CAJ | 학술논문

目的探讨肝脏枯否细胞(KC)在肝移植后早期免疫反应中的可能作用.方法将KC和(或)异体PBMC共培养,收集细胞上清液,培养结束时分别收获培养的KC和PBMC.检测HLA-G在细胞表面的表达;测定上清液中NO、IFN-γ、IL-10和TGF-β1的浓度;MTT试验观察KC对淋巴细胞增殖的影响.结果实验组及对照组中KC和PBMC表面均未检测到HLA-G的表达.与不含KC实验组相比,含KC实验组中,NO、IL-10和TGF-β1的产量显著升高,而IFN-γ呈相对偏低趋势;对照组中未能检测到IL-10和IFN-γ的分泌,仅含KC的对照组中含少量NO及TGF-β1,且显著低于实验组.MTT实验发现,不含KC实验组OD值显著高于含KC实验组及对照组.结论体外KC接触异体PBMC后早期,各细胞膜表面均无HLA-G表达,但参与了NO及Th2/Th3样细胞因子的分泌调节,并能抑制淋巴细胞增殖反应,可能促进肝脏移植早期免疫耐受的形成.
목적 탐토간장고부세포(KC)재간이식후조기면역반응중적가능작용.방법 장KC화(혹)이체PBMC공배양,수집세포상청액,배양결속시분별수획배양적KC화PBMC.검측HLA-G재세포표면적표체;측정상청액중NO、IFN-γ、IL-10화TGF-β1적농도;MTT시험관찰KC대림파세포증식적영향.결과 실험조급대조조중KC화PBMC표면균미검측도HLA-G적표체.여불함KC실험조상비,함KC실험조중,NO、IL-10화TGF-β1적산량현저승고,이IFN-γ정상대편저추세;대조조중미능검측도IL-10화IFN-γ적분비,부함KC적대조조중함소량NO급TGF-β1,차현저저우실험조.MTT실험발현,불함KC실험조OD치현저고우함KC실험조급대조조.결론 체외KC접촉이체PBMC후조기,각세포막표면균무HLA-G표체,단삼여료NO급Th2/Th3양세포인자적분비조절,병능억제림파세포증식반응,가능촉진간장이식조기면역내수적형성.
Objective To explore the possible role and mechanism of the Kupffer cells (KCs) in liver allo-geneic transplantation at the early stage. Methods In vitro cell contact coculture system was established. Culture supernatants were collected respectively on the 1st, 2nd, 4th, 6th d after cocul-ture and the KCs and PBMCs were harvested on the 6th day after culture. The expression of HLA-G on the membrane of the KCs and PBMCs was detected with immunochemistry. Nitrate reduction test was used to determine the concentration of nitric oxide. IFN-γ, IL-10, TGF-β1 cytokine levels in the supernatants were also measured with ELISA. The proliferation of lymphocytes was evaluated with MTT. Results six days later, no HLA-G molecules were detected on the membrane of the KCs and PBMCs. In the experimental group containing KCs, the levels of NO, IL-10 and TGF-β1 was signifi-cantly increased(P<0. 05), while the levels of IFN-γ was relatively lower(P<0. 05) as compared to the experimental group without KCs. No IL-10 and IFN-γ were detected in the control group, and on-ly few NO and TGF-β1 was found in the control group with KCs. MTT test showed that the value of optical density was lower in the experimental group with KCs than that in any other group(P<0. 05).Conclusion No HLA-G is expressed on the membrane of KCs and PBMCs after contact coculture.KCs may participate in regulating production of NO and Th2/Th3-like cytokines and suppressing the proliferation of lymphocytes, through which KCs probably take part in inducing immunotolerance of liver transplantation in early stage.