Telomerase activity of HIV-1-specific CD8+ T cells: constitutive up-regulation in controllers and selective increase by blockade of PD ligand 1 in progressors - PubMed (original) (raw)

. 2008 Nov 1;112(9):3679-87.

doi: 10.1182/blood-2008-01-135442. Epub 2008 Aug 26.

Danlei Mou, Thai Duong Hong Cung, Katie L Williams, Michael T Waring, Jinghe Huang, Florencia Pereyra, Alicja Trocha, Gordon J Freeman, Eric S Rosenberg, Bruce D Walker, Xu G Yu

Affiliations

Telomerase activity of HIV-1-specific CD8+ T cells: constitutive up-regulation in controllers and selective increase by blockade of PD ligand 1 in progressors

Mathias Lichterfeld et al. Blood. 2008.

Abstract

Exhaustion of virus-specific T cells may play an important role in the pathophysiology of chronic viral infections. Here, we analyzed telomere length and telomerase activity in HIV-1-specific CD8+ T cells from progressors or controllers to determine underlying molecular pathways of T-cell exhaustion and senescence. Telomere lengths of HIV-1-specific CD8+ T cells from progressors were significantly shorter compared with autologous cytomegalovirus (CMV)/Epstein-Barr virus (EBV)-specific CD8+ T cells or bulk CD8+ T cells, while telomere lengths from controllers significantly exceeded those of autologous bulk CD8+ T cells and reached a similar level as HIV-1-specific CD8+ T cells collected during primary HIV-1 infection. Telomere length stabilization in controllers corresponded to high levels of constitutive telomerase activity, which was associated with preservation of cytotoxic and proliferative properties. Conversely, limited constitutive telomerase activity was observed in HIV-1-specific CD8+ T cells from progressors, although an increase in both telomere length and telomerase activity was achieved in antigenic-peptide-stimulated cells from progressors after blocking the PD-1/PD ligand 1 (PD-L1) pathway. Collectively, these data suggest a causal role of telomere shortening for the functional deficiencies of HIV-1-specific CD8+ T cells in chronic progressive infection, while high constitutive telomerase activities appears to contribute to maintenance of polyfunctional HIV-1-specific CD8+ T cells from HIV-1 controllers.

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Figures

Figure 1

Figure 1

Assessment of telomere length in HIV-1–specific, CMV/EBV-specific, and bulk CD8+ T cells from different patient populations with HIV-1. (A) Proportion of studied antigen-specific tetramer+ CD8+ T cells in total CD8+ T-cell populations from individuals with chronic progressive, chronic long-term nonprogressive, or acute HIV-1 infection, as determined by flow cytometry. Horizontal bars reflect medians. (B) Telomere length of HIV-1–specific, CMV/EBV-specific, and autologous bulk CD8+ T cells from the corresponding individuals. Horizontal bars reflect means.

Figure 2

Figure 2

Comparative analysis of telomerase activity in HIV-1–specific CD8+ T-cell populations from individuals with progressive or controlled HIV-1. (A) Relative telomerase activity (RTA) of HIV-1–specific CD8+ T cells, as well as corresponding CMV/EBV-specific and bulk CD8+ T cells, from HIV progressors or controllers measured by RQ-TRAP. Horizontal bars reflect medians. (B) Visualization of telomeric extension of substrate primers by RQ-TRAP following exposure to protein extractions collected from the indicated sorted HIV-1–specific CD8+ T-cell populations from progressors or controllers. Left panel to the ladder reflect samples from progressors; right panel reflect samples from controllers; the last 2 lanes reflect the heat-inactivated samples and lysis buffer only control. (C) Correlation between telomere length and telomerase activity of the analyzed HIV-1–specific CD8+ T cells.

Figure 3

Figure 3

Correlation between telomere length/telomerase activity of HIV-1–specific CD8+ T cells and their proliferative and cytotoxic activity. (A) Representative dot plots indicating gating of proliferating (CFSEdim) and nonproliferating (CFSEbright) tetramer+ HIV-1–specific CD8+ T cells. Numbers in quadrants reflect proportion of tetramer+ CD8+ T cells. (B) Comparison of telomere lengths between proliferating and corresponding nonproliferating tetramer+ HIV-1–specific CD8+ T cells. Horizontal bars reflect medians. (C) Representative dot plots reflecting caspase-3 expression in peptide-pulsed target cells following exposure to HIV-1–specific CD8+ T cells from individuals with progressive or controlled viremia. Numbers in quadrants reflect proportion of target cells. (D) Proportion of caspase-3–positive target cells after coincubation with PBMCs from HIV-1 controllers (n = 5) or HIV-1 progressors (n = 5). Horizontal bars reflect means. (E) Correlation between cytotoxic activity of HIV-1–specific CD8+ T cells (measured by proportion of caspase-3–positive target cells) and corresponding telomere length.

Figure 4

Figure 4

Augmentation of telomere length and telomerase activity of HIV-1–specific CD8+ T cells after antibody-mediated PD-L1 blockade. (A) Dot plots reflecting proliferative activity of HIV-1–specific CD8+ T cells in the presence or absence of PD-L1–blocking antibodies. CFSEdim proliferating HIV-1–specific CD8+ T cells after antigenic stimulation and bulk CFSEbright CD8+ T cells were sorted according to the gates indicated. Numbers in quadrants reflect proportion of gated lymphocytes. (B) Telomere length of HIV-1–specific CD8+ T cells proliferating in the presence or absence of PD-L1–blocking antibodies as well as of the corresponding nonproliferating bulk CD8+ T cells. (C,D) Telomerase activity of bulk CD8+ T cells and sorted HIV-1–specific CD8+ T cells that were stimulated with antigenic peptides in the presence or absence of PD-L1–blocking antibodies, as determined by RQ-TRAP with subsequent analysis by gel electrophoresis (panel C; vertical lines have been inserted to indicate repositioned gel lanes) and after normalization to a reference cell line as described in “Methods” (D). Horizontal bars in panels B and D reflect means.

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