Tan Hyon-Xhi, Gilbertson Brad P, Jegaskanda Sinthujan, Alcantara Sheilajen, Amarasena Thakshila, Stambas John, McAuley Julie L, Kent Stephen J, De Rose Robert
Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia; Laboratory of Infectious Diseases, NIAID, NIH, Bethesda, MD 20892, United States.
Vaccine. 2016 Feb 24;34(9):1172-9. doi: 10.1016/j.vaccine.2016.01.030. Epub 2016 Jan 28.
Influenza viruses are promising mucosal vaccine vectors for HIV but their use has been limited by difficulties in engineering the expression of large amounts of foreign protein. We developed recombinant influenza viruses incorporating the HIV-1 p24 gag capsid into the NS-segment of PR8 (H1N1) and X31 (H3N2) influenza viruses with the use of multiple 2A ribosomal skip sequences. Despite the insertion of a sizable HIV-1 gene into the influenza genome, recombinant viruses were readily rescued to high titers. Intracellular expression of p24 capsid was confirmed by in vitro infection assays. The recombinant influenza viruses were subsequently tested as mucosal vaccines in BALB/c mice. Recombinant viruses were attenuated and safe in immunized mice. Systemic and mucosal HIV-specific CD8 T-cell responses were elicited in mice that were immunized via intranasal route with a prime-boost regimen. Isolated HIV-specific CD8 T-cells displayed polyfunctional cytokine and degranulation profiles. Mice boosted via intravaginal route induced recall responses from the distal lung mucosa and developed heightened HIV-specific CD8 T-cell responses in the vaginal mucosa. These findings demonstrate the potential utility of recombinant influenza viruses as vaccines for mucosal immunity against HIV-1 infection.
流感病毒是用于HIV的有前景的粘膜疫苗载体，但由于在工程化表达大量外源蛋白方面存在困难，其应用受到限制。我们利用多个2A核糖体跳跃序列，开发了将HIV-1 p24 gag衣壳整合到PR8（H1N1）和X31（H3N2）流感病毒NS片段中的重组流感病毒。尽管在流感病毒基因组中插入了一个相当大的HIV-1基因，但重组病毒仍能很容易地拯救到高滴度。通过体外感染试验证实了p24衣壳的细胞内表达。随后在BALB/c小鼠中测试了重组流感病毒作为粘膜疫苗的效果。重组病毒在免疫小鼠中减毒且安全。通过鼻内途径采用初免-加强方案免疫的小鼠引发了全身性和粘膜性HIV特异性CD8 T细胞应答。分离出的HIV特异性CD8 T细胞表现出多功能细胞因子和脱颗粒特征。通过阴道途径加强免疫的小鼠诱导了远端肺粘膜的回忆反应，并在阴道粘膜中产生了增强的HIV特异性CD8 T细胞应答。这些发现证明了重组流感病毒作为针对HIV-1感染的粘膜免疫疫苗的潜在效用。
