Evaluation of Cellular Phenotypes Implicated in Immunopathogenesis and Monitoring Immune Reconstitution Inflammatory Syndrome in HIV/Leprosy Cases

BACKGROUND: It is now evident that HAART-associated immunological improvement often leads to a variety of new clinical manifestations, collectively termed immune reconstitution inflammatory syndrome, or IRIS. This phenomenon has already been described in cases of HIV coinfection with Mycobacterium leprae, most of them belonging to the tuberculoid spectrum of leprosy disease, as observed in leprosy reversal reaction (RR). However, the events related to the pathogenesis of this association need to be clarified. This study investigated the immunological profile of HIV/leprosy patients, with special attention to the cellular activation status, to better understand the mechanisms related to IRIS/RR immunopathogenesis, identifying any potential biomarkers for IRIS/RR intercurrence. METHODS/PRINCIPAL FINDINGS: Eighty-five individuals were assessed in this study: HIV/leprosy and HIV-monoinfected patients, grouped according to HIV-viral load levels, leprosy patients without HIV coinfection, and healthy controls. Phenotypes were evaluated by flow cytometry for T cell subsets and immune differentiation/activation markers. As expected, absolute counts of the CD4+ and CD8+ T cells from the HIV-infected individuals changed in relation to those of the leprosy patients and controls. However, there were no significant differences among the groups, whether in the expression of cellular differentiation phenotypes or cellular activation, as reflected by the expression of CD38 and HLA-DR. Six HIV/leprosy patients identified as IRIS/RR were analyzed during IRIS/RR episodes and after prednisone treatment. These patients presented high cellular activation levels regarding the expression of CD38 in CD8+ cells T during IRIS/RR (median: 77,15%), dropping significantly (p<0,05) during post-IRIS/RR moments (median: 29,7%). Furthermore, an increase of cellular activation seems to occur prior to IRIS/RR. CONCLUSION/SIGNIFICANCE: These data suggest CD38 expression in CD8+ T cells interesting tool identifying HIV/leprosy individuals at risk for IRIS/RR. So, a comparative investigation to leprosy patients at RR should be conducted

Flow Cytometry Evaluation of the T-Cell Receptor Vβ Repertoire Among HIV-1 Infected Individuals Before and After Antiretroviral Therapy

Submitted by Sandra Infurna ([email protected]) on 2019-12-12T10:59:01Z No. of bitstreams: 1 MarizaG_Morgado_etal_IOC_2005.pdf: 156927 bytes, checksum: 768faed9fa4a8f3bb039ed71b2bcfc26 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2019-12-12T11:09:45Z (GMT) No. of bitstreams: 1 MarizaG_Morgado_etal_IOC_2005.pdf: 156927 bytes, checksum: 768faed9fa4a8f3bb039ed71b2bcfc26 (MD5)Made available in DSpace on 2019-12-12T11:09:45Z (GMT). No. of bitstreams: 1 MarizaG_Morgado_etal_IOC_2005.pdf: 156927 bytes, checksum: 768faed9fa4a8f3bb039ed71b2bcfc26 (MD5) Previous issue date: 2005Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Departamento de Micro-Imuno-Parasitologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Departamento de Doenças Infecciosas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.HIV-1 infection leads to serious impairment of the immune system and perturbations in the T cell receptorVβ repertoire are also described. Immune reconstitution can be potentially achieved in response to HAART. In the present study 10 patients were investigated for the Vβ pattern expression before and after six months of HAART. TCR were analyzed for T CD4+ and CD8+ subsets, separately, by flow cytometry, using a monoclonal antibody set of 24 different Vβ chains. Compared to eight Brazilian healthy controls, no differences in Vβ pattern of expression was observed for patients before or on antiretroviral therapy. Some chains such as Vβ 3, 14, 16, 20 and 21.3 were over utilized by both T subsets, independently of HIV infection and/or antiretroviral treatment, differing from the ones described for individuals of other nationalities. However, when each patient was taken individually, particular alterations were detected for the Vβ gene usage, compared to controls, for all individuals. After treatment, significantVβ usage changes were observed for seven patients. One or more chains on both T subsets were engaged in this process, defining a preferential oligoclonal profile for TCR repertoire distribution, after HAART. Although no pattern of specific Vβ changes was detected in the circulating T cells, we cannot exclude that differential immune responses to HIV or other important antigens are being focused by these cells

The percentage of HLA-DR+ in CD3+ T lymphocytes (<i>upper panel</i>) and the percentage of CD38+ in CD8+ T lymphocytes (<i>lower panel</i>) in samples from HIV/leprosy patients from group 1 (VL80 copies/ml patients; leprosy-infected individuals; and healthy controls.

