FR2804126A1 - Determining a protocol for deferred use of stored immunocompetent cells, useful e.g. for strengthening the immune system, using information on the subject's state of health - Google Patents

Abstract

Process for determining the parameters (P) of a protocol for deferred use of immunocompetent cells (A) taken from a personal library, of a human or animal. The library contains all the immunological information stored in (A), sampled successively and stored as preserved batches in one or more locations. Process for determining the parameters (P) of a protocol for deferred use of immunocompetent cells (A) taken from a personal library, of a human or animal comprises: (a) a set of measurements (M) of physical and/or biological characteristics, performed on a sample of body fluid and/or hair before, or during, sampling of (A); (b) characteristic information (CI) about the subject's state, deduced from (M); (c) treatment of (CI) in an information system to determine (P); and (d) storing treated CI is a database for management of (A). Independent claims are also included for the following: (1) process for managing batches of (A), for subsequent use, which uses determined (P); (2) system for determining (P); and (3) system for managing batches of (A).

Description

 "METHOD AND SYSTEM FOR MANAGING LOTS OF IMMUN-COMPETENT CELLS TAKEN FROM HUMAN OR ANIMAL SUBJECTS FOR DIFFERENT USES" The present invention relates to a method for managing batches of immunocompetent cells, in particular leukocytes, lymphocytes or monocytes, taken from human or animal subjects for deferred use. It also relates to a management system for implementing the method according to the invention.

Scientific and clinical studies have highlighted the therapeutic virtues of the self-use of lymphocyte and monocytic derivatives which contributes in particular to an increase in cellular immunity.

One promising application of this therapeutic method concerns the possibility of strengthening the immunity of a patient at a time in his life when this strengthening proves necessary or vital or to maintain this immunity throughout his life.

However, an important difficulty to be overcome lies in the availability of a patient's lymphocytes or monocytes over periods of time which could be between several months and several decades. Cryogenic storage techniques are already known which are now widely used in many fields of human and animal biology. In particular, banks have been set up for the conservation and storage of biological elements.

Document W089 / 04168 discloses a process for the isolation and preservation of hematopoietic cells from fetal and neonatal blood. This process is aimed in particular at the therapeutic use of fetal and neonatal cells for hematopoietic reconstitutions or in gene therapy, and can be implemented for the cryopreservation of fetal or neonatal blood cells for the purpose of autologous reconstitution.

Immunocompetent cells (lymphocytes, phagocytic cells: monocytes, macrophages) play a central role in the immune system. In particular, lymphocytes store information during life and are the support of the memory of cellular and humoral immunity. These immunocompetent cells constitute in a way a library, lymphocyte in particular, which enriches itself during the life, as the individual meets foreign organisms, during viral, parasitic or bacterial infections. It is thanks to it that the body can minimize the impact of infections during life. The mechanism of action of cellular immunity is known. Information is stored in particular in the walls of lymphocytes, as shown by the transfer factor and numerous publications. This mechanism also contributes to defense against malignant cells.

On the one hand, this memory is partially erased over time, as shown by the need to practice vaccination reminders to maintain effective protection. Regarding humoral immunity, antibody levels drop, quickly for IgM, more slowly for IgG and IgA.

On the other hand, errors are introduced over time and immunity often becomes less effective over the years. It is because of this degradation that infections like the flu are much more serious in the elderly. It would therefore be advantageous to be able to keep the information acquired throughout life.

The document WO9953030 discloses a method for managing batches of immuno-competent cells, comprising, for a given human subject: - packaging and storage of batches of immuno-competent cells in one or more storage centers, - a constitution and a enrichment of a personal library of immunocompetent cells from successively sampled batches, this personal library preserving a sum of immune information stored in the immuno-competent cells sampled, and, - in response to a request for treatment for the benefit of said human subject: - a processing of all or part of the immune information accumulated in said personal library, and - a localization of one or more stored batches of immuno-competent cells, followed by a transfer of this or these batches to a cell processing requesting.

This process thus provides patients with a guarantee of long-term storage of their lymphocytes, with the prospect of being able to dispose of them at any time, in particular to strengthen their immune system. It then becomes possible to restore people to their previous immunity and to transmit cellular immunity under rational and reliable management conditions, and also to provide access to their genetic code corresponding to the instant of blood collection.

