JP5036945B2 - Method and apparatus for determining protocol parameters for deferred use of immunocompetent cells - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a method for determining parameters of a protocol for deferred use (delayed use) of immunocompetent cells (immunocompetent cells), and immunocompetent cells collected from a human or animal subject (subject) for the purpose of deferred use. The present invention relates to a method for managing lots (particularly leukocytes, lymphocytes or mononuclear leukocytes). The present invention also relates to an apparatus for determining parameters of a protocol for deferring use of immunocompetent cells, and a lot management apparatus for immunocompetent cells using the parameter determination apparatus.
[0002]
[Prior art]
Scientific and clinical studies have demonstrated the therapeutic effect of automatic use of lymphocyte and leukocyte derivatives, which in particular contributes to increased cellular immunity.
[0003]
A promising application of this method of treatment relates to the possibility of enhancing a patient's immunity when it is necessary or important during the lifetime of the patient or to maintain immunity throughout the patient's lifetime.
[0004]
However, an important problem that must be solved is that the patient's lymphocytes and white blood cells are available for months to decades. Cryopreservation techniques that are used extensively in many fields of human or animal biology are known. In particular, banks have been established for the preservation and storage of biological elements.
[0005]
Document WO89 / 04168 discloses a method for separating and storing hematopoietic cells from fetal and neonatal blood. This method is specifically intended for the therapeutic use of fetal and neonatal cells for hematopoietic remodeling or in gene therapy, and for cryopreservation of fetal or neonatal blood cells for the purpose of autologous remodeling can do.
[0006]
Immune competent cells (lymphocytes, phagocytic cells, mononuclear leukocytes, macrophages) play a central role in the immune system. In particular, lymphocytes preserve information during life and are memory carriers for cellular and humoral immunity. These immunocompetent cells constitute certain libraries (especially lymphocyte libraries) that are enriched in the living process as the solid encounters foreign organisms during infection by viruses, parasites or bacteria . It is this library that allows living organisms to reduce the effects of infection in the process of living. The mechanism of cellular immunity is known. As described in many publications, information is stored especially in the walls of lymphocytes. This mechanism also contributes to protection against malignant cells.
[0007]
On the one hand, this memory partially disappears over time, indicating that it is necessary to repeat vaccine injections to preserve effective protection. For humoral immunity, the antibody ratio declines rapidly for IgM and more slowly for IgG and IgA.
[0008]
On the other hand, errors are introduced over time and the effectiveness of immunity often decreases year by year. It is because of such deterioration that infections such as epidemic colds become very heavy for the elderly. Therefore, it would be convenient if the information acquired during the lifetime could be saved.
[0009]
The document WO9953030 discloses a method for managing a lot of immunocompetent cells, which for each given human subject,
-Preparing and storing lots of immunocompetent cells in one or more storage centers;
Constructing and enriching a personal library of immunocompetent cells from a sequentially collected lot (this personal library stores the sum of the immune information stored in the collected immunocompetent cells), and
-In response to a processing request for said human subject,
- treating all or part of the immune information stored in the personal library, and,
- localized one or more immunocompetent cells lots stored, to transfer the lot to the cell processing center,
Is included.
[0010]
This method therefore guarantees the patient to store the patient's lymphocytes for an extended period of time, making them available at any time, particularly to strengthen the patient's immune system. Thus, it is possible to re-provide people's previous immunity, transmit cellular immunity with reasonable and reliable control conditions, and provide access to the corresponding genetic code at the time of blood collection.
[0011]
[Problems to be solved by the invention]
However, in addition to the requirement to preserve immunocompetent cells for deferred use, an important issue is raised regarding the definition of an appropriate protocol for this deferred use. That is, it has been found that mere reinfusion of immunocompetent cells previously collected from human subjects can potentially lead to serious immunity problems. Furthermore, it was found that the potential quality and effectiveness of the collected immunocompetent cells strongly depend on the general health condition of the human subject, which has an adverse effect on the performance of conventional methods.
