymphopenia in the peripheral blood, is a strong predictor for outcome. Importantly, the same observation was published very recently by Tan et ah, 2020, for COVID-19 infection. Tan et al. categorized patients based on lymphocyte numbers and found that lymphocyte count was a reliable indicator for outcome. These studies strongly implicate peripheral blood and lymphocyte mediated viral pathogenesis in the progressively worsening cases of COVID-19 and SARS.

[0006] There is currently limited information on blood derived viremia/viral counts in COVID-19 patients during the course of infection. Some studies documented rare and relatively low level of viral detection in blood (Wang et ah, 2020), but the actual timing of the blood specimens in regards to the course of disease was not documented. Similarly, a recent rigorous study (including multiple site and repeated nucleic acid based, and also viral culture testing) in 9 symptomatic patients with mild disease course did not find live SARS-CoV-2 virus in blood. (Wolfel et ah, 2020) One of the first reports, however, from Chen et ah, 2020, indicated that the 2019-nCoV (i.e. SARS-CoV-2) RNA was readily detected in the blood in 6 of 57 patients. Importantly, all of the 6 patients with detectable viral RNA in the blood progressed to severe symptom stage, suggesting a strong correlation between serum viral RNA and disease severity (p-value = 0.0001). These findings are consistent with those during the SARS epidemic. Median concentration of serum SARS RNA was found to be 26 to 30 fold higher at hospital admission in those patients requiring ICU care, compared to those who did not. (Ng et ah, 2003) Therefore, current evidence indicates that patients progressing towards worsening clinical status develop significant viremia in the blood, consistent with the hypothesis of Lin, et ah, 2020.

BRIEF SUMMARY

I. Methods of the Disclosure

[0007] The present disclosure is directed to methods for treating or preventing a microbial infection in an individual. In specific embodiments, the methods comprise the steps of: (a) separating blood withdrawn from an individual into plasma and cellular material; (b) treating the plasma with light radiation to produce light-treated plasma; (c) optionally filtering the light-treated plasma to produce light-treated and, optionally, filtered plasma; and (d) reinfusing the cellular material and the light-treated and, optionally, filtered plasma back into the individual. In specific embodiments, the methods comprise the steps of: (a) separating blood withdrawn from an individual into plasma and cellular material; (b) treating the plasma with light radiation to produce light-treated plasma; (c) filtering the light-treated plasma to produce light-treated and filtered plasma; and (d) reinfusing the cellular material and the light-treated and filtered plasma back into the individual. In specific embodiments, the methods comprise the steps of: (a) separating blood withdrawn from an individual into plasma and cellular material; (b) treating the plasma with light radiation to produce light-treated plasma; and (c) reinfusing the cellular material and the light-treated plasma back into the individual. In specific embodiments, the methods are effective to direct a substantially continuous flow of blood from the individual and of the cellular material and light-treated and, optionally, filtered plasma back into the individual. In specific embodiments, the methods are effective to reduce microbial load in the individual. In specific embodiments, the methods are effective to treat, prevent, delay onset of, and/or reduce severity of one or more symptoms of a microbial infection in the individual.

[0008] In specific embodiments, the light radiation is ultraviolet light (UV) radiation and/or visible light radiation. In specific embodiments, the UV light radiation is UVA, UVB, and/or UVC light radiation.

[0009] In specific embodiments, the step of treating the plasma with light radiation further comprises the steps of: (i) directing the plasma through a light transparent reservoir, and (ii) treating the plasma passing through the light transparent reservoir with light using a light irradiation unit.

[0010] In specific embodiments, the step of treating the plasma with light radiation further comprises the steps of (i) adding a photosensitizer to the plasma (ii) directing the plasma and photosensitizer through a light transparent reservoir, (iii) treating the plasma and photosensitizer passing through the light transparent reservoir with light using a light irradiation unit to produce light-treated plasma and photosensitizer, and (iv) optionally, removing the photosensitizer from the light-treated plasma.

[0011] In specific embodiments, the photosensitizer is an organic photosensitizer and/or an inorganic photosensitizer. In specific embodiments, the organic photosensitizer is selected from the group consisting of: riboflavin, amotosalen, methylene blue, rose bengal, porphyrins, and phthalocyanines. In specific embodiments, the inorganic photosensitizer is selected from the group consisting of: T1O2, ZnO, ZnS, CdS, F_e20₃, and WO3.

[0012] In specific embodiments, the step of treating the plasma with light radiation is effective to inactivate microbes in the plasma. In specific embodiments, the step of treating the plasma with light radiation is effective to inactivate at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbes in the plasma.

[0013] In specific embodiments, the step of filtering the light-treated plasma, if performed, is effective to remove microbial particles from light-treated plasma. In specific embodiments, the step of filtering the light-treated plasma is effective to remove at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbial particles from light- treated plasma. In specific embodiments, the step of filtering the light-treated plasma is effective to exclude removal of antibodies in the light-treated plasma.