However, beyond the need to preserve immunocompetent cells for the purposes of deferred use, there are important questions relating to the definition of an appropriate protocol for this deferred use. Indeed, it has been shown that a simple reinjection of immuno-competent cells previously taken from a human subject could lead to potentially serious immune problems. In addition, it has been shown that the quality and potential efficacy of the immuno-competent cells collected can be highly dependent on the general state of health of the human subject, with adverse consequences on the performance of current methods.

The aim of the present invention is to remedy these drawbacks by proposing a method for the management of batches of immuno-competent cells sampled for deferred use, which completely integrates for human subjects and by extension for animal subjects, the dimension of identity and state of health.

This objective is achieved with a process for managing batches of immunocompetent cells taken from human or animal subjects with a view to deferred use, comprising for each of these human or animal subjects: - packaging and storage of batches of immuno-competent cells successively sampled, in one or more storage centers, and - a constitution and an enrichment of a personal library of immuno-competent cells from the lots sampled, this personal library accumulating a sum of immune information stored in immunocompetent cells collected.

According to the invention, this method further comprises - a collection of information characteristic of the state of said human or animal subject, carried out before or during the collection of immuno-competent cells, and - a processing of said characteristic information to determine parameters of a protocol for deferred use of immunocompetent cells from the personal library of said human or animal subject.

With the method according to the invention, information characterizing the state of the human or animal subject, in particular his state of health and his psychological state, can be processed both to determine the advisability of taking immuno-competent cells and for select parameters of a deferred use protocol.

The information characterizing the state of the animal or human subject is preferably obtained from a treatment of a blood sample taken from the human or animal subject. They may include bioelectronic information resulting from a processing of respective measurements of the pH, of the oxidation reduction Rh2 and of the resistivity p of the blood previously drawn, according to the bioelectronic method of Vincent. The information characteristic of the state of the human or animal subject may also include information originating from a processing of images of sensitive crystallization of the blood previously taken from said human or animal subject, and / or information obtained by a capillarity study. elements of the hair system of said human or animal subject.

The information characteristic of the state of human or animal subjects and the immune information stored in the immunocompetent cells of said human or animal subjects can advantageously be introduced into an expert system used to determine parameters of the deferred use protocols. This expert system can be designed to provide an interpretation of information characteristic of the state and immune information of a human or animal subject with respect to a particular gene.

The processing of information characteristic of the state of a human or animal subject can be arranged to provide a determination of respective optimal proportions of different immuno-competent cells with a view to their deferred use, and in particular a determination of an optimal ratio between T4 and T8 lymphocytes. When the method according to the invention is implemented in a therapeutic protocol including a reinjection of lymphocytes onto a human or animal subject, the immuno-competent cells previously removed and stored can be subjected to an ex-vivo treatment prior to their reinjection . The method according to the invention can also be implemented in a therapeutic protocol including a reinjection of T lymphocytes with specific cytotoxic activity after ex-vivo growth, or in a gene therapy protocol.

It is also advantageously possible to provide that the method according to the invention further comprises, before the step of cryopreservation of a batch of immunocompetent cells, an initial step of cryogenization of said batch designed to cause the disappearance of the antibodies initially present in said lot.

This process can also comprise, before any reuse of a batch of immunocompetent cells, a step to verify the disappearance of the antibodies in this batch.

In a particular form of implementation of the method according to the invention, this further comprises, during a sequence of conditioning a batch of immuno-competent cells previously removed, an immunomagnetic selection step for lymphocytes or purified monocytes.

According to another aspect of the invention, a system is proposed for managing batches of immunocompetent cells belonging to human or animal subjects from which these batches have been removed for deferred use, implementing the method according to the invention, this system comprising for each of these human or animal subjects: - means for conditioning and for preserving batches of immuno-competent cells successively sampled, in one or more storage centers, and - means for constitute and to enrich a personal library of immunocompetent cells from the lots collected, this personal library accumulating a sum of immune information stored in the immunocompetent cells collected.

