The study of correlation between the mRNA expression of Toll-like receptors and cytokines of intracellular CD_4~+ T cell in splenic macrophages of allergic asthma mice model / 中华检验医学杂志

ABSTRACT

Objective To understand the expression of mRNA for two Toll-like receptors (TLRs) (TLR2 and TLR4) and two T helper (Th)1 and Th2 cytokines [ interleukin (IL)-4, interferon (IFN)-gamma] in splenic macrophages of ovalbumin-induced allergic asthma mice model. Methods The expression of mRNA for TLB2、TLR4 and the house-keeping gene HPRT in both splenic macrophages of BALB/c allergic asthma mice and the control mice were analysed by real-time polymerase chain reaction (PCR). We examined the threshold cycle times (Ct) of cDNA of TLR2、TLR4 and HPRT gene to get Ct2、Ct4 and CtH respectively. The value of relative quantity of mRNA level was calculated by the ratio of Ct2/ CtH、Ct4/CtH. After stimulating cells with PMA and Ionomycin with anti-CD3/CD8 in the presence of Brefeldin A, intracellular levels of Th1 cytokines (IFN-?) and Th2 cytokines (IL-4) were determined for CD3+ CD8- and CD3+ CD8+ T cells. Results We observed Ct2/CtH (1. 26?0. 04,1. 35?0. 11, P 0.05) in splenic macrophages between the control mice and allergic asthma mice; the expression of TLR2 mRNA of allergic asthma mice was significantly reduced. We examined IFN-?[IFN-?(20. 83?1. 32)%, (31.78?2. 54) %, IL4(2. 04?0. 24)% , (5. 91?0.83)% P