[0014] In specific embodiments, the microbial infection is a bacterial infection, a fungal infection, a protozoan infection, a viral infection, or a combination thereof. In specific embodiments, the microbial infection is a viral infection. In specific embodiments, the viral infection is an infection caused by a Coronaviridae family vims. In specific embodiments, the Coronaviridae family virus is a betacoronoavims. In specific embodiments, the betacoronavims is SARS-CoV-2. In specific embodiments, the viral infection is an infection caused by a Filoviridae family vims. In specific embodiments, the Filoviridae family vims is an ebolavims. [0015] In specific embodiments, the step of reinfusing the cellular material and the light- treated and, optionally, filtered, plasma into the individual is effective to reduce viral load and/or lymphocyte reinfection rate in the individual. In specific embodiments, the step of reinfusing the cellular material and the light-treated and, optionally, filtered, plasma into the individual is effective to reduce viral load and/or lymphocyte reinfection rate in the individual by at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99%.

[0016] In specific embodiments, the method precedes, runs concurrently with, or follows an additional antimicrobial therapy administered to the individual.

II. Systems of the Disclosure

[0017] The present disclosure is directed to systems for treating a microbial infection in an individual in need thereof. In specific embodiments, the systems comprise: (a) a first conduit, (b) a plasmapheresis unit, (c) a light irradiation unit, (d) optionally, a filtration unit, and (e) a second conduit. In specific embodiments, the systems comprise: (a) a first conduit, (b) a plasmapheresis unit, (c) a light irradiation unit, (d) a filtration unit, and (e) a second conduit. In specific embodiments, the systems comprise: (a) a first conduit, (b) a plasmapheresis unit, (c) a light irradiation unit, and (d) a second conduit.

[0018] In specific embodiments, first conduit is configured to withdraw blood from an individual. In specific embodiments, the plasmapheresis unit is configured to separate the blood into plasma and cellular material. In specific embodiments, the light irradiation unit is configured to treat the plasma with light radiation. In specific embodiments, the filtration unit is configured to filter the light-treated plasma. In specific embodiments, the second conduit is configured to reinfuse the combined cellular material and light-treated and, optionally, filtered plasma back into the individual. In specific embodiments, the first and second conduits provide for substantially continuous flow of blood from the individual and of the cellular material and light-treated and, optionally, filtered plasma back into the individual In specific embodiments, the systems are configured to reduce microbial load in the individual.

[0019] In specific embodiments, the light irradiation unit is configured to treat the plasma with UV light radiation and/or visible light radiation. In specific embodiments, light irradiation unit is configured to treat the plasma with UVA, UVB, and/or UVC radiation. [0020] In specific embodiments, the systems comprise a light transparent reservoir through which the plasma is directed. In specific embodiments, the light irradiation unit is further configured to treat the plasma with light radiation as the plasma passes through the light transparent reservoir.

[0021] In specific embodiments, the systems comprise a third conduit configured to direct a photosensitizer into the plasma prior to the light treatment of the plasma by the light irradiation unit. In specific embodiments, the systems comprise a light transparent reservoir through which the plasma and the photosensitizer are directed. In specific embodiments, the light irradiation unit is further configured to treat the plasma and the photosensitizer with light radiation as the plasma and the photosensitizer pass through the light transparent reservoir to produce light-treated plasma and photosensitizer. In specific embodiments, the systems comprise a photosensitizer removal unit configured to remove the photosensitizer from the light- treated plasma.

[0022] In specific embodiments, the third conduit is configured to direct an organic photosensitizer and/or an inorganic photosensitizer into the plasma. In specific embodiments, the third conduit is configured to direct an organic photosensitizer selected from the group consisting of: riboflavin, amotosalen, methylene blue, rose bengal, porphyrins, and phthalocyanines, into the plasma. In specific embodiments, the third conduit is configured to direct an inorganic photosensitizer selected from the group consisting of: T1O2, ZnO, ZnS, CdS, Fe₂0₃, and WO3, into the plasma.

[0023] In specific embodiments, the light irradiation unit is further configured to inactivate microbes in the plasma during the treatment of the plasma with light radiation. In specific embodiments, the light irradiation unit is further configured to inactivate at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbes in the plasma during the treatment of the plasma with light radiation.

[0024] In specific embodiments, the filtration unit (that may or may not be optional in the system) is further configured to remove microbial particles from light-treated plasma during the filtration of the light-treated plasma. In specific embodiments, the filtration unit is further configured to remove at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbial particles from light-treated plasma during the filtration of the light-treated plasma. In specific embodiments, the filtration unit is further configured to exclude removal of antibodies in the light- treated plasma. In specific embodiments, the system is further configured to reduce viral load and/or lymphocyte reinfection rate in the individual. In specific embodiments, the system is further configured to reduce viral load and/or lymphocyte reinfection rate in the individual by at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99%.

[0025] In specific embodiments the microbial infection is a bacterial infection, fungal infection, protozoan infection, and/or a viral infection. In specific embodiments, the microbial infection is a viral infection. In specific embodiments, the viral infection is an infection caused by a Coronaviridae family vims. In specific embodiments, the Coronaviridae family virus is a betacoronoavims. In specific embodiments, the betacoronavirus is SARS-CoV-2. In specific embodiments, the viral infection is an infection caused by a Filoviridae family virus. In specific embodiments, the Filoviridae family vims is an Ebola vims. In specific embodiments, the system reduces viral load and/or lymphocyte reinfection rate in the individual. In specific embodiments, the system reduces viral load and/or lymphocyte reinfection rate in the individual by at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99%.

