负载布鲁杆菌WboA^(-/-)S_2株对小鼠骨髓源性肥大细胞的活化作用研究

Studies on the activation of murine bone marrow-derived mast cells pulsed with Brucella WobA^(-/-)S_2

研究猪种布鲁杆菌WboA-/-S2株对骨髓源性肥大细胞(BMMC)的激活作用。用粗糙型布鲁杆菌WboA-/-S2株感染BMMC,并以光滑型S2株作为对照,采用RT-PCR法观察BMMC载菌后不同时间点TLR4和TLR8表达的变化;用ELISA法检测感染后BMMC上清中TNF-α、IL-6、IL-1和IL-12的含量。结果显示负载WboA-/-S2株和S2株12h时BMMC TLR4的表达量均明显高于正常对照组,其中负载WboA-/-S2株的BMMC TLR4表达又明显高于S2株负载组,24h时二者表达均明显减弱;在负载12hWboA-/-S2株组BMMC TLR8表达明显高于正常对照组,而S2株组的BMMC TLR8与正常对照组相比条带无明显变化,24h时二者表达均减弱。负载WboA-/-S2株后3h、6h、12h、24hBMMC所分泌的TNF-α和IL-6都明显高于S2株负载组(P<0.05),而在负载两菌的BMMC上清中均未检测到IL-1和IL-12。结果表明WboA-/-S2株较S2株更易被BMMC识别,激活BMMC的能力也明显强于S2株,并可快速诱导BMMC分泌大量细胞因子。

This study investigated the effects of B. suis Whoa / S2 on the activation of bone marrow derived mast cells （BM MC）. The BMMC were pulsed by B. suis WhoA ＂ Se in vitro and B. Suis Szacted as the control. The expression of TLR4 and TLR8 after infection were observed by RT-PCR at different time points. The levels of TNFa, IL-6, II.-1 and IIA2 in the ser- um at different time points of infection were detected by ELISA. The expression of TLR4 at 12h of pulsation with both WboA Sz and Szwere significantly higher than that of the normal group, besides, with the WhoA -Se-pulsed group being significantly higher than the S2-pulsed group. Then, the expression of TLR4 in both of the groups was significantly decreased at 24h post pulsation; the expression of TI.R8 at 12h of infection with WhoA/ S2 was significantly higher than that of the normal group. However, the S2 pulsed group showed no significant difference as compared with the normal group. The expression of TI.R8 in both of the groups decreased at 24h post-pulsation. The levels of both TNF-α and IL 6 in the serum of WboA Se pulsed BM MC were significantly higher than those in the Sz pulsed group at 3h, 6h, 12h, 24h （P〈0.05）, but IL-1 and 1L-12 could not be found in the serum of BMMC in both of the groups. Our results indicate that B. suis WhoA ＂ S2 is more easily recognized by BMMC and is much more powerful to activate BMMC than B. suis Se, besides, it can rapidly induce BMMC to secrete large number of cytokines.