Kita M, Tong L J, Imanishi J
Département de Microbiologie, Université médicale de la Préfecture de Kyoto, Japon.
C R Seances Soc Biol Fil. 1993;187(4):561-8.
The induction of the cytokine mRNA after infection with Herpes simplex virus (HSV) was studied using RT-PCR method capable of detecting low levels of mRNA. Total RNA was prepared from spleen lymphocytes 3 h after infection with HSV-1 (+GC virulent variant and -GCr attenuated variant of Miyama strain) by acid guanidinium thiocyanate-phenol-chloroform (AGPC) method. cDNA was synthesized by M-MLV reverse transcriptase, and amplified using the specific oligonucleotide primers for IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, TNF-alpha, IFN-alpha, IFN-beta and IFN-gamma by PCR method. After HSV-1 infection, IFN-alpha, IFN-beta, IFN-gamma, IL-1 beta, IL-4, IL-6 and TNF-alpha mRNA were significantly induced, but IL-2, IL-3 and IL-5 mRNA were not induced. Although IFN-alpha, IFN-gamma and IL-6 mRNA were more strongly induced by infection with +GC virulent variant than -GCr attenuated variant, there was no significant difference in the expression of other cytokine mRNA between two variants. These results demonstrate that cytokine mRNA in addition to IFN was induced by HSV infection, and suggest that cytokines as well as IFNs may play a role in the defense mechanism against HSV infection.
采用能够检测低水平mRNA的逆转录聚合酶链反应（RT-PCR）方法，研究单纯疱疹病毒（HSV）感染后细胞因子mRNA的诱导情况。通过酸性异硫氰酸胍-酚-氯仿（AGPC）法，在感染1型单纯疱疹病毒（宫山株的+GC强毒株和-GCr减毒株）3小时后，从脾淋巴细胞中制备总RNA。用M-MLV逆转录酶合成cDNA，并通过PCR方法，使用针对白细胞介素-1（IL-1）、白细胞介素-2（IL-2）、白细胞介素-3（IL-3）、白细胞介素-4（IL-4）、白细胞介素-5（IL-5）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、干扰素-α（IFN-α）、干扰素-β（IFN-β）和干扰素-γ（IFN-γ）的特异性寡核苷酸引物进行扩增。1型单纯疱疹病毒感染后，干扰素-α、干扰素-β、干扰素-γ、白细胞介素-1β、白细胞介素-4、白细胞介素-6和肿瘤坏死因子-α的mRNA被显著诱导，但白细胞介素-2、白细胞介素-3和白细胞介素-5的mRNA未被诱导。尽管与-GCr减毒株相比，+GC强毒株感染更强烈地诱导了干扰素-α、干扰素-γ和白细胞介素-6的mRNA，但两种毒株之间其他细胞因子mRNA的表达没有显著差异。这些结果表明，除干扰素外，细胞因子mRNA也由单纯疱疹病毒感染诱导，并提示细胞因子以及干扰素可能在抗单纯疱疹病毒感染的防御机制中发挥作用。
