Department of Molecular Biology, Norman Bethune Medical College, Jilin University, Changchun 130021, China.
J Immunol Methods. 2012 Dec 14;386(1-2):78-84. doi: 10.1016/j.jim.2012.09.002. Epub 2012 Sep 12.
Complete Freund's adjuvant (CFA) containing the inactivated mycobacterium has been conventionally used to induce typical arthritis in rats. In comparison, incomplete Freund's adjuvant (IFA), lacking the mycobacterium, can only induce less severe arthritis. However, the key components responsible for the arthritogenic effect of the whole mycobacterium are rarely known. Although mycobacterial heat-shock protein 65 (MHSP65) specific humoral and cellular immune responses were detected in rats with arthritis induced by CFA, MHSP65 alone cannot induce arthritis. In this study, we replaced the whole mycobacterium in CFA with recombinant MHSP65 (rMHSP65), prepared rMHSP65-IFA emulsion by mixing rMHSP65 and IFA, and investigated whether rMHSP65-IFA could induce arthritis in rats as CFA did. We found that intradermal injection of the rMHSP65-IFA emulsion induced arthritic lesions in testing animals to the same degree as those induced by CFA, manifested by severe swelling in hind paws, and synovial thickening, cartilage erosion and lymphocytes infiltration in ankle joints. Notably, the rMHSP65-IFA recipe also induced the production of anti-dsDNA and -rMHSP65 antibodies in rats. These results thus demonstrate that rMHSP65 can be used to substitute the inactivated mycobacteria in CFA to induce typical arthritis in rats.
弗氏完全佐剂（CFA）含有灭活的分枝杆菌，常用于诱导大鼠典型关节炎。相比之下，缺乏分枝杆菌的弗氏不完全佐剂（IFA）只能诱导较轻的关节炎。然而，导致整个分枝杆菌致关节炎作用的关键成分知之甚少。尽管在 CFA 诱导的关节炎大鼠中检测到分枝杆菌热休克蛋白 65（MHSP65）特异性体液和细胞免疫反应，但 MHSP65 本身不能诱导关节炎。在这项研究中，我们用重组 MHSP65（rMHSP65）代替 CFA 中的整个分枝杆菌，通过混合 rMHSP65 和 IFA 制备 rMHSP65-IFA 乳剂，并研究 rMHSP65-IFA 是否可以像 CFA 那样诱导大鼠关节炎。我们发现，皮内注射 rMHSP65-IFA 乳剂可使试验动物产生与 CFA 相同程度的关节炎病变，表现为后爪严重肿胀，踝关节滑膜增厚、软骨侵蚀和淋巴细胞浸润。值得注意的是，rMHSP65-IFA 配方还诱导大鼠产生抗 dsDNA 和-rMHSP65 抗体。这些结果表明，rMHSP65 可替代 CFA 中的灭活分枝杆菌诱导大鼠典型关节炎。
