CAJ | 학술논문

目的探讨B7-H4分子在C3H10 T1/2细胞(C3H10)移植治疗小鼠实验性变态反应性脑脊髓炎(EAE)中的作用.方法 ①用慢病毒将B7-H4靶向shRNA转染至C3H10细胞(C3H10-shRNA)并观察其对小鼠脾脏细胞活化增殖的调节作用;②利用MOG35-55和完全弗氏佐剂制备小鼠EAE模型,将50只小鼠分为正常对照组、EAE模型组、C3H10细胞组、C3H10-NC细胞组、C3H10-shRNA细胞组,通过HE染色、Fast-blue染色观察病理学改变,用免疫组化法检测B7-H4表达,结合神经功能评分观察不同干预方式对EAE损伤组织结构和功能的影响.结果 ①下调表达B7-H4的C3H10细胞可使其对小鼠脾细胞的免疫抑制作用减弱;②C3H10移植于EAE小鼠可减少、延缓发病,而移植C3 H 10-shRNA细胞的EAE小鼠发病时间、神经缺损评分和炎性细胞浸润、脱髓鞘改变及B7-H4的阳性细胞数均介于EAE组与其他对照组之间.结论 C3H10细胞移植治疗EAE的安全、有效,B7-H4分子可能通过影响C3H10的免疫调节作用等生物学行为而影响移植疗效.
목적 탐토B7-H4분자재C3H10 T1/2세포(C3H10)이식치료소서실험성변태반응성뇌척수염(EAE)중적작용.방법 ①용만병독장B7-H4파향shRNA전염지C3H10세포(C3H10-shRNA)병관찰기대소서비장세포활화증식적조절작용;②이용MOG35-55화완전불씨좌제제비소서EAE모형,장50지소서분위정상대조조、EAE모형조、C3H10세포조、C3H10-NC세포조、C3H10-shRNA세포조,통과HE염색、Fast-blue염색관찰병이학개변,용면역조화법검측B7-H4표체,결합신경공능평분관찰불동간예방식대EAE손상조직결구화공능적영향.결과 ①하조표체B7-H4적C3H10세포가사기대소서비세포적면역억제작용감약;②C3H10이식우EAE소서가감소、연완발병,이이식C3 H 10-shRNA세포적EAE소서발병시간、신경결손평분화염성세포침윤、탈수초개변급B7-H4적양성세포수균개우EAE조여기타대조조지간.결론 C3H10세포이식치료EAE적안전、유효,B7-H4분자가능통과영향C3H10적면역조절작용등생물학행위이영향이식료효.
Objective To investigate the role of the negative co-stimulatory molecule B7-H4 in mesenchymal stem cell line C3H10 T1/2 (C3H10) in the treatment of experimental allergic encephalitis (EAE).Methods ①The lentiviral vectors with mouse B7-H4 target shRNA were transfected into mouse mesenchymal stem cell (C3H10-shRNA) ; the flow cytometry was used to detect the proliferation of splenocyte that was cocultured with C3H10 cell.②EAE mice models were immunized with peptide MOG35-55 in complete Freund's adjuvant (CFA); C57BL/6 mice(n =50) were divided into Sham-operated group,EAE model group,C3H10 cell group,C3H10-NC cell group,and C3H10-shRNA cell group.Through HE and luxol fast blue staining to observe the pathologic changes of mice.Immunohistochemistry was used to label B7-H4.Combining with the neural function defect score,the effect of transplantation of mice with different cells was evaluated.Results ① By knocking down B7-H4 on C3H10,the inhibitory effect of C3H10 on splenocyte proliferation was partly revised;②Transplantation of C3H10 into EAE mice can decrease and delay the attack.The onset time of the attack,the neural function defect score,the inflammatory cell infiltration,the demyelination and the number of B7-H4 positive cells of the C3H10-shRNA cell group was between EAE model group and other control groups.Conclusions The treatment of EAE by C3H10 transplantation is safe and effective.The co-inhibitor molecule B7-H4 expressed on C3H10 cell involved in the effect of the treatment of EAE by C3H10 transplantation.