Elite controllers (EC) are people living with HIV (PLWH) able to control HIV replication without antiretroviral therapy and have been proposed as a model of functional cure. Significant evidence suggests that despite viral control, EC subjects may have altered levels of systemic inflammation. We have performed a comprehensive characterization of the systemic inflammation profile in EC compared to non-controllers PLWH either with or without ART-mediated control of viral replication. 40 participants were included: 10 EC, 10 non-controllers PLWH on ART (onART), 10 non-controllers PLWH ART-naïve (offART), and 10 uninfected controls (UC) as reference. Plasmatic levels of 92 surrogate markers of inflammation were assessed using the proximity extension assay (Olink proteomics, Sweden). An additional panel of nine proteins was also assayed by comercial ELISA. Differential expression analysis, principal component analysis (PCA), and clustering analysis were carried out using the Metaboanalyst software and the R software. Pathway enrichment analysis (PEA) was carried out using the STRING platform. Compared to UC, offART group showed the highest disturbance in the inflammatory markers with 30 different proteins differentially expressed (DE) (29/30 upregulated; p <0.05). In contrast, onART group presented a profile similar to UC with only 4 proteins DE (3/4 upregulated). Interestingly, EC group presented a more disturbed inflammatory profile than onART group, with 10 different proteins DE with respect to UC group (10/10 upregulated; p <0.05), of which 3 were DE only in EC group. Of note, among these 3 proteins were CCL4 (a ligand for HIV-correceptor CCR5) and CCL13 (a ligand for HIV-correceptor CCR2). Regarding clinical status, a substantial comorbidity burden was observed in both groups with controlled viremia (EC and onART), with metabolic and cardiovascular conditions being the most prevalent. Our study shows that HIV control mechanisms, ART-mediated or spontaneous, are linked to distinct inflammatory profiles. Despite successful viral suppression, both EC and onART individuals maintain persistent inflammatory signatures compared to uninfected controls, which are associated with a significant burden of non-AIDS comorbidities. These findings underscore the need for clinical monitoring and personalized strategies to mitigate inflammation-driven morbidity in all PLWH phenotypes.

BackgroundConventional HIV-1 management relies on clinical phenotypes, assuming a uniform inflammatory status and overlooking biological heterogeneity. We aimed to employ a high-throughput proteomic approach to stratify people living with HIV (PLWH) based on plasma inflammatory profiles and evaluate their association with comorbidity burden.MethodsThis cross-sectional study included 30 PLWH (10 treatment-naïve, 10 ART-suppressed, and 10 elite controllers (EC)) and 10 seronegative controls. Unsupervised hierarchical clustering was applied to 87 inflammatory proteins (analyzed by proximity extension assay and ELISA). A Random Forest model stabilized with 1,000 trees identified the variables defining the biotypes. Comorbidity burden was assessed within the virally suppressed groups (ART-suppressed and EC).ResultsClustering revealed two distinct biological profiles independent of clinical classifications: a high-inflammation biotype (n=11) and a low-inflammation biotype (n=19). Within the treatment-naïve, elite controller, and ART-suppressed groups, 60%, 30%, and 20% of participants, respectively, were classified as belonging to the high-inflammation biotype. PDL1, CD40, TNF, LAP-TGFbeta1, and SLAMF1 were the top biological drivers of this separation. A trend for a higher comorbidity burden was observed in the high-inflammation biotype compared to the low-inflammation group (p=0.06).ConclusionsPlasma inflammatory profiling stratifies PLWH into distinct biological biotypes that bypass conventional classifications. The observed differences in comorbidity burden between inflammatory biotypes warrant further investigation in larger prospective studies to clarify their clinical relevance and potential implications for long-term health outcomes.