[0026] The foregoing has outlined rather broadly the features and technical advantages of the present disclosure in order that the detailed description that follows may be better understood. Additional features and advantages will be described hereinafter which form the subject of the claims herein. It should be appreciated by those skilled in the art that the conception and specific embodiments disclosed may be readily utilized as a basis for modifying or designing other structures for carrying out the same purposes of the present designs. It should also be realized by those skilled in the art that such equivalent constructions do not depart from the spirit and scope as set forth in the appended claims. The novel features which are believed to be characteristic of the designs disclosed herein, both as to the organization and method of operation, together with further objects and advantages will be better understood from the following description when considered in connection with the accompanying figures. It is to be expressly understood, however, that each of the figures is provided for the purpose of illustration and description only and is not intended as a definition of the limits of the present disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] For a more complete understanding of the present disclosure, reference is now made to the following descriptions taken in conjunction with the accompanying drawing, in which. [0028] FIG. 1 illustrates a circuit diagram showing a particular embodiment of the flow circuit of plasmapheresis and extracorporeal light irradiation of plasma.

[0029] FIG. 2 illustrates a flow diagram for a method for plasmapheresis and extracorporeal light irradiation of plasma in accordance with a certain embodiment of the system.

DETAILED DESCRIPTION

I. Definitions

[0030] As used herein, the term “microbial infection” refers to the presence of one or more microbes in the body. In particular cases, the microbial infection is of a pathogenic microbe. Examples of microbes include viruses of any kind, bacteria of any kind, fungi of any kind, protozoa of any kind, and so forth.

[0031] As used herein, the term “to reinfuse” or “reinfusing” refers to the return of something to the body after being withdrawn, including at least by administration directly into a vessel, such as a vein.

[0032] As used herein, the term “microbial load” refers to the amount of an active microbe in an organism, typically in the bloodstream, stated as active microbe particles per unit volume.

[0033] As used herein, the term “active microbe” refers to a microbe with the ability to infect and/or cause disease at maximum severity.

[0034] As used herein, the term “to inactivate microbes” refers to destroying microbe particles, killing microbes, or rendering live microbes less able or completely unable to infect and/or cause disease.

[0035] As used herein, the term “filtering” refers to removing unwanted material from a liquid, including at least by means of passing the liquid through a size-exclusion membrane.

[0036] As used herein, the term “lymphocyte reinfection rate” refers to the number of microorganisms infecting/attaching to circulating lymphocytes in the blood over a defined period of time. [0037] As used herein, the term “light” refers to electromagnetic radiation which includes radio waves, microwaves, infrared, visible light, ultraviolet light, X-rays, and gamma rays.

II. General Embodiments

[0038] Embodiments of the disclosure include systems, compositions, and methods useful for treating, preventing, delaying onset of, and/or reducing severity of a microbial infection in any individual in need thereof. Particular embodiments of the disclosure include systems, compositions, and methods useful for identifying or targeting the individual in need thereof. In certain embodiments, the individual in need thereof is or is not at risk of being infected with a microbe, has or has not been exposed to an individual infected with a microbe, has or has not been tested for the a microbial infection, has or has not tested positive for a microbial infection, and/or is or is not symptomatic of a microbial infection. An individual may be subject to methods of the disclosure upon determination that the individual has the microbial infection, upon determination that the individual has been exposed to one or more certain microbes or is at risk for infection, or upon routine preventative health maintenance for the individual. In certain embodiments, the individual in need thereof is a mammal. In certain embodiments, the mammal may be of any kind, including humans, dogs, cats, horses, pigs, sheep, and goats, for example.

[0039] In certain embodiments the microbial infection is a bacterial, fungal, protozoan, and/or a viral infection. In specific embodiments, the microbial infection is a bacterial infection. Examples of bacteria include, but are not limited to, Actinomyces, Bacillus, Bacteroides, Bordetella, Bartonella, Borrelia, Brucella, Campylobacter, Capnocytophaga, Chlamydia, Corynebacterium, Coxiella, Dermatophilus, Enterococcus, Ehrlichia, Escherichia, Francisella, Fusobacterium, Haemobartonella, Haemophilus, Helicobacter, Klebsiella, L-form bacteria, Leptospira, Listeria, Mycobacteria, Mycoplasma, Neisseria, Neorickettsia, Nocardia, Pasteurella, Peptococcus, Peptostreptococcus, Pneumococcus, Proteus, Pseudomonas, Rickettsia, Rochalimaea polypeptides, Salmonella, Shigella, Staphylococcus, group A streptococcus, group B streptococcus, Treponema, and Yersinia.

[0040] In specific embodiments, the microbial infection is a fungal infection. Examples of fungi include, but are not limited to, Absidia, Acremonium, Alternaria, Aspergillus, Basidiobolus, Bipolaris, Blastomyces, Candida, Coccidioides, Conidiobolus, Cryptococcus, Curvalaria, Epidermophyton, Exophiala, Geotrichum, Histoplasma, Madurella, Malassezia, Microsporum, Moniliella, Mortierella, Mucor, Paecilomyces, Penicillium, Phialemonium, Phialophora, Prototheca, Pseudallescheria, Pseudomicrodochium, Pythium, Rhino sporidium, Rhizopus, Scolecobasidium, Sporothrix, Stemphylium, Trichophyton, Trichosporon, and Xylohypha.