[0012]
It is an object of the present invention to improve the above mentioned difficulties and to provide a method for determining parameters of a protocol for deferred use of immunocompetent cells obtained from a human or animal-based personal library. library is accumulated sum of immune information stored in the immunocompetent cells are sequentially collected adjustment in the form of lots stored in one or more storage centers, the individual library for each person entity, and Extensiblely fully integrates the dimensions of identity and health status for each animal subject.
[0013]
[Means for Solving the Problems]
According to the present invention, this determination method comprises:
Multiple measurements of physical and / or biological characteristics performed on said human or animal-based body fluids and / or hair samples before or during collection of immunocompetent cells;
-Collection of information characteristic of the human or animal-based state obtained from the measurement,
The processing of the characteristic information in an information device to determine parameters of the protocol for deferred use; and
-Storing the processed information in a cell management database;
And
Execution of a sequence for collecting immunocompetent cells from a human or animal subject is adjusted according to the result of processing the characteristic information.
[0014]
According to the method of the present invention, information characterizing the human or animal-based state (especially its health and psychological state) can be used to determine the opportunity for collection of immunocompetent cells as well as parameters for deferred use protocols. Can be processed at the same time.
[0015]
Information characterizing the state of the human or animal subject is preferably obtained by processing a blood sample collected from the human or animal subject. These pieces of information may include bioelectronic information obtained by processing the measured values of blood pH, redox potential Rh2, and resistance ρ collected in advance according to Vincent's bioelectronic method.
[0016]
Information characteristic of the state of the human or animal subject is also information obtained by processing a perceptible crystallized image of blood previously collected from the human or animal subject and / or human or animal subject hair. It can contain information obtained by studying capillary action of the elements.
[0017]
Information characteristic of human or animal-based status and immune information stored in human or animal-based immune competent cells is preferably introduced into the expert system used to determine the parameters of the deferred protocol Is done. The expert system can be configured to provide an interpretation of the information and immune information characteristic of a human or animal-based condition for a particular gene.
[0018]
The processing of information characteristic of the human or animal-based state is particularly useful for determining the optimal proportion of different immunocompetent cells taking into account the deferred use of immunocompetent cells, in particular for lymphocyte T4 and lymphocyte T8. It can be configured to determine the optimal ratio.
[0019]
When the method of the invention is used in a therapeutic protocol involving lymphocyte reinfusion for human or animal subjects, previously harvested and stored immune competent cells are subjected to in vitro treatment prior to reinfusion. The methods of the invention can also be used in therapeutic or gene therapy protocols involving reinfusion of lymphocyte T with specific cytotoxic effects after in vitro growth.
[0020]
The treatment protocol in which the method of the invention is used generally includes the step of confirming the innocuous nature of lymphocytes prior to reinfusion. This confirmation step involves a test of lymphocyte oxidative stress before reinfusion, in which the lymphocytes are attacked by free radicals. In accelerated testing for a period of about 3 to 6 months, poorly preserved lymphocytes exposed to free radicals produced quantifiable oxidative stress.
[0021]
Oxidative stress tests can preferably be used to confirm different treatment methods by in vitro treatment and their sufficiency against the human or animal subject. That would be too aggressive lymphocyte products are very large amount of generating free radical. In the case of in vitro treatments including cytotoxin treatments, this test makes it possible to select the treatment most suitable for the patient.
[0022]
Another embodiment where it is particularly promising to use the methods of the present invention relates to therapeutic protocols, which include in vitro treatment between lymphocytes and vaccine prior to reinfusion. It was confirmed that no antibody was produced.
[0023]
The methods of the invention can also be used in therapeutic protocols that include in vitro treatments for allergy desensitization of lymphocytes prior to reinfusion. The product thus obtained is then mixed with the stored lymphocytes before reinfusion.
[0024]
It should be noted that the treatment protocol in which the method of the invention is used can include lymphocyte reinfusion with lymphatic vessels. The aim is to reduce the dosage to be prescribed, in particular to improve patient tolerance and improve the speed of response.