Elite controllers (ECs) represent a unique subset of people living with HIV (PLWHs), who can suppress viral replication without requiring antiretroviral therapy (ART). However, despite this viral control, ECs exhibit increased incidences of various comorbid conditions and heightened systemic inflammation, which has been linked to monocyte activation. In this study, we performed an in-depth phenotypic analysis of monocytes in a cohort of long-term ECs (LTECs) and compared them to non-controller patients with ART-mediated control of HIV replication and to non-controller patients with uncontrolled viral replication. A total of 67 participants were included: 22 LTECs, 15 non-controllers on ART (onART), 10 non-controllers without ART (offART), and 20 uninfected controls (UCs) as a reference group. Monocyte phenotypes were analyzed using spectral flow cytometry with a 13-marker panel. The data were analyzed using two approaches: (a) FCS Express software v.7 to define different subsets of monocytes and assess the levels of expression of eight different monocyte functional markers and (b) R software v.4.1.1 for unsupervised multidimensional analysis, including batch correction, dimensionality reduction, and clustering analysis. Monocyte phenotypic profiling was conducted using three different approaches: (1) assessment of monocyte subsets (classical, intermediate, and non-classical monocytes); (2) evaluation of the levels of expression of eight monocyte functional markers, and (3) characterization of monocyte clusters defined through the dimensionality reduction of flow cytometry data (56 different clusters). The monocyte phenotype of the onART group closely resembled that of the UC group. In contrast, LTECs exhibited important alterations in the monocyte phenotype compared to that of the UCs, including (a) an increased proportion of intermediate monocytes and a decreased proportion of classical monocytes (p < 0.01), (b) altered expressions of functional markers across monocyte subsets (p < 0.05), and (c) alterations in sixteen different monocyte clusters (twelve decreased and four increased, p < 0.05). Many of these alterations were also observed when comparing the LTEC and onART groups. Our findings suggest that monocyte-driven mechanisms may contribute to HIV control in LTECs; however, some of these alterations could also promote systemic inflammation and immune activation. These observations provide a compelling rationale for considering therapeutic interventions in this unique population of PLWHs.

BACKGROUND:Identifying characteristics associated with functional cure of HIV-1 infection is of pivotal importance. Here, we describe a unique case of functional cure in an exceptional elite controller, comparing immunological, virological, and clinical characteristics with those of other elite controllers. METHODS:In this case report, we compared the case (a heterosexual Indian woman aged 48 years) with her partner (an Indian man aged 58 years) and a cohort of elite controllers (people with HIV with at least three consecutive plasma HIV viral loads below detection during at least 12 months of follow-up, in the absence of antiretroviral therapy; cohort 1) and long-term elite controllers (elite controllers with immunological [no statistically significant decrease of CD4 cell count] and virological control during a minimum of 10 years and maintained at follow-up; cohort 2), from reference hospitals in Madrid, Spain. A group of HIV-negative donors was also included. The case was first diagnosed with sexually transmitted HIV-1 infection in 2002, after her partner was diagnosed with HIV-1 infection and Pneumocystis jiroveci pneumonia. We did a comprehensive analysis of virological and host features. Virological assays were reservoir quantification, quantitative viral outgrowth assay, and next-generation sequencing of viral RNA and DNA; host assays were HLA-I typing, host restriction factors, plasma inflammatory markers, HIV-1 specific T-cell response, and immunophenotypic analysis of T cells, monocytes, and natural killer cells. FINDINGS:The case maintained undetectable plasma viral load and stable CD4 cell count in the absence of combination antiretroviral therapy for 22 years and never presented clinical manifestations or symptoms related to HIV infection. Western blot reactivity was limited to glycoprotein 41 and glycoprotein 160. HIV-1 DNA was undetectable in resting memory CD4 cells and peripheral T follicular helper cells. Full-length individual proviral sequencing assays revealed only 0·82 HIV genomes per million CD4 cells; all were defective with large internal deletions. Replication-competent viruses were not detected. Compared with reference cohorts 1 (n=9) and 2 (n=12), the case had increased expression of several microRNAs and mRNAs with anti-HIV action; preservation of T-cell homoeostasis with low expression of markers of cell exhaustion (PD1, Tim3, CTLA4, and TIGIT), apoptosis (CD28 and CD95), and senescence (CD28 and CD57); natural killer cells with low inhibitory receptor expression and higher expression of regulatory natural killer cells; and decreased plasma expression of several inflammatory markers (D-dimer, TNFR1, and VCAM-1). INTERPRETATION:Functional cure is an uncommon but feasible outcome of HIV-1 infection. Our findings encourage the search for therapeutic strategies that could enable people with HIV-1 to reach this level of functional cure. FUNDING:Carlos III Institute of Health (Spain).

