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Table 6 IFNγ ELISPOT assay specificity

From: Validation of a HLA-A2 tetramer flow cytometric method, IFNgamma real time RT-PCR, and IFNgamma ELISPOT for detection of immunologic response to gp100 and MelanA/MART-1 in melanoma patients

TIL

TIL1520

TIL1235

TIL specificity

gp100209

MART-1

Peptide specificity

Relevant

Relevant

Irrelevant

Relevant

Irrelevant

Irrelevant

Peptide

gp100209

gp100pool

MART-1

MART-1

gp100209

gp100pool

1

44

41

1

8

2

2

2

42

31

1

9

0

1

3

37

40

1

10

2

1

4

43

32

1

10

1

1

5

38

52

2

7

1

1

Mean (n = 5)

41

39

1.1

9

1.3

1.3

SD

3.1

8.5

0.4

1.2

0.8

0.6

  1. TIL cells (80 cells/well) were spiked in two different lots of PBMC (105 cells/well), stimulated with peptides, and analyzed for the number of IFNγ secreting cells per well (average value from triplicate wells). Two analysts each performed five assays. The numbers of IFNγ secreting cells from 2 PBMC lots by two analysts were found to be comparable. Data (secreting cells, Mean from 5 tests, and SD) from PBMC lot 1 by analyst 1 is shown as an example.