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Kennedy Epitope (KE)-dependent Retrograde Transport of Efficiently Cleaved HIV-1 Envelopes (Envs) and its Effect on Env Cell Surface Expression and Viral Particle Formation

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A Correction to this article was published on 26 February 2024

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Abstract

Efficiently cleaved HIV-1 Envs are the closest mimics of functional Envs as they specifically expose only bNAb (broadly neutralizing antibody) epitopes and not non-neutralizing ones, making them suitable for developing vaccine immunogens. We have previously identified several efficiently cleaved Envs from clades A, B, C and B/C. We also described that truncation of the CT (C-terminal tail) of a subset of these Envs, but not others, impairs their ectodomain conformation/antigenicity on the cell surface in a CT conserved hydrophilic domain (CHD) or Kennedy epitope (KE)-dependent manner. Here, we report that those Envs (4 − 2.J41 and JRCSF), whose native-like ectodomain conformation/antigenicity on the cell surface is disrupted upon CT truncation, but not other Envs like JRFL, whose CT truncation does not have an effect on ectodomain integrity on the cell surface, are also defective in retrograde transport from early to late endosomes. Restoration of the CHD/KE in the CT of these Envs restores wild-type levels of distribution between early and late endosomes. In the presence of retrograde transport inhibitor Retro 2, cell surface expression of 4 − 2.J41 and JRCSF Envs increases [as does in the presence of Rab7a DN and Rab7b DN (DN: dominant negative)] but particle formation decreases for 4 − 2.J41 and JRCSF Env pseudotyped viruses. Our results show for the first time a correlation between CT-dependent, CHD/KE regulated retrograde transport and cell surface expression/viral particle formation of these efficiently cleaved Envs. Based on our results we hypothesize that a subset of these efficiently cleaved Envs use a CT-dependent, CHD/KE-mediated mechanism for assembly and release from late endosomes.

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Acknowledgements

We thank Dr. Bimal Chakrabarti, ex-Program Director, THSTI-IAVI HIV Vaccine Design Program, Translational Health Science and Technology Institute, India for helpful discussions.

Funding

This work was supported primarily by IAVI intramural research, as well as by the Department of Biotechnology (DBT), Govt. of India intramural research program and its generous donors. IAVI’s work is made possible by generous support from many donors including: the Bill & Melinda Gates Foundation; the Ministry of Foreign Affairs of Denmark; Irish Aid; the Ministry of Finance of Japan; the Ministry of Foreign Affairs of the Netherlands; the Norwegian Agency for Development Cooperation (NORAD); the United Kingdom Department for International Development (DFID), and the United States Agency for International Development (USAID). The full list of IAVI donors is available at www.iavi.org. This study is made possible by the generous support of the American people through USAID. The contents are the responsibility of the International AIDS Vaccine Initiative and do not necessarily reflect the views of USAID or the United States Government.

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  1. Hilal Ahmad Parray and Adarsh Kumar Chiranjivi contributed equally.

Authors and Affiliations

  1. Infection and Immunology, Translational Research Program, 3rd Milestone, Faridabad-Gurgaon Expressway, PO box #04, Faridabad, 121001, Haryana, India

    Supratik Das, Hilal Ahmad Parray, Adarsh Kumar Chiranjivi, Prince Kumar, Abhishek Goswami, Deepak Kumar Rathore & Sweety Samal

  2. Translational Health Science and Technology Institute, NCR Biotech Science Cluster, 3rd Milestone, Faridabad-Gurgaon Expressway, PO box #04, Faridabad, 121001, Haryana, India

    Supratik Das & Manish Bansal

  3. Department of Biosciences and Bioengineering, Indian Institute of Technology – Roorkee, Roorkee, Uttarakhand, 247667, India

    Rajesh Kumar

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Contributions

S.D. designed, directed, carried out a majority of experiments and wrote the manuscript. H.A.P. carried out the p24 ELISA experiments. AKC carried out the western blot experiment for Fig. 6. P.K. carried out the western blot analysis of Fig. 2B. A.G. assisted with experiments. M.B. generated reagents. D.K.R. carried out analysis of FACS data. R.K. revised the manuscript. S.S. designed Fig. 7.All authors reviewed the manuscript.

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Correspondence to Supratik Das.

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Das, S., Parray, H.A., Chiranjivi, A.K. et al. Kennedy Epitope (KE)-dependent Retrograde Transport of Efficiently Cleaved HIV-1 Envelopes (Envs) and its Effect on Env Cell Surface Expression and Viral Particle Formation. Protein J (2023). https://doi.org/10.1007/s10930-023-10161-1

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