[0041] In specific embodiments, the microbial infection is a protozoan infection. Examples of protozoa include, but are not limited to, Babesia, Balantidium, Besnoitia, Cryptosporidium, Eimeria, Encephalitozoon, Entamoeba, Giardia, Hammondia, Hepatozoon, Isospora, Leishmania, Micro sporidia, Neospora, Nosema, Pentatrichomonas, Plasmodium. Examples of helminth parasites include, but are not limited to, Acanthocheilonema, Aelurostrongylus, Ancylostoma, Angiostrongylus, Ascaris, Brugia, Bunostomum, Capillaria, Chabertia, Cooperia, Crenosoma, Dictyocaulus, Dioctophyme, Dipetalonema, Diphyllobothrium, Diplydium, Dirofilaria, Dracunculus, Enterobius, Filaroides, Haemonchus, Lagochilascaris, Loa, Mansonella, Muellerius, Nanophyetus, Necator, Nematodirus, Oesophagostomum, Onchocerca, Opisthorchis, Ostertagia, Parafilaria, Paragonimus, Parascaris, Physaloptera, Protostrongylus, Setaria, Spirocerca Spirometra, Stephanofilaria, Strongyloides, Strongylus, Thelazia, Toxascaris, Toxocara, Trichinella, Trichostrongylus, Trichuris, Uncinaria, Wuchereria, Pneumocystis, Sarcocystis, Schistosoma, Theileria, Toxoplasma, and Trypanosoma.

[0042] In other embodiments, the microbial infection is a viral infection. Examples of viruses include adenovirus, alphavirus, calicivirus, coronavirus, distemper virus, Ebola virus, enterovirus, flavivirus, hepatitis virus, herpesvirus, infectious peritonitis virus, leukemia virus, Marburg virus, Norwalk virus, orthomyxovirus, papilloma virus, parainfluenza virus., the, paramyxovirus, parvovirus, pestivirus, picoma virus, pox virus, rabies virus, reovirus polypeptides, retrovirus, rotavirus, and vaccinia virus.

[0043] In specific embodiments, the viral infection is caused by a Coronaviridae family virus, including at least SARS-CoV-2, although in some cases the coronavirus is SARS-CoV or MERS, for example. In specific embodiments, the Coronaviridae family virus is a betacoronoavirus. In specific embodiments, the betacoronavirus is SARS-CoV-2. In specific embodiments, the viral infection is caused by a Filoviridae family virus, including at least an Ebola virus. Ebola is among the viruses which are highly sensitive to UV-C radiation, (Lytle et ah, 2005) and a critical step in Ebola virus infection is its replication in monocytes and vascular endothelial cells that leads to extreme viral loads in patient blood. (Ksiazek et ah, 1999)

[0044] Examples of specific SARS-CoV-2 viruses include the following listed in the NCBI GenBank® Database, and these GenBank® Accession sequences are incorporated by reference herein in their entirety: (a) LC534419 and LC534418 and LC528233 and LC529905 (examples of different strains from Japan); (b) MT281577 and MT226610 and NC_045512 and MN996531 and MN908947 (examples of different strains from China); (c) MT281530 (Iran); (d) MT126808 (Brazil); (e) MT020781 (Finland); (f) MT093571 (Sweden); (g) MT263074 (Peru); (h) MT292582 and MT292581 and MT292580 and MT292579 (examples of different strains from Spain); (i) examples from the United States, such as MT276331 (TX); MT276330 (FL); MT276328 (OR) MT276327 (GA); MT276325 (WA); MT276324 (CA); MT276323 (RI); MT188341 (MN); and (j) MT276598 (Israel). In particular embodiments, the disclosure encompasses treatment or prevention of infection of any of these or similar viruses, including viruses whose genome has at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or 99.9% identity to any of these viruses. In particular embodiments, the disclosure encompasses treatment or prevention of infection of any of these or similar viruses, including viruses whose genome has its entire sequence that is greater than 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or 99.9% identity to any of these viruses. As one specific example, the present disclosure includes methods of treatment or prevention of infection of a vims having a genome sequence of represented by GenBank® Accession No. NC_045512; origin Wuhan, China.

[0045] Embodiments of the disclosure include systems and methods useful to treat, prevent, delay onset of, or reduce severity of the symptoms of a microbial infection (including viral infection) in an individual in need thereof. In specific embodiments, an individual in need thereof may have one or more symptoms of a microbial infection. In specific embodiments, an individual in need thereof may have one or more symptoms of an infection by a vims of the Coronaviridae family, such as SARS-CoV-2. Common initial signs and symptoms of SARS- CoV-2 may include fever, cough, shortness of breath or difficulty breathing, tiredness, aches, chills, sore throat, loss of smell, loss of taste headache, diarrhea, or vomiting. As the viral infection progresses, the individual may develop pneumonia or acute respiratory distress syndrome (ARDS). [0046] Embodiments of the disclosure also include systems and methods useful to decrease the microbial load of an individual with a microbial infection (including a viral infection). In specific embodiments, virus inactivation is performed at the end of the viremic phase ( e.g ., 7-10 days after onset of symptoms, determined by the trend of lymphocyte count, and possible viral load assessment) of a viral infection in an individual with a progressing disease (e.g., an individual with persistent or worsening clinical condition). In specific embodiments, inactivation of said virus stops lymphocyte reinfection and/or reduces the lymphocyte reinfection rate in the individual. In specific embodiments, inactivation of said virus reduces extrapulmonary spread of SARS-CoV-2, including immune system damage. In specific embodiments, slowing the lymphocyte reinfection rate adequately attenuate the viral infection, until effective seroconversion can occur.

