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目的研究IL-4调控树突状细胞特异性非整合素(dendritic-cell specific ICAM-3 grabbing non-integrin,DC-SIGN)的表达对树突状细胞免疫学功能的影响,以探讨DC-SIGN表达与结核病发生的关系。方法用不同剂量的IL-4(5、10、50、100、200ng/ml)调控DCs表面DC-SIGN的表达水平;根据加入IL-4剂量将DCs分为5ng/ml组和50ng/ml组,分别加入100ng/ml的脂多糖(lipopolysaccharide,LPS)、结核杆菌H37Rv菌悬液共培养。应用流式细胞仪检测各组DCs表型(CD11c、DC-SIGN、CD83、CD86、HLA-DR),ELISA法检测细胞培养液中IL-12、IL-10含量,混合淋巴细胞培养法检测各组DCs刺激T淋巴细胞增殖的能力。结果①IL-4能调控DC-SIGN的表达水平,并呈剂量依赖性,当加入IL-4的剂量由5ng/ml增加到50ng/ml时,DC-SIGN的表达水平显著增加(P<0.05)。当IL-4的剂量由50ng/ml增至200ng/ml时,DC-SIGN的表达水平虽仍随IL-4剂量增加而增加,但差异无统计学意义(P>0.05)。②表达不同水平DC-SIGN的DCs成熟度无显著差异(P>0.05)。③表达不同水平DC-SIGN的DCs加入LPS、H37Rv诱导后,培养液中IL-12含量无显著差异(P>0.05),加入LPS诱导后培养液中IL-10含量也无显著差异(P>0.05),但DC-SIGNhigh-DCs加入H37Rv诱导后培养液中IL-10含量明显高于DC-SIGNlow-DCs(P<0.05)。④表达不同水平DC-SIGN的DCs加入LPS诱导后刺激同种异体T淋巴细胞增殖能力差异无统计学意义(P>0.05)。DC-SIGNhigh-DCs加入H37Rv诱导成熟后与DCs加入LPS诱导成熟后刺激同种异体T淋巴细胞增殖的能力相当(P>0.05),但DC-SIGNlow-DCs加入H37Rv诱导后刺激同种异体T淋巴细胞增殖能力明显低于其他3组(P<0.05)。结论DC-SIGNhigh-DCs加入H37Rv诱导后分泌IL-10水平明显高于DC-SIGNlow-DCs,但DC-SIGNlow-DCs加入H37Rv诱导后刺激T淋巴细胞增殖的能力明显降低。DC-SIGN表达过高或过低均不利于机体抗结核免疫应答。
Objective To investigate the effect of dendritic-cell specific ICAM-3 grabbing non-integrin (DC-SIGN) on the immunological function of dendritic cells (DCs) Expression and the relationship between tuberculosis. Methods DC-SIGN expression on DCs was induced by different doses of IL-4 (5, 10, 50, 100, 200 ng / ml). DCs were divided into 5ng / ml and 50ng / ml groups , Were added 100ng / ml of lipopolysaccharide (lipopolysaccharide, LPS), Mycobacterium tuberculosis H37Rv bacterial suspension co-culture. The phenotypes of DCs (CD11c, DC-SIGN, CD83, CD86 and HLA-DR) in each group were detected by flow cytometry. The contents of IL-12 and IL-10 in the culture medium were measured by ELISA. Group DCs stimulate the proliferation of T lymphocytes. Results ① IL-4 could regulate the expression of DC-SIGN in a dose-dependent manner. When the dosage of IL-4 was increased from 5ng / ml to 50ng / ml, the expression of DC-SIGN was significantly increased (P <0.05) . When IL-4 dose increased from 50ng / ml to 200ng / ml, the expression level of DC-SIGN still increased with the increase of IL-4 dose, but the difference was not statistically significant (P> 0.05). ② There was no significant difference in the maturation of DCs expressing different levels of DC-SIGN (P> 0.05). (3) DCs expressing different levels of DC-SIGN were added to LPS, there was no significant difference of IL-12 content in culture medium after H37Rv induction (P> 0.05). However, the content of IL-10 in DC-SIGNhigh-DCs induced by H37Rv was significantly higher than that of DC-SIGNlow-DCs (P <0.05). ④ The DCs expressing different levels of DC-SIGN had no significant difference in the proliferation of allogeneic T lymphocytes when added with LPS (P> 0.05). The ability of DC-SIGNhigh-DCs to induce the proliferation of allogeneic T lymphocytes was similar to that of DCs (P> 0.05) when they were added into H37Rv and induced by DCs added with LPS. However, DC-SIGNlow-DCs stimulated allogeneic T lymphocytes Cell proliferation was significantly lower than the other three groups (P <0.05). Conclusions The level of IL-10 secreted by DC-SIGNhigh-DCs induced by H37Rv is significantly higher than that of DC-SIGNlow-DCs. However, the ability of DC-SIGNlow-DCs to induce proliferation of T lymphocytes was significantly decreased after H37Rv treatment. DC-SIGN expression is too high or too low are not conducive to the body’s anti-TB immune response.