Chattopadhyay Subhasis, Chakraborty Nitya G
Division of Hematology/Oncology, Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030-1628, USA.
Hum Immunol. 2005 Aug;66(8):884-91. doi: 10.1016/j.humimm.2005.06.002. Epub 2005 Jul 20.
The generation of tumor-associated, but self-antigen specific cytotoxic T lymphocytes (CTL) response is possible by vaccination even in patients at the advanced stages of the disease. The in vivo expansion of such CTLs is now the most important objective of the immunotherapy. In human melanoma, we show here that MART-1(27-35)-specific CTLs generated with purified CD8+ cells survive and maintain their activity longer in culture than those CTLs generated by using total peripheral blood lymphocytes (PBL) taken either from patients or from normal donors. When PBL are grown under Th1 conditioning the quantity and quality of CTL with total PBL are comparable with that of the CTLs generated with purified CD8+ cells. For patients either autologous melanoma tumor cells or MART-1(27-35) peptide pulsed autologous DC were used to generate CTL responses. For normal donors MART-1(27-35) peptide pulsed autologous DC were used. For both normal donors or patients, polarization of PBL with Th1 conditioning with interleukin (IL)-12 (250 U/ml) and anti IL-4 antibody 1 mug/ml for 7 days before CTL generation, induced better and longer living CTL response and prevented the expansion of CD4+ T cells that have downregulatory activity. We show that continuous presence of Th1 conditioning in cultures with total PBL generated significantly higher number of antigen-specific CTLs as detected by MART-1 HLA-A2 tetramer staining. The antigen specificity of such CTLs were determined by IFN-gamma secretion in response to target cells bearing the specific antigen. Our observations indicate that Th1 conditioning results in a longer lasting CTL response in vitro and points toward a newer approach for vaccine strategy.
即使是处于疾病晚期的患者，通过疫苗接种也有可能产生肿瘤相关但自身抗原特异性的细胞毒性T淋巴细胞（CTL）反应。这种CTL在体内的扩增是目前免疫治疗的最重要目标。在人类黑色素瘤中，我们在此表明，用纯化的CD8+细胞产生的MART-1(27-35)特异性CTL在培养中比使用从患者或正常供体获取的全外周血淋巴细胞（PBL）产生的CTL存活时间更长且活性维持得更好。当PBL在Th1条件下培养时，全PBL产生的CTL的数量和质量与用纯化的CD8+细胞产生的CTL相当。对于患者，使用自体黑色素瘤肿瘤细胞或MART-1(27-35)肽脉冲的自体树突状细胞（DC）来产生CTL反应。对于正常供体，使用MART-1(27-35)肽脉冲的自体DC。对于正常供体或患者，在产生CTL前7天，用白细胞介素（IL）-12（250 U/ml）和抗IL-4抗体1 μg/ml进行Th1条件极化的PBL，可诱导出更好且存活时间更长的CTL反应，并防止具有下调活性的CD4+T细胞扩增。我们表明，在全PBL培养物中持续存在Th1条件，通过MART-1 HLA-A2四聚体染色检测到产生的抗原特异性CTL数量显著更高。这种CTL的抗原特异性通过对携带特定抗原的靶细胞产生的γ干扰素分泌来确定。我们的观察结果表明，Th1条件在体外可导致更持久的CTL反应，并指向一种新的疫苗策略方法。