Elite controllers (ECs), a unique group of people with HIV (PWH), exhibit remarkable control of viral replication in the absence of antiretroviral therapy. In this study, we comprehensively characterized the NK cell repertoire in ECs after long-term viral control. Phenotypic profiling of NK cells revealed profound differences compared with other PWH, but marked similarities to uninfected individuals, with a distinctive prevalence of NKG2C+CD57+ memory-like NK cells. Functional analyses indicated that ECs had limited production of functional molecules upon NK stimulation and consequently reduced natural cytotoxicity against non-HIV target cells. Importantly, ECs showed an exceptional ability to kill primary HIV-infected cells by the antibody-dependent cell cytotoxicity adaptive mechanism, which was achieved by a specific memory-like NK population expressing CD16, NKG2A, NKG2C, CD57, and CXCR3. In-depth single-cell RNA-seq unveiled a unique transcriptional signature in these NK cells linked to increased cell metabolism, migration, chemotaxis, effector functions, cytokine secretion, and antiviral response. Our findings underscore a pivotal role of NK cells in the immune control of HIV and identify specific NK cells as emerging targets for immunotherapies.

Elite controllers (ECs) are people living with HIV (PLWH) able to control HIV replication without antiretroviral therapy and have been proposed as a model of a functional HIV cure. Much evidence suggests that this spontaneous control of HIV has a cost in terms of T cell homeostasis alterations. We performed a deep phenotypic study to obtain insight into T cell homeostasis disturbances in ECs maintaining long-term virologic and immunologic control of HIV (long-term elite controllers; LTECs). Forty-seven PLWH were included: 22 LTECs, 15 non-controllers under successful antiretroviral therapy (onART), and 10 non-controllers not receiving ART (offART). Twenty uninfected participants (UCs) were included as a reference. T cell homeostasis was analyzed by spectral flow cytometry and data were analyzed using dimensionality reduction and clustering using R software v3.3.2. Dimensionality reduction and clustering yielded 57 and 54 different CD4 and CD8 T cell clusters, respectively. The offART group showed the highest perturbation of T cell homeostasis, with 18 CD4 clusters and 15 CD8 clusters significantly different from those of UCs. Most of these alterations were reverted in the onART group. Interestingly, LTECs presented several disturbances of T cell homeostasis with 15 CD4 clusters and 13 CD8 clusters different from UC. Moreover, there was a specific profile of T cell homeostasis alterations associated with LTECs, characterized by increases in clusters of naïve T cells, increases in clusters of non-senescent effector CD8 cells, and increases in clusters of central memory CD4 cells. These results demonstrate that, compared to ART-mediated control of HIV, the spontaneous control of HIV is associated with several disturbances in CD4 and CD8 T cell homeostasis. These alterations could be related to the existence of a potent and efficient virus-specific T cell response, and to the ability to halt disease progression by maintaining an adequate pool of CD4 T cells.

