流感病毒血凝素茎干部通用疫苗的构建及免疫原性研究

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Abstract In order to construct a Hemagglutinin-stem based universal vaccine and investigate its immunogenicity, the hemagglutinin (HA) gene of A/Puerto Rico/8/1934(H1N1) virus was used as template in this study to amplify the stem region, namely HA2(main protein of HA stem region) and HA-stem (whole stem region). The amplicons were inserted into pCAGGS vector, and the constructed recombinant plasmids were transiently transfected into HEK-293T cells. Then, the proteins expression was identified by RT-PCR and indirect immunofluorescence (IFA) assay. 6~8 weeks female BALB/c mice (Mus musculus) were divided into 2 recombinant plasmids group, pCAGGS group, and PBS group. The mice were intramuscularly inoculated twice at 0 and 30 d . Orbital blood was sampled at 0 and 60 d post immunization, and subjected to HI antibody titration, neutralizing antibody titration and INF-γ detection. The results showed that recombinant plasmids pCAGGS-HA2 and pCAGGS-HA-stem were successfully constructed, and could normally express target proteins in HEK-293T cells. pCAGGS-HA2 and pCAGGS-HA-stem induced antibody against A/Puerto Rico/8/1934(H1N1)in mice with HI titre 5 log2, and neutralizing antibody 1∶27 and 1∶24, respectively. While HI titre and neutralizing antibody against A/Chicken/Hebei/4/2008(H9N2) was 3 log2 and 6 log2, 1∶24 and 1∶22. The INF-γ level in pCAGGS-HA2 and pCAGGS-HA-stem group was (2362.40±98.24)ng/Land (2497.06±120.44) ng/L.The antibody titre and INF-γ level induced by pCAGGS-HA2 and pCAGGS-HA-stem was significantly higher than those induced by control group(P＜0.05). It indicates that pCAGGS-HA2 and pCAGGS-HA-stem can induce cross reaction with heterologous influenza virus, and may be a hopeful candidate universal influenza virus.

Key words： Influenza virus Hemagglutinin (HA) Stem region Universal vaccine

Received: 04 May 2016 Published: 01 October 2016

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	MA Li-Zhao
	LIU Huan
	LI Gui-Mei
	HUANG Juan
	DAN Hu

Cite this article:

MA Li-Zhao,LIU Huan,LI Gui-Mei, et al. Construction and Immunogenicity of Universal Influenza Vaccine Based on Hemagglutinin Stem Region[J]. , 2016, 24(11): 1764-1770.

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http://journal05.magtech.org.cn/Jwk_ny/EN/ OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2016/V24/I11/1764

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