Yano N, Endoh M, Naka R, Takemura F, Nomoto Y, Sakai H
Department of Internal Medicine, Tokai University School of Medicine, Kanagawa, Japan.
J Clin Immunol. 1996 Jan;16(1):71-9. doi: 10.1007/BF01540975.
Previously we reported disease-specific interaction between interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) in patients with IgA nephropathy (IgAN), suggesting the existence of unusual T cell behavior in this disease. In the present study, we investigated characteristic synthesis of interferon-gamma (IFN-gamma) and expression of IFN-gamma receptor (IFN-gamma R) in the peripheral blood mononuclear cells (PBMC) from patients with IgAN and other chronic proliferative glomerulonephritis (PGN). Heparinized peripheral blood samples were obtained from 38 patients with chronic mesangial proliferative glomerulonephritis (CGN; including 24 with IgA nephropathy) and 20 healthy controls. PBMC were isolated by gradient centrifugation and fragments were cultured in Iscove's modified Dulbecco's medium (IMDM) supplemented with 10% fetal calf serum (FCS) for 72 hr. IFN-gamma concentrations in supernatants were evaluated by the enzyme-linked immunosorbent assay (ELISA). Other parts of PBMC pellets were reacted with anti-human IFN-gamma R monoclonal antibody and FITC-labeled anti-mouse second antibody for analysis of IFN-gamma R expression on these cells by FACScan. The remaining PBMC were fractionated into CD4+ T cells, CD8+ T cells, B cells, NK, cells and macrophages using the MACS cell sorting system. The isolated cells were evaluated for IFN-gamma or IFN-gamma R mRNA expression by the semiquantitative RT-PCR method. In vitro IFN-gamma synthesis was enhanced in patients with CGN, and NK cells were revealed to be responsible for such enhancement. On the other hand, the expression of IFN-gamma R on macrophages was suppressed in CGN patients. These results suggest that impairment of regulation of the IFN-gamma system might be involved in the development of CGN.
此前我们报道了在IgA肾病（IgAN）患者中，γ干扰素（IFN-γ）与白细胞介素-4（IL-4）之间存在疾病特异性相互作用，提示该疾病中存在异常的T细胞行为。在本研究中，我们调查了IgAN患者及其他慢性增殖性肾小球肾炎（PGN）患者外周血单个核细胞（PBMC）中γ干扰素（IFN-γ）的特征性合成及IFN-γ受体（IFN-γR）的表达。从38例慢性系膜增殖性肾小球肾炎（CGN；包括24例IgA肾病患者）患者及20名健康对照者采集肝素化外周血样本。通过梯度离心分离PBMC，将细胞碎片在添加10%胎牛血清（FCS）的Iscove改良杜尔贝科培养基（IMDM）中培养72小时。采用酶联免疫吸附测定（ELISA）评估上清液中IFN-γ的浓度。PBMC沉淀的其他部分与抗人IFN-γR单克隆抗体及异硫氰酸荧光素（FITC）标记的抗小鼠二抗反应，通过流式细胞仪（FACScan）分析这些细胞上IFN-γR的表达。使用磁性激活细胞分选系统（MACS）将剩余的PBMC分离为CD4⁺T细胞、CD8⁺T细胞、B细胞、自然杀伤（NK）细胞及巨噬细胞。采用半定量逆转录聚合酶链反应（RT-PCR）方法评估分离细胞中IFN-γ或IFN-γR mRNA的表达。CGN患者体外IFN-γ合成增强，且自然杀伤细胞被证实是这种增强的原因。另一方面，CGN患者巨噬细胞上IFN-γR的表达受到抑制。这些结果提示IFN-γ系统调节受损可能参与了CGN的发病过程。