Background Human immunodeficiency virus (HIV) infection causes sustained chronic immune activation which is associated with earlier and more frequent onset of comorbidities. Coinfections, such as those caused by hepatitis C virus (HCV), amplify this immune status. However, direct-acting antivirals (DAAs) transformed HIV/HCV management by eradicating HCV quickly and effectively, bypassing the systemic proinflammatory and immunomodulatory effects of interferon regimens. HCV infection significantly impacts the immunopathogenesis of HIV, and eradication of HCV with DAAs improves, but does not entirely normalize, the levels of markers of systemic inflammation, endothelial dysfunction or T-cell activation and exhaustion. This study aimed to determine whether HIV-infected individuals versus individuals with HIV/HCV coinfection, in the era of interferon-free therapies, exhibit an increased incidence of comorbidities and non-AIDS-related events. Methods A retrospective analysis was conducted at a Spanish tertiary hospital, involving 229 HIV/HCV-coinfected patients and 229 HIV-infected patients, all with effectively controlled HIV. Coinfected patients underwent HCV clearance using DAAs and had no history interferon treatment. The incidences of hypertension, diabetes mellitus, cardiovascular disease, kidney disease, liver disease, non-AIDS cancer and death were compared between the groups. Univariate logistic regression models and subsequent multivariate adjustment for all factors potentially impacting outcomes were used to assess the risk of clinical event onset. Propensity score (PS) analyses were also conducted to support the multivariate model results. Results Univariate logistic regression analyses revealed that, compared to HIV/HCV coinfected patients, HIV monoinfected patients presented a greater risk for hypertension (Odds Ratio [OR] = 1.93; 95% Confidence Interval [CI] = 1.03–3.74; p = 0.040), dyslipidemia (OR = 1.83; 95%CI = 1.13–3.01; p = 0.014) and kidney disease (OR = 3.14; 95%CI = 1.19–9.80; p = 0.019) onset. Monoinfection was also associated with a lower risk for developing liver disease (OR = 0.12; 95%CI = 0.01–0.67; p = 0.012) and death (OR = 0.29; 95%CI = 0.06–0.96; p = 0.043). Multivariate models and PS showed that previous exposure to HCV was not associated with the onset of any clinical events studied. Conclusions Successful HCV elimination using DAAs improved the outlook regarding comorbidities and survival across both patient cohorts. Early HCV detection and DAA therapy could enhance clinical results. These findings provide an optimistic perspective for those living with HIV/HCV coinfection and underscore the importance of continuing efforts toward early detection and DAA treatment initiation.

Elite controllers (ECs) are an exceptional group of people living with HIV (PLWH) that control HIV replication without therapy. Among the mechanisms involved in this ability, natural killer (NK)-cells have recently gained much attention. We performed an in-deep phenotypic analysis of NK-cells to search for surrogate markers associated with the long term spontaneous control of HIV. Forty-seven PLWH (22 long-term EC [PLWH-long-term elite controllers (LTECs)], 15 noncontrollers receiving antiretroviral treatment [ART] [PLWH-onART], and 10 noncontrollers cART-na & iuml;ve [PLWH-offART]), and 20 uninfected controls were included. NK-cells homeostasis was analyzed by spectral flow cytometry using a panel of 15 different markers. Data were analyzed using FCSExpress and R software for unsupervised multidimensional analysis. Six different subsets of NK-cells were defined on the basis of CD16 and CD56 expression, and the multidimensional analysis revealed the existence of 68 different NK-cells clusters based on the expression levels of the 15 different markers. PLWH-offART presented the highest disturbance of NK-cells homeostasis and this was not completely restored by long-term ART. Interestingly, long term spontaneous control of HIV (PLWH-LTEC group) was associated with a specific profile of NK-cells homeostasis disturbance, characterized by an increase of CD16(dim)CD56(dim) subset when compared to uninfected controls (UC) group and also to offART and onART groups (p < 0.0001 for the global comparison), an increase of clusters C16 and C26 when compared to UC and onART groups (adjusted p-value < 0.05 for both comparisons), and a decrease of clusters C10 and C20 when compared to all the other groups (adjusted p-value < 0.05 for all comparisons). These findings may provide clues to elucidate markers of innate immunity with a relevant role in the long-term control of HIV.

