Morphological study of macrophages infected with constructed recombinant adenovirus carrying gp120 gene of Chinese HIV-1 strain |
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Authors: | WANG Tong MA Wen-xin GUO Jia-hui CHEN Zhi-peng CUI Yi-zhi |
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Institution: | Institute of Life and Health Engineering, Jinan University, Guangzhou 510632, China |
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Abstract: | AIM: To construct a recombinant adenovirus carrying gp120 gene of Chinese HIV-1 strain,which can infect mouse bone marrow-derived macrophages (BMM). METHODS: Co-transfection of shuttle and backbone plasmids of AdMax system into 293Ad5+ cells was performed, followed by viral packaging, propagation and purification. These viruses were subject to Karber TCID50 titration. The expression of gp120 protein in 293Ad5+ cells was determined by ELISA. The viral titration was validated by a multiplicity of infection (MOI) test with BMM. RESULTS: The titers of the outcome viruses, including AdMax-HIV-1 gp120 (Ad-gp120) and its vector control Ad-GFP, were 108.3 and 108.1 TCID50/mL, respectively. Both recombinant adenoviruses infected BMM with similar capacity of 293Ad5+ cell infection, which validated the TCID50 titration.The gp120 protein was positive in 293Ad5+ cell lysates. BMM activation was observed morphologically after Ad-gp120 infection as compared with Ad-GFP-infected cells. CONCLUSION: Functional adenovirus containing HIV-1 gp120 of prevalent strains in China was successfully constructed. Infection of Ad-gp120 causes BMM activation. |
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Keywords: | Recombinant adenovirus HIV envelope protein gp120 Bone marrow-derived macrophages |
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