Pairing processes for preparing alloreactive cytotoxic T cells
Provided in certain embodiments are methods for pairing patient cells and donor cells to prepare cytotoxic T cells. Such cytotoxic T cells could be administered to the patient for treating certain disorders, such as a cancer (for example, brain cancer).
1. A method of preparing alloreactive cytotoxic T cells comprising:
providing stimulator information, wherein the stimulator information comprises patient cell antigen information determined through one or more of serotyping or a high or intermediate resolution molecular sequencing method of major histocompatibility complex (MHC) information;
providing responder information, wherein the responder information comprises donor cell antigen information determined through one or more of serotyping or a high or intermediate resolution molecular sequencing method of MHC information;
comparing the stimulator information to the responder information;
identifying presence or absence of a partial mismatch between the stimulator information and the responder information by analyzing and comparing HLA alleles of donor and patient, the partial mismatch comprising 1-6 antigen units or 1-10 amino acid differences in corresponding HLA molecules of the donor and the patient;
selecting a patient cell and donor cell pair to prepare alloreactive cytotoxic T cells based on the presence of the partial mismatch; and
combining cells of the patient with cells of the donor to generate the alloreactive cytotoxic T cells in an alloreactive cytotoxic T cell reaction based on the patient cell and donor cell pair,
whereby the alloreactive cytotoxic T cells are produced, the alloreactive cytotoxic T cells being activated against the cells of the patient which present the antigen.
2. The method of claim 1 , wherein the patient cell antigen information is determined at least partly through a high or intermediate resolution molecular sequencing method of MHC information.
3. The method of claim 1 , wherein the patient cell antigen information is determined at least partly through a high or intermediate resolution molecular DNA sequencing method.
4. The method of claim 1 , wherein the patient cell antigen information is determined at least partly through a high or intermediate resolution molecular RNA sequencing method.
5. The method of claim 1 , wherein the patient cell antigen information is determined at least partly through a high or intermediate resolution molecular protein sequencing method.
6. The method of claim 1 , wherein the donor cell antigen information is determined at least partly through a high or intermediate resolution molecular DNA sequencing method.
7. The method of claim 1 , wherein the high or intermediate resolution molecular sequencing method comprises one or more of a sequence based typing (SBT), sequence specific primer (SSP), restriction fragment length polymorphism (RFLP), or sequence specific oligonucleotide (SSO) method.
8. The method of claim 1 ,
wherein providing patient cell information comprises providing patient cell information which includes HLA class I or HLA class II antigen information; and
wherein the donor cell information is determined through a high or intermediate resolution molecular sequencing method of HLA class I or HLA class II antigen information of a responder and a stimulator.
9. The method of claim 1 ,
wherein providing patient cell information comprises providing patient cell antigen information which includes HLA class I antigen information; and
wherein the donor cell information is determined through a high or intermediate resolution molecular sequencing method of HLA class I antigen-type information.
10. The method of claim 9 , wherein the cell antigen information includes HLA class I alpha helix, beta helix, or peptide binding groove information.
11. The method of claim 1 ,
wherein providing patient cell information comprises providing patient cell antigen information which includes HLA II antigen information; and
wherein the donor cell information is determined through a high or intermediate resolution molecular DNA sequencing method of HLA class II antigen-type information.
12. The method of claim 1 , wherein identifying the presence or absence of a partial mismatch comprises identifying eplet partial mismatch information.
13. The method of claim 1 , wherein either or both of the responder information or the stimulator information are generated from one or more of monocytes, antigen presenting cells, dendritic cells, lymphocytes, lymphoblasts, or T cells.
14. The method of claim 1 , wherein identifying the presence or absence of a partial mismatch comprises identifying the presence or absence of a partial mismatch by employing a computer algorithm configured to identify the presence or absence of the partial mismatch by analyzing one or both of extent or type of HLA mismatch between the donor and the patient.
15. The method of claim 14 , wherein the computer algorithm is configured to provide number and type of mismatched eplets between the donor and the patient.
16. The method of claim 14 , further comprising the step of training the algorithm with a training set of data, the training set of data comprising number and type of amino acid sequence differences between the donor and the patient and functional data collected from in vitro tests.
17. The method of claim 16 , wherein one or more of cytotoxic T cell activation or cytotoxic T cell activity are compiled in the training set.
18. The method of claim 1 , further comprising the step of inactivating the cells of the patient.
19. The method of claim 18 , further comprising the step of detecting presence or absence of cytotoxic T cell activation.
20. The method of claim 1 , further comprising the steps of:
preparing the cells for administration to the patient; and
administering the alloreactive T cells to the patient.
21. The method of claim 20 , wherein the cells of the patient which present the antigen comprise cancer cells.
22. The method of claim 21 , wherein the cancer cells are brain cancer cells.
23. The method of claim 21 , wherein the cancer cells are brain glioma cells, brain stem glioma cells, or leptomeningeal gliomatosus of carcinomatosus cells.
24. The method of claim 20 , wherein the step of administering the alloreactive T cells to the patient comprise administering the alloreactive T cells to an immune-privileged site of the patient.