PRODUCTION OF CYTOKINES BY MOUSE B-CELLS - B-LYMPHOMAS AND NORMAL B-CELLS PRODUCE INTERLEUKIN-10
PRODUCTION OF CYTOKINES BY MOUSE B-CELLS - B-LYMPHOMAS AND NORMAL B-CELLS PRODUCE INTERLEUKIN-10
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DOI：
10.1093/intimm/2.9.821
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发表时间：
1990-09-01
影响因子：
4.4
通讯作者：
HOWARD, M
中科院分区：
文献类型：
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作者：
OGARRA, A;STAPLETON, G;HOWARD, M
我们检查了一组小鼠 Ly-1+ B 淋巴瘤和纯化的正常小鼠腹膜 B 细胞，根据 Ly-1 表面抗原的表达情况将其分为亚群，以了解它们产生细胞因子的能力。在可能的情况下，我们结合使用了细胞因子检测方法，以补偿敏感性和特异性的差异以及抑制剂掩盖活性的可能性。所有测试的淋巴瘤均显示组成型表达TGF-β。和CSIF/IL-10。此外，不同水平的IL-6、TNF-α。 TNF-β和G-CSF在大多数淋巴瘤中均得到证实，并且一种淋巴瘤(CH12)的变体另外产生不同水平的IL-3、IL-4和GM-CSF，FACS纯化的正常Ly-1+和Ly-1-腹膜B细胞也显示表达编码CSIF/IL-10、IL-6、TNF-α的RNA。用 LPS 刺激后，TNF-β 和 G-CSF 水平非常低。使用特异性免疫测定法检测 LPS 刺激的 Ly-1+ 和 Ly-1- B 细胞上清液中的 IL-6 和 CSIF/IL-10 支持了这些数据。淋巴瘤或 B 细胞制剂均不产生 IL-1.α.、IL-2、IL-5、IL-7 或 IFN-.gamma。我们正常 B 细胞群的纯度通过 FACS 上的表型分析以及纯化后某些 mRNA 转录物的消失进行评估，例如 CD4、c-fms、GM-CSF 和 IFN-.gamma，其中大部分可在 LPS 刺激中检测到。腹膜细胞总数。这表明我们的 B 细胞纯化方法已将污染 T 细胞 (CD4)、巨噬细胞 (c-fms、GM-CSF) 和 NK 细胞 (IFN-γ) 减少到我们的测定中检测不到的水平。 LPS 刺激的 Ly-1+ 和 Ly-1- B 细胞不表达 IL-3、IL-4、IL-5 和 GM-CSF，减少了人们对污染腹膜肥大细胞可能导致观察到的细胞因子产生的担忧。因此，我们相信我们的数据为 LPS 刺激的正常 B 细胞产生细胞因子子集提供了强有力的支持。 Ly-1+ B淋巴瘤和正常Ly-1+和Ly-1- B细胞似乎都能够表达IL-6、TNF-α、TNF-β和CSIF/IL-10。因此，虽然其他细胞因子仅由 B 淋巴瘤表达，例如 B 淋巴瘤。 IL-3、IL-4和GM-CSF(尽管限于CH12淋巴瘤)可能是转化的结果，IL-6、TNF-α、TNF-β和CSIF/IL-10可能是正常免疫应答中重要的B细胞衍生的免疫调节分子，包括B细胞介导的抗原呈递、B细胞的自分泌生长或B细胞介导的免疫抑制。
We have examined a panel of murine Ly-1+ B lymphomas and purified normal murine peritoneal B cells separated into subsets on the basis of expression of the Ly-1 surface antigen, for their ability to produce cytokines. Where possible, we have used a combination of cytokine detection methods in order to compensate for differences in sensitivity and specificity, and the possibility of inhibitors masking an activity. All the lymphomas tested were shown to constitutively express TGF-.beta. and CSIF/IL-10. In addition, varying levels of IL-6, TNF-.alpha. and TNF-.beta., and G-CSF, were demonstrable in most of the lymphomas, and variants of one lymphoma (CH12) additionally produced varying levels of IL-3, IL-4, and GM-CSF, FACS purified normal Ly-1+ and Ly-1- peritoneal B cells, were also shown to express RNA encoding CSIF/IL-10, IL-6, TNF-.alpha. and TNF-.beta., and very low levels of G-CSF, following stimulation with LPS. These data were supported by the detection of IL-6 and CSIF/IL-10 in supernatants from LPS-stimulated Ly-1+ and Ly-1- B cells using specific immunoassays. None of the lymphomas or B cell preparations produced IL-1.alpha., IL-2, IL-5, IL-7, or IFN-.gamma.. The purity of our normal B cell populations was assessed by phenotypic analysis on the FACS and also by the disappearance of certain mRNA transcripts after purification, e.g., CD4, c-fms, GM-CSF, and IFN-.gamma., most of which could be detected in LPS-stimulated total peritoneal cell populations. This suggested that our B cell purification method had reduced, to a level undetectable in our assays, contaminating T cells (CD4), macrophages (c-fms, GM-CSF), and NK cells (IFN-.gamma.). Absence of IL-3, IL-4, IL-5, and GM-CSF expression by LPS-stimulated Ly-1+ and Ly-1- B cells reduced the concern that contaminating peritoneal mast cells could account for the observed cytokine production. We therefore believe our data provide strong support for production of a subset of cytokines by LPS-stimulated normal B cells. Both they Ly-1+ B lymphomas and normal Ly-1+ and Ly-1- B cells appear capable of expressing IL-6, TNF-.alpha., TNF-.beta., and CSIF/IL-10. Thus, while other cytokines expressed only by the B lymphomas, e.g. IL-3, IL-4, and GM-CSF (although restricted to the CH12 lymphomas) may be a result of transformation, IL-6, TNF-.alpha., TNF-.beta., and CSIF/IL-10 may be important B cell derived immunoregulatory molecules in normal immune responses including B cell mediated antigen presentation, autocrine growth of B cells, or B cell mediated immunosuppression.