Longitudinal Changes in Cd4+, Cd8+ T Cell Phenotype and Activation Marker Expression Following Antiretroviral Therapy Initiation among Patients with Cryptococcal Meningitis

Alice Bayiyana, Samuel Okurut, Rose Nabatanzi, Godfrey Zziwa, David R Boulware, Fredrick Lutwama, David Meya, Alice Bayiyana, Samuel Okurut, Rose Nabatanzi, Godfrey Zziwa, David R Boulware, Fredrick Lutwama, David Meya

Abstract

Despite improvement in the prognosis of HIV/AIDS (human immunodeficiency virus/acquired immune deficiency syndrome) patients on antiretroviral therapy (ART), cryptococcal meningitis (CM) still causes 10-15% mortality among HIV-infected patients. The immunological impact of ART on the CD4+ and CD8+ T cell repertoire during cryptococcal co-infection is unclear. We determined longitudinal phenotypic changes in T cell subsets among patients with CM after they initiated ART. We hypothesized that ART alters the clonotypic phenotype and structural composition of CD4+ and CD8+ T cells during CM co-infection. For this substudy, peripheral blood mononuclear cells (PBMC) were isolated at four time points from CM patients following ART initiation during the parent study (ClinicalTrials.gov number, NCT01075152). Phenotypic characterization of CD4+ and CD8+ T cells was done using T cell surface marker monoclonal antibodies by flow cytometry. There was variation in the expression of immunophenotypic markers defining central memory (CD27+CD45R0+), effector memory (CD45R0+CD27-), immune activation (CD38+ and Human Leucocyte Antigen DR (HLA-DR+), and exhaustion (Programmed cell death protein one (PD-1) in the CD4+ T cell subset. In comparison to the CD4+ T cell population, the CD8+ central memory subset declined gradually with minimal increase in the effector memory subset. Both CD4+ and CD8+ T cell immune exhaustion and activation markers remained elevated over 12 weeks. The relative surge and decline in the expression of T cell surface markers outlines a variation in the differentiation of CD4+ T cells during ART treatment during CM co-infection.

Keywords: ART; HIV; T cell phenotypes; cryptococcal meningitis.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Gating strategy: Flow cytometry analysis of cryptococcal-specific CD4+ and CD8+ T cell memory and phenotypes evaluated in a 6 h assay: (A) We first used a singlet gate to exclude doublets by gating on forward scatter-area (FSC-A) against forward scatter-height (FSC-H). (B) Lymphocytes were then selected using a forward scatter-area (FSC-A) against side scatter-area (SSC-A) gate. (C) Conventional T cells were selected by gating on CD3+ cells from the total lymphocyte population, which were further divided to (D) CD4+ and CD8+ T cells. Memory phenotypes were assessed by variable expression of CD45R0 and CD27 on the CD4+ (E) and CD8+ (I) T cell populations; while T cell activation was assessed by expression of CD38 and HLA-DR on the CD4+ (G) and CD8+ (H) T cell subsets. T cell exhaustion was assessed by expression of PD-1 (F) on both populations. CM represents cryptococcal meningitis. QuaradntDEM.
Figure 2
Figure 2
Recovery of CD4+ and CD8+ T cell populations.
Figure 3
Figure 3
The CD4+ TCM subset frequency remained stable yet the CD8+ TCM subset declined.
Figure 4
Figure 4
CD4+ TEM subset expression shows significant decline contrary to the CD8+ TEM subsets.
Figure 5
Figure 5
Immune activation levels remained elevated with of the T cell subsets.
Figure 6
Figure 6
Immune exhaustion levels remained elevated for CD4+ and CD8+ T cell subpopulations.

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