We have previously shown a clear differential genetic evolution of the hemagglutinin (HA) of human AH1N1 and AH1N2 viruses, isolated in southern France between 2001 and 2004. However, our analysis revealed that one single AH1N2 isolate, detected in 2003 (A/Lyon/0838/2003), had its HA clustering within the HAs of the AH1N1 subtypes. To determine if this virus was a new reassortant, the nucleotide sequences of its eight RNA gene segments were compared with those of five representative strains of the AH1N1, AH1N2 and AH3N2 viruses, isolated during the same time-period. According to the sequences obtained, the neuraminidase (NA) gene segment of the A/Lyon/0838/2003 (H1N2) virus was genetically closely related to those of the AH1N2 viruses, whereas the six internal genes appeared to be clustering with those of the AH1N1 viruses. This suggested that the A/Lyon/0838/2003 (H1N2) virus was the result of a second reassortment event that had occurred during the winter of 2002-2003; the N2 gene segment of an AH1N2 virus being introduced into an AH1N1 genetic background. Subsequently, we analysed the extremities of each gene segment of the viruses from the different subtypes, and experimentally reproduced, in vitro, AH1N2 reassortant viruses through co-infection of Madin-Darby canine kidney (MDCK) cells with both AH1N1 and AH1N2 viruses, isolated in the 2002-2003 influenza season. While the comparison of the AH1N1 and the AH1N2 gene segment extremities revealed no major differences, we successfully reproduced an AH1N2 reassortant virus similar to the A/Lyon/0838/2003 (H1N2) virus. This result provided an experimental evidence of the compatibility between their respective surface H1 and N2 glycoproteins, and suggests that similar events may occur silently amongst human subtypes.

译文

:我们之前已经显示了在2001年至2004年之间在法国南部分离出的人类AH1N1和AH1N2病毒的血凝素(HA)的明显遗传差异。然而,我们的分析表明,在2003年检测到了一个单独的AH1N2分离株(A / Lyon / 0838/2003),其HA群集在AH1N1亚型的HA中。为了确定这种病毒是否是新的重配体,将其八个RNA基因区段的核苷酸序列与在同一时期分离出的五个代表性AH1N1,AH1N2和AH3N2病毒株的核苷酸序列进行了比较。根据获得的序列,A / Lyon / 0838/2003(H1N2)病毒的神经氨酸酶(NA)基因片段在遗传上与AH1N2病毒密切相关,而六个内部基因似乎与A1L2 AH1N1病毒。这表明A / Lyon / 0838/2003(H1N2)病毒是2002-2003年冬季发生的第二次重排事件的结果;将AH1N2病毒的N2基因片段引入AH1N1遗传背景。随后,我们分析了来自不同亚型的病毒的每个基因片段的末端,并通过联合感染Madin-Darby犬肾(MDCK)细胞与AH1N1和AH1N2病毒,在体外实验复制了AH1N2重配病毒在2002-2003流感季节。虽然AH1N1和AH1N2基因片段末端的比较没有发现主要差异,但我们成功地复制了类似于A / Lyon / 0838/2003(H1N2)病毒的AH1N2重配病毒。该结果提供了它们各自的表面H1和N2糖蛋白之间相容性的实验证据,并表明相似的事件可能在人类亚型中悄然发生。

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