鸡Th1样淋巴因子mRNA的体外表达

In Vitro Expression of Chicken Th1-like Lymphokines Genes

应用 RT-PCR技术研究了鸡脾细胞在体外受到有丝分裂原 Con A的刺激后 ,其 Th1样淋巴因子的体外表达动态。结果表明 ,鸡脾细胞在 Con A诱导培养 1、2、3、4、8、1 2、2 4、4 8h后 ,均表达α-干扰素 m RNA,且未经有丝分裂原刺激的脾细胞以及未诱导但培养了 3、8、4 8h的脾细胞 ,也表达α-干扰素 m RNA。γ-干扰素在上述诱导培养时 ,其 m RNA均发生表达 ;未经有丝分裂原刺激的脾细胞培养 3、8h时 ,也表达 γ-干扰素m RNA,但培养 4 8h时则未检测到 ;未受有丝分裂原刺激的脾细胞不表达 γ-干扰素 m RNA。鸡白细胞介素 -2m RNA仅在刺激 2、3、4、8、1 2、2 4、4 8h时表达 ,在其他情况下则未检测到。本研究还发现 1种与已报道的鸡α-干扰素序列不同的基因 ,但其体外的表达动态类似于鸡α-干扰素。上述研究表明 ,鸡 Th1样淋巴因子的体外表达类似于哺乳动物 。

Reverse transcription-polymerase chain reaction(RT-PCR) was employed in order to prove the in vitro expression of chicken Th1-like lymphokines in ConA-stimulated splenocytes.It was demonstrated that the IFN-α mRNA was expressed in vitro when the ConA-stimulated splenocytes were cultured from 0 h to 48 h and the unstimulated splenocytes were cultured for 3 h,8 h and 48 h.The kinetics of chicken IFN-γ mRNA expression of ConA-stimulated splenocytes was similar to that of IFN-α but the IFN-γ mRNA was undetectable when the unstimulated splenocytes were cultured for 0 h and 48 h.Chicken IL-2 mRNA was detected from 2 h to 48 h in ConA-stimulated splenocytes but could not be detected in other conditions.Interestingly a possible lymphokine gene related to IFN-α was found in chicken splenocytes and it was shown that its expression kinetics in vitro was just the same as IFN-α.The results indicated that in vitro expression of Th1-like lymphokines of chicken were similar to that of mammals with minor deviation.