Single Cell Profiling Reveals PTEN Overexpression in Influenza-Specific B cells in Aging HIV-infected individuals on Anti-retroviral Therapy

Lesley R De Armas,Sarah Andrews,Nicola Cotugno,Rajendra Pahwa,Li Pan,Savita Pahwa,Stefano Rinaldi,Paolo Palma,Adrian B Mcdermott,Suresh Pallikkuth

doi:10.1038/s41598-019-38906-y

Abstract

Memory B cells (MBC) respond to secondary antigen challenge to protect against infection and to boost immunity following vaccinations. Despite effective treatment, chronic HIV infection disturbs MBCs by reducing numbers and altering functionality due to hyper-activation and increased apoptosis leading to suboptimal antibody responses against common infectious agents. We used single cell gene expression analysis to evaluate antigen-specific memory B cells in peripheral blood of virally-suppressed HIV-infected individuals and healthy controls stratified by serum H1N1 antibody response 3 weeks post-administration of the seasonal trivalent inactivated influenza vaccine. We used a fluorescent probe to isolate influenza H1N1-specific B cells and a multiplexed and targeted RT-PCR approach to measure expression levels of 96 genes involved in B cell activation and function. Gene profiling revealed a 4-gene predictive signature containing the phosphoinositide-3 kinase (PI3K) inhibitor, PTEN, for identifying antigen-specific MBC from HIV-infected individuals compared to healthy controls. Gene co-expression analysis showed that in addition to overexpression of PTEN, there was increased co-expression of type I interferon-associated genes with PTEN on single cell level in HIV compared to controls. This study highlights the persistent defects in MBC from HIV-infected individuals and points to the PI3K signaling pathway as a target for potential immune intervention.

Highlights

Response Status Non-Responder Non-Responder Non-Responder Responder Responder Responder Non-Responder Non-Responder Non-Responder Responder Responder Responder reverted with ART initiation, Memory B cells (MBC) impairment remains[6] due to chronic immune activation attributed to persistence of HIV antigen in lymph nodes and other sanctuary sites[7,8,9,10]
We used a single cell, targeted multiplex gene expression platform and predictive modeling to show that following in vivo stimulation with the seasonal flu vaccine, influenza-specific MBC exhibit divergent gene signatures in HIV-infected, ART-suppressed individuals compared to age-matched healthy controls (HC)
Reduced memory B cell responses to influenza vaccination in HIV-infected individuals. 12 individuals were selected from a cohort of HIV-infected and healthy control adult volunteers participating in an influenza vaccination study (FLORAH cohort)[15] to evaluate gene profiles of ex vivo H1N1specific B cells (Table 1)

Summary

Introduction

Response Status Non-Responder Non-Responder Non-Responder Responder Responder Responder Non-Responder Non-Responder Non-Responder Responder Responder Responder reverted with ART initiation, MBC impairment remains[6] due to chronic immune activation attributed to persistence of HIV antigen in lymph nodes and other sanctuary sites[7,8,9,10]. We used a single cell, targeted multiplex gene expression platform and predictive modeling to show that following in vivo stimulation with the seasonal flu vaccine, influenza-specific MBC exhibit divergent gene signatures in HIV-infected, ART-suppressed individuals compared to age-matched healthy controls (HC).

Results

Conclusion