Zhou Ying-Bo, Yuan Xiang, Wang Qi-Kai, Zhang Hong, Wang Guo-Sheng, Li Xiang-Pei, Wang Yiping, Harris David, Li Xiao-Mei
Department of Rheumatology and Immunology, the First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, PR China.
Westmead Institute for Medical Research, University of Sydney, Westmead, 2145 NSW, Australia.
Immunol Lett. 2020 Oct;226:62-70. doi: 10.1016/j.imlet.2020.07.005. Epub 2020 Jul 21.
Overexpression of CD40 has been reported in patients with primary Sjögren's syndrome (pSS). The increased CD40 expression promote autoimmune response and enhance inflammation in pSS. The aim of this study is to block CD40-CD154 interaction with CD40 DNA vaccine to slow the disease progression of SS in non-obese diabetic (NOD) mice.
Female NOD mice were treated with CD40 DNA vaccine, empty vector and normal saline. The salivary flow rate was measured, whereas lymphocytes infiltration in the salivary glands was assessed by histopathology. Expression of CD40 and B220 in salivary were examined by immunohistochemistry. Splenic lymphocyte phenotypes were analyzed by flow cytometry. CD40, IL-1β, TNF-α and IL-6 levels in the salivary glands were detected by PCR. Serum anti-CD40 antibody was measured by ELISA. Serum anti-nuclear antibody (ANA) was monitored by immunofluorescence.
NOD mice treated with CD40 DNA vaccine showed higher levels of anti-CD40 antibody compared with the controls. The expression of CD40 in the salivary glands of NOD mice in CD40 DNA vaccine group was decreased. The infiltration of lymphocytes was reduced in the salivary glands and saliva secretion was increased in the treatment group. The expression level of TNF-α and IL-6 in salivary glands were declined. The splenic dendritic cell and plasma cell populations were reduced and the level of ANA was decreased in NOD mice with CD40 DNA vaccine treatment.
CD40 DNA vaccine inhibits the immune response and reduce inflammation in epithelial tissues SS in non-obese diabetic (NOD) mice, suggesting that CD40 DNA vaccine could be a new therapeutic approach in treatment of pSS.
原发性干燥综合征(pSS)患者中已报道CD40过表达。CD40表达增加会促进自身免疫反应并加剧pSS中的炎症。本研究的目的是用CD40 DNA疫苗阻断CD40-CD154相互作用,以减缓非肥胖糖尿病(NOD)小鼠干燥综合征的疾病进展。
对雌性NOD小鼠分别用CD40 DNA疫苗、空载体和生理盐水进行处理。测量唾液流速,通过组织病理学评估唾液腺中的淋巴细胞浸润情况。用免疫组织化学法检测唾液中CD40和B220的表达。通过流式细胞术分析脾淋巴细胞表型。用PCR检测唾液腺中CD40、IL-1β、TNF-α和IL-6的水平。用ELISA法检测血清抗CD40抗体。通过免疫荧光监测血清抗核抗体(ANA)。
与对照组相比,用CD40 DNA疫苗处理的NOD小鼠显示出更高水平的抗CD40抗体。CD40 DNA疫苗组中NOD小鼠唾液腺中CD40的表达降低。治疗组唾液腺中淋巴细胞浸润减少,唾液分泌增加。唾液腺中TNF-α和IL-6的表达水平下降。CD40 DNA疫苗处理的NOD小鼠中脾树突状细胞和浆细胞数量减少,ANA水平降低。
CD40 DNA疫苗可抑制非肥胖糖尿病(NOD)小鼠上皮组织干燥综合征中的免疫反应并减轻炎症,提示CD40 DNA疫苗可能是治疗pSS的一种新的治疗方法。