Characterization and growth factor requirements of SJL lymphomas. II. Interleukin 5 dependence of the in vitro cell line, cRCS-X, and influence of other cytokines.
Characterization and growth factor requirements of SJL lymphomas. II. Interleukin 5 dependence of the in vitro cell line, cRCS-X, and influence of other cytokines.
复制标题
SJL 淋巴瘤的特征和生长因子需求。
DOI：
10.1002/eji.1830190222
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发表时间：
1989
影响因子：
5.4
通讯作者：
Thorbecke,GJ
中科院分区：
文献类型：
--
作者：
Lasky,JL;Thorbecke,GJ
SJL 小鼠的滤泡 B 细胞淋巴瘤 [网状细胞肉瘤 (RCS)] 的生长依赖于同基因 Ly-1,2-T 细胞。这些 T 细胞响应 γ 照射的 RCS 细胞的刺激而产生许多淋巴因子，包括白细胞介素 (IL) 2、干扰素-γ (IFN-γ)、IL 4 和 IL 5，其中一些可能是肿瘤生长所必需的。先前的研究表明，如果在培养物中补充 Y 照射的淋巴结 (LN) 细胞或人 B 细胞生长因子 (BCGF) 制剂，RCS 细胞系 cRCS-X 可以在体外无限期维持。在本研究中，通过琼脂糖中[3H]胸苷掺入和集落形成的短期测定更详细地分析了该细胞系的生长要求。重组鼠 IL 5 引起 cRCS-X 细胞的剂量依赖性增殖，类似于 BCGF 诱导的增殖。添加 IL 5 后，最佳浓度的 BCGF 诱导的 cRCS-X 细胞的集落形成水平并未进一步增加，这表明这两个因子通过共同的机制发挥作用。在两项测定中，IL 1 和 IFN-γ 均增强 BCGF 或 IL 5 诱导的 cRCS-X 增殖。 IL 1 加 BCGF、IFN-γ 加 BCGF、IL 1 加 IL 5 的作用明显具有协同作用。在 [3H] 胸苷掺入测定中，cRCS-X 细胞与 IL 1 预孵育增强了它们响应 BCGF 或 IL 5 的增殖能力，但细胞因子添加的相反顺序显示 IL 1 没有影响。IFN-gM 则没有观察到这种效应。事实上，IL 1 和 IFN-gM 似乎通过不同的机制影响 BCGF 诱导的 cRCS-X 生长，并且它们的综合作用大于单独添加 IL 1 或 IFN-gM 的作用。研究的其他细胞因子，包括 IL 2、IL 3、IL 4、IL 6、肿瘤坏死因子-γ、粒细胞单核细胞集落刺激因子或转化生长因子-β，无论是单独使用还是与 BCGF 或 IL 5 联合使用，均对 cRCS-X 细胞没有任何可检测到的影响。与 IL 5 一样，SJL γ 照射的 LN 细胞在剂量依赖性方式。 IL 1 或 IL 1 加 IFN-γ 的组合在 cRCS-X CFU 的诱导中与 LN 细胞明显协同，表明 LN 细胞贡献 IL 5。在 LN 细胞和 IL 1 存在的情况下，最佳剂量的 BCGF 诱导的 CFU 水平进一步增强。然而，当 IL 1 和 IFN-γ 都添加到培养基中时，没有观察到这种情况，在这种情况下，最佳剂量的 BCGF 诱导的 CFU 水平为添加最佳剂量的 γ 照射的 LN 细胞不会进一步增强。结合之前的观察，这些数据支持这样的假设：RCS 体内和体外生长依赖于并促进同基因 RCS 特异性辅助 T 细胞产生的 IL 5 和 IFN-γ 以及巨噬细胞产生的 IL 1。
Follicular B cell lymphomas of SJL mice [reticulum cell sarcoma (RCS)] are dependent on syngeneic Ly‐1,2‐T cells for their growth. These T cells produce a number of lymphokines in response to stimulation with γ‐irradiated RCS cells including interleukin (IL) 2, interferon‐γ (IFN‐γ), IL 4 and IL 5, some of which may be required for growth of the tumor. Previous studies have shown that an RCS cell line, cRCS‐X, can be maintainedin vitroindefinitely, if the cultures are supplemented with Y‐irradiated lymph node (LN) cells or with a preparation of human B cell growth factor (BCGF). In the present studies, the growth requirements of this cell line were analyzed in more detail in short‐term assays of both [3H]thymidine incorporation and colony formation in agarose. Recombinant murine IL 5 caused dose‐dependent proliferation of cRCS‐X cells similar to that induced with BCGF. The level of colony formation by cRCS‐X cells induced by optimal concentrations of BCGF was not increased further by the addition of IL 5, suggesting that the two factors act via a common mechanism. IL 1 and IFN‐y each enhanced cRCS‐X proliferation induced by BCGF or IL 5 in both assays. The effects of IL 1 plus BCGF, IFN‐γ plus BCGF, and IL 1 plus IL 5 were clearly synergistic. Preincubation of cRCS‐X cells with IL 1 enhanced their ability to proliferate in response to BCGF or IL 5 in [3H]thymidine incorporation assays, but the reverse sequence of cytokine addition showed no effect of IL 1. No such effect was seen with IFN‐gM. Indeed, IL 1 and IFN‐gM appeared to affect BCGF‐induced cRCS‐X growth by different mechanisms and their combined effects were greater than that of IL 1 or IFN‐gM added separately. None of the other cytokines studied, including IL 2, IL 3, IL 4, IL 6, tumor necrosis factor‐γ, granulocyte monocyte‐colony‐stimulating factor or transforming growth factor‐β, had any detectable effect on cRCS‐X cells, either alone or in combination with BCGF or IL 5.Like IL 5, SJL γ‐irradiated LN cells induced cRCS‐X colony‐forming units (CPU) in a dose‐dependent manner. IL 1, or the combination of IL 1 plus IFN‐γ, clearly synergized with LN cells in the induction of cRCS‐X CFU, suggesting that LN cells contribute IL 5. The level of CFU induced by an optimal dose of BCGF was enhanced further in the presence of LN cells and IL 1. However, this was not observed when both IL 1 and IFN‐γ were added to the medium, in which case the level of CFU induced by an optimal dose of BCGF was not further enhanced by the addition of an optimal dose of γ‐irradiated LN cells. In combination with previous observations, these data support the hypothesis that RCS growthin vivoandin vitrois dependent on and promoted by IL 5 and IFN‐γ produced by syngeneic RCS‐specific helper T cells, and IL 1 produced by macrophages.