III. Systems of the Disclosure

[0047] In specific embodiments, the systems and methods useful to attenuate the microbial infection (including viral infection) include a plasmapheresis circuit combined with extracorporeal antiviral light irradiation of the plasma of the individual. This treatment option has not yet been considered against SARS-CoV-2 in COVID-19, (Cunningham el ah, 2020) or even any other light sensitive virus induced disease. In specific embodiments, the plasmapheresis circuit is a closed-loop system. In specific embodiments, the plasmapheresis circuit includes a light transparent reservoir and a light generator. In specific embodiments, the plasma of the individual is exposed to light as the plasma passes through the light transparent reservoir. In specific embodiments, the light irradiation inactivates microbes in the plasma. In specific embodiments, a filter eliminates microbial particles from the light-treated plasma. In specific embodiments, the optionally employed filter selectively eliminates microbial particles but not antimicrobial immunoglobulins (e.g., reinfusion of the individual’s own irradiated and filtered plasma, containing the already developed neutralizing antibodies, but devoid of infective microbes). In specific embodiments, the closed-loop plasmapheresis circuit reinfuses the potentially, but not necessarily filtered, light-treated plasma back into the individual. Importantly, this methodology would allow for the preservation of serum proteins and antibodies during irradiation (Chin et ah, 1997) and is readily available in advanced medical settings.

[0048] Turning to the figures, FIG. 1 illustrates a circuit diagram for the flow of blood components through a plasmapheresis and light irradiation system in accordance with a certain embodiment of the disclosure. In certain embodiments, the system 100 withdraws blood 115 from an individual 105 through a first conduit 110 and reinfuses blood components - cellular material 135 and light-treated and, optionally, filtered plasma 185 - back into the individual 105 through a second conduit 190.

[0049] In one embodiment of operation, blood 115 is withdrawn from the individual. The blood 115 flows through a first conduit 110 to the plasmapheresis unit 120, which separates the blood into blood components - plasma 125 and cellular material 130. Optionally, a third conduit 140 may direct a photosensitizer 145, into the plasma. From the plasmapheresis unit 120, the plasma 125 and, optionally, photosensitizer 145, are directed through a light transparent reservoir 160. While the plasma 125, and optionally, photosensitizer 145, pass through the light transparent reservoir 160, the plasma 125 and, optionally, photosensitizer 145, are exposed to light radiation 155 generated by a light irradiation unit 150 to produce light-treated plasma 165 and, optionally, photosensitizer 145. From the light transparent reservoir 160, the light-treated plasma 165 and, optionally, photosensitizer 145 are optionally directed through a photosensitizer removal unit 170 and/or a filtration unit 180 to produce light-treated and, optionally, filtered plasma 175. After light-treatment and, optionally, filtration, the blood components - cellular material 135 and light-treated and, optionally, filtered plasma 185 - are reinfused back into the individual 105 through a second conduit 190.

[0050] FIG. 2 illustrates a flow diagram for a method of plasmapheresis and light irradiation of plasma method in accordance with a certain embodiment of the disclosure. The method generally separates blood withdrawn from an individual into blood components - plasma and cellular material - and reinfuses the blood components - cellular material and light-treated and, optionally, filtered plasma - back into the individual. The method first separates blood withdrawn from an individual into blood components - plasma and cellular material. Optionally, a photosensitizer is directed into the plasma. The plasma, and, optionally, photosensitizer, are treated with light radiation by first directing the plasma, and, optionally, photosensitizer, through a light transparent reservoir, and then exposing the plasma, and, optionally, photosensitizer, passing through the light transparent reservoir to light radiation generated by a light irradiation unit to produce light-treated plasma, and, optionally, photosensitizer. The photosensitizer is optionally removed from the light-treated plasma. The light-treated plasma is optionally filtered to produce light-treated and, optionally, filtered plasma. Lastly, the cellular material and light-treated and, optionally, filtered plasma is reinfused back into the individual.

IV. Combination Treatments

[0051] In specific embodiments, an additional viral therapy or preventative may be provided in combination with the disclosed treatment. In specific embodiments, the additional viral therapy or preventative is for a Coronaviridae family infection (including SARS-CoV-2) selected from the group consisting of Azithromycin, AC-55541, Apicidin, AZ3451, AZ8838, Bafilomycin Al, CCT 365623, Daunorubicin, E-52862, Entacapone, GB110, H-89, Haloperidol, Indomethacin, JQ1, Loratadine, Merimepodib, Metformin, Midostaurin, Migalastat, Mycophenolic acid, PB28, PD-144418, Ponatinib, Remdesivir (GS-5734), Ribavirin, RS-PPCC, Ruxolitinib, RVX-208, S-verapamil, Silmitasertib , TMCB, UCPH-101, Valproic Acid, XL413, ZINC 1775962367, ZINC4326719, ZINC4511851, ZINC95559591, 4E2RCat, ABBV-744, Camostat, Captopril, CB5083, Chloramphenicol, Chloroquine (and/or Hydroxychloroquine), CPI-0610, Dabrafenib, DBeQ, dBET6, IHVR-19029, Linezolid, Lisinopril, Minoxidil, ML240, MZ1, Nafamostat, Pevonedistat, PS3061, Rapamycin (Sirolimus), Sanglifehrin A, Sapanisertib (INK128/M1N128), FK-506 (Tacrolimus), Tematin 4 (DA3), Tigecycline, Tomivosertib (eFT- 508), Verdinexor, WDB002, Zotatifin (eFT226), and a combination thereof. This list is not limiting for the additional viral therapy or preventative, however.