[0025]
Also within the scope of the present invention, a treatment protocol for blood transfusion from a donor subject to a receptor subject can be provided, which includes replacing donor-dominant lymphocytes with receptor-dominant lymphocytes.
[0026]
Also, the method of the present invention preferably further comprises an initial step of freezing the lot to eliminate the antibody initially present in the lot prior to the cryopreservation step of the lot of immunocompetent cells.
[0027]
The method can further include the step of confirming the disappearance of antibodies in the lot prior to reuse of the lot of immunocompetent cells.
[0028]
In a particular embodiment of the method of the invention, the method comprises the step of immunomagnetically sorting purified lymphocytes or mononuclear leukocytes during a preparation sequence of a previously collected lot of immunocompetent cells. In addition.
[0029]
The determination method of the present invention can be preferably carried out by a method of managing a lot of immunocompetent cells collected from a human or animal subject for the purpose of deferred use, and this method is performed for each human or animal subject. ,
Preparing and storing sequentially collected lots of immunocompetent cells in one or more storage centers; and
Constructing and enriching a personal library of immunocompetent cells from the collected lots (this personal library accumulates the sum of the immune information stored in the collected immunocompetent cells),
including.
[0030]
According to another aspect of the invention, the lot management method further includes:
-Collection of information characteristic of the human or animal-based condition, performed before or during collection of immunocompetent cells; and
Processing the characteristic information to determine the parameters of the protocol for deferred use of immunocompetent cells obtained from the human or animal-based personal library;
Including
Execution of a sequence for collecting immunocompetent cells from a human or animal subject is adjusted according to the result of processing the characteristic information.
[0031]
According to yet another aspect of the present invention, the present invention provides an apparatus for determining the parameters of a protocol for deferred use of immune competent cells obtained from a human or animal-based personal library while performing the method of the present invention. As provided, this personal library accumulates a sum of immune information stored in immunocompetent cells that have been sequentially collected and adjusted in the form of lots stored in one or more preservation centers.
[0032]
This parameter determination device
Means for measuring physical and / or biological characteristics of said human or animal-based body fluid and / or hair sample before or during collection of immunocompetent cells;
Means for collecting information characteristic of the human or animal-based state obtained from the measurement;
The characteristic information is used to determine the parameters of the protocol for deferred use and to control the execution of a sequence of collecting immune competent cells from a human or animal subject according to the results of processing the characteristic information. Means for processing;
Means for storing the processed information in a cell management database;
It is characterized by having.
[0033]
According to still another aspect of the present invention, the present invention provides a method for carrying out the method of the present invention, and comprising the step of immunocompetent cell belonging to a human or animal subject from which a lot of immunocompetent cells has been collected for the purpose of deferred use. Provide a device to manage, for each human or animal subject:
Means for preparing and storing lots of sequentially collected immunocompetent cells in one or more storage centers; and
Means for constructing and enriching a personal library of immunocompetent cells from the collected lots (this personal library accumulates the sum of the immune information stored in the collected immunocompetent cells);
It has.
[0034]
According to the invention, this device further comprises:
-Means for collecting information characteristic of said human or animal-based condition before or during collection of immunocompetent cells; and
-To determine the parameters of the protocol for deferred use of immunocompetent cells obtained from the human or animal-based personal library, and to obtain immunocompetent cells from human or animal subjects according to the results of processing characteristic information. Means for processing said characteristic information to control execution of a sequence to be collected;
It has.
[0035]
Other features and advantages of the present invention will become more apparent according to the following description. The accompanying drawings show non-limiting examples.
[0036]
DETAILED DESCRIPTION OF THE INVENTION
Referring to FIG. 1, the management method of the present invention generally includes an initial sequence that characterizes and identifies the state of a human or animal subject, and a plurality of sequences before and after collecting immunocompetent cells from the subject, This is followed by a plurality of sequences in which cell lots are stored frozen and stored, and one or more sequences in which the stored cells are deferred.
[0037]
In the sequence to be characterized and identified, samples of blood and other body fluids and secretions such as saliva or urine are taken from a human or animal subject. Hair samples can also be taken. A sample of blood can be processed by any suitable biological method that provides information characterizing effective identity.