Objective: To determine whether HIV-infected individuals versus individuals with HIV/HCV coinfection, in the era of interferon-free therapies, exhibit an increased incidence of comorbidities and non-AIDS-related events. Methods: A retrospective analysis was conducted by collecting data from clinical records of Spanish patients at a tertiary hospital involving HIV/HCV-coinfected and HIV-infected patients, all with effectively controlled HIV. Coinfected patients underwent HCV clearance using direct-acting antivirals (DAAs) and had no history of interferon treatment. The incidences of hypertension, diabetes mellitus, cardiovascular disease, kidney disease, liver disease, non-AIDS cancer, and death were compared between the groups. Multivariate adjustments for all factors potentially impacting outcomes were used to assess the risk of clinical event onset. Propensity score (PS) analyses were also conducted to support the multivariate model results. Results: Data were available from 229 HIV/HCV-coinfected patients and 229 HIV-infected patients. Both cohorts were comparable in terms of age, gender distribution, follow-up, and HIV-related characteristics. Multivariate models and PS showed that previous exposure to HCV was not associated with the onset of any clinical events studied. Significant differences between HIV/HCV-coinfected and HIV-infected were not found for survival according to the log-rank test (p = 0.402). Conclusions: Successful HCV elimination using DAAs improved the outlook regarding comorbidities and survival across HIV/HCV-coinfected cohorts. Early HCV detection and DAA therapy could enhance clinical results. These findings provide an optimistic perspective for those living with HIV/HCV coinfection and underscore the importance of continuing efforts toward early detection and DAA treatment initiation.

Human leukocyte antigen (HLA) class I/killer cell immunoglobulin-like receptor (KIR) genotypes influence human immunodeficiency virus type 1 (HIV-1) disease progression and viral load, but their role in primary infection is uncertain. Inconsistent results from previous studies suggest that the inoculum size and transmission route-parenteral versus sexual-may influence this association. We conducted a genome-wide association study in a population of people with HIV-1 and HIV-1-exposed seronegative individuals exposed to the virus through the sexual route. Our data do not support any role of the HLA/KIR system in susceptibility to sexually transmitted HIV-1 infection. The genetics basis of HIV-1 viral load and disease progression are distinct from the genetics of HIV resistance, a paradox worth exploring. HLA-I/KIR genotypes impact HIV-1 progression, but their role in primary infection is debated. A new genome-wide association study shows no role in sexual transmission susceptibility. The genetic basis of HIV-1 progression differs from resistance, presenting a paradox worth exploring. Graphical abstract

IRF5–TNPO3 polymorphisms have previously been related to immune response, and TNPO3 plays a role in human immunodeficiency virus (HIV)‐1 infection after nuclear import. Therefore, we analyzed the genetic association between IRF5–TNPO3 polymorphisms and the HIV elite control in long‐term nonprogressors (LTNPs). We performed a retrospective cohort study on 183 LTNPs, who were antiretroviral therapy‐naïve with CD4+ ≥ 500 cells/mm3, viral load ≤10 000 copies/mL, and asymptomatic over 10 years after HIV seroconversion. The primary outcome variable was HIV elite control (undetectable viral load in at least 90% of the measurements for at least 1 year). Seven IRF5–TNPO3 polymorphisms were genotyped using Agena Bioscience's MassARRAY platform. We found a significant association between specific IRF5–TNPO3 genotypes and HIV elite control: rs2004640 TT (aOR = 2.05; p = 0.041), rs10954213 AA (aOR = 1.95; p = 0.035), rs2280714 TT (aOR = 2.02; p = 0.031), and rs10279821 CC (aOR = 2.12; p = 0.017). We also found a significant association between IRF5‐TNPO3 haplotype TATC composed of the favorable significant polymorphisms (rs2004640, rs10954213, rs2280714, and rs10279821) and the HIV elite control (aOR = 1.59; p = 0.048). IRF5–TNPO3 rs2004640, rs10954213, rs2280714, and rs10279821 polymorphisms were related to HIV elite control in LTNPs. Our data provide new knowledge about the impact of IRF5–TNPO3 polymorphisms on HIV pathogenesis to understand the phenomenon of natural HIV control.

