Mouse CD40-transfected cell lines cannot exhibit the binding and RANTES-stimulating activity of exogenous heat shock protein 70
Mouse CD40-transfected cell lines cannot exhibit the binding and RANTES-stimulating activity of exogenous heat shock protein 70
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DOI：
10.1016/j.molimm.2006.06.002
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发表时间：
2007-02-01
影响因子：
3.6
通讯作者：
Tani, Fumito
中科院分区：
文献类型：
--
作者：
Tao, Yufeng;Nomura, Masayo;Tani, Fumito
在这里，我们证明了诱导型小鼠Hsp 72显着结合淋巴瘤巨噬细胞样P388 D1细胞。为了检测小鼠CD 40是否可以以类似于人CD 40的方式在外源性施用的HSP 70的信号传导中发挥作用，我们建立了人293细胞和鼠前B细胞系Ba/F3的小鼠CD 40转染子。在293衍生的转染子上表达的一小部分小鼠CD 40是具有信号转导的C末端结构域的成熟形式，而大多数表达的抗原显示出比我们预期的小的分子大小。流式细胞术显示，小鼠热休克蛋白72，但既不是它的缺失变体，也不是相关的大肠杆菌DnaK，绑定到293衍生的转染子，无论CD 40的表达。表达的CD 40分子与可溶性的CD 40 L结合，但这种结合不受过量HSP 70的抑制。CD 40 L，而不是任何HSP 70重组蛋白，刺激转染子中趋化因子RANTES的产生。此外，没有RANTES的生产诱导的HSP 70-RCMLA复合物的转染，虽然它结合到293-衍生的细胞在CD 40-非依赖性的方式。通过使用表达成熟形式的小鼠CD 40的Ba/F3衍生的转染子，检测到小鼠CD 40和HSP 70重组蛋白之间没有相互作用。目前的结果表明，小鼠CD 40表达的转染子不同于其人类同源物的结合外源性给药的HSP 70。(c)2006爱思唯尔有限公司保留所有权利。
Here we demonstrate the inducible mouse Hsp72 binds markedly to lymphoid neoplastic macrophage-like P388D1 cells. To examine whether mouse CD40 can play a role in signaling exogenously administered HSP70 in a fashion similar to that of human CD40, we established mouse CD40-transfectants of both human 293 cells and murine-pro-B cell line Ba/F3. A small portion of mouse CD40 expressed on 293-derived transfectants was the mature form with a signal-transducible C-terminal domain, whereas a majority of expressed antigen showed the molecular size smaller than we expect. Flow cytometry showed that mouse Hsp72, but neither its deletion variants nor the related Escherichia coli DnaK, bound to the 293-derived transfectants regardless of CD40 expression. CD40 molecules expressed on the transfectants showed the binding of soluble form of CD40L but this binding was not inhibited by excess amount of HSP70. CD40L, but not any HSP70 recombinant proteins, stimulated the production of chemokine RANTES in the transfectants. Furthermore, no RANTES production was induced by HSP70-RCMLA complex in the transfectants, although it binds to 293-derived cells in a CD40-independent manner. No interaction between mouse CD40 and HSP70 recombinant proteins was detected by using the Ba/F3-derived transfectants that express the mature form of mouse CD40. The present results imply that mouse CD40 expressed on the transfectants differs from its human homolog in the binding of exogenously administered HSP70. (c) 2006 Elsevier Ltd. All rights reserved.