[0038]
For example, Vincent's bioelectronic method provides an interesting characterization of the subject's current health condition, including measuring the pH of a blood sample, measuring the redox potential rH2, and measuring the resistance ρ.
[0039]
Another useful biological assessment method consists of processing and interpreting perceptible crystallized images of blood, providing accurate data on various physiological problems along with other characteristic information about the subject's health. To do.
[0040]
State characterization information obtained by bioelectronic methods or perceptible crystallization image methods can be validated by information processed by capillary action studies on hair samples.
An evaluation method that uses the photobiological action of visible incoherent polarized light (in vitro or in vivo) on immunocompetent cells can also be used for biological evaluation of the subject in question.
[0041]
All information characteristic of the state is collected and processed to generate data for identification, which can be input to an expert system where specific rules are executed.
Following the characterization and identification sequence described above, the acquisition sequence is performed provided that the identification data meets the physiological requirements necessary for this acquisition sequence.
[0042]
Blood is collected and separated to obtain various constituent cells, such as lymphocytes and mononuclear leukocytes. The various cells obtained by separation are identified, sorted, prepared, processed if necessary, and divided into n lots. These n lots of immunocompetent cells are then conditioned, frozen and processed at various storage sites (1,...) Positioned to ensure the best integrity and safety of the cells of these lots. Store in i, ..., n).
[0043]
It should be noted that immunomagnetic sorting can be used to obtain purified lymphocytes and mononuclear leukocytes. Further, preferably, a cryoprotective agent can be used during cryopreservation of immunocompetent cells (particularly lymphocytes).
Immunocompetent cells are preferably stored at a temperature between -80 ° C and -120 ° C. That is, storage in liquid nitrogen at −196 ° C. often results in problems of infrastructure and logic calculations.
[0044]
Deferred use of lymphocytes can cause problems when storing antibodies. This is because as time passes, there is a reaction to the subject's own antibody against the subject and there is antibody rejection. An initial step of cryogenic liquefaction can be provided to destroy the antibodies in the lot of immunocompetent cells. Before reusing this lot of cells, a step of confirming effective disappearance of the antibody is performed.
[0045]
Ensuring the structural and functional integrity of frozen storage cells is important to ensure their reliability for deferred use.
Thus, immunocompetent cells or hematopoietic component lots, if appropriate storage of the hematopoietic system components over a long period is guaranteed, can be stored for long variable period to obtain Oyobi decades from days . Furthermore, the principle of not storing all lots of a patient in one place contributes substantially to the safety of the supply.
[0046]
Through the processing sequence, data relating to each basic element stage is collected, integrated, and stored in a cell management database (which also receives data from the expert system, eg, data for identifying the subject). Conserved immunocompetent cells belonging to a human or animal subject constitute an individual (individual) immune library of the subject associated with a cell management database, which can be stored at multiple storage sites or one or more preparations. -It can be placed in the management center that controls the frozen storage center.
[0047]
When the management center receives a re-use request for a human or animal subject, refer to the cell management database (R) and identify the appropriate cell lot identification data (this is preferably deferred from the storage site by express transport). The identification of the individual cell lot is performed by receiving (D) from the database (transferred towards a center specially prepared for use). This center can also be the first collection center.
[0048]
The lot is then brought to room temperature and the immunocompetent cells are cultured and / or treated in vitro (ex vivo). By using these data for identification purposes obtained by querying the cell management database and processing these data for the purpose of determining the optimal proportion of lymphocytes T4 and T8 for reinfusion, for example, Determine the protocol parameters. This reuse protocol is then used in a reuse process applied to human or animal subjects.
[0049]
As shown in FIG. 2, the management method according to the present invention uses a personal cell library and a personal database (data obtained from sequential characterization sequences and an expert system) during the life of a human or animal subject. Including the data to be generated). For example, at time To, a human or animal subject undergoes a process SCo (providing information characterizing the subject's physiological identity and health status) that characterizes and identifies the condition. If this characterization sequence leads to a correct assessment, an immunocompetent cell collection sequence for human or animal subjects is performed at time Tco. This process of characterizing and identifying the state implements the subject's test “screening” and biological assessment operations.
