Yasumura S, Hirabayashi H, Schwartz D R, Toso J F, Johnson J T, Herberman R B, Whiteside T L
Pittsburgh Cancer Institute, Pennsylvania.
Cancer Res. 1993 Mar 15;53(6):1461-8.
Human cytotoxic T-lymphocyte (CTL) lines with specificity restricted for autologous squamous cell carcinoma of the head and neck (SCCHN) were established from peripheral blood lymphocytes obtained at the time of surgery and again at two different times after surgery from a patient with cancer of the tongue. The CTL lines were cultured in the presence of interleukin (IL) 2, IL4, and autologous tumor (AuTu) cell monolayers. All three lines were CD3+CD8+CD11b-HLA-DR+ T-cell receptor alpha/beta+. They were tested in 4-h51Cr release assays against SCCHN cell lines (n = 5) and a variety of nonsquamous human tumor (n = 5) and normal (n = 5) cell targets and was found to lyse only AuTu (PCI-50) and three allogenic SCCHN cell lines. Lysis of AuTu and the three allogenic SCCHN targets by the established CTL lines appeared to be major histocompatibility complex class I restricted, since it was blocked by monoclonal antibodies to class I histocompatibility complex antigens. The CTL lines proliferated in vitro in response to autologous PCI-50 or an allogenic SCCHN cell line (PCI-1). The lines have been maintained in culture in the presence of AuTu monolayers and retained cytotoxicity against AuTu for over 20 weeks. The AuTu (PCI-50) cell line was tested for in vitro sensitivity to cytotoxic or cytostatic effects of various effector cells, including the CTL lines. PCI-50 targets were resistant to lysis by resting human mononuclear cells but sensitive to IL2-activated natural killer cells in 4-h 51Cr release assays. In comparison with IL2-activated natural killer cells, the CTL line mediated lower levels of lysis against AuTu. Growth of PCI-50 cells in culture was significantly inhibited by a combination of gamma-interferon and IL2 or by high concentrations of tumor necrosis factor alpha. While supernants of IL2-activated natural killer cells were growth inhibitory, those of the CTL line were not. On the other hand, lysis of AuTu targets by the CTL line was increased by preincubation of the tumor cells with tumor necrosis factor alpha or gamma-interferon. These cytokines augmented expression of HLA-class I, HLA-class II, and intercellular adhesion molecule I, but not squamous cell carcinoma-associated antigens, E7 and A9, on PCI-50 cells. The CTL lines described are the first with restricted specificity for autologous SCCHN ever reported and their availability will facilitate studies of the AuTu T-cell response in head and neck cancer.
从一名舌癌患者手术时及术后两个不同时间获取的外周血淋巴细胞中建立了对自体头颈部鳞状细胞癌（SCCHN）具有特异性限制的人细胞毒性T淋巴细胞（CTL）系。CTL系在白细胞介素（IL）-2、IL-4和自体肿瘤（AuTu）细胞单层存在的情况下进行培养。所有三个系均为CD3⁺CD8⁺CD11b⁻HLA-DR⁺T细胞受体α/β⁺。在4小时⁵¹Cr释放试验中，它们针对SCCHN细胞系（n = 5）、多种非鳞状人类肿瘤（n = 5）和正常（n = 5）细胞靶标进行测试，发现仅能裂解AuTu（PCI-50）和三个同种异体SCCHN细胞系。已建立的CTL系对AuTu和三个同种异体SCCHN靶标的裂解似乎受主要组织相容性复合体I类限制，因为它被针对I类组织相容性复合体抗原的单克隆抗体阻断。CTL系在体外对自体PCI-50或同种异体SCCHN细胞系（PCI-1）产生增殖反应。这些系在AuTu单层存在的情况下在培养中得以维持，并对AuTu保持细胞毒性超过20周。对AuTu（PCI-50）细胞系进行了针对各种效应细胞（包括CTL系）的细胞毒性或细胞抑制作用的体外敏感性测试。在4小时⁵¹Cr释放试验中，PCI-50靶标对静息人单核细胞的裂解具有抗性，但对IL-2激活的自然杀伤细胞敏感。与IL-2激活的自然杀伤细胞相比，CTL系介导的对AuTu的裂解水平较低。γ干扰素和IL-2的组合或高浓度的肿瘤坏死因子α可显著抑制PCI-50细胞在培养中的生长。虽然IL-2激活的自然杀伤细胞的上清液具有生长抑制作用，但CTL系的上清液则没有。另一方面，通过用肿瘤坏死因子α或γ干扰素预孵育肿瘤细胞，CTL系对AuTu靶标的裂解增加。这些细胞因子增强了PCI-50细胞上HLA-I类、HLA-II类和细胞间黏附分子I的表达，但未增强鳞状细胞癌相关抗原E7和A9的表达。所描述的CTL系是首次报道的对自体SCCHN具有特异性限制的细胞系，它们的可用性将有助于对头颈部癌中AuTu T细胞反应的研究。
