β-1,4-半乳糖基转移酶6在淋巴细胞迁移中的分子机制研究  

Molecular mechanism ofβ-1,4-galactosyltransferase 6 in regulating lymphocyte migration

作　　者：骈亚亚 高振祥[1] 聂晶晶 胡继红[1] Pian Yaya;Gao Zhenxiang;Nie Jingjing;Hu Jihong(National Center for Clinical Laboratories,Beijing Hospital,National Center of Gerontology,Institute of Geriatric Medicine of Chinese Academy of Medical Sciences,Beijing 100730,China)

机构地区：[1]北京医院/国家老年医学中心/国家卫生健康委临床检验中心/中国医学科学院/老年医学研究院,100730

出　　处：《中华微生物学和免疫学杂志》2020年第3期206-210,共5页Chinese Journal of Microbiology and Immunology

基　　金：北京医院博士启动基金项目(bj-2018-027);北京市科协金桥工程种子资金项目(ZZ19059);北京市东城区优秀人才培养资助项目(2019DCT-M-11)。

摘　　要：目的:研究β-1,4-半乳糖基转移酶6(β4galt6)在炎症条件下调节淋巴细胞迁移的分子机制。方法:CRISPR/Cas9敲除小鼠胰岛血管内皮细胞MS1上的β4galt6基因;黏附试验比较淋巴细胞与野生细胞株和基因敲除细胞株的黏附能力;RT-PCR及流式细胞术比较野生细胞株和基因敲除细胞株表面黏附分子的表达情况;Transwell模型比较淋巴细胞穿过野生细胞株和基因敲除细胞株的迁移能力。结果:基因敲除细胞株β4galt6 KO构建成功;淋巴细胞在炎症条件下与野生细胞株MS1的黏附显著高于与基因敲除细胞株β4galt6 KO;野生细胞株MS1表面细胞间黏附分子1(ICAM1)、血管内皮细胞黏附分子1(VCAM1)、P-selectin的表达显著高于基因敲除细胞株β4galt6 KO;淋巴细胞穿过野生细胞株MS1的能力显著高于基因敲除细胞株β4galt6 KO。结论:在炎症条件下,β4galt6能够促进淋巴细胞的迁移。Objective To study the molecular mechanism ofβ-1,4-galactosyltransferase 6(β4galt6)in regulating lymphocyte migration under inflammatory conditions.Methods CRISPR/Cas9 system was used to knock out theβ4galt6 gene of mouse islet vascular endothelial cells(MS1).Adhesion assay was performed to compare the adhesion ability of lymphocytes to wild-type cells and gene knockout cells.Expression of adhesion molecules on the surface of wild-type cells and gene knockout cells were compared using RT-PCR and flow cytometry.Transwell model was used to compare the transmigration ability of lymphocytes across wild-type cells and gene knockout cells.Results Theβ4galt6 gene knockout cell line,β4galt6 KO,was successfully constructed.The percentage of lymphocytes adhereing to wild-type MS1 cells was significantly higher than that toβ4galt6 KO cells under inflammatory conditions.The expression of adhesion molecules including intercellular adhesion molecule 1(ICAM1),vascular cell adhesion molecule 1(VCAM1)and P-selectin on the surface of wild-type MS1 cells was significantly higher than that onβ4galt6 KO cells.Moreover,the percentage of lymphocytes passing through wild-type MS1 cells was significantly higher than that throughβ4galt6 KO cells.Conclusion Under inflammatory conditions,β4galt6 could promote the migration of lymphocytes.

关 键 词：β-1 4-半乳糖基转移酶6 淋巴细胞 迁移 炎症 

分 类 号：R3[医药卫生—基础医学]