[0050]
At a later point in time Tj, another state characterization sequence is performed on the human or animal subject, which generates data indicative of physiological troubles that inhibit cell harvesting. In this case, treatment treatments can be proposed to improve the diagnosed trouble, and other characterization sequences are finally performed at time Tn until a correct evaluation is obtained that allows the collection sequence i of immune competent cells. To do.
[0051]
Data obtained from the sequential state characterization sequence is collected, processed by an expert system, and stored in a cell management database. On the other hand, immunocompetent cells enrich the main individual cell library.
If an immunocompetent cell reuse process is prescribed for a human or animal subject, a deferred use protocol can be used, for example, using data stored in the database, along with an optimal percentage of each type of cell. decide. Next, the selected immunocompetent cells are extracted from the personal cell library and treated in vitro as necessary.
Once these immunocompetent cells are ready for use, a reuse process according to a pre-determined personal protocol is performed at time Tu.
[0052]
Patients or subjects that follow this program will use a stock of immunocompetent cells or hematopoietic components that is intended to enhance the immune system, for the purpose of gene therapy, or as needed. For large-scale use, it is possible to provide the patient or subject for a predetermined period of time, for example continuous automatic use (especially automatic use in the form of automatic injection).
[0053]
The management method according to the present invention includes, for example, software installed in a data processing device (which can be connected to a data processing site located in a blood pheresis / storage / rapid processing center) located in a lot management center. Embodied in form.
Besides the many applications aimed at the field of human therapy, the method according to the invention also has important applications in the field of animal mass production (especially as an alternative to antibiotic therapy). This method can also be used in the field of racehorses for the purpose of providing immune protection and for pet animals as an anti-aging factor and for its action on the immune system.
[0054]
Of course, the present invention is not limited to the embodiments described above, and various modifications can be made to these embodiments without departing from the scope of the present invention. That is, at the collection and reuse stage, additional technical stages can be provided depending on medical requirements and safety constraints. Communication means used between different operation and management centers can be of any nature.
Furthermore, the data collected by the management method of the present invention can be preferably processed for statistical purposes, for use in the field of prevention and insurance.
[Brief description of the drawings]
FIG. 1 is a schematic diagram of a management method of the present invention.
FIG. 2 is a time chart showing main steps of the management method of the present invention.

Claims (20)

A method for determining the parameters of a protocol for deferred use of immunocompetent cells obtained from a human or animal-based personal library, which is collected sequentially in the form of lots stored in one or more preservation centers. Accumulating the immune information stored in the adjusted immunocompetent cells, this method features:
Multiple measurements of physical and / or biological characteristics performed on said human or animal-based body fluids and / or hair samples before or during collection of immunocompetent cells;
-Collection of information characteristic of the human or animal-based state obtained from the measurement,
The processing of the characteristic information in an information device to determine parameters of the protocol for deferred use; and
-Storing the processed information in a cell management database;
A method comprising comprising:   2. The method according to claim 1, wherein the information characterizing the state of the human or animal subject is obtained by processing a blood sample collected from the human or animal subject.   Information characteristic of the human or animal-based state includes bioelectronic information obtained by processing pre-collected blood pH, redox potential Rh2, and resistance ρ measurements. A method according to claim 2.   4. The information according to claim 1, wherein the information characteristic of the state of the human or animal subject includes information obtained by processing a sensitive crystallized image of blood previously collected from the human or animal subject. Based on the method.   5. The method according to claim 1, wherein the information characteristic of the human or animal-based state includes information obtained by studying capillary phenomenon of the human or animal-based hair element.   Information characteristic of the condition of the human or animal subject and the immune information stored in the immunocompetent cells of the human or animal subject should be introduced into the expert system used to determine the parameters of the deferred protocol. A method based on any one of claims 1 to 5 characterized in that The expert system, with respect to one gene, a method based on claim 6, characterized in that it is configured to provide an interpretation of the characteristic the information and the immune information on the state of the human or animal entity.   