重组猪源CD40L体外诱导猪肺泡巨噬细胞IL-1β和TNF-α的分泌

Recombinant porcine CD40L induced IL-1β and TNF-α secretion from porcine alveolar macrophages in vitro

本试验旨在研究重组猪源CD40L蛋白(recombinant porcine CD40L,rpCD40L)的生物学活性。利用Alexa Fluor 488标记rpCD40L后,在体外与CD40+原代猪肺泡巨噬细胞(porcine alveolar macrophages,PAMs)作用,荧光显微镜观察rpCD40L与PAMs的结合,并通过qPCR、ELISA检测作用12 h和24 h后PAMs IL-1β和TNF-αm RNA与蛋白水平的变化。结果显示,荧光标记的rpCD40L与CD40+PAMs作用后,PAMs呈现绿色荧光,rpCD40L与PAMs作用后在基因和蛋白水平促进了IL-1β和TNF-α的表达。结果表明,课题组前期获得的rpCD40L能够与CD40+PAMs结合,且诱导PAMs IL-1β和TNF-α的分泌。

This experiment was design to study the biological activity of recombinant porcine CD40 L protein(rp CD40 L).After being labeled with Alexa Fluor 488,rp CD40 L was combined with CD40+porcine alveolar macrophages(PAMs)in vitro.The binding of rp CD40 L to PAMs was observed by fluorescence microscope,and the fold change of IL-1βand TNF-αof PAMs were detected after 12 h and 24 h by q PCR and ELISA.In result,when the fluorescence-labeled rp CD40 L acted with CD40+PAMs liberate green fluorescence,it promote the expression of IL-1βand TNF-αboth at gene and protein level at time dependent manner.This resultshowed that rp CD40 L obtained previously by our research group has the capability of binding to CD40+PAMs and thus induce the expression of PAMs IL-1βand TNF-αsecretion.