He Jing, Wang Hui, Zhao Jin-xia, Li Zhan-guo
Department of Rheumatology and Immunology, Peking University People's Hospital, Beijing 100044, China.
Zhonghua Yi Xue Za Zhi. 2008 Aug 26;88(33):2360-3.
To explore the possibility to establish Sjögren's syndrome models by immunizing mice with alpha-fodrin.
Twenty-four 4-week-old BALB/C mice were randomly divided into 4 equal groups to undergo subcutaneous injection of alpha-Fodrin, submaxillary gland homogenate and glutathione S-transferase (GST) or phosphate-buffered saline (PBS) (negative control groups) on days 0, 14, 35, and 56 respectively. The drinking amount of water was measured. Blood samples were collected every 2 - 3 weeks. Munofluorescence assays and ELISA were used to examine the presence of anti-Fodrin, anti-type 3 muscarinic acetylcholine receptor polypeptide (M3RP), anti-SSA, anti-SSB, rheumatoid factor (RF), and antinuclear antibody (ANA). Immunochemistry was used to detect the levels of interferon (IFN)-gamma, interleukin (IL)-2, and IL-10. One mouse was killed from each group every 2 - 3 weeks. The salivary glands were examined.
(1) No auto-immune antibody was found in the serum samples of the mice before immunization. Antibodies against alpha-Fodrin and M2RP, and ANA were positive in the serum samples of the alpha-Fodrin and submaxillary gland homogenate groups since the 35th day after immunization, and were all negative in the 2 control groups. However, no antibodies against SSA, SSB and RF were found in all 4 groups. (2) Lymphocytic infiltration could be seen in the salivary glands of the immunized animals since 50th days after the first immunization of alpha-Fodrin and submaxillary gland homogenate. Immunohistochemistry showed alpha-Fodrin expression in the submaxillary glands of the alpha-Fodrin and submaxillary gland homogenate groups, but not in the PBS and GST controls. (3) The serum IFN-alpha levels of the alpha-Fodrin and submaxillary gland homogenate groups were (81.6 +/- 7.1) and (90.5 +/- 4.9) pg/ml respectively, both significantly higher than those of the GST and PBS groups [(30.1 +/- 5.9) and (19.3 +/- 6.4) pg/ml respectively, both P < 0.05]. The serum IL-2 levels of the alpha-Fodrin and submaxillary gland homogenate groups were (18.7 +/- 2.3) and (19.8 +/- 0.9) pg/ml respectively, both significantly higher than those of the GST and PBS groups [(4.9 +/- 1.1) and (3.5 +/- 1.6) pg/ml respectively, both P < 0.05]. No difference was found in the level of serum IL-10 among the 4 groups. (4) There was no significant difference in the volume of water volume drunk among the 4 groups.
(1) It is possible to establish mouse mice SS models by immunization with alpha-Fodrin or submaxillary gland homogenate that are reminiscent of human SS. (2) The appearance of multiple antibodies may be related to the antigen epitope spreading. (3) The pathogenesis of SS may be related to Th1 type response.
探讨通过用α- fodrin免疫小鼠建立干燥综合征模型的可能性。
将24只4周龄的BALB/C小鼠随机分为4组，分别于第0、14、35和56天皮下注射α- fodrin、颌下腺匀浆和谷胱甘肽S-转移酶（GST）或磷酸盐缓冲盐水（PBS）（阴性对照组）。测量饮水量。每2 - 3周采集血样。采用免疫荧光分析和酶联免疫吸附测定法检测抗fodrin、抗3型毒蕈碱型乙酰胆碱受体多肽（M3RP）、抗SSA、抗SSB、类风湿因子（RF）和抗核抗体（ANA）的存在情况。采用免疫化学方法检测干扰素（IFN）-γ、白细胞介素（IL）-2和IL-10的水平。每2 - 3周从每组处死1只小鼠，检查唾液腺。
（1）免疫前小鼠血清样本中未发现自身免疫抗体。自免疫后第35天起，α- fodrin组和颌下腺匀浆组血清样本中抗α- fodrin和M2RP抗体以及ANA呈阳性，2个对照组均为阴性。然而，4组中均未发现抗SSA、抗SSB和RF抗体。（2）自首次注射α- fodrin和颌下腺匀浆免疫后第50天起，免疫动物的唾液腺中可见淋巴细胞浸润。免疫组织化学显示α- fodrin组和颌下腺匀浆组颌下腺中有α- fodrin表达，而PBS组和GST对照组中无表达。（3）α- fodrin组和颌下腺匀浆组血清IFN-α水平分别为（81.6±7.1）和（90.5±4.9）pg/ml，均显著高于GST组和PBS组[分别为（30.1±5.9）和（19.3±6.4）pg/ml，P均<0.05]。α- fodrin组和颌下腺匀浆组血清IL-2水平分别为（18.7±2.3）和（19.8±0.9）pg/ml，均显著高于GST组和PBS组[分别为（4.9±1.1）和（3.5±1.6）pg/ml，P均<0.05]。4组血清IL-10水平无差异。（4）4组饮水量无显著差异。
（1）用α- fodrin或颌下腺匀浆免疫小鼠有可能建立类似于人类干燥综合征的小鼠模型。（2）多种抗体的出现可能与抗原表位扩展有关。（3）干燥综合征的发病机制可能与Th1型反应有关。