<p>Bars represent median values. * <i>p</i><0.05 – Mann-Whitney <i>U</i> test for each group compared with HIV/leprosy patients from group 1; † <i>p</i><0.05 – Mann-Whitney <i>U</i> test for each group compared with HIV/leprosy patients from group 2; ‡ <i>p</i><0.05 – Mann-Whitney <i>U</i> test for each group compared with HIV/leprosy patients from group 3. VL: HIV viral load. LD: Limit of detection.</p

Risk factors for increased immune reconstitution in response to Mycobacterium tuberculosis antigens in tuberculosis HIV-infected, antiretroviral-naïve patients

Abstract Background Little is known regarding the restoration of the specific immune response after combined antiretroviral therapy (cART) and anti-tuberculosis (TB) therapy introduction among TB-HIV patients. In this study, we examined the immune response of TB-HIV patients to Mycobacterium tuberculosis (Mtb) antigens to evaluate the response dynamics to different antigens over time. Moreover, we also evaluated the influence of two different doses of efavirenz and the factors associated with immune reconstitution. Methods This is a longitudinal study nested in a clinical trial, where cART was initiated during the baseline visit (D0), which occurred 30 ± 10 days after the introduction of anti-TB therapy. Follow-up visits were performed at 30, 60, 90 and 180 days after cART initiation. The production of IFN-γ upon in vitro stimulation with Mtb antigens purified protein derivative (PPD), ESAT-6 and 38 kDa/CFP-10 using ELISpot was examined at baseline and follow-up visits. Results Sixty-one patients, all ART-naïve, were selected and included in the immune reconstitution analysis; seven (11.5%) developed Immune Reconstitution Inflammatory Syndrome (IRIS). The Mtb specific immune response was higher for the PPD antigen followed by 38 kDa/CFP-10 and increased in the first 60 days after cART initiation. In multivariate analysis, the variables independently associated with increased IFN-γ production in response to PPD antigen were CD4+ T cell counts <200 cells/mm3 at baseline, age, site of tuberculosis, 800 mg efavirenz dose and follow-up CD4+ T cell counts. Moreover, the factors associated with the production of IFN-γ in response to 38 kDa/CFP-10 were detectable HIV viral load (VL) and CD4+ T cell counts at follow-up visits of ≥200 cells/mm3. Conclusions These findings highlight the differences in immune response according to the specificity of the Mtb antigen, which contributes to a better understanding of TB-HIV immunopathogenesis. IFN-γ production elicited by PPD and 38 kDa/CFP-10 antigens have a greater magnitude compared to ESAT-6 and are associated with different factors. The low response to ESAT-6, even during immune restoration, suggests that this antigen is not adequate to assess the immune response of immunosuppressed TB-HIV patients

Median and interquartile range (IQR) of T cell immunophenotyping percentage values or absolute counts obtained from HIV/leprosy individuals according to viral load (VL) values and from HIV-monoinfected, leprosy patients and healthy controls.

<p>*<i>p</i><0,05 – Mann-Whitney <i>U</i> test: groups <i>versus</i> Group 1 HIV/leprosy individuals (VL†</p><p><i>p</i><0,05 – Mann-Whitney <i>U</i> test: groups <i>versus</i> Group 2 HIV/leprosy individuals (VL≥80<10,000 copies/ml);</p>‡<p><i>p</i><0,05 – Mann-Whitney <i>U</i> test: groups <i>versus</i> Group 3 HIV/leprosy individuals (VL≥10,000 copies/ml);</p><p>LD: limit of detection.</p

Impact of HIV-Related Immune Impairment of Yellow Fever Vaccine Immunogenicity in People Living with HIV—ANRS 12403

The yellow fever (YF) vaccine is one of the safest and most effective vaccines currently available. Still, its administration in people living with HIV (PLWH) is limited due to safety concerns and a lack of consensus regarding decreased immunogenicity and long-lasting protection for this population. The mechanisms associated with impaired YF vaccine immunogenicity in PLWH are not fully understood, but the general immune deregulation during HIV infection may play an important role. To assess if HIV infection impacts YF vaccine immunogenicity and if markers of immune deregulation could predict lower immunogenicity, we evaluated the association of YF neutralization antibody (NAb) titers with the pre-vaccination frequency of activated and exhausted T cells, levels of pro-inflammatory cytokines, and frequency of T cells, B cells, and monocyte subsets in PLWH and HIV-negative controls. We observed impaired YF vaccine immunogenicity in PLWH with lower titers of YF-NAbs 30 days after vaccination, mainly in individuals with CD4 count 3. At the baseline, those individuals were characterized by having a higher frequency of activated and exhausted T cells and tissue-like memory B cells. Elevated levels of those markers were also observed in individuals with CD4 count between 500 and 350 cells/mm3. We observed a negative correlation between the pre-vaccination level of CD8+ T cell exhaustion and CD4+ T cell activation with YF-NAb titers at D365 and the pre-vaccination level of IP-10 with YF-NAb titers at D30 and D365. Our results emphasize the impact of immune activation, exhaustion, and inflammation in YF vaccine immunogenicity in PLWH