According to the invention, the system further comprises - means for collecting information characteristic of the state of said human or animal subject, before or during the collection of immuno-competent cells, and - means for processing said characteristic information in view of determining parameters of a protocol for deferred use of immunocompetent cells from the personal library of said human or animal subject.

Other features and advantages of the invention will appear in the description below. In the appended drawings given by way of nonlimiting examples: - Figure 1 is a block diagram of the management method according to the invention; and - Figure 2 is a timing diagram representing the main steps of the management method according to the invention.

The management method according to the invention generally comprises, with reference to FIG. 1, a first sequence of characterization and identification of the state of the human or animal subject, successive sequences of sampling of immuno-competent cells from said subject followed by sequences of cryopreservation and storage of batches of cells, and by one or more sequences of deferred use of conserved cells. During the characterization and identification sequence, samples of blood and other fluids and secretions such as saliva or urine are taken from the human or animal subject. Hair samples can also be taken. The blood sample can be processed by any suitable biological process to provide information characterizing the effective identity.

For example, Vincent's bioelectronic method, which includes a pH measurement, a measurement of the redox potential rH2, and a measurement of the p resistivity of the blood sample, provides an interesting characterization of the state of subject's present health.

Another useful method of biological evaluation is the processing and interpretation of sensitive blood crystallization images, and provides other characteristic state information regarding the subject's state of health with specific data on various disorders. physiological.

Characteristic state information provided either by a bioelectronic method or by a sensitive crystallization image method can be corroborated by information processed from a capillarity study carried out on a hair sample.

One can also use for the biological evaluation of the subject concerned an evaluation method implementing the photobiological effects of the incoherent polarized light visible on immunocompetent cells (in-vitro and in-vivo).

All the characteristic state information is gathered and processed to generate identity data which can be entered into an expert system for which specific rules are implemented.

Following the characterization and identification sequence which has just been described, a sampling sequence is carried out on the condition that the identity data satisfy physiological criteria to authorize this sampling sequence. The blood is collected and separated in order to obtain different cellular components such as lymphocytes or monocytes. The different cells obtained by separation are identified, selected, prepared and if necessary processed and divided into a plurality of n batches. These n batches of immunocompetent cells are then conditioned, treated by cryogenics, preserved and stored in various storage sites 1, ..., i, ..., n located to ensure the best level of integrity and security for these batches of cells.

It should be noted that an immunomagnetic selection method can be used for the purified lymphocytes and monocytes. Furthermore, cryo-protectants can advantageously be used during cryo-preservation of immunocompetent cells, in particular lymphocytes.

The immunocompetent cells are preferably stored at a temperature of between -80 ° C. and -120 ° C. Indeed, storage in liquid nitrogen at -196 ° C. often poses problems of infrastructure and logistics. The use of delayed lymphocytes can pose problems if there is conservation of antibodies because over time, there may be on the subject concerned a reaction of its own antibodies and a rejection of these. An initial cryogenization phase can be provided, aimed at removing the antibodies, within a batch of immunocompetent cells. Prior to any reuse of this batch of cells, a step of verifying the effective disappearance of these antibodies is carried out.

It is important to ensure the structural and functional integrity of the cryopreserved cells, in order to guarantee their reliability with a view to deferred use.

The batches of immunocompetent cells or components of the hematopoietic system can thus be stored over large variable periods of time, which can range from a few days to several decades, provided that appropriate storage of the components of the hematopoietic system over the long term be guaranteed. Furthermore, the principle of not storing all the batches of a patient on the same site contributes significantly to the security of the supply.

Throughout these processing sequences, data relating to each elementary step is collected, collected and stored in a cell management database which also receives data from the expert system including the subject's identity data. The stored immuno-competent cells belonging to a human or animal subject constitute their personal immune library which is linked to the cell management database which can be located within a management center controlling the plurality of storage sites and one or more preparation and cryopreservation centers.

When the management center receives a request for reuse for the benefit of a human or animal subject, an identification of the personal batches of cells is carried out by consulting (R) the cell management database and receiving (D) from said database of identification data associated with an appropriate batch of cells which is transmitted, preferably by express transport, from its storage site to a center specifically equipped for deferred use. This center can also be the center from which the initial sample was taken.

