Stevens E J, Jacknin L, Robbins P F, Kawakami Y, el Gamil M, Rosenberg S A, Yannelli J R
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
J Immunol. 1995 Jan 15;154(2):762-71.
PBLs were isolated from 13 patients with metastatic melanoma. Mixed lymphocyte tumor cell cultures (ML TCs) were established (15 times) by using irradiated HLA-matched (one class I locus) allogeneic melanoma tumor cell lines (13 times) or autologous melanoma tumor cell lines (two times) in medium containing 120 IU/ml IL-2 and 100 IU/ml IL-4. PBLs grew to levels that could be assessed for functional reactivity 9 of 15 times. In seven of nine cases, CD3+CD8+ CTLs grew from MLTCs that were tumor specific; five were restricted by HLA-A2 and two were restricted by HLA-A24. Four of the tumor-specific CTL lines lysed autologous fresh tumor cells. Tumor-specific CTLs from two of three patients had cytolytic activity identical with tumor-infiltrating lymphocytes (TIL) derived from tumor biopsies removed earlier and grown in high concentrations (6000 IU/ml) of IL-2. Three of the HLA-A2-restricted tumor-specific CTLs were shown to recognize 293 cells transfected with HLA-A2.1 cDNA and the gene encoding the melanoma Ag, MART-1. In addition, these CTLs recognized the T2 cell line pulsed exogenously with the peptide MART-1(27-35), which is the nine-amino acid immunodominant epitope of the MART-1 Ag recognized on melanoma tumor cells by nearly all HLA-A2-restricted TIL. Thus, we have demonstrated the ability to generate tumor-specific CTLs from PBLs that are similar in their reactivity to TIL. This technique obviates the need for autologous tumor tissue and suggests that PBLs contain sufficient CTL precursors for use in generating antitumor CTLs for cellular immunotherapy trials.
从13例转移性黑色素瘤患者中分离出外周血淋巴细胞（PBL）。通过在含有120 IU/ml白细胞介素-2（IL-2）和100 IU/ml白细胞介素-4（IL-4）的培养基中，使用经照射的HLA匹配（一个I类基因座）的同种异体黑色素瘤肿瘤细胞系（13次）或自体黑色素瘤肿瘤细胞系（2次），建立混合淋巴细胞肿瘤细胞培养物（MLTC）（共15次）。15次中有9次PBL生长到可评估功能反应性的水平。在9例中的7例中，肿瘤特异性的CD3 + CD8 +细胞毒性T淋巴细胞（CTL）从MLTC中生长出来；其中5例受HLA - A2限制，2例受HLA - A24限制。4个肿瘤特异性CTL系裂解自体新鲜肿瘤细胞。3例患者中有2例的肿瘤特异性CTL具有与早期切除的肿瘤活检组织中分离并在高浓度（6000 IU/ml）IL-2中培养所获得的肿瘤浸润淋巴细胞（TIL）相同的细胞溶解活性。3个受HLA - A2限制的肿瘤特异性CTL被证明可识别转染了HLA - A2.1 cDNA和编码黑色素瘤抗原MART - 1基因的293细胞。此外，这些CTL识别用肽MART - 1(27 - 35)外源脉冲处理的T2细胞系，该肽是几乎所有受HLA - A2限制的TIL在黑色素瘤肿瘤细胞上识别的MART - 1抗原的九氨基酸免疫显性表位。因此，我们已经证明了从PBL中产生肿瘤特异性CTL的能力，其反应性与TIL相似。该技术无需自体肿瘤组织，并表明PBL含有足够的CTL前体，可用于为细胞免疫治疗试验生成抗肿瘤CTL。
