Ability of HIV-1 Nef to downregulate CD4 and HLA class I differs among viral subtypes

Retrovirology

Zabrina L Brumme⁷  Mark A Brockman⁷  Thumbi Ndung’u¹  Jeff N Martin²  P Richard Harrigan⁷  Bruce D Walker⁶  Philip JR Goulder⁵  Peter W Hunt²  David R Bangsberg⁶  Conrad Muzoora⁴  Rosemary McCloskey⁷  Ryan Danroth⁸  Bemuluyigza Baraki⁸  Guinevere Q Lee⁷  Eric Martin⁷  Saleha Omarjee¹  Philip Mwimanzi⁸  Chanson J Brumme⁷  Anh Q Le⁸  Xiaomei T Kuang³  Helen Byakwaga²  Jaclyn K Mann¹ 

[1] KwaZulu-Natal Research Institute for Tuberculosis and HIV, University of KwaZulu-Natal, Durban, South Africa;University of California San Francisco, San Francisco, CA, USA;Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada;Mbarara University of Science and Technology, Mbarara, Uganda;Department of Paediatrics, University of Oxford, OX1 3SY, United Kingdom;Massachusetts General Hospital and Harvard University, Boston, MA, USA;British Columbia Centre for Excellence in HIV/AIDS, Vancouver, BC, Canada;Faculty of Health Sciences, Simon Fraser University, Burnaby, BC, Canada

关键词: HLA class I;    CD4;    Pathogenesis;    Viral diversity;    Nef;    HIV/AIDS;   

Others  :  807205
DOI  :  10.1186/1742-4690-10-100

 received in 2013-04-27, accepted in 2013-09-09,  发布年份 2013

【 摘 要 】

Background

The highly genetically diverse HIV-1 group M subtypes may differ in their biological properties. Nef is an important mediator of viral pathogenicity; however, to date, a comprehensive inter-subtype comparison of Nef in vitro function has not been undertaken. Here, we investigate two of Nef’s most well-characterized activities, CD4 and HLA class I downregulation, for clones obtained from 360 chronic patients infected with HIV-1 subtypes A, B, C or D.

Results

Single HIV-1 plasma RNA Nef clones were obtained from N=360 antiretroviral-naïve, chronically infected patients from Africa and North America: 96 (subtype A), 93 (B), 85 (C), and 86 (D). Nef clones were expressed by transfection in an immortalized CD4+ T-cell line. CD4 and HLA class I surface levels were assessed by flow cytometry. Nef expression was verified by Western blot. Subset analyses and multivariable linear regression were used to adjust for differences in age, sex and clinical parameters between cohorts. Consensus HIV-1 subtype B and C Nef sequences were synthesized and functionally assessed. Exploratory sequence analyses were performed to identify potential genotypic correlates of Nef function. Subtype B Nef clones displayed marginally greater CD4 downregulation activity (p = 0.03) and markedly greater HLA class I downregulation activity (p < 0.0001) than clones from other subtypes. Subtype C Nefs displayed the lowest in vitro functionality. Inter-subtype differences in HLA class I downregulation remained statistically significant after controlling for differences in age, sex, and clinical parameters (p < 0.0001). The synthesized consensus subtype B Nef showed higher activities compared to consensus C Nef, which was most pronounced in cells expressing lower protein levels. Nef clones exhibited substantial inter-subtype diversity: cohort consensus residues differed at 25% of codons, while a similar proportion of codons exhibited substantial inter-subtype differences in major variant frequency. These amino acids, along with others identified in intra-subtype analyses, represent candidates for mediating inter-subtype differences in Nef function.

Conclusions

Results support a functional hierarchy of subtype B > A/D > C for Nef-mediated CD4 and HLA class I downregulation. The mechanisms underlying these differences and their relevance to HIV-1 pathogenicity merit further investigation.

【 授权许可】

   
2013 Mann et al.; licensee BioMed Central Ltd.

【 图 表 】

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