The transmitted batch is then repackaged at room temperature and the immunocompetent cells are cultured and / or subjected to ex-vivo treatment. Parameters of a reuse protocol are determined from identity data obtained by interrogating the cell management database and by processing this data in order for example to determine an optimal ratio between T4 and T8 lymphocytes for a reinjection. This reuse protocol is then implemented in a reuse process applied to the human or animal subject.

The management method according to the invention allows the constitution, throughout the life of a human or animal subject, of both a personal cellular library and a personal database containing data resulting from successive characterization sequences. and data generated by using the expert system, with reference to FIG. 2. By way of example, at a time To, a human or animal subject is subjected to a state characterization and identification process SCo which provides information characterizing the physiological identity and state of health of the subject. If this characterization sequence leads to a correct evaluation, a sequence of sampling of immuno-competent cells is then carried out at a time TCo on the human or animal subject. This state characterization and identification process carries out a test screening and biological evaluation of the subject concerned.

At a later instant Tj, another sequence of characterization of state is carried out on the human or animal subject and this sequence leads to data revealing a physiological disorder preventing any removal of cells. Therapeutic treatment can then be proposed in order to remedy the diagnosed disorder and another characterization sequence is carried out subsequently at a time Tn until a correct evaluation is obtained authorizing a sequence i for sampling immunocompetent cells.

The data from successive state characterization sequences are collected and processed in the expert system and stored in the cell management database, while the immunocompetent cells collected enrich the subject's personal cell library. When a reuse process for immunocompetent cells is prescribed for a human or animal subject, a deferred use protocol is determined using data stored in the database with, for example, optimal proportions between each cell type. The selected immunocompetent cells are then extracted from the personal cell library and, if necessary, treated ex-vivo. When these immunocompetent cells are ready for use, a reuse process according to the predetermined personal protocol is undertaken at time Tu.

It can be expected that a patient or subject following this program has a stock of immunocompetent cells or components of the hematopoietic system, which allows him, for example, to provide for successive self-uses, in particular in the form of self- injections, over a period of time, for the purpose of strengthening your immune system or for the purpose of gene therapy, or even for heavy use if necessary.

The management method according to the invention is for example implemented in the form of a software package installed on data processing and management systems, which can be implemented in batch management centers and can be connected to the all of the data processing sites located in the cytopheresis, storage and rapid logistics centers.

In addition to the numerous applications envisaged in the field of human therapy, the method according to the invention can also find particularly important applications in the field of mass animal production, in particular as an alternative to antibiotic therapy. They can also be used in the field of race horses to provide them with immune protection, as well as for pets as an anti-aging factor and for its action on the immune system.

Of course, the invention is not limited to the examples which have just been described and numerous modifications can be made to these examples without departing from the scope of the invention. Thus, it is possible to provide, within the framework of the sampling and reuse phases, additional technical steps according to medical imperatives and security constraints. The means of communication implemented between the various operational and management centers can be of any kind.

Furthermore, the information collected within the framework of the management method according to the invention can advantageously be processed for statistical purposes, with applications in the field of prevention and insurance.

Claims (24)