Changes in the NK cell repertoire related to initiation of TB treatment and onset of immune reconstitution inflammatory syndrome in TB/HIV co-infected patients in Rio de Janeiro, Brazil-ANRS 12274

Submitted by Janaína Nascimento ([email protected]) on 2019-12-13T14:36:39Z No. of bitstreams: 1 ve_Giacoia-Gripp_Carmem_etal_INI_2019.pdf: 6220733 bytes, checksum: f410b79bb31706b3a1ef853889ee7b8a (MD5)Approved for entry into archive by Janaína Nascimento ([email protected]) on 2019-12-13T14:59:57Z (GMT) No. of bitstreams: 1 ve_Giacoia-Gripp_Carmem_etal_INI_2019.pdf: 6220733 bytes, checksum: f410b79bb31706b3a1ef853889ee7b8a (MD5)Made available in DSpace on 2019-12-13T14:59:57Z (GMT). No. of bitstreams: 1 ve_Giacoia-Gripp_Carmem_etal_INI_2019.pdf: 6220733 bytes, checksum: f410b79bb31706b3a1ef853889ee7b8a (MD5) Previous issue date: 2019Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Aids e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Aids e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Aids e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Micobacterioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Micobacterioses. Rio de Janeiro, RJ, Brasil.Nova Iguaçu General Hospital. HIV Clinical Research Center. Rio de Janeiro, RJ, Brazil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa em Imunização e Vigilância em Saúde. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Plataforma de Pesquisa Clínica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Micobacterioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Aids e Imunologia Molecular. Rio de Janeiro, RJ, Brasil / Nova Iguaçu General Hospital. HIV Clinical Research Center. Rio de Janeiro, RJ, Brazil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Micobacterioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Aids e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Pasteur Institute. Unit of Lymphocyte Cell Biology. Paris, France.Tuberculosis (TB) is the most common comorbidity and the leading cause of death among HIV-infected individuals. Although the combined antiretroviral therapy (cART) during TB treatment improves the survival of TB/HIV patients, the occurrence of immune reconstitution inflammatory syndrome (IRIS) in some patients poses clinical and scientific challenges. This work aimed to evaluate blood innate lymphocytes during therapeutic intervention for both diseases and their implications for the onset of IRIS. Natural killer (NK) cells, invariant NKT cells (iNKT), γδ T cell subsets, and in vitro NK functional activity were characterized by multiparametric flow cytometry in the following groups: 33 TB/HIV patients (four with paradoxical IRIS), 27 TB and 25 HIV mono-infected subjects (prior to initiation of TB treatment and/or cART and during clinical follow-up to 24 weeks), and 25 healthy controls (HC). Concerning the NK cell repertoire, several activation and inhibitory receptors were skewed in the TB/HIV patients compared to those in the other groups, especially the HCs. Significantly higher expression of CD158a (p = 0.025), NKp80 (p = 0.033), and NKG2C (p = 0.0076) receptors was detected in the TB/HIV IRIS patients than in the non-IRIS patients. Although more NK degranulation was observed in the TB/HIV patients than in the other groups, the therapeutic intervention did not alter the frequency during follow-up (weeks 2-24). A higher frequency of the γδ T cell population was observed in the TB/HIV patients with inversion of the Vδ2+/Vδ2- ratio, especially for those presenting pulmonary TB, suggesting an expansion of particular γδ T subsets during TB/HIV co-infection. In conclusion, HIV infection impacts the frequency of circulating NK cells and γδ T cell subsets in TB/HIV patients. Important modifications of the NK cell repertoire were observed after anti-TB treatment (week 2) but not during the cART/TB follow-up (weeks 6-24). An increase of CD161+ NK cells was related to an unfavorable outcome. Despite the low number of cases, a more preserved NK cell profile was detected in IRIS patients previous to treatment, suggesting a role for these cells in IRIS onset. Longitudinal evaluation of the NK repertoire showed the impact of TB treatment and implicated these cells in TB pathogenesis in TB/HIV co-infected patients