Interleukin 7 receptor (IL7R) is vital in the adaptive immune response against human immunodeficiency viruses (HIV). We assessed IL7RA polymorphisms (SNPs) in antiretroviral therapy (ART)-naive HIV patients for their association with spontaneous HIV infection control. We conducted a retrospective cohort study involving 667 ART-naive patients categorized by HIV progression (ordinal variable): 150 rapid progressors, 334 moderate/typical progressors, 86 long-term nonprogressors elite controllers (LTNPs-EC), and 97 LTNPs-non-EC. We genotyped three IL7RA SNPs using Agena Bioscience's MassARRAY platform. The association between IL7RA SNPs and spontaneous HIV infection control was evaluated using ordinal logistic regression. Individuals carrying the rs10491434 G allele have a higher likelihood of spontaneous HIV infection control (adjusted odds ratio [aOR]=1.33; p=0.023). Moreover, the IL7RA GCT haplotype, consisting of three specific SNPs (rs6897932, rs987106, and rs10491434), demonstrated an association with the control of untreated HIV infection (aOR=1.34; p=0.050). Remarkably, the rs10491434 SNP and the IL7RA GCT haplotype exhibited similar aOR values, suggesting that rs10491434 may be primarily responsible for the observed effect of the haplotype. IL7RA rs10491434 G allele is associated with a higher likelihood of spontaneous HIV infection control, indicating its significant role in the pathogenesis of HIV, possibly influencing infection course and viral replication control.

The immune system of people living with HIV (PLWH) is persistently exposed to antigens leading to systemic inflammation despite combination antiretroviral treatment (cART). This inflammatory milieu promotes T-cell activation and exhaustion. Furthermore, it produces diminished effector functions including loss of cytokine production, cytotoxicity, and proliferation, leading to disease progression. Exhausted T cells show overexpression of immune checkpoint molecules (ICs) on the cell surface, including programmed cell death protein 1 (PD-1), cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4), T-cell immunoglobulin and mucin-domain containing-3 (TIM-3), T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT), and lymphocyte activation gene-3 (LAG-3). The ICs also play a crucial role in T-cell exhaustion by reducing the immune response to cancer antigens. Immunotherapy based on immune checkpoint inhibitors (ICIs) has changed the management of a diversity of cancers. Additionally, the interest in exploring this approach in the setting of HIV infection has increased, including AIDS-defining cancers and non-AIDS-defining cancers in PLWH. To date, research on this topic suggests that ICI-based therapies in PLWH could be a safe and effective approach. In this review, we provide an overview of the current literature on the potential role of ICI-based immunotherapy not only in cancer remission in PLWH but also as a therapeutic intervention to restore immune response against HIV, revert HIV latency, and attain a functional cure for HIV infection.

Background and Purpose Reperfusion therapy is the standard of care for ischaemic stroke; however, there is a need to identify new therapeutic targets able to ameliorate cerebral damage. Neutrophil β 1 adrenoceptors (β1AR) have been linked to neutrophil migration during exacerbated inflammation. Given the central role of neutrophils in cerebral damage during stroke, we hypothesize that β1AR blockade will improve stroke outcomes. Experimental Approach Rats were subjected to middle cerebral artery occlusion–reperfusion to evaluate the effect on stroke of the selective β1AR blocker metoprolol (12.5 mg·kg −1 ) when injected i.v. 10 min before reperfusion. Key Results Magnetic resonance imaging and histopathology analysis showed that pre‐reperfusion i.v. metoprolol reduced infarct size. This effect was accompanied by reduced cytotoxic oedema at 24 h and vasogenic oedema at 7 days. Metoprolol‐treated rats showed reduced brain neutrophil infiltration and those which infiltrated displayed a high proportion of anti‐inflammatory phenotype (N2, YM1 + ). Additional inflammatory models demonstrated that metoprolol specifically blocked neutrophil migration via β1AR and excluded a significant effect on the glia compartment. Consistently, metoprolol did not protect the brain in neutrophil‐depleted rats upon stroke. In patients suffering an ischaemic stroke, β1AR blockade by metoprolol reduced circulating neutrophil–platelet co‐aggregates. Conclusions and Implications Our findings describe that β1AR blockade ameliorates cerebral damage by targeting neutrophils, identifying a novel therapeutic target to improve outcomes in patients with stroke. This therapeutic strategy is in the earliest stages of the translational pathway and should be further explored.

