T helper 2 transcriptional profile predicts single-cell HIV envelope-specific polyfunctional CD4+ T cells correlated with reduced risk of infection in RV144 trial

Despite the critical role antigen-specific T cells play in responding to viral infections, their aggregate frequencies in peripheral blood have not correlated with clinical protection during HIV infection. However, a subset of HIV-specific CD4+ T cells, termed polyfunctional T cells, can produce multiple effector cytokines simultaneously. In the RV144 HIV vaccine trial, polyfunctional T cells correlated with reduced risk of HIV infection. Little is known about what differentiates polyfunctional T cells from other vaccine-elicited T cells. Therefore, we developed a novel live-cell multiplexed cytokine capture assay, to identify and transcriptionally profile vaccine-specific polyfunctional CD4+ T cells. We applied these methods to samples from the HVTN 097 clinical trial of the same vaccine regimen as RV144. We discovered two surface receptors that were enriched among polyfunctional CD4+ T cells and a Th2-biased signature (IL-4, IL-5, and IL-13) that specifically predicted the envelope-specific polyfunctional CD4+ T cells that were correlated with reduced risk of HIV infection in RV144. By linking single-cell transcriptional and functional profiles, we may be able to further define the role of vaccine-elicited polyfunctional T cells in contributing to effective immunity.

Download Full-text

A single-cell reference atlas delineates CD4+ T cell subtype-specific adaptation during acute and chronic viral infections

10.1101/2021.09.20.458613 ◽

2021 ◽

Author(s):

Massimo Andreatta ◽

Zachary Sherman ◽

Ariel Tjitropranoto ◽

Michael C Kelly ◽

Thomas Ciucci ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Immune Responses ◽

Single Cell ◽

Cd4 T Cells ◽

Viral Infections ◽

Cd4 T Cell ◽

Chronic Infections ◽

Chronic Viral Infections ◽

Memory States

CD4+ T cells are critical orchestrators of immune responses against a large variety of pathogens, including viruses. The multifaceted roles of CD4+ T cells, including their help to innate cells, CD8+ T and B cells and their support for long-lived immunity rely on a profound functional heterogeneity. While multiple CD4+ T cell subtypes and their key transcriptional regulators have been identified, there is a lack of consistent definition for CD4+ T cell transcriptional states. In addition, the progressive changes affecting CD4+ T cell subtypes during and after immune responses remain poorly defined. Taking advantage of single-cell transcriptomics, efficient computational methods, and robust animal models, we characterize the transcriptional landscape of CD4+ T cells responding to self-resolving and chronic viral infections. We build a comprehensive atlas of virus-specific CD4+ T cells and their evolution over time, and identify six major distinct cell states that are consistently observed in acute and chronic infections. During the course of acute infection, T cell composition progressively changes from effector to memory states, with subtype-specific gene modules and kinetics. Conversely, T cells in persistent infections fail to transition from effector to memory states, and acquire distinct, chronicity-associated transcriptional programs. By single-cell T cell receptor (TCR) sequencing analysis, we characterize the clonal structure of virus-specific CD4+ T cells across individuals and T cell subtypes. We find that virus-specific CD4+ T cell responses are mainly private across individuals and that most T cells differentiate into all subtypes independently of their TCR, in both acute and chronic infections. Finally, we show that our CD4+ T cell atlas can be used as a reference to accurately interpret cell states in external single-cell datasets. Overall, this study describes a previously unappreciated level of adaptation of the transcriptional states of CD4+ T cells responding to viruses and provides a new computational resource for CD4+ T cell analysis, available online at https://spica.unil.ch.

Download Full-text

Faculty Opinions recommendation of Transcription factor FOXO3a controls the persistence of memory CD4(+) T cells during HIV infection.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1104534.569097 ◽

2008 ◽

Author(s):

Vicente Planelles

Keyword(s):

T Cells ◽

Transcription Factor ◽

Hiv Infection ◽

Cd4 T Cells ◽

Memory Cd4 T Cells

Download Full-text

Faculty Opinions recommendation of Autocrine production of beta-chemokines protects CMV-Specific CD4 T cells from HIV infection.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1279973.747077 ◽

2009 ◽

Author(s):

Scott Sieg ◽

Nicholas Funderburg

Keyword(s):

T Cells ◽

Hiv Infection ◽

Cd4 T Cells

Download Full-text

Circulating integrin α 4 β 7 + CD4 T cells are enriched for proliferative transcriptional programs in HIV infection

FEBS Letters ◽

10.1002/1873-3468.14163 ◽

2021 ◽

Author(s):

Yashavanth S. Lakshmanappa ◽

Jamin W. Roh ◽

Niharika N. Rane ◽

Ashok R. Dinasarapu ◽

Daphne D. Tran ◽

...

Keyword(s):

T Cells ◽

Hiv Infection ◽

Cd4 T Cells

Download Full-text

Identification of novel genes involved in apoptosis of HIV-infected macrophages using unbiased genome-wide screening

BMC Infectious Diseases ◽

10.1186/s12879-021-06346-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Simon X. M. Dong ◽

Frederick S. Vizeacoumar ◽

Kalpana K. Bhanumathy ◽

Nezeka Alli ◽

Cristina Gonzalez-Lopez ◽

...

