Effect of CCK-8 on B7.1 and B7.2 expressions and costimulatory activity of LPS-activated murine macrophages
Effect of CCK-8 on B7.1 and B7.2 expressions and costimulatory activity of LPS-activated murine macrophages
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发表时间：
2007
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通讯作者：
Zhuang Gui-jun
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作者：
Zhuang Gui-jun
目的探讨CCK-8对脂多糖(LPS)激活的巨噬细胞B7.1、B7.2表达及T淋巴细胞共刺激活性的影响。方法分离小鼠腹腔巨噬细胞，与脂多糖和/或CCK-8(10-12~10-6)mol·M-1共同孵育一定时间后，用流式细胞术检测小鼠腹腔巨噬细胞B7.1、B7.2的表达。用免疫磁珠从小鼠脾中分离出CD4+T细胞，与1/4的巨噬细胞共同培养，经LPS、CCK-8和/或抗B7.1抗体处理后，结果抗B7.2抗体、CCK1R拮抗剂CR1409、CCK2R拮抗剂CR2945作用于巨噬细胞24 h后，加入ConA刺激T细胞增殖，β法测定[3 H]-TdR掺入率。结果CCK-8呈剂量依赖性抑制巨噬细胞B7.1、B7.2的表达及共刺激活性，以10-7mol·L-1~10-9mol·L·-1抑制作用最强。CR1409和CR2945均能逆转CCK-8的协同刺激作用，其中CR1409的作用更为显著。结论CCK-8通过下调巨噬细胞B7.1和B7.2的表达，抑制巨噬细胞的共刺激活性，CCK1R和CCK2R参与了CCK1R和CCK2R的调节作用。CCK1R可能是CCK-8调节共刺激作用的主要受体。
Aim To investigate in vitro effects of CCK-8 on the expressions of B7.1 and B7.2 and the costim-ulatory activity for T lymphocytes in LPS-activated macrophages.Methods Mouse peritoneal macrophages were isolated and incubated with LPS and/or CCK-8(10-12~10-6)mol·M-1 for indicated times.The B7.1 and B7.2 expressions of murine peritoneal macrophages were analyzed by flow cytometry.CD4+ T cells were isolated from mouse spleen by using immunomagnetic beads, and cultured with 1/4 numbers of macrophages which were pretreated with LPS, CCK-8 and/or anti-B7.1 antibody, anti-B7.2 antibody, CCK1R antagonist CR1409, CCK2R antagonist CR2945 for 24 h. ConA was added into the culture medium to stimulate CD4+T cell proliferation.The proliferation was determined by measuring[3H]-TdR incorporation in a β-scintillation counter.Results LPS-induced B7.1 and B7.2 expressions and costimulatory activity of peritoneal macrophages were inhibited by CCK-8 in a dose-dependent manner, with the maximal effects occurred at the concentrations from 10-7 mol·L-1 to 10-9 mol·L-1. Both CR1409 and CR2945 reversed the effect of CCK-8 on costimulation, and the role of CR1409 was more significant. Anti-B7.1 antibody and anti-B7.2 antibody inhibited the modulatory role of LPS on costimulatory activity.Conclusion CCK-8 inhibited LPS-induced macrophage costimulatory activity by down-regulating B7.1 and B7.2 expressions, which was mediated by CCK1R and CCK2R. CCK1R might be the major receptor responsible for the modulation of CCK-8 on costimulation.