The role of HCV on the HIV reservoir is controversial since the reduction on HIV-DNA levels after HCV eradication with IFNα/RBV treatment seems to be the result of drugs instead of HCV clearance. We assessed whether HCV eradication can decrease HIV-DNA content in HIV/HCV-coinfected patients treated with direct-acting antivirals, DAAs (IFNα/RBV-free regimens). Cell-associated HIV-DNA was measured by ddPCR in 25 HIV-monoinfected and 25 HIV/HCV-coinfected patients. There were no differences in HIV-DNA levels between groups neither at baseline nor at 12 weeks after DAAs treatment completion. Our results indicate that HCV does not appear to influence the HIV reservoir size and suggest the lack of an anti-HIV action for DAAs.

Abstract Previously, we identified a stabilization of the HIV reservoir size in HIV + monoinfected and HIV+/HCV + coinfected patients who eliminated HCV with DAAs as well as a decrease in HIV reservoir size in HIV+/HCV- spontaneously clearers. However, variations in the distribution of memory CD4 + T cell subpopulations in coinfected patients after HCV clearance remains unclear. To analyze the expression pattern of T cell maturation (CD45RA and CD27) and senescence (CD28 and CD57) markers by flow cytometry in those patients. Longitudinal study (52 weeks follow-up) of 49 HIV+/HCV+, 38 HIV+/HCV- and 59 HIV + subjects, all of them aviremic. A generalized increase in expression of CD28 was observed for CD8 + and in CD4 + CD27 + cells. The proportion of both CD4 + TEM TH1 and TH2 cells decreased significantly in HIV+/HCV- while remained stable in HIV + and HIV+/HCV + individuals. The general decline in cell senescence of cytotoxic lymphocytes may be due to reduced antigenic stimulation of HIV-specific and HCV-specific lymphocytes. The dynamic of HIV viral reservoir in HIV+/HCV- patients matches the dynamic of CD4 + cells with a TH1 phenotype, which could mean a decrease in the number of circulating TFH1 lymphocytes after an improvement in generalized inflammation state.

We have analyzed BNT162b2 vaccine-induced immune responses in naive subjects and individuals recovered from coronavirus disease 2019 (COVID-19), both soon after (14 days) and later after (almost 8 months) vaccination. Plasma spike (S)-specific immunoglobulins peak after one vaccine shot in individuals recovered from COVID-19, while a second dose is needed in naive subjects, although the latter group shows reduced levels all along the analyzed period. Despite how the neutralization capacity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mirrors this behavior early after vaccination, both groups show comparable neutralizing antibodies and S-specific B cell levels late post-vaccination. When studying cellular responses, naive individuals exhibit higher SARS-CoV-2-specific cytokine production, CD4+ T cell activation, and proliferation than do individuals recovered from COVID-19, with patent inverse correlations between humoral and cellular variables early post-vaccination. However, almost 8 months post-vaccination, SARS-CoV-2-specific responses are comparable between both groups. Our data indicate that a previous history of COVID-19 differentially determines the functional T and B cell-mediated responses to BNT162b2 vaccination over time.

