Tim-3 expression on PD-1+ HCV-specific human CTLs is associated with viral persistence, and its blockade restores hepatocyte-directed in vitro cytotoxicity
Rachel H. McMahan, … , David R. Gretch, Hugo R. Rosen
Rachel H. McMahan, … , David R. Gretch, Hugo R. Rosen
Published November 15, 2010
Citation Information: J Clin Invest. 2010;120(12):4546-4557. https://doi.org/10.1172/JCI43127.
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Research Article Virology

Tim-3 expression on PD-1+ HCV-specific human CTLs is associated with viral persistence, and its blockade restores hepatocyte-directed in vitro cytotoxicity

Abstract

Having successfully developed mechanisms to evade immune clearance, hepatitis C virus (HCV) establishes persistent infection in approximately 75%–80% of patients. In these individuals, the function of HCV-specific CD8+ T cells is impaired by ligation of inhibitory receptors, the repertoire of which has expanded considerably in the past few years. We hypothesized that the coexpression of the negative regulatory receptors T cell immunoglobulin and mucin domain–containing molecule 3 (Tim-3) and programmed death 1 (PD-1) in HCV infection would identify patients at risk of developing viral persistence during and after acute HCV infection. The frequency of PD-1–Tim-3– HCV-specific CTLs greatly outnumbered PD-1+Tim-3+ CTLs in patients with acute resolving infection. Moreover, the population of PD-1+Tim-3+ T cells was enriched for within the central memory T cell subset and within the liver. Blockade of either PD-1 or Tim-3 enhanced in vitro proliferation of HCV-specific CTLs to a similar extent, whereas cytotoxicity against a hepatocyte cell line that expressed cognate HCV epitopes was increased exclusively by Tim-3 blockade. These results indicate that the coexpression of these inhibitory molecules tracks with defective T cell responses and that anatomical differences might account for lack of immune control of persistent pathogens, which suggests their manipulation may represent a rational target for novel immunotherapeutic approaches.

Authors

Rachel H. McMahan, Lucy Golden-Mason, Michael I. Nishimura, Brian J. McMahon, Michael Kemper, Todd M. Allen, David R. Gretch, Hugo R. Rosen

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Figure 4

PD-1hiTim-3hi CTLs are associated with diminished antiviral cytokine production and chronic infection.

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PD-1hiTim-3hi CTLs are associated with diminished antiviral cytokine pro...
(A) PD-1hiTim-3lo, PD-1loTim-3hi, PD-1loTim-3lo, and PD-1hiTim-3hi pentamer-positive T cells from acute→chronic, acute→resolved, long-term chronic, and remote resolved patients as well as intrahepatic T cells from long-term chronic (liver) patients. The differences between long-term chronic and remote resolved patients were highly statistically significant (see Results). (B and C) PBMCs from acute→chronic patients were stimulated with the HLA-A1–restricted NS31436 epitope or PMA and ionomycin for 5 hours in the presence of the CD107a antibody. Cell surface staining for CD3, CD8, HLA-A11436 pentamer, Tim-3, and PD-1 was carried out, followed by intracellular cytokine staining for IFN-γ and TNF-α. Flow cytometric analysis was used to determine the proportions of the PD-1hiTim-3hi, PD-1loTim-3lo, and PD-1–Tim-3– cells producing IFN-γ, TNF-α, and CD107a after stimulation. (B) Histograms from a representative patient with high pentamer frequency (2.2%). (C) Percent cells producing IFN-γ, TNF-α, and CD107a according to PD-1 and Tim-3 expression following stimulation with the NS31436 peptide or PMA and ionomycin. Pentamer frequencies for patients acute 4 and acute 6 are shown in Supplemental Figure 3.