Sneha Priya Rangasamy, Veena Menon, Kalisz Irene, Whitney Stephen, Priyanka Dhopeshwarkar, LaBranche Celia C, Sri Teja Mullapudi, Montefiori David C, Pal Ranajit, Mahalingam Sundarasamy, Kalyanaraman Vaniambadi S
From the Laboratory of Molecular Virology and Cell Biology, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India.
Advanced Bioscience Laboratories Inc., Rockville, Maryland 20850, and.
J Biol Chem. 2015 Apr 3;290(14):9195-208. doi: 10.1074/jbc.M114.621185. Epub 2015 Feb 17.
Human immunodeficiency virus type 1 (HIV-1) isolates from India mainly belong to clade C and are quite distinct from clade C isolates from Africa in terms of their phylogenetic makeup, serotype, and sensitivity to known human broadly neutralizing monoclonal antibodies. Because many of these properties are associated with the envelope proteins of HIV-1, it is of interest to study the envelope proteins of Indian clade C isolates as part of the ongoing efforts to develop a vaccine against HIV-1. To this end, we purified trimeric uncleaved gp145 of a CCR5 tropic Indian clade C HIV-1 (93IN101) from the conditioned medium of 293 cells. The purified protein was shown to be properly folded with stable structure by circular dichroism. Conformational integrity was further demonstrated by its high affinity binding to soluble CD4, CD4 binding site antibodies such as b12 and VRC01, quaternary epitope-specific antibody PG9, and CD4-induced epitope-specific antibody 17b. Sera from rabbits immunized with gp145 elicited high titer antibodies to various domains of gp120 and neutralized a broad spectrum of clade B and clade C HIV-1 isolates. Similar to other clade B and clade C envelope immunogens, most of the Tier 1 neutralizing activity could be absorbed with the V3-specific peptide. Subsequent boosting of these rabbits with a clade B HIV-1 Bal gp145 resulted in an expanded breadth of neutralization of HIV-1 isolates. The present study strongly supports the inclusion of envelopes from Indian isolates in a future mixture of HIV-1 vaccines.
来自印度的1型人类免疫缺陷病毒(HIV-1)分离株主要属于C亚型,在系统发育组成、血清型以及对已知人类广泛中和单克隆抗体的敏感性方面,与来自非洲的C亚型分离株有很大不同。由于这些特性中的许多都与HIV-1的包膜蛋白相关,因此作为开发抗HIV-1疫苗的持续努力的一部分,研究印度C亚型分离株的包膜蛋白很有意义。为此,我们从293细胞的条件培养基中纯化了一株CCR5嗜性的印度C亚型HIV-1(93IN101)的三聚体未切割gp145。通过圆二色性显示纯化的蛋白折叠正确且结构稳定。其与可溶性CD4、CD4结合位点抗体(如b12和VRC01)、四级表位特异性抗体PG9以及CD4诱导表位特异性抗体17b的高亲和力结合进一步证明了构象完整性。用gp145免疫的兔血清产生了针对gp120各个结构域的高滴度抗体,并中和了广泛的B亚型和C亚型HIV-1分离株。与其他B亚型和C亚型包膜免疫原类似,大多数1级中和活性可被V3特异性肽吸收。随后用B亚型HIV-1 Bal gp145对这些兔子进行加强免疫,导致HIV-1分离株的中和广度扩大。本研究有力支持在未来的HIV-1疫苗混合物中纳入来自印度分离株的包膜。