Impact of multi-targeted antiretroviral treatment on gut T cell depletion and HIV reservoir seeding during acute HIV infection.

BackgroundLimited knowledge exists on early HIV events that may inform preventive and therapeutic strategies. This study aims to characterize the earliest immunologic and virologic HIV events following infection and investigates the usage of a novel therapeutic strategy.Methods and findingsWe prospectively screened 24,430 subjects in Bangkok and identified 40 AHI individuals. Thirty Thais were enrolled (8 Fiebig I, 5 Fiebig II, 15 Fiebig III, 2 Fiebig IV) of whom 15 completed 24 weeks of megaHAART (tenofovir/emtricitabine/efavirenz/raltegravir/maraviroc). Sigmoid biopsies were completed in 24/30 at baseline and 13/15 at week 24. At baseline, the median age was 29 years and 83% were MSM. Most were symptomatic (87%), and were infected with R5-tropic (77%) CRF01_AE (70%). Median CD4 was 406 cells/mm(3). HIV RNA was 5.5 log(10) copies/ml. Median total blood HIV DNA was higher in Fiebig III (550 copy/10(6) PBMC) vs. Fiebig I (8 copy/10(6) PBMC) (p = 0.01) while the median %CD4+CCR5+ gut T cells was lower in Fiebig III (19%) vs. Fiebig I (59%) (p = 0.0008). After 24 weeks of megaHAART, HIV RNA levels of <50 copies were achieved in 14/15 in blood and 13/13 in gut. Total blood HIV DNA at week 0 predicted reservoir size at week 24 (p<0.001). Total HIV DNA declined significantly and was undetectable in 3 of 15 in blood and 3 of 7 in gut. Frequency of CD4+CCR5+ gut T cells increased from 41% at baseline to 64% at week 24 (p>0.050); subjects with less than 40% at baseline had a significant increase in CD4+CCR5+ T cells from baseline to week 24 (14% vs. 71%, p = 0.02).ConclusionsGut T cell depletion and HIV reservoir seeding increases with progression of AHI. MegaHAART was associated with immune restoration and reduced reservoir size. Our findings could inform research on strategies to achieve HIV drug-free remission

Switch to dolutegravir is well tolerated in Thais with HIV infection

INTRODUCTION: Dolutegravir (DTG) is recommended as part of first-line antiretroviral therapy (ART) for people living with HIV(PLHIV). We sought to determine the rate of adverse events (AEs) and discontinuations among Thais treated during acute HIV infection (AHI) and switched to DTG-based regimens. METHODS: Thai participants in the SEARCH010/RV254 cohort who initiated ART during AHI and switched to DTG for at least 48 weeks were prospectively observed and included in the analysis. Rates and characteristics of DTG-related AEs and discontinuations were described. RESULTS: A total of 313 Thai participants were included in the analysis. The median age was 29 years, 96% were male, 64% had a Bachelor's degree or higher and 16% had a body mass index (BMI) <18.5 kg/m2 . Participants were on ART for a median of 124 weeks before switching to DTG. The median (IQR) body weight increased from 63 (56 to 70) kg before to 65 (58 to 73) kg (p < 0.0001) after 48 weeks of DTG. Forty-nine (16%) developed DTG-related AEs, corresponding to an incidence of 16.6 per 100 person-years. Neuropsychiatric symptoms were most frequently encountered (n = 25, 8%), followed by laboratory abnormalities (n = 16, 5%). Six (2%) discontinued DTG, corresponding to an incidence of 2.4 per 100 person-years. All discontinuations were due to increased liver enzymes in the presence of hepatitis C virus coinfection. In the multivariate analysis, incident hepatitis C virus infection was the only risk factor for discontinuing DTG (hazard ratio 59.4, 95% CI 8.5 to 297.9, p < 0.0001). Neither low BMI nor concurrent abacavir therapy was associated with discontinuation. CONCLUSIONS: DTG was well tolerated with few discontinuations in this cohort of young men. Incident hepatitis C virus infection was a driver of liver-related AEs leading to discontinuations. In populations at risk, regular testing for hepatitis C virus during ART is recommended to anticipate possible AEs, guide management and improve safety

Altered Memory Circulating T Follicular Helper-B Cell Interaction in Early Acute HIV Infection