Accumulation of mutations in epitopes of cytolytic-T-lymphocytes immune response (CTL) in HIV-reservoir seems to be one of the reasons for shock-and-kill strategy failure. Ten non-controller patients on successful cART (TX) and seven elite controllers (EC) were included. HIV-Gag gene from purified resting memory CD4+ T-cells was sequenced by Next-Generation-Sequencing. HLA class-I alleles were typed to predict optimal HIV-Gag CTL epitopes. For each subject, the frequency of mutated epitopes in the HIV-Gag gene, the proportion of them considered as CTL-escape variants as well as their effect on antigen recognition by HLA were assessed. The proportion (%) of mutated HIV-Gag CTL epitopes in the reservoir was high and similar in EC and TX (86%[50-100] and 57%[48-82] respectively, p=0.315). Many of them were predicted to negatively impact antigen recognition. Moreover, the proportion of mutated epitopes considered to be CTL-escape variants was also similar in TX and EC (77%[49-92] vs. 50%[33-75] respectively, p=0.117). Thus, the most relevant finding of our study was the high and similar proportions of HIV-Gag CTL-escape mutations in the reservoir of both HIV-noncontroller patients with cART (TX) and patients with spontaneous HIV-control (EC). Our findings suggest that escape mutations of CTL-response may be another obstacle to eliminate the HIV reservoir and constitute a proof of concept that challenges HIV cure strategies focused on the reactivation of reservoirs. Due to the small sample size that could impact the robustness of the study, further studies with larger cohorts of elite controller patients are needed to confirm these results.

Long-term elite controllers (LTECs) are a fascinating small subset of HIV individuals with viral and immunological HIV control in the long term that have been designated as models of an HIV functional cure. However, data on the LTEC phenotype are still scarce, and hence, the metabolomics and lipidomics signatures in the LTEC-extreme phenotype, LTECs with more than 10 years of viral and immunological HIV control, could be pivotal to finding the keys for functional HIV remission. Metabolomics and lipidomics analyses were performed using high-resolution mass spectrometry (ultra-high-performance liquid chromatography–electrospray ionization–quadrupole time of flight [UHPLC-(ESI) qTOF] in plasma samples of 13 patients defined as LTEC-extreme, a group of 20 LTECs that lost viral and/or immunological control during the follow-up study (LTEC-losing) and 9 EC patients with short-term viral and immunological control (less than 5 years; no-LTEC patients). Long-term viral and immunological HIV-1 control was found to be strongly associated with elevated tricarboxylic acid (TCA) cycle function. Interestingly, of the nine metabolites identified in the TCA cycle, α-ketoglutaric acid (p = 0.004), a metabolite implicated in the activation of the mTOR complex, a modulator of HIV latency and regulator of several biological processes, was found to be a key metabolite in the persistent control. On the other hand, a lipidomics panel combining 45 lipid species showed an optimal percentage of separation and an ability to differentiate LTEC-extreme from LTEC-losing, revealing that an elevated lipidomics plasma profile could be a predictive factor for the reignition of viral replication in LTEC individuals.

Human immunodeficiency virus-1 (HIV-1) elite controllers are heterogeneous due to different immunovirological features. We aimed to identify plasma biomarkers associated with loss of spontaneous HIV-1 control in long-term elite controllers (HIV-LTECs). We performed a retrospective study in 60 HIV-LTECs [36 true-LTECs and 24 LTECs losing control (LTECs-LC)]. We selected a plasma sample from true-LTECs (towards the middle of the follow-up period) and two samples from LTECs-LC (one far from the loss of control and another close to loss of control). Plasma biomarkers were evaluated using multiplex immunoassays. The partial least squares-discriminant analysis provided the variable importance in projection (VIP), and the adjusted Generalized Linear Model provided the adjusted arithmetic mean ratio (aAMR). At the moment of the first LTECs-LC samples, the only plasma biomarker with a VIP≥1.5 was sTNF-R1, which showed higher values in LTECs-LC than true-LTECs [aAMR=1.62 (95%CI=1.20-2.19); p=0.001]. After a median of 3.9 (IQR=4.5) years of follow-up from the first sample, we also had access to a second plasma sample from 10 LTECs-LC patients. At the moment of this second LTECs-LC sample, the only plasma biomarker with VIP≥1.5 was also sTNF-R1, which showed higher values in LTECs-LC than true-LTECs [aAMR=1.93 (95%CI=1.41-2.65); p<0.001]. The difference between the first and second samples of LTECs-LC was significant (Δx= 6.58 (95%=0.3; 12.88); p=0.040). In conclusion, high plasma values of sTNF-R1 appear to discriminate HIV-LTECs that lose the natural control of HIV-1, helping to define a specific phenotype that may be useful for the clinical management of these patients.