[0052] Alternatively, the disclosed treatment may precede, follow, or both an additional viral treatment or preventative by intervals ranging from minutes to weeks to months. In embodiments where the disclosed treatment and the additional agent are provided separately to an individual, one would generally ensure that a significant period of time did not expire between the time of each delivery, such that the disclosed treatment and the additional agent would still be able to exert an advantageously combined effect against the infectious disease process. In such instances, it is contemplated that one may deliver both modalities within about 12-24 h of each other and, in some embodiments, within about 6-12 h of each other. In some situations, it may be desirable to extend the time period for treatment significantly, however, where several days (2, 3, 4, 5, 6 or 7) to several weeks (1, 2, 3, 4, 5, 6, 7 or 8) to several months (from 1-12), any subrange therein, and so forth, lapse between the respective administrations.

[0053] Various combinations may be employed, for example, wherein the disclosed treatment is “A” and the secondary additional viral treatment or preventative is “B”: A/B/A B/A/B B/B/A A/A/B A/B/B B/A/A A/B/B/B B/A/B/B B/B/B/A B/B/A/B A/A/B/B A/B/A/B A/B/B/A B/B/A/A

B/A/B/A B/A/A/B A/A/A/B B/A/A/A A/B/A/A A/A/B/A

[0054] Administration of the disclosed treatment to a patient will follow general protocols for the administration of drugs, taking into account the toxicity, if any, of the molecule. It is expected that the treatment cycles would be repeated as necessary. It also is contemplated that various standard therapies, as well as surgical intervention, may be applied in combination with the described therapy.

III. Examples

[0055] The following examples are included to demonstrate exemplary embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow present techniques discovered by the inventors to function well in the practice of the disclosure, and thus can be considered to constitute exemplary modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure.

EXAMPLE 1

EXTRACORPOREAL UV LIGHT-PLASMAPHERESIS TREATMENT FOR COVID-19

[0056] An open label clinical trial is performed with patients with laboratory-confirmed COVID-19 and acute respiratory distress syndrome (ARDS) who meet the following criteria: severe pneumonia with rapid progression and continuously high viral load. Patients receive plasmapheresis and UV light radiation treatment of plasma according to the present disclosure. The treatment interval was day 1, 2, 4, 8, and 9 for a total of 5 sessions, each session for 4 hours. The standard plasmapheresis protocol consisted of removal, separation, and treatment of 1.5 plasma volume during each session, with 100% of the plasma volume reinfused into the patient along with blood cells. Patients are evaluated before and after treatment for the following: changes of body temperature, Sequential Organ Failure Assessment (SOFA) score (range 0-24, with higher scores indicating more severe illness), P_A02/F102, viral load, serum antibody titer, routine blood biochemical index, ARDS, and ventilator and extracorporeal membrane oxygenation (ECMO) supports. For at least a portion of the patients, following plasmapheresis, body temperature normalizes, SOFA score decreases, PA02/F102 increases, viral load decreases and becomes negative, SARS-CoV-2-specific ELISA and neutralizing antibody titers increase, ARDS resolves, and patients are weaned from mechanical ventilation. All patients are discharged from the hospital and remain in stable condition.

EXAMPLE 2

EXTRACORPOREAL VISIBLE LIGHT-METHYLENE BLUE-PLASMAPHERESIS

TREATMENT FOR COVID-19

[0057] An open label clinical trial is performed with patients with laboratory-confirmed COVID-19 and acute respiratory distress syndrome (ARDS) who meet the following criteria: severe pneumonia with rapid progression and continuously high viral load. Patients receive plasmapheresis and visible light radiation treatment of plasma and methylene blue photosensitizer according to the present disclosure. The treatment interval was day 1, 2, 4, 8, and 9 for a total of 5 sessions, each session for 4 hours. The standard plasmapheresis protocol consisted of removal, separation, and treatment of 1.5 plasma volume during each session, with 100% of the plasma volume reinfused into the patient along with blood cells. Patients are evaluated before and after treatment for the following: changes of body temperature, Sequential Organ Failure Assessment (SOFA) score (range 0-24, with higher scores indicating more severe illness), P_A02/F102, viral load, serum antibody titer, routine blood biochemical index, ARDS, and ventilator and extracorporeal membrane oxygenation (ECMO) supports. For at least a portion of the patients, following plasmapheresis, body temperature normalizes, SOFA score decreases, P_A02/F102 increases, viral load decreases and becomes negative, SARS-CoV-2-specific ELISA and neutralizing antibody titers increase, ARDS resolves, and patients are weaned from mechanical ventilation. All patients are discharged from the hospital and remain in stable condition.

[0058] Although the present disclosure and its advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the design as defined by the appended claims. Moreover, the scope of the present application is not intended to be limited to the particular embodiments of the process, machine, manufacture, composition of matter, means, methods and steps described in the specification. As one of ordinary skill in the art will readily appreciate from the present disclosure, processes, machines, manufacture, compositions of matter, means, methods, or steps, presently existing or later to be developed that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein may be utilized according to the present disclosure. Accordingly, the appended claims are intended to include within their scope such processes, machines, manufacture, compositions of matter, means, methods, or steps.