<div><p>The RV254 cohort of HIV-infected very early acute (4thG stage 1 and 2) (stage 1/2) and late acute (4thG stage 3) (stage 3) individuals was used to study T helper- B cell responses in acute HIV infection and the impact of early antiretroviral treatment (ART) on T and B cell function. To investigate this, the function of circulating T follicular helper cells (cTfh) from this cohort was examined, and cTfh and memory B cell populations were phenotyped. Impaired cTfh cell function was observed in individuals treated in stage 3 when compared to stage 1/2. The cTfh/B cell cocultures showed lower B cell survival and IgG secretion at stage 3 compared to stage 1/2. This coincided with lower IL-10 and increased RANTES and TNF-α suggesting a role for inflammation in altering cTfh and B cell responses. Elevated plasma viral load in stage 3 was found to correlate with decreased cTfh-mediated B cell IgG production indicating a role for increased viremia in cTfh impairment and dysfunctional humoral response. Phenotypic perturbations were also evident in the mature B cell compartment, most notably a decrease in resting memory B cells in stage 3 compared to stage 1/2, coinciding with higher viremia. Our coculture assay also suggested that intrinsic memory B cell defects could contribute to the impaired response despite at a lower level. Overall, cTfh-mediated B cell responses are significantly altered in stage 3 compared to stage 1/2, coinciding with increased inflammation and a reduction in memory B cells. These data suggest that early ART for acutely HIV infected individuals could prevent immune dysregulation while preserving cTfh function and B cell memory.</p></div

Impact of early acute HIV infection on blood memory B cell subsets.

<p>PBMCs from week 0 stage 1/2 (n = 17) and stage 3 (n = 16) patients were analyzed by flow cytometry for resting memory (RM) B cells CD19⁺CD10^-CD21⁺CD27⁺ and (A) compared with each other and (B) with plasma viral load. (C) Comparisons were also done with PBMCs from week 72 of ART (W72) (n = 19 and n = 18 respectively), chronic HIV infected patients (n = 14) and HIV negative patients (n = 14). PBMCs from week 0 stage 1/2 (n = 17) and stage 3 (n = 16) patients were analyzed by flow cytometry for (D) Total B cells which were identified as CD19⁺; (E) resting naïve (RN) B cells CD19⁺CD10^-CD21⁺CD27^-IgD⁺ (F) activated memory (AM) B cells CD19⁺CD10^-CD21^-CD27⁺ cells, (G) Tissue-like (TL) B cells CD19⁺CD10^-CD21^-CD27^-. For (A) and (E)-(H), bars represent mean ±SD. Symbols on the graphs represent stage 1/2 individuals (black circles) and stage 3 individuals (black squares). Statistics were carried out using the Mann-Whitney non-parametric test. For (B) symbols represent stage 1/2 individuals (black circles) and stage 3 individuals (black squares) and correlation data was analyzed using Spearman r test. * P < 0.05. For (C), symbols represent individuals from week 0 stage 1/2 (black circles), week 0 stage 3 (black squares), week 72 ART stage 1/2 (upward pointing black triangles), week 72 ART stage 3 (downward pointing black triangles), chronic individuals (black diamonds) and HIV negative (open circles). Statistics were carried out using Kruskal-Wallis multiple comparison test. * P< 0.05.</p

cTfh-mediated memory B cell help is impaired in late stage acute HIV-infected individuals.

<p>PBMCs from week 0 stage 1/2 (n = 9) and stage 3 (n = 4–7) individuals were sorted. cTfh (CD3⁺CD4⁺CD45RA^-CXCR5⁺CXCR3^-) or the less efficient T-helper cells (CD3⁺CD4⁺CD45RA^-CXCR5⁺CXCR3⁺) denoted CXCR5⁺CXCR3⁺ were placed in culture with autologous resting memory B cells (CD19⁺CD10^-CD21⁺CD27⁺) in the presence of SEB. Quantification of cTfh-mediated B cell help was carried out by measuring (A) total and (B) HIV-specific IgG ELISA in 7 day culture supernatant. Absolute numbers of live (C) B and (D) T cells were quantified. (E) Correlation analysis was carried out between the absolute number of B cells and HIV-specific IgG. (F) HIV-specific IgG ELISA in 7 day culture supernatant from cultures of CXCR5⁺CXCR3⁺ T cells and memory B cells was also quantified. For (A)-(D) and (F) bars represent mean ±SD. Symbols on the graphs represent stage 1/2 individuals (black circles) and stage 3 individuals (black squares) and statistics were carried out using the Mann-Whitney non-parametric test. For (E) symbols represent stage 1/2 individuals (black circles) and stage 3 individuals (black squares) and correlation data was analyzed using Spearman r test. * P< 0.05. Results were significant if P < 0.05.</p

IL-10 and TNF-α levels in cocultures of cTfh and B cells from late stage acute individuals influence HIV-specific IgG production.

