Characterization and induction of antitumor lymphokine derived from human lymph node cell

本研究旨在探讨淋巴结细胞是否可以通过细胞因子介导对肿瘤细胞发挥抗增殖活性。采用人肿瘤克隆形成试验（HTCA）研究了宫颈癌患者局部淋巴结细胞的可溶性抗肿瘤活性。一个显着的抗增殖活性的淋巴结细胞（LNC）对宫颈癌细胞系，HeLa细胞，被证明与植物血凝素（PHA）或刀豆球蛋白A（ConA）的刺激，但不与白细胞介素-2（IL-2）。此外，一个显着的抗增殖活性的LNC从宫颈癌和子宫内膜癌患者对子宫内膜癌细胞系，RL 95 -2，也证明了当它们被PHA刺激。这种抗增殖活性主要存在于非粘附细胞中，可能是T细胞。LNC产生干扰素γ（IFN γ）和肿瘤坏死因子（TNF），并且证明HeLa细胞对重组人IFN γ敏感，RL 95 -2细胞对重组人TNF敏感。这些结果表明，抗肿瘤活性分别归因于IFN γ和TNF。此外，关于HeLa细胞，通过使用中和实验证实了IFN γ负责活性。这些结果表明，人LNC能够发挥抗增殖活性介导的细胞因子通过适当的刺激。在这两项研究中，LNC不能通过用IL-2刺激来显示抗增殖活性，但是在某些情况下，外周血淋巴细胞通过用IL-2刺激HTCA而发挥抗增殖活性（提交的论文）。此外，当LNC被IL-2刺激较长时间时，它们产生细胞因子，如TNF α、TNF β和IFN γ（正在进行的研究）。IL-2刺激LNC可能是一种有前途的方法，因此目前正在深入研究中。

This study was designed to investigate whether lymph node cells could exert an antiproliferative activity on tumor cells that are mediated through cytokines. The soluble antitumor activity of regional lymph node cells obtained from patients with cervical cancer was investigated by using a human tumor clonogenic assay (HTCA). A significant antiproliferative activity of the lymph node cells (LNCs) against a cervical cancer cell line, HeLa cells, was demonstrated by stimulation with either phytohemagglutinin (PHA) or concanavalin A(ConA), but not with interleukin-2(IL-2). In addition, a significant antiproliferative activity of the LNCs obtained from cervical cancer and endometrial cancer patients against a endometrial cancer cell line, RL95-2, was also demonstrated when they were stimulated by PHA. This antiproliferative activity was found mainly in nonadherent cells, possibly T-cells. LNCs produced both interferon gamma(IFNgamma) and tumor necrosis factor (TNF), and it was demonstrated that HeLa cells were sensitive to recombinant human IFNgamma and RL95-2 cells were sensitive to recombinant human TNF. These findings suggested the antitumor activities were attributed to IFNgamma and TNF, respectively. Moreover, about HeLa cells, it was confirmed that IFNgamma was responsible for the activity by using neutralizing experiments. These results indicate that human LNCs were able to exert an antiproliferative activity mediated through the cytokines by appropriate stimulation. In the two studies, LNCs could not demonstrate an antiproliferative activity by stimulation with IL-2, but peripheral blood lymphocytes, in some cases, exerted antiproliferative activity by stimulation with IL-2 on HTCA (submitted paper). In addition, LNCs produced cytokines, such as TNFalpha, TNFbeta, and IFNgamma when they were stimulated by IL-2 for longer duration (ongoing study). IL-2 stimulation on LNCs might be a promising procedure, so it is presently under intensive research investigation.