REFERENCES

[0059] All patents and publications mentioned in this specification are indicative of the level of those skilled in the art to which the disclosure pertains. All patents and publications herein are incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference in their entirety.

PUBLICATIONS

[0060] Chang L, Yan Y, Wang L. Coronavims Disease 2019: Coronaviruses and Blood Safety. TransfusMed Rev 2020.

[0061] Chen W, Lan Y, Yuan X, et al. Detectable 2019-nCoV viral RNA in blood is a strong indicator for the further clinical severity. Emerg Microbes Infect 2020; 9(1): 469-73.

[0062] Chin S, Jin R, Wang XL, et al. Virucidal treatment of blood protein products with UVC radiation. Photochem Photobiol 1997; 65(3): 432-5.

[0063] Cunningham AC, Goh HP, Koh D. Treatment of COVID-19: old tricks for new challenges. Crit Care2020; 24(1): 91.

[0064] Eickmann M, Gravemann U, Handke W, et al. Inactivation of Ebola virus and Middle East respiratory syndrome coronavims in platelet concentrates and plasma by ultraviolet C light and methylene blue plus visible light, respectively. Transfusion 2018; 58(9): 2202-7.

[0065] Gu J, Gong E, Zhang B, et al. Multiple organ infection and the pathogenesis of SARS. J Exp Med2005; 202(3): 415-24.

[0066] Ksiazek TG, Rollin PE, Williams AJ, et al. Clinical virology of Ebola hemorrhagic fever (EHF): virus, virus antigen, and IgG and IgM antibody findings among EHF patients in Kikwit, Democratic Republic of the Congo, 1995. J Infect Dis 1999; 179 Suppl 1: S177-87.

[0067] Li L, Wo J, Shao J, et al. SARS-coronavirus replicates in mononuclear cells of peripheral blood (PBMCs) from SARS patients. J Clin Virol 2003; 28(3): 239-44.

[0068] Lin L, Lu L, Cao W, Li T. Hypothesis for potential pathogenesis of SARS-CoV-2 infection-a review of immune changes in patients with viral pneumonia. Emerg Microbes Infect 2020; 9(1): 727-32.

[0069] Lytle CD, Sagripanti JL. Predicted inactivation of viruses of relevance to biodefense by solar radiation. J Virol 2005; 79(22): 14244-52.

[0070] Ng EK, Hui DS, Chan KC, et al. Quantitative analysis and prognostic implication of SARS coronavirus RNA in the plasma and serum of patients with severe acute respiratory syndrome. Clin Chem 2003; 49(12): 1976-80.

[0071] Tan L, Wang Q, Zhang D, et al. Lymphopenia predicts disease severity of COVID-19: a descriptive and predictive study. Signal Transduct Target Ther 2020; 5: 33.

[0072] Wang W, Xu Y, Gao R, et al. Detection of SARS-CoV-2 in Different Types of Clinical Specimens. JAMA 2020.

[0073] Wilder-Smith A, Chiew CJ, Lee VJ. Can we contain the COVID-19 outbreak with the same measures as for SARS? Lancet Infect Dis 2020.

[0074] Wolfel R, Corman VM, Guggemos W, et al. Virological assessment of hospitalized patients with COVID-2019. Nature 2020.

Claims

CLAIMS What is claimed is:

1. A method for treating or preventing a microbial infection in an individual, the method comprising the steps of:

(a) separating blood withdrawn from an individual into plasma and cellular material;

(b) treating the plasma with light radiation to produce light-treated plasma;

(c) optionally, filtering the light-treated plasma to produce light-treated and, optionally, filtered plasma; and

(d) reinfusing the cellular material and the light-treated and, optionally, filtered plasma back into the individual; wherein the method is effective to direct a substantially continuous flow of blood from the individual and a substantially continuous flow of the cellular material and light- treated and, optionally, filtered plasma back into the individual; and wherein the method is effective to reduce microbial load in the individual.

2. The method of claim 1, wherein the method is effective to treat, prevent, delay onset of, and/or reduce severity of one or more symptoms of a microbial infection in the individual.

3. The method of claim 1 or 2, wherein the light radiation is UV light radiation, visible light radiation, or a combination thereof.

4. The method of claim 1 or 2, wherein the light radiation is UVA radiation, UVB radiation, UVC radiation, or a combination thereof.

5. The method of any one of claims 1 to 4, wherein the step of treating the plasma with light radiation further comprises the steps of: (i) directing the plasma through a light transparent reservoir, and (ii) treating the plasma passing through the light transparent reservoir with light using a light irradiation unit to produce light-treated plasma.

6. The method of any one of claims 1 to 4, wherein the step of treating the plasma with light radiation further comprises the steps of (i) adding a photosensitizer to the plasma (ii) directing the plasma and photosensitizer through a light transparent reservoir, (iii) treating the plasma and photosensitizer passing through the light transparent reservoir with light using a light irradiation unit to produce light-treated plasma and photosensitizer, and (iv) optionally, removing the photosensitizer from the light treated plasma.

7. The method of claim 6, wherein the photosensitizer is an organic photosensitizer, an inorganic photosensitizer, or a combination thereof.

8. The method of claim 7, wherein the photosensitizer is a compound selected from the group consisting of: riboflavin, amotosalen, methylene blue, rose bengal, a porphyrin, and a phthalocyanine.

9. The method of claim 7, wherein the photosensitizer is a compound selected from the group consisting of: Ti02, ZnO, ZnS, CdS, Fe203, and W03.

