Indirect antigen-specific T cell recognition assay

1. A method for analyzing simultaneously multiple human antigen-specific cell populations of a sample, the sample comprising B cells and antigen-specific cells, the method comprising

a) separation of B cells from said sample

b) dividing the B cells into n sub-samples

c) differentially labeling the B cells of said sub-samples, wherein at least n-1sub-samples are labeled

d) pulsing of the B cells of each sub-sample with single or multiple peptides

e) pooling of the labeled and peptide-pulsed B cells with cells of said sample comprising said antigen-specific cells of step a)

f) co-cultivation of the cells of step e)

g) flow cytometry analysis of the B cells with regard to their cell number and/or CD83 expression loaded with single or multiple peptides of step d), wherein the potency of the antigen specific cells is determined by

i) B cell numbers if extensive killing of target B cells occurs,

ii) B cell numbers and ratio of CD83 expression on survived target B cells if moderate killing of target B cells occurs, or

iii) ratio of CD83 expression on target B cells if no cytotoxic activity occurs.

2. The method of claim 1 , wherein in step b), additionally to the separation of the B cells, said antigen-specific cells of said sample are separated into CD4 T cells, CD8 T cells or NK cells.

3. The method of claim 1 , wherein the B cells in step c) are labeled with single or multiple fluorescent dyes in different concentrations and/or combinations thereof.

4. The method of claim 1 further comprising estimating the prognosis of adoptively T cell therapy, wherein the potency of the antigen specific cells indicates the prognosis of the adoptive T cell therapy.