<p>Cocultures of cTfh and autologous resting memory B cells from week 0 stage 1/2 (n = 9) and stage 3 (n = 7) individuals were analyzed for the presence of cytokines including (A) RANTES, (B) TNF-α and (C) IL-10. Correlation analysis was carried out between (D) TNF-α; (E) IL-10 and HIV-specific IgG. For (A)-(C) bars represent mean ±SD. Symbols on the graphs represent stage 1/2 individuals (black circles) and Stage 3 individuals (black squares) and statistics were carried out using the Mann-Whitney non-parametric test. For (D) and (E) symbols represent stage 1/2 individuals (black circles) and stage 3 individuals (black squares) and correlation data was analyzed using Spearman r test. * P< 0.05.</p

Elevated plasma viral load is associated with reduction in the cTfh-dependent humoral response.

<p>(A) Viral load was quantified in the plasma from stage 1/2 (n = 24) and from stage 3 (n = 30) HIV-infected individuals at day 0 prior to ART initiation. Correlation analysis was carried out between plasma viral load at day 0 and cTfh and memory 7 day culture supernatant (B) -Total IgG, (C) -HIV-specific IgG, (D) absolute number of B cells, (E) IL-10, (F) RANTES and (G) TNF-α from stage 1/2 (n = 8) and stage 3 (n = 5–7). For (A) bars represent mean SD, symbols on the graphs represent stage 1/2 individuals (black circles) and stage 3 individuals (black squares) and statistics were carried out using the Mann-Whitney non-parametric test. For (B)-(G) symbols represent stage 1/2 individuals (black circles) and stage 3 individuals (black squares) and correlation data was analyzed using Spearman r test.* P < 0.05.</p

Impact of multi-targeted antiretroviral treatment on gut T cell depletion and HIV reservoir seeding during acute HIV infection.

BackgroundLimited knowledge exists on early HIV events that may inform preventive and therapeutic strategies. This study aims to characterize the earliest immunologic and virologic HIV events following infection and investigates the usage of a novel therapeutic strategy.Methods and findingsWe prospectively screened 24,430 subjects in Bangkok and identified 40 AHI individuals. Thirty Thais were enrolled (8 Fiebig I, 5 Fiebig II, 15 Fiebig III, 2 Fiebig IV) of whom 15 completed 24 weeks of megaHAART (tenofovir/emtricitabine/efavirenz/raltegravir/maraviroc). Sigmoid biopsies were completed in 24/30 at baseline and 13/15 at week 24. At baseline, the median age was 29 years and 83% were MSM. Most were symptomatic (87%), and were infected with R5-tropic (77%) CRF01_AE (70%). Median CD4 was 406 cells/mm(3). HIV RNA was 5.5 log(10) copies/ml. Median total blood HIV DNA was higher in Fiebig III (550 copy/10(6) PBMC) vs. Fiebig I (8 copy/10(6) PBMC) (p = 0.01) while the median %CD4+CCR5+ gut T cells was lower in Fiebig III (19%) vs. Fiebig I (59%) (p = 0.0008). After 24 weeks of megaHAART, HIV RNA levels of &lt;50 copies were achieved in 14/15 in blood and 13/13 in gut. Total blood HIV DNA at week 0 predicted reservoir size at week 24 (p&lt;0.001). Total HIV DNA declined significantly and was undetectable in 3 of 15 in blood and 3 of 7 in gut. Frequency of CD4+CCR5+ gut T cells increased from 41% at baseline to 64% at week 24 (p&gt;0.050); subjects with less than 40% at baseline had a significant increase in CD4+CCR5+ T cells from baseline to week 24 (14% vs. 71%, p = 0.02).ConclusionsGut T cell depletion and HIV reservoir seeding increases with progression of AHI. MegaHAART was associated with immune restoration and reduced reservoir size. Our findings could inform research on strategies to achieve HIV drug-free remission