EXPRESSION OF IL-10R ON LYMPH NODES AND SPLEEN CELLS OF BORRELIA-INFECTED MICE

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。我们先前报道了促炎症细胞因子干扰素-γ(IFN-？)抗炎细胞因子IL-10对脂化外表面蛋白A(OspA)和合成的脂六肽Pam3Cys刺激产生的白细胞介素6(IL-6)在抗病C57BL/6J小鼠的淋巴结层细胞中的抑制作用明显高于对疾病敏感的C3H/HeJ小鼠。IL-10对C57小鼠LN细胞中这些细胞因子的有效下调能力与C57小鼠LN中IL-10受体(IL-10R)的表达显著高于C3H小鼠相关。相反，两种品系小鼠的脾细胞均表现出相似的IL-10R表达水平，这与IL-10抑制干扰素产生的能力有关。和IL-6在两种品系小鼠脾细胞培养中的作用相同。在这项研究中，我们研究了LN细胞中的特异性淋巴细胞群，这可能是IL-10抑制小鼠致炎细胞因子的靶点。用伯氏杆菌感染C3H和C57小鼠1周后，体外分离淋巴细胞和脾细胞，用流式细胞仪检测CD3+、CD4+、CD8+和CD19+淋巴细胞上IL-10R的表达。C3H和C57小鼠LN和脾细胞中的IL-10R主要为CD19+B细胞。然而，C3H小鼠的CD19+细胞比C57小鼠的CD19+细胞表达更多的IL-10R，这表明CD19+细胞很可能不是IL-10下调促炎细胞因子的靶点，因为IL-10抑制C57小鼠LN细胞的促炎细胞因子的效率更高。CD3+、CD4+和CD8+T淋巴细胞均未表达明显的IL-10R。这些发现可能提示巨噬细胞或树突状细胞等其他携带IL-10R的细胞可能是IL-10抑制莱姆病小鼠LN和脾中炎性细胞因子的靶点。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We previously reported that the proinflammatory cytokines interferon-gamma (IFN-?) and interleukin-6 (IL-6), produced in response to lipidated outer surface protein A (L-OspA) and the synthetic lipohexapeptide Pam3Cys, were more efficiently inhibited by the anti-inflammatory cytokine IL-10 in lymph node (LN) cells of disease-resistant C57BL/6J than in those of disease-susceptible C3H/HeJ mice. The ability of IL-10 to efficiently down-modulate these cytokines in LN cells of C57 mice as compared with those of C3H mice correlated with the expression of significantly more IL-10 receptor (IL-10R)-bearing cells in LN of C57 than in those of C3H mice. On the contrary, spleen cells from both strains of mice showed similar levels of IL-10R bearing cells, correlating with the ability of IL-10 to dampen the production of IFN-? and IL-6 with the same efficiency in spleen cell cultures of both strains of mice. In this study, we investigated the specific lymphocyte population in LN cells that may be the target of IL-10 inhibition of proinflammatory cytokines in mice. LN and spleen cells were obtained ex vivo after one-week of infection of C3H and C57 mice with B. burgdorferi and assessed for IL-10R expression on CD3+ CD4+, CD8+ and CD19+ lymphocytes by flow cytometry. The majority of IL-10R bearing cells were of the CD19+ phenotype (B cells) in LN and spleen cells from C3H and C57 mice. However, CD19+ cells from C3H mice expressed more IL-10R than those of C57 mice, suggesting that CD19+ cells are most likely not the targets of IL-10 down-modulation of proinflammatory cytokines, given the greater efficiency by which IL-10 inhibits proinflammatory cytokines in LN cells of C57 mice. Neither CD3+, CD4+ nor CD8+ T lymphocytes expressed a significant percentage of IL-10R. These findings may suggest that other IL-10R bearing cells such as macrophages or dendritic cells may be targets of IL-10 inhibition of inflammatory cytokines in the LN and spleen of mice with Lyme disease.