Impaired Upregulation of the Costimulatory Molecules, CD27 and CD28, on CD4+ T Cells from HIV Patients Receiving ART Is Associated with Poor Proliferative Responses.

HIV patients beginning antiretroviral therapy (ART) with advanced immunodeficiency often retain low CD4+ T cell counts despite virological control. We examined proliferative responses and upregulation of costimulatory molecules, following anti-CD3 stimulation, in HIV patients with persistent CD4+ T cell deficiency on ART. Aviremic HIV patients with nadir CD4+ T cell counts <100 cells/µL and who had received ART for a median time of 7 (range 1-11) years were categorized into those achieving low (<350 cells/µL; n = 13) or normal (>500 cells/µL; n = 20) CD4+ T cell counts. Ten healthy controls were also recruited. CD4+ T cell proliferation (Ki67) and upregulation of costimulatory molecules (CD27 and CD28) after anti-CD3 stimulation were assessed by flow cytometry. Results were related to proportions of CD4+ T cells expressing markers of T cell senescence (CD57), activation (HLA-DR), and apoptotic potential (Fas). Expression of CD27 and/or CD28 on uncultured CD4+ T cells was similar in patients with normal CD4+ T cell counts and healthy controls, but lower in patients with low CD4+ T cell counts. Proportions of CD4+ T cells expressing CD27 and/or CD28 correlated inversely with CD4+ T cell expression of CD57, HLA-DR, and Fas. After anti-CD3 stimulation, induction of CD27hiCD28hi expression was independent of CD4+ T cell counts, but lower in HIV patients than in healthy controls. Induction of CD27hiCD28hi expression correlated with induction of Ki67 expression in total, naïve, and CD31+ naïve CD4+ T cells from patients. In HIV patients responding to ART, impaired induction of CD27 and CD28 on CD4+ T cells after stimulation with anti-CD3 is associated with poor proliferative responses as well as greater CD4+ T cell activation and immunosenescence.

Short Communication: Few Liver-Infiltrating Cells Express CXCR3 in HIV/HCV Patients Commencing Antiretroviral Therapy.

Coinfections with Hepatitis C virus and human immunodeficiency virus accelerate the progression of both conditions and hamper effective treatment. Here we describe expression of CXCR3 on liver-infiltrating cells and peripheral T cells from coinfected patients commencing antiretroviral therapy (ART) in Indonesia. CXCR3 was expressed by small number of intrahepatic inflammatory cells, mostly in the portal areas. The number of cells did not change on ART and was markedly lower than the number of CD4+ and CD8+ cells in the liver. Data suggest that CXCR3 may contribute to liver infiltration but demonstrate a dynamic situation, changing as the immune system recovers on ART.

Naive and Memory CD4⁺ T Cells Are Differentially Affected in Indonesian HIV Patients Responding to ART.

While most HIV patients beginning antiretroviral therapy (ART) with advanced immunodeficiency recover CD4(+) T cell numbers, the profiles and functions of the newly acquired CD4(+) T cells have not been monitored in a resource-limiting setting. In this study, HIV patients (n = 31) from Jakarta, Indonesia, were studied 9 months after commencing ART with nadir CD4(+) T cell counts <200 cells/µL. All patients were hepatitis C virus (HCV) seropositive, but asymptomatic. Twelve healthy age-matched controls from the same community were included. CD4(+) T cell subsets, immune activation (HLA-DR), and expression of the interleukin (IL)-7 receptor α chain (CD127) were quantitated by flow cytometry. Proliferation (expression of Ki67) was measured following in vitro stimulation (5 days) with anti-CD3 antibody or IL-7. Fifty-two percent of patients recovered CD4(+) T cell counts >200 cells/µL over 12 months. At 9 months, patients had fewer naive and CD31(+)-naive CD4(+) T cells, more effector memory (EM) CD4(+) T cells, and higher HLA-DR expression on CD4(+) T cells than controls. CD127 expression was low on all CD4(+) T cell subsets except for naive cells, where it was similar to controls. Similarly, after anti-CD3 antibody or IL-7 stimulation, patients had lower Ki67 expression than controls in all subsets, except naive CD4(+) T cells where it was normal or elevated. Overall in the first year of ART, patients had fewer naive and more EM CD4(+) T cells. Ongoing immune activation and, antigen-driven stimulation and differentiation of naive T cells may reduce the naive T cell pool, while driving the maturation and accumulation of memory cells with proliferative defects.