Keyword(s):

T Cells ◽

Hiv Infection ◽

Respiratory Chain ◽

Cd4 T Cells ◽

Chain Complex ◽

Respiratory Chain Complex ◽

Complex Ii ◽

Genome Wide ◽

Latently Infected ◽

Induced Apoptosis

Abstract Background Macrophages, besides resting latently infected CD4+ T cells, constitute the predominant stable, major non-T cell HIV reservoirs. Therefore, it is essential to eliminate both latently infected CD4+ T cells and tissue macrophages to completely eradicate HIV in patients. Until now, most of the research focus is directed towards eliminating latently infected CD4+ T cells. However, few approaches have been directed at killing of HIV-infected macrophages either in vitro or in vivo. HIV infection dysregulates the expression of many host genes essential for the survival of infected cells. We postulated that exploiting this alteration may yield novel targets for the selective killing of infected macrophages. Methods We applied a pooled shRNA-based genome-wide approach by employing a lentivirus-based library of shRNAs to screen novel gene targets whose inhibition should selectively induce apoptosis in HIV-infected macrophages. Primary human MDMs were infected with HIV-eGFP and HIV-HSA viruses. Infected MDMs were transfected with siRNAs specific for the promising genes followed by analysis of apoptosis by flow cytometry using labelled Annexin-V in HIV-infected, HIV-exposed but uninfected bystander MDMs and uninfected MDMs. The results were analyzed using student’s t-test from at least four independent experiments. Results We validated 28 top hits in two independent HIV infection models. This culminated in the identification of four target genes, Cox7a2, Znf484, Cstf2t, and Cdk2, whose loss-of-function induced apoptosis preferentially in HIV-infected macrophages. Silencing these single genes killed significantly higher number of HIV-HSA-infected MDMs compared to the HIV-HSA-exposed, uninfected bystander macrophages, indicating the specificity in the killing of HIV-infected macrophages. The mechanism governing Cox7a2-mediated apoptosis of HIV-infected macrophages revealed that targeting respiratory chain complex II and IV genes also selectively induced apoptosis of HIV-infected macrophages possibly through enhanced ROS production. Conclusions We have identified above-mentioned novel genes and specifically the respiratory chain complex II and IV genes whose silencing may cause selective elimination of HIV-infected macrophages and eventually the HIV-macrophage reservoirs. The results highlight the potential of the identified genes as targets for eliminating HIV-infected macrophages in physiological environment as part of an HIV cure strategy.

Download Full-text

Mucosal stromal fibroblasts markedly enhance HIV infection of CD4+ T cells

PLoS Pathogens ◽

10.1371/journal.ppat.1006163 ◽

2017 ◽

Vol 13 (2) ◽

pp. e1006163 ◽

Cited By ~ 32

Author(s):

Jason A. Neidleman ◽

Joseph C. Chen ◽

Nargis Kohgadai ◽

Janis A. Müller ◽

Anders Laustsen ◽

...

Keyword(s):

T Cells ◽

Hiv Infection ◽

Cd4 T Cells ◽

Stromal Fibroblasts

Download Full-text

The Class II Phosphatidylinositol 3 kinase C2β Is Required for the Activation of the K+ Channel KCa3.1 and CD4 T-Cells

Molecular Biology of the Cell ◽

10.1091/mbc.e09-05-0390 ◽

2009 ◽

Vol 20 (17) ◽

pp. 3783-3791 ◽

Cited By ~ 46

Author(s):

Shekhar Srivastava ◽

Lie Di ◽

Olga Zhdanova ◽

Zhai Li ◽

Santosha Vardhana ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Activation ◽

Cd4 T Cells ◽

Critical Role ◽

Class Ii ◽

K Channel ◽

Channel Activity ◽

Phosphatidylinositol 3 Kinase ◽

Phosphatidylinositol 3

The Ca2+-activated K+ channel KCa3.1 is required for Ca2+ influx and the subsequent activation of T-cells. We previously showed that nucleoside diphosphate kinase beta (NDPK-B), a mammalian histidine kinase, directly phosphorylates and activates KCa3.1 and is required for the activation of human CD4 T lymphocytes. We now show that the class II phosphatidylinositol 3 kinase C2β (PI3K-C2β) is activated by the T-cell receptor (TCR) and functions upstream of NDPK-B to activate KCa3.1 channel activity. Decreased expression of PI3K-C2β by siRNA in human CD4 T-cells resulted in inhibition of KCa3.1 channel activity. The inhibition was due to decreased phosphatidylinositol 3-phosphate [PI(3)P] because dialyzing PI3K-C2β siRNA-treated T-cells with PI(3)P rescued KCa3.1 channel activity. Moreover, overexpression of PI3K-C2β in KCa3.1-transfected Jurkat T-cells led to increased TCR-stimulated activation of KCa3.1 and Ca2+ influx, whereas silencing of PI3K-C2β inhibited both responses. Using total internal reflection fluorescence microscopy and planar lipid bilayers, we found that PI3K-C2β colocalized with Zap70 and the TCR in peripheral microclusters in the immunological synapse. This is the first demonstration that a class II PI3K plays a critical role in T-cell activation.

Download Full-text