CAJ | 학술논문

目的：探讨结核杆菌 H37Ra 对诱导共刺激分子(ICOS)的配体 B7-H2表达的影响。方法体外培养肺泡Ⅱ型细胞 A549并且从人外周血中分离 B 淋巴细胞,用逆转录聚合酶链反应(RT-PCR)方法分析 hGL50和 B7-H2在两种细胞中不同的表达。A549细胞分别用结核杆菌 H37Ra,脂多糖(LPS)和白细胞介素4(IL-4)诱导4、8、12小时,用 RT-PCR 方法分析 B7-H2的表达。用免疫荧光方法分析 B7-H2分子在 A549细胞表面的表达。用流式细胞方法分析 H37Ra、干扰素γ(IFN-γ)和 IL-4诱导4小时和24小时后 A549细胞 B7-H2和 CD40分子表达情况。结果B7-H2 mRNA 在 A549细胞表达,hGL50 mRNA 在 B 细胞高表达。与 LPS、IL-4相比,结核杆菌 H37Ra 可以诱导A549细胞高表达 B7-H2。免疫荧光染色可以看到 A549细胞高表达 B7-H2分子。流式细胞结果显示,与 IFN-γ和IL-4相比,结核杆菌 H37Ra 可以显著诱导 A549细胞表面表达 B7-H2分子。结论 ICOS 配体 B7-H2表达于肺泡Ⅱ型上皮细胞 A549,结核杆菌 H37Ra 诱导可从 mRNA 和蛋白水平显著提高 B7-H2的表达。
목적：탐토결핵간균 H37Ra 대유도공자격분자(ICOS)적배체 B7-H2표체적영향。방법체외배양폐포Ⅱ형세포 A549병차종인외주혈중분리 B 림파세포,용역전록취합매련반응(RT-PCR)방법분석 hGL50화 B7-H2재량충세포중불동적표체。A549세포분별용결핵간균 H37Ra,지다당(LPS)화백세포개소4(IL-4)유도4、8、12소시,용 RT-PCR 방법분석 B7-H2적표체。용면역형광방법분석 B7-H2분자재 A549세포표면적표체。용류식세포방법분석 H37Ra、간우소γ(IFN-γ)화 IL-4유도4소시화24소시후 A549세포 B7-H2화 CD40분자표체정황。결과B7-H2 mRNA 재 A549세포표체,hGL50 mRNA 재 B 세포고표체。여 LPS、IL-4상비,결핵간균 H37Ra 가이유도A549세포고표체 B7-H2。면역형광염색가이간도 A549세포고표체 B7-H2분자。류식세포결과현시,여 IFN-γ화IL-4상비,결핵간균 H37Ra 가이현저유도 A549세포표면표체 B7-H2분자。결론 ICOS 배체 B7-H2표체우폐포Ⅱ형상피세포 A549,결핵간균 H37Ra 유도가종 mRNA 화단백수평현저제고 B7-H2적표체。
Objective To investigate the effects of tuberculosis bacili H37Ra on the inducible costimulatory molecule(ICOS)ligand B7-H2 expression.Methods Alveolar type Ⅱ cells A549 were cultured in vitro and B lymphocytes were isolated from human peripheral blood.HGL50 and B7-H2 expression in two different cells were analyzed by reverse transcription-polymerase chain reaction(RT-PCR).A549 cells were stimulated with tuberculosis bacili H37Ra,lipopolysaccharides (LPS)and interleukin 4 (IL-4 ),respectively,for 4 h,8 h and 12 h,and the expression of B7-H2 was analyzed by RT-PCR.B7-H2 expression on the surface of A549 cells was also detected with immunofluorescence analysis.B7-H2 and CD40 molecular expression were determined on A549 cells after being stimulated by H37Ra,interferon γ(IFN-γ)and IL-4 for 4 h and 24 h by flow cytometry analysis.Results B7-H2 mRNA was expressed in A549 cells,and hGL50 mRNA was highly expressed in B cell.Compared with LPS and IL-4, tuberculosis bacili H37Ra induced higher expression of B7-H2 in A549 cells.Immunofluorescence staining showed the expression of B7-H2 in A549 cells.Flow cytometry showed that tuberculosis bacili H37R could significantly stimulate the B7-H2 in A549 cell surface expression,compared with IFN-γand IL-4.Conclusion B7-H2 is expressed in alveolar type Ⅱ epithelial cell A549,and tuberculosis bacili H37Ra stimulation significantly increases the expression of B7-H2 in mRNA and protein levels.