NY-ESO-1 is highly expressed in poor-prognosis multiple myeloma and induces spontaneous humoral and cellular immune responses

Abstract

The presence of a metaphase cytogenetic abnormality (CA) is the key negative predictor of outcome in patients with multiple myeloma (MM). Gene expression profiling (GEP) of such patients showed increased expression of NY-ESO-1 compared to patients with normal cytogenetics (60% versus 31%; P = .004). NY-ESO-1 was also highly expressed in relapsing MM especially patients with CA (100% versus 60.7%; P < .001). GEP findings were confirmed at the protein level by immunostaining of marrow biopsies for NY-ESO-1. We detected spontaneous NY-ESO-1-specific antibodies by enzyme-linked immunosorbent assay in 33% of patients with NY-ESO-1+ MM, especially in CA patients (9 of 13; 70%), but in none of the NY-ESO-1- patients with MM (n = 27) or healthy donors (n = 21). Spontaneous NY-ESO-1(157-165)-specific T cells (0.2%-0.6% of CD8+ T cells) were found in the peripheral blood of NY-ESO-1+ MM with HLA-A*0201/NY-ESO-1(157-165) tetramers. These NY-ESO-1-specific T cells, when expanded, killed primary MM cells (50% lysis, effector-target [E/T] ratio, 10:1). Our data demonstrate that NY-ESO-1 is frequently expressed in MM with CA and is capable of eliciting spontaneous humoral and T-cell immunity. The pool of NY-ESO-1-specific cytotoxic T cells expands easily on NY-ESO-1 peptide stimulation and is functionally active. NY-ESO-1 should therefore be an ideal tumor target antigen for immunotherapy of patients with poor-prognosis MM.

Figure 1.

NY-ESO-1 gene expression. Gene expression profiling shows that NY-ESO-1 is expressed at increased frequency in patients with abnormal cytogenetics (a positive score is more than mean + 3 SD [90] of the GEP score of normal human PCs). SMM indicates smoldering MM.

Figure 2.

BM biopsy stained with B9.8 shows strong expression of NY-ESO-1 protein in myeloma plasma cells. Images (left panel, original magnification × 10, numerical aperture 0.3; right panel, original magnification × 20, numerical aperture 0.46) were visualized with an Olympus microscope, type BH2-RFCA. Images were acquired with a SPOT camera (Diagnostics Instruments, Sterling Heights, MI) and SPOT Advanced version 3.5.6 software without image manipulation.

Figure 3.

Expansion of NY-ESO-1–specific CTLs. (A) NY-ESO-1–specific T cells are spontaneously present and can be progressively expanded by stimulation with APCs pulsed with NY-ESO-1_157-C165V analog peptide. Note that there is equivalent staining with NY-ESO-1_157-165 wild-type and NY-ESO-1_157-C165V analog peptide-loaded A2 tetramers. (B) NY-ESO-1–specific CTLs kill both primary, the HLA-A2⁺ U266 myeloma cell line, and autologous PHA-blasts pulsed with NY-ESO-1_157-C165V, and not control PHA-blasts pulsed with a MAGE-3_112-120 A2-binding peptide or K562 cells. (C) NY-ESO-1–specific CTLs produce IFN-γ and not IL-4 (Tc1 type), contain cytolytic granules, and are of memory-effector type. VLA indicates very late antigen; FITC, fluorescein isothiocyanate.