Interleukin-7 signalling defects in naive CD4+ T cells of HIV patients with CD4+ T-cell deficiency on antiretroviral therapy are associated with T-cell activation and senescence.

OBJECTIVE: To examine the relationship of defects in interleukin (IL)-7-induced naive CD4 T-cell homeostasis with residual immune activation and CD4 T-cell senescence in HIV patients receiving antiretroviral therapy (ART) who exhibit persistent CD4 T-cell deficiency. DESIGN: IL-7 induced proliferation of, and IL-7 receptor signalling in, total and naive CD4 T cells of HIV patients who had low (<350 cells/µl) or normal (>500 cells/µl) CD4 T-cell counts on ART was examined and related to markers of CD4 T-cell activation and senescence and innate immune activation. METHODS: Total, naive (CD45RA CD27) and CD31 naive CD4 T cells from aviremic HIV patients (n=39) with nadir CD4 T-cell counts less than 100 cells/µl, who had received ART for a median time of 7 (range 1-11) years, were assessed for CD127 expression, proliferation (Ki67), signal transducer and activator of transcription 5 phosphorylation (pSTAT5) and CD127 modulation following IL-7 stimulation. Changes were related to proportions of CD4 T cells expressing HLA-DR or CD57 and plasma levels of sCD14, CXCL9 and CXCL10. RESULTS: Patients with CD4 T-cell deficiency exhibited lower expression of CD127 on total, naive and CD31 naive CD4 T cells. Downregulation of CD127 after culture with IL-7 correlated inversely with CD4 T-cell counts and directly with Ki67 expression. Induction of pSTAT5 in CD4 T-cell subsets was greater in patients with normal CD4 T-cell counts. CD127 expression correlated inversely with proportions of CD4CD57 T cells, and pSTAT5 induction correlated inversely with CD4 T-cell expression of HLA-DR and CD57. CONCLUSION: Defects of IL-7 signalling in HIV patients with persistent CD4 T-cell deficiency receiving ART are associated with CD4 T-cell activation and senescence.

CD4+ T-cell deficiency in HIV patients responding to antiretroviral therapy is associated with increased expression of interferon-stimulated genes in CD4+ T cells.

Most patients with human immunodeficiency virus (HIV) who remain CD4(+) T-cell deficient on antiretroviral therapy (ART) exhibit marked immune activation. As CD4(+) T-cell activation may be mediated by microbial translocation or interferon-alpha (IFN-α), we examined these factors in HIV patients with good or poor CD4(+) T-cell recovery on long-term ART. Messenger RNA levels for 3 interferon-stimulated genes were increased in CD4(+) T cells of patients with poor CD4(+) T-cell recovery, whereas levels in patients with good recovery did not differ from those in healthy controls. Poor CD4(+) T-cell recovery was also associated with CD4(+) T-cell expression of markers of activation, senescence, and apoptosis, and with increased serum levels of the lipopolysaccharide receptor and soluble CD14, but these were not significantly correlated with expression of the interferon-stimulated genes. Therefore, CD4(+) T-cell recovery may be adversely affected by the effects of IFN-α, which may be amenable to therapeutic intervention.

Thymic tissue is not evident on high-resolution computed tomography and [¹8F]fluoro-deoxy-glucose positron emission tomography scans of aviraemic HIV patients with poor recovery of CD4⁺ T cells.

