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该文研究了卵白蛋白诱导的WHBE(white hair black eyes)兔外周血来源树突状细胞(dendritic cells,DCs)甘露糖受体(mannose receptor,MR)的表达及功能,以阐明其对变应原易感的机制。分离WHBE兔外周血单核细胞,以20 ng/m L粒细胞–巨噬细胞集落刺激因子(granulocytemacrophage colony stimulating factor,GM-CSF)和20 ng/m L IL-4(interleukin-4)在体外联合诱导培养6 d,获得的细胞显示典型的树突状细胞形态特征。流式细胞术检测DCs表面分子CD86和人白细胞DR抗原(human leukocyte antigen-DR,HLA-DR)的表达,结果显示,WHBE兔和日本大耳白(Japanese white,JW)兔外周血DCs经卵白蛋白(ovalbumin,OVA)诱导后,CD86和HLA-DR表达水平均升高,其中,WHBE兔CD86水平高于JW兔。荧光定量PCR检测结果显示,OVA诱导后,两个品系兔外周血DCs的MR m RNA相对水平均显著升高(P<0.05),且WHBE兔显著高于JW兔(P<0.05)。抗原摄取实验发现,DCs经OVA刺激24 h后,抗原摄取能力低于对照组,MR抑制剂能显著降低DCs的抗原摄取率(P<0.05,P<0.01)。WHBE兔DCs抗原摄取率和平均荧光强度(mean flourscence indensity,MFI)均高于JW兔相同处理组,其中MFI值呈现极显著性差异(P<0.01)。该研究结果表明,OVA诱发后,WHBE兔DCs可能通过其表面共刺激分子CD86、抗原识别受体MR的高表达,使抗原递呈功能增强,介导Th2型细胞过度活化,导致气道变态反应性炎症。WHBE兔MR的表达及功能特点可能是影响WHBE兔DCs对变应原敏感性的关键因素。
The aim of this study was to investigate the expression and function of mannose receptor (MR) in dendritic cells (DCs) of white blood cells (WHBE) derived from ovalbumin (WHBE) The original susceptible mechanism. WHBE rabbit peripheral blood mononuclear cells were isolated and cultured in vitro with 20 ng / mL granulocyte macrophage colony stimulating factor (GM-CSF) and 20 ng / mL IL-4 The combination of induction and culture for 6 days, the obtained cells showed the typical dendritic cell morphology. Flow cytometry was used to detect the expression of DCs CD86 and HLA-DR on peripheral blood of WHBE rabbits and Japanese white rabbits (JW) After ovalbumin (OVA) induction, the expression levels of CD86 and HLA-DR were increased, of which the level of CD86 in WHBE rabbits was higher than that in JW rabbits. The results of real-time PCR showed that the relative levels of MR m RNA in DCs induced by OVA were significantly increased (P <0.05), and those in WHBE rabbits were significantly higher than those in JW rabbits (P <0.05). Antigen uptake assay showed that the antigen uptake capacity of DCs stimulated with OVA for 24 h was lower than that of the control group. MR inhibitors could significantly reduce the antigen uptake rate of DCs (P <0.05, P <0.01). The antigen uptake rate and mean flourscence intensity (MFI) of DCs in WHBE rabbits were all higher than those in the same group of JW rabbits. The MFI showed a very significant difference (P <0.01). The results show that after induced by OVA, WHBE rabbit DCs may be through its surface costimulatory molecules CD86, antigen recognition receptor MR expression, enhanced antigen presentation, mediated Th2-type over-activation of cells, leading to airway allergy Sexual inflammation. The expression and functional characteristics of WHBE rabbit MR may be the key factors that affect the sensitivity of WHBE rabbit DCs to allergen.