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目的研究不同结核菌株诱导人DC分泌I型干扰素的水平差异。方法应用免疫磁珠分选术从健康人外周血单个核细胞(PBMC)中分选CD14+单核细胞,在GM-CSF和IL-4刺激下诱导分化为DC,随后应用不同结核分枝杆菌(H37Ra、H37Rv)诱导,应用ELISA测定不同时间段(0 h、1 h、2 h、4 h、8 h、16 h、24h和48 h)上清中的I型干扰素含量。结果 IFN-β分泌水平在感染16 h后达高峰;结核菌刺激16 h后,强毒株(H37Rv)和弱毒株(H37Ra)诱导DC产生IFN-β的量分别为(4708.7±548.5)pg/ml、(3507.6±343.1)pg/ml,差异有统计学意义(P<0.05);刺激24 h后,H37Rv和H37Ra诱导DC产生IFN-β的量分别为(3696.3±458.4)pg/ml、(2597.1±341.6)pg/ml,差异有统计学意义(P<0.05)。IFN-α分泌水平在感染48h后达到高峰;结核菌刺激24 h和48 h后,H37Rv和H37Ra诱导DC产生IFN-α的量分别为(376.5±73.2)pg/ml vs(305.3±64.1)pg/ml,(678.3±115.2)pg/ml vs(596.6±99.3)pg/ml,差异无统计学意义(P>0.05)。结论IFN-β分泌水平在16 h后达到高峰,结核菌强毒株能够诱导更高水平的IFN-β;IFN-α分泌水平在48 h后达到高峰,不同毒力结核菌株之间差异不明显。
Objective To study the differences in the level of type I interferon secretion induced by different strains of tuberculosis. Methods CD14 + monocytes were isolated from healthy human peripheral blood mononuclear cells (PBMCs) by immunomagnetic bead sorting and induced to differentiate into DCs stimulated by GM-CSF and IL-4. Then different Mycobacterium tuberculosis H37Ra and H37Rv). The levels of type I interferon in the supernatants of different time periods (0 h, 1 h, 2 h, 4 h, 8 h, 16 h, 24 h and 48 h) were determined by ELISA. RESULTS: The levels of IFN-β secreted by the virulent strain (H37Rv) and attenuated strain (H37Ra) after induction of 16 h reached the peak at 16 h after infection (4708.7 ± 548.5 pg / ml, and (3507.6 ± 343.1) pg / ml respectively (P <0.05). The amounts of IFN-β induced by H37Rv and H37Ra were (3696.3 ± 458.4) pg / ml after 24 h stimulation 2597.1 ± 341.6) pg / ml, the difference was statistically significant (P <0.05). The levels of IFN-α secreted by H37Rv and H37Ra after 24 and 48 h stimulation were (376.5 ± 73.2) pg / ml vs (305.3 ± 64.1) pg / ml, (678.3 ± 115.2) pg / ml vs (596.6 ± 99.3) pg / ml, the difference was not statistically significant (P> 0.05). Conclusions The level of IFN-βis peaked after 16 h, and the virulent strain of M. tuberculosis can induce a higher level of IFN-β. The level of IFN-αis peaked at 48 h, but there is no significant difference between different strains of virulent TB .