Some previously immunodeficient HIV patients responding to antiretroviral therapy display poor recovery of CD4⁺ T cells. Evaluation of the contribution of thymic function requires sensitive detection and quantitation of metabolically active thymic tissue. We describe patients with low but detectable thymopoiesis assessed as circulating CD4⁺ naive T cells expressing CD31. High-resolution computed tomography and PET scans found no residual thymic tissue even though metabolic activity was demonstrable by PET in lymph nodes.

Vaccine-induced IgG2 anti-HIV p24 is associated with control of HIV in patients with a 'high-affinity' FcgammaRIIa genotype.

OBJECTIVES: We have previously shown that vaccination with a recombinant fowlpox virus carrying the genes for HIV Gag-Pol and interferon-gamma (IFN-gamma) was associated with partial control of HIV replication after antiretroviral therapy (ART) was ceased but not with increased anti-HIV T-cell responses. Because IFN-gamma enhances IgG2 production, and IgG2 antibodies to HIV antigens and the 'high-affinity' polymorphism of FcgammaRIIa (the major Fc receptor for IgG2) have been associated with a favourable outcome of HIV infection, we examined the association of IgG2 antibodies to HIV p24 and 'high-affinity' polymorphisms of FcgammaRIIa with control of HIV replication in these patients. METHODS: Plasma from weeks 0 (cessation of ART 1 week after the last vaccination), 9 and 20 was available from patients who had received the full construct vaccine, a partial construct (without IFN-gamma) or placebo. IgG2 and IgG1 anti-p24 and anti-gp41 were assayed and all patients were genotyped for the FcgammaRIIa 131 R/H polymorphism that affects IgG2 binding. RESULTS: At week 0, IgG2 anti-p24 was present in five of nine full construct patients but none of 14 partial construct or placebo patients and was associated with a smaller increase in plasma HIV RNA over 20 weeks. Patients with IgG2 anti-p24 and the 'high-affinity' polymorphism of FcgammaRIIa exhibited lower HIV replication than other patients at week 20. CONCLUSION: The role of IgG2 anti-HIV antibodies and FcgammaRIIa in the control of HIV replication should be investigated further. Inclusion of an IFN-gamma gene in DNA vaccine constructs might be a means of enhancing IgG2 antibody production.

CD31 (PECAM-1) is a marker of recent thymic emigrants among CD4+ T-cells, but not CD8+ T-cells or gammadelta T-cells, in HIV patients responding to ART.

Some severely immunodeficient HIV patients experience poor recovery of CD4(+) T-cell counts on antiretroviral therapy (ART). Evaluation of the function of thymopoiesis in T-cell production in individual patients requires a simple marker of T-cells that have recently emigrated from the thymus. Here, we address whether expression of CD31 on CD4(+) T-cells, CD8(+) T-cells, regulatory T-cells and gammadelta T-cells correlates with other indicators of thymus function. Adult HIV-1 patients (n=27) with nadir CD4(+) T-cell counts <100 per mul and a sustained virological response to ART and healthy controls (n=23) were studied. CD31 expression was assessed by flow cytometry, T-cell receptor excision circles content by real-time PCR and thymic volume by spiral computed tomography. Proportions of CD4(+) T-cells expressing CD45RA and CD31 declined with age in HIV patients (P=0.03) and healthy controls (P<0.0001), and correlated directly with other markers of thymus function. In controls, proportions of CD8(+) T-cells expressing CD45RA and CD31 declined with age (P=0.003) and correlated directly with some markers of thymus function, but this was not seen in HIV patients. Proportions of CD45RA(+) CD31(+) gammadelta T-cells were higher in patients than controls (P=0.007) and did not correlate with thymus volume. In controls, proportion of gammadelta T-cells co-expressing CD45RA and CD31 increased with age (P=0.002). These data support the use of CD31 as a marker of recent thymic origin in CD4(+) T-cells, but not CD8(+) T-cells in HIV patients receiving ART. In such patients, CD31 expression is unlikely to indicate thymic origin in gammadelta T-cells.