<U> CLAIMS </U> 1. Method for determining parameters of a protocol for the deferred use of immunocompetent cells from a personal library of a human or animal subject, this personal library accumulating a sum of immune information stored in the immune cells successively collected and packaged in the form of batches kept in one or more storage centers, characterized in that it comprises - a set of measurements of physical and / or biological characteristics carried out on samples of fluid and / or of hair of said human or animal subject before or during the collection of immunocompetent cells, - collection of information characteristic of the state of said human or animal subject resulting from said measurements, - processing of said characteristic information in an information system to determine parameters of said deferred use protocol s, and - storage of said processed information in a cell management database. 2. Method according to claim 1, characterized in that the information characterizing the state of the animal or human subject is obtained from a treatment of a blood sample taken from the human or animal subject. 3. Method according to claim 2, characterized in that the information characteristic of the state of the human or animal subject comprises bioelectronic information resulting from a processing of respective measurements of the pH, of the redox potential Rh2 and of the resistivity p of the blood previously taken (Vincent's bioelectronic method). 4. Method according to any one of the preceding claims, characterized in that the information characteristic of the state of the human or animal subject includes information originating from a processing of images of sensitive crystallization of the blood previously drawn from said human subject or animal. 5. Method according to any one of the preceding claims, characterized in that the information characteristic of the state of the human or animal subject comprises information resulting from a study of capillarity of elements of the hair system of said human or animal subject. 6. Method according to any one of the preceding claims, characterized in that the information characteristic of the state of human or animal subjects and the immune information stored in the immuno-competent cells of said human or animal subjects are introduced into an expert system used to determine parameters of deferred use protocols. 7. Method according to claim 6, characterized in that this expert system is designed to provide an interpretation of said information characteristic of the state and of said immune information of a human or animal subject with respect to a particular gene. 8. Method according to any one of the preceding claims, characterized in that the processing of information characteristic of the state of a human or animal subject is arranged to provide a determination of respective optimal proportions of different immuno-competent cells in view of their deferred use. 9. Method according to claim 8, characterized in that the processing of information characteristic of the state of a human subject provides a determination of an optimal ratio between T4 and T8 lymphocytes for their deferred use. 10. Method according to any one of the preceding claims, implemented in a therapeutic protocol including a reinjection of lymphocytes onto a human or animal subject, characterized in that the immuno-competent cells previously removed and preserved are subjected to an ex treatment -vivo prior to their reinjection. 11. The method of claim 10, implemented in a therapeutic protocol including a reinjection of T lymphocytes with specific cytotoxic activity after ex-vivo growth. 12. Method according to any one of the preceding claims, implemented in a gene therapy protocol. 13. Method according to any one of the preceding claims, characterized in that it further comprises, before the stage of cryopreservation of a batch of immunocompetent cells, an initial stage of cryogenization of said batch designed to cause disappearance antibodies initially present in said batch. 14. The method of claim 13, characterized in that it further comprises, prior to any reuse of a batch of immuno-competent cells, a step to verify the disappearance of the antibodies in this batch. 15. Method according to any one of the preceding claims, characterized in that it further comprises, during a sequence of conditioning a batch of immuno-competent cells previously removed, an immunomagnetic selection step for lymphocytes or purified monocytes. 16. System for determining parameters of a protocol for deferred use of immunocompetent cells from a personal library of a human or animal subject, this personal library accumulating a sum of immune information stored in the immune cells competent successively sampled and packaged in the form of batches kept in one or more storage centers, implementing the method according to any one of the preceding claims, characterized in that it comprises - means for measuring physical characteristics and / or biological samples of fluid and / or hair of said human or animal subject before or during the collection of immuno-competent cells, - means for collecting information characteristic of the state of said human or animal subject, said information characteristics resulting from said measurements, - means for trai ter said characteristic information in order to determine parameters of said deferred use protocol, and - means for storing said processed information in a cell management database. 17. System according to claim 16, characterized in that it is intended to process information characteristic of the state of the animal or human subject obtained from a treatment of a blood sample taken from the human or animal subject . 18. The system as claimed in claim 17, characterized in that it is intended to process bio electronic information resulting from a processing of respective measurements of the pH, of the Rh2 redox potential and of the resistivity of the blood previously drawn ( Vincent's bioelectronic method). 19. System according to any one of claims 16 to 18, characterized in that it is further provided for processing information originating from a processing of images of sensitive crystallization of the blood previously taken from said human or animal subject. 20. System according to any one of claims 16 to 19, characterized in that it is further provided for processing information originating from a capillarity study of elements of the hair system of said human or animal subject. 21. System according to any one of claims 16 to 20, characterized in that it further comprises means for controlling and enriching an expert system from information characteristic of the state of human or animal subjects and of immune information stored in the immuno-competent cells of said human or animal subjects, in order to determine parameters of the protocols for deferred use. 22. System according to claim 21, characterized in that the processing means are provided to provide an interpretation of said information characteristic of the state and of said immune information of a human or animal subject with respect to a particular gene. 23. System according to any one of claims 16 to 22, characterized in that it further comprises means for providing, from information characteristic of the state of the human or animal subject, a determination of respective optimal proportions of different immuno-competent cells with a view to their deferred use. 24. The system of claim 23, characterized in that the determination means comprise means for determining an optimal ratio between T4 and T8 lymphocytes for their deferred use.