摘要
目的明确重症肌无力(MG)患者胸腺组织CC趋化因子配体21(CCL21)的分布;通过CCL21基因转染MG患者细胞角蛋白8/18(CK8/18)阳性胸腺上皮细胞(TEC),探讨CCL21过表达对CK8/18阳性TEC中抗原提呈功能相关分子表达的影响。方法采用免疫组织化学检测CCL21、CK8/18阳性细胞在MG患者胸腺的表达及分布,采用实时荧光定量PCR(qRT-PCR)检测MG患者胸腺CCL21、CCL19及CC趋化因子受体7(CCR7)的mRNA水平;p CMV-CCL21质粒转染MG患者分离的CK8/18阳性TEC,qRT-PCR检测转染前后TEC功能性基因CD80、细胞间黏附分子1(ICAM-1)、CD86、HLA-DR、HLA-A的mRNA水平。结果 MG胸腺组织中CK8/18阳性细胞明显多于正常对照,且在皮质及髓质区均有分布,CCL21蛋白、CCR7mRNA的水平显著增高。p CMV-CCL21质粒转染TEC后,HLA-A、HLA-DR、ICAM和CD80 mRNA水平显著增加。结论 CCL21过表达,可引起MG患者CK8/18阳性胸腺上皮细胞抗原呈递功能相关基因的表达增高。
Objective To identify the distribution of chemokine （C-C motif） ligand 21 （CCL2!） in the thymus of patients with myasthenia gravis （MG） and explore the effects of up-regulation of CCL21 on the expressions of antigen presentation-related genes in cytokeratin 8/18 （CK8/18） positive thymic epithelial cells （TECs） after transfected with CCL21 genes. Methods The expressions and distributions of CK8/18 and CCL21 in the thymus tissue of MG patients were detected by immunohistochemistry. The mRNA levels of CCL21, CCL19 and their receptor chemokine （C-C motif） receptor 7 （CCR7） in the thymus tissue of MG patients were determined by real-time quantitative PCR （qRT-PCR）. Primary cultured CK8/18^＋ TECs were transfected with pCMV-CCL21, and the relative mRNA expressions of function-associated genes （CD80, ICAM-1, CD86, HLA-DR, HLA-A） in CK8/18＋ TECs before and after the transfection were investigated by qRT-PCR. Results Immunohistochemical results showed that the number of CK8/18 positive cells in the hyperplastic thymus tissues of MG patients was significantly more than that in the normal controls, and the protein expression of CCL21 was also much higher in the hyperplastic thymus tissues. The qRT-PCR showed that the expressions of CCL21 and CCR7 mRNA increased significantly in hyperplastic thymus tissues of MG patients compared with those in normal controls, while there was no difference in the expression of CCL19. Furthermore, CK8/18 positive cells were found mainly located in cortico-medullary junction and medulla area. The relative mRNA expression levels of HI_A-A, HI_A-DR, ICAM and CD80 rose significantly in CK8/18^＋ TECs after transfected with pCMV-CCL21. Conclusion The over-expression of CCL21 could increased the expressions of antigen presentation-related genes in CK8/18^＋ TECs in MG patients.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2015年第7期957-960,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81172874)