10. The method of any one of claims 1 to 9, wherein the step of treating the plasma with light radiation is effective to inactivate microbes in the plasma.

11. The method of any one of claims 1 to 10, wherein the step of treating the plasma with light radiation is effective to inactivate at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbes in the plasma.

12. The method of any one of claims 1 to 11, wherein the step of filtering the light-treated plasma is effective to remove microbial particles from light-treated plasma.

13. The method of any one of claims 1 to 12, wherein the step of filtering the light-treated plasma is effective to remove at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbial particles from light-treated plasma.

14. The method of claims 1 to 13, wherein the step of filtering the light-treated plasma is effective to exclude removal of antibodies in the light-treated plasma.

15. The method of any one of claims 1 to 14, wherein the microbial infection is a bacterial infection, a fungal infection, a protozoan infection, a viral infection, or a combination thereof.

16. The method of any one of claims 1 to 15, wherein the microbial infection is a viral infection.

17. The method of claim 16, wherein the viral infection is an infection caused by a Coronaviridae family virus.

18. The method of claim 17, wherein the Coronaviridae family virus is a betacoronoavims.

19. The method of claim 18, wherein the betacoronavirus is SARS-CoV-2, SARS-CoV, or MERS-CoV.

20. The method of claim 19, wherein the betacoronavirus is SARS-CoV-2.

21. The method of claim 16, wherein the viral infection is an infection caused by a Filoviridae family virus.

22. The method of claim 21, wherein the Filoviridae family vims is an ebolavirus.

23. The method of any one of claims 10 to 22, wherein the step of reinfusing the cellular material and the light-treated and, optionally, filtered plasma into the individual is effective to reduce microbial load and/or cell reinfection rate in the individual by at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99%.

24. The method of any one of claims 1 to 23, wherein the method precedes, runs concurrently with, or follows an additional antimicrobial therapy administered to the individual.

25. A system for treating a microbial infection in an individual in need thereof, comprising:

(a) a first conduit configured to withdraw blood from an individual;

(b) a plasmapheresis unit configured to separate the blood into plasma and cellular material;

(c) a light irradiation unit configured to expose the plasma to light radiation;

(d) optionally, a filtration unit configured to filter the light-treated plasma; and

(e) a second conduit configured to reinfuse the combined cellular material and light- treated and, optionally, filtered plasma back into the individual; wherein the first and second conduits provide for substantially continuous flow of blood from the individual and of the cellular material and light-treated and, optionally, filtered plasma back into the individual; and wherein the system is configured to reduce microbial load in the individual.

26. The system of claim 25, wherein the light irradiation unit is configured to treat the plasma with UV radiation, visible light radiation, or a combination thereof.

27. The system of claim 25 or 26, wherein the light irradiation unit is configured to treat the plasma with UVA radiation, UVB radiation, UVC radiation, or a combination thereof.

28. The system of any one of claims 25 to 27, further comprising a light transparent reservoir through which the plasma is directed, wherein the light irradiation unit is further configured to expose the plasma to light radiation as the plasma passes through the light transparent reservoir to produce light-treated plasma.

29. The system of any one of claims 25 to 28, further comprising: a third conduit configured to direct a photosensitizer into the plasma prior to the light treatment of the plasma by the light irradiation unit; a light transparent reservoir through which the plasma and the photosensitizer are directed, wherein the light irradiation unit is further configured to treat the plasma and the photosensitizer with light radiation as the plasma and the photosensitizer pass through the light transparent reservoir to produce light-treated plasma and photosensitizer; and, optionally, a photosensitizer removal unit configured to remove the photosensitizer from the light-treated plasma.

30. The system of claim 29, wherein the third conduit is configured to direct an organic photosensitizer, an inorganic photosensitizer, or a combination thereof, into the plasma.

31. The system of claim 29 or 30, wherein the third conduit is configured to direct an organic photosensitizer selected from the group consisting of: riboflavin, amotosalen, methylene blue, rose bengal, a porphyrin, and a phthalocyanine, into the plasma.

32. The system of claim 29 or 30, wherein the third conduit is configured to direct an inorganic photosensitizer selected from the group consisting of: T1O2, ZnO, ZnS, CdS, Fe₂0₃, and WO3, into the plasma.

33. The system of any one of claims 25 to 32, wherein the light irradiation unit is further configured to inactivate microbes in the plasma during the treatment of the plasma with light radiation.

34. The system of any one of claims 25 to 33, wherein the light irradiation unit is further configured to inactivate at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbes in the plasma during the treatment of the plasma with light radiation.

35. The system of any one of claims 25 to 34, wherein the filtration unit is further configured to remove microbial particles from light-treated plasma during the filtration of the light- treated plasma.

36. The system of any one of claims 25 to 35, wherein the filtration unit is further configured to remove at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99% of microbial particles from light-treated plasma during the filtration of the light-treated plasma.

37. The system of any one of claims 25 to 36, wherein the filtration unit is further configured to exclude removal of antibodies in the light-treated plasma.

38. The system of any one of claims 25 to 37, wherein the system is further configured to reduce microbial load and/or cell reinfection rate in the individual.

39. The system of any one of claims 25 to 38, wherein the system is further configured to reduce microbial load and/or cell reinfection rate in the individual by at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, 95%, 96%, 97%, 98%, or 99%.