The processing of information characteristic of a human or animal-based condition is configured to determine an optimal proportion of each different immunocompetent cell taking into account the deferred use of immunocompetent cells. A method based on one of 1 to 7.   9. The method according to claim 8, wherein the processing of the information characteristic of the human subject state determines an optimal ratio of lymphocyte T4 and lymphocyte T8 in consideration of deferred use of immune competent cells. A method for managing a lot of immunocompetent cells collected from a human or animal subject for the purpose of deferral use while performing the parameter determination method according to any one of claims 1 to 9 , comprising: Or for each animal subject,
Preparing and storing sequentially collected lots of immunocompetent cells in one or more storage centers; and
Constructing and enriching a personal library of immunocompetent cells from the collected lots that accumulates the sum of the immune information stored in the collected immunocompetent cells;
Including
The features of this method are further:
-Collection of information characteristic of the human or animal-based condition, performed before or during collection of immunocompetent cells; and
Processing the characteristic information to determine the parameters of the protocol for deferred use of immunocompetent cells obtained from the human or animal-based personal library;
A method comprising comprising: An apparatus for determining parameters of a protocol for deferred use of immunocompetent cells obtained from a human or animal-based personal library while performing a method according to any of claims 1 to 9 , wherein the personal library comprises one or more Accumulated sum of immune information stored in immunocompetent cells collected and adjusted sequentially in the form of lots stored in the preservation center of
The features of this device are:
Means for measuring physical and / or biological characteristics of said human or animal-based body fluid and / or hair sample before or during collection of immunocompetent cells;
Means for collecting information characteristic of the human or animal-based state obtained from the measurement;
Means for processing the characteristic information to determine parameters of the protocol for deferred use;
Means for storing the processed information in a cell management database;
A device comprising: 12. The apparatus according to claim 11 , wherein the apparatus is configured to process information characteristic of a human or animal-based state obtained by processing a blood sample collected from a human or animal-based body. 13. An apparatus according to claim 12 , wherein the apparatus is configured to process bioelectronic information obtained by processing pre-collected blood pH, redox potential Rh2, and resistance measurements. Further based on either of the claims 11 to 13, characterized in that it is configured to process the information obtained by treating the sensitive crystallization image blood previously collected from a human or animal main device. 15. An apparatus according to any one of claims 11 to 14 , further configured to process information obtained by studying capillarity of human or animal-based hair elements. Control and enrich the expert system based on information characteristic of human or animal-based conditions and immune information stored in human- or animal-based immune competent cells to determine deferred protocol parameters 16. The apparatus according to any one of claims 11 to 15 , further comprising means. 17. The apparatus according to claim 16 , wherein the processing means is configured to provide an interpretation of the information and the immune information characteristic of a human or animal-based condition for a gene. Claim 11, characterized in that it further comprises means for determining based on the optimal ratio of each of the different immune competent cells in consideration of the deferred use of immunocompetent cells characteristic information on the state of the human or animal entity To 17 devices. 19. The apparatus according to claim 18 , wherein the determining means includes means for determining an optimal ratio of lymphocyte T4 and lymphocyte T8 in consideration of deferred use of immune competent cells. An apparatus for managing a lot of immunocompetent cells belonging to a human or animal subject from which a lot of immunocompetent cells have been collected for deferred use while performing the method according to claim 10 , comprising: For each human or animal subject:
Means for preparing and storing lots of sequentially collected immunocompetent cells in one or more storage centers; and
Means for constructing and enriching a personal library of immunocompetent cells that accumulates the sum of the immune information stored in the collected immunocompetent cells from the collected lots;
With
The features of this device are further:
-Means for collecting information characteristic of said human or animal-based condition before or during collection of immunocompetent cells; and
Means for processing the characteristic information to determine parameters of a protocol for deferred use of immunocompetent cells obtained from the human or animal-based personal library;
A device comprising:

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