Monitoring CD8 T cell responses to NY-ESO-1: correlation of humoral and cellular immune responses - PubMed (original) (raw)

. 2000 Apr 25;97(9):4760-5.

doi: 10.1073/pnas.97.9.4760.

Y Nagata, S Gnjatic, H Wada, E Stockert, J Karbach, P R Dunbar, S Y Lee, A Jungbluth, D Jäger, M Arand, G Ritter, V Cerundolo, B Dupont, Y T Chen, L J Old, A Knuth

Affiliations

Monitoring CD8 T cell responses to NY-ESO-1: correlation of humoral and cellular immune responses

E Jäger et al. Proc Natl Acad Sci U S A. 2000.

Abstract

NY-ESO-1, a member of the cancer-testis family of antigens, is expressed in a subset of a broad range of different human tumor types. Patients with advanced NY-ESO-1-expressing tumors frequently develop humoral immunity to NY-ESO-1, and three HLA A2-restricted peptides were defined previously as targets for cytotoxic CD8(+) T cells in a melanoma patient with NY-ESO-1 antibody. The objectives of the present study were (i) to develop enzyme-linked immunospot (ELISPOT) and tetramer assays to measure CD8(+) T cell responses to NY-ESO-1, (ii) to determine the frequency of CD8(+) T cell responses to NY-ESO-1 in a series of HLA-A2 patients with NY-ESO-1 expressing tumors, (iii) to determine the relation between CD8(+) T cell and humoral immune responses to NY-ESO-1, and (iv) to compare results of NY-ESO-1 ELISPOT assays performed independently in two laboratories with T cells from the same patients. NY-ESO-1 ELISPOT and tetramer assays with excellent sensitivity, specificity, and reproducibility have been developed and found to correlate with cytotoxicity assays. CD8(+) T cell responses to HLA-A2-restricted NY-ESO-1 peptides were detected in 10 of 11 patients with NY-ESO-1 antibody, but not in patients lacking antibody or in patients with NY-ESO-1-negative tumors. The results of ELISPOT assays were concordant in the two laboratories, providing the basis for standardized monitoring of T cell responses in patients receiving NY-ESO-1 vaccines.

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Figures

Figure 1

Figure 1

CD8+ T cell reactivity to HLA-A2-restricted peptides in ELISPOT IFN-γ assays. (A) CD8+ T cells from NW731, a patient with NY-ESO-1-positive melanoma and NY-ESO-1 antibody. (B) CD8+ T cells from NW681, a patient with NY-ESO-1-positive melanoma and no NY-ESO-1 antibody. CD8+ T cells were presensitized with NY-ESO-1 p157–165, Melan-A/MART-1 p26–35, MAGE-3 p271–279, or flu matrix p58–66 and tested against T2 cells pulsed with the peptide panel shown at the bottom of the figure (including NY-ESO-1 p157–167). Effector-to-target cell ratios in these assays were 1:1 (solid bars) and 0.2:1 (hatched bars).

Figure 2

Figure 2

Effect of presensitization of CD8+ T cells with HLA-A2-restricted NY-ESO-1 p157–165 and flu matrix p58–66 on ELISPOT assays. Results with CD8+ T cells from three patients with NY-ESO-1 expressing stage IV melanoma and NY-ESO-1 antibody are shown. Fifty thousand CD8+ T cells, with or without presensitization with NY-ESO-1 p157–165 or flu matrix p58–66, were cocultured with 50,000 T2 cells alone or pulsed with NY-ESO-1 p157–165 or flu matrix p58–66. (Upper) Photographs of wells in an ELISPOT assay. (Lower) Histograms with no presensitization (open bars), presensitization with NY-ESO-1 p157–165 (solid bar), or presensitization with flu matrix p58–66 (hatched bar). Each bar represents average of spots in duplicate wells.

Figure 3

Figure 3

Cytotoxicity tests with CD8+ T cells from four HLA-A2 patients with NY-ESO-1-positive melanomas. Patients NW29 and NW889 had NY-ESO-1 antibody, and patients NW28 and NW681 did not. Cytotoxicity was measured by standard 51Cr release assay, and values represent averages of duplicate wells (▵, T2 cells; ●, T2 cells pulsed with NY-ESO-1 p157–165).

Figure 4

Figure 4

Tetramer analysis of CD8+ T cells from two stage IV melanoma patients, NW29 and NW731, with NY-ESO-1-positive tumors and NY-ESO-1 antibody. Tests with unsensitized PBLs (Upper) and CD8+ T cells presensitized with NY-ESO-1 p157–165 (Lower) for each patient are shown. Samples were double-stained with phycoerythrin-labeled tetramers and anti-Tricolor-CD8 mAb. Tetramers were prepared with NY-ESO-1 p157–165, Melan-A p26–35, and MAGE-3 p271–279 as indicated at the bottom of each grid. Values indicate percentage of tetramer-positive CD8+ T cells.

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