Institut National de Santé et de Recherche Médicale, Unité 892, CLCC René Gauducheau, Saint Herblain, France.
Clin Cancer Res. 2010 Sep 15;16(18):4607-15. doi: 10.1158/1078-0432.CCR-10-1485. Epub 2010 Jul 29.
NY-ESO-1 (ESO), a tumor-specific antigen of the cancer/testis group, is presently viewed as an important model antigen for the development of generic anticancer vaccines. The ESO(119-143) region is immunodominant following immunization with a recombinant ESO vaccine. In this study, we generated DRB10101/ESO(119-143) tetramers and used them to assess CD4 T-cell responses in vaccinated patients expressing DRB10101 (DR1).
We generated tetramers of DRB1*0101 incorporating peptide ESO(119-143) using a previously described strategy. We assessed ESO(119-143)-specific CD4 T cells in peptide-stimulated postvaccine cultures using the tetramers. We isolated DR1/ESO(119-143) tetramer(+) cells by cell sorting and characterized them functionally. We assessed vaccine-induced CD4(+) DR1/ESO(119-143) tetramer(+) T cells ex vivo and characterized them phenotypically.
Staining of cultures from vaccinated patients with DR1/ESO(119-143) tetramers identified vaccine-induced CD4 T cells. Tetramer(+) cells isolated by cell sorting were of T(H)1 type and efficiently recognized full-length ESO. We identified ESO(123-137) as the minimal optimal epitope recognized by DR1-restricted ESO-specific CD4 T cells. By assessing DR1/ESO(119-143) tetramer(+) cells using T cell receptor (TCR) β chain variable region (Vβ)-specific antibodies, we identified several frequently used Vβ. Finally, direct ex vivo staining of patients' CD4 T cells with tetramers allowed the direct quantification and phenotyping of vaccine-induced ESO-specific CD4 T cells.
The development of DR1/ESO(119-143) tetramers, allowing the direct visualization, isolation, and characterization of ESO-specific CD4 T cells, will be instrumental for the evaluation of spontaneous and vaccine-induced immune responses to this important tumor antigen in DR1-expressing patients.
NY-ESO-1（ESO）是癌症/睾丸组的一种肿瘤特异性抗原，目前被视为开发通用抗癌疫苗的重要模型抗原。用重组 ESO 疫苗免疫后，ESO（119-143）区域具有免疫优势。在这项研究中，我们生成了 DRB10101/ESO（119-143）四聚体，并使用它们来评估表达 DRB10101（DR1）的接种患者中的 CD4 T 细胞反应。
我们使用先前描述的策略生成了包含肽 ESO（119-143）的 DRB1*0101 四聚体。我们使用四聚体在肽刺激的接种后培养物中评估 ESO（119-143）特异性 CD4 T 细胞。我们通过细胞分选分离 DR1/ESO（119-143）四聚体（+）细胞，并对其功能进行了表征。我们评估了体外疫苗诱导的 CD4（+）DR1/ESO（119-143）四聚体（+）T 细胞，并对其表型进行了表征。
用 DR1/ESO（119-143）四聚体对接种患者的培养物进行染色，鉴定了疫苗诱导的 CD4 T 细胞。通过细胞分选分离的四聚体（+）细胞为 T（H）1 型，能够有效识别全长 ESO。我们确定 ESO（123-137）是受 DR1 限制的 ESO 特异性 CD4 T 细胞识别的最小最佳表位。通过使用 T 细胞受体（TCR）β链可变区（Vβ）特异性抗体评估 DR1/ESO（119-143）四聚体（+）细胞，我们鉴定了几个常用的 Vβ。最后，直接用四聚体对患者的 CD4 T 细胞进行体外染色，可直接定量和表型分析疫苗诱导的 ESO 特异性 CD4 T 细胞。
DR1/ESO（119-143）四聚体的开发允许直接可视化、分离和鉴定 ESO 特异性 CD4 T 细胞，这对于评估 DR1 表达患者中这种重要肿瘤抗原的自发和疫苗诱导免疫反应将是非常有帮助的。
