Despite the effectiveness of combined antiretroviral therapy (ART), HIV infection remains a chronic condition characterised by persistent inflammation and immune activation, likely associated with viral persistence and other factors such as cytomegalovirus (CMV) co-infection. Long-acting (LA) injectable formulations, a newer class of ART with improved and sustained bioavailability, may help modulate the HIV-associated immunoinflammatory state. Therefore, we analysed dynamic changes in lymphoid immune-activation and -senescence markers in people with HIV (PWH) switching to LA injectable cabotegravir and rilpivirine (CAB/RPV-LA), compared with PWH who continued oral ART by choice. The relationship between CMV-specific immune responses and phenotypic T-cell alterations was also evaluated. T-cell immune-activation (CD38 + HLA-DR+) and -senescence (CD28-CD57+) were measured at baseline (T0), at 4 (T4), 28 (T28), 48 (T48) and at 72 (T72) weeks after switching to CAB/RPV-LA. A control group (CG) of PWH (PWH-CG) on daily oral ART was studied at T0 and at 48 weeks (T48). Furthermore, CMV-specific cellular and humoral immune responses were assessed. Thirty-seven PWH switching to CAB/RPV-LA and nine PWH-CG were enrolled. In PWH-LA, total CD4 levels remained stable over time, while CD8 levels decreased significantly at T72 versus T0 and T4. In CG, an increasing CD4 and decreasing CD8 trend was observed. The CD4/CD8 ratio remained stable in both PWH-LA and -CG and showed a nonsignificant increase at T48 in PWH-CG. Immune-activation decreased in PWH-LA: CD4 activation levels were significantly lower at T48 and T72 versus earlier time points and CD8 activation was reduced at T72 versus T0 and T4. Immune-senescence declined in both compartments, with a significant reduction in CD8 senescence at T28, T48 and T72 compared to earlier time-points. Delta analysis confirmed a greater reduction in immune-activation and -senescence in PWH-LA compared to PWH-CG. Regarding CMV immune response, anti-CMV antibody levels remained stable with only a minor decline over time, whereas CMV-specific T-cell responses transiently increased at T28 before returning to baseline at T48 and T72. Switching to CAB/RPV-LA is associated with reduced T-cell activation and senescence, particularly in CD8 T-cell, suggesting improved immune homeostasis beyond viral suppression alone. However, despite these promising immunological changes, CMV coinfection remains a key driver of residual immune dysfunction.

The aim of this study was to examine the dynamic changes in monocyte/macrophage- and dendritic cell (DC)-related immune activation in people with HIV (PWH) before switching to long-acting (LA) injectable cabotegravir (CAB) plus rilpivirine (RPV) (T0), after six (T6m) and twelve months (T12m). A total of 30 aviremic PWH switching to LA CAB plus RPV, and 32 healthy donors (HDs) were enrolled. At each time point, in comparison to HDs, PWH exhibited higher classical and intermediate monocyte counts and lower slanDC and pDC counts. Compared to HDs, in PWH non-classical monocyte counts were higher only at T0. PWH exhibited elevated sCD163 and sCD14 plasmatic levels at each time point, in comparison to HDs. No virological failure was observed. At T12m, no differences in the total cell-associated HIV-1 DNA levels compared to T0 were found. The longitudinal evaluation showed a significant reduction in non-classical and intermediate monocyte counts at T12m compared to T0 and an increase in classical monocyte and pDC counts. Furthermore, a reduction in sCD14 plasma levels was observed. The constant drug concentrations provided by CAB plus RPV injections appear to be effective for a sustained and deep virological suppression that may reduce monocyte/macrophage activation in PWH.

Most studies on HPV vaccination have focused on the antibody response and on bivalent and quadrivalent vaccines. For these reasons, an evaluation of T-cell responses in PWH on effective ART to the Gardasil 9® vaccine according to prior HPV infection and baseline CD4 T-cell count was performed. We prospectively enrolled PWH on effective ART. T-cell responses were assessed by flow cytometry at pre-vaccination (T0), on the same day of the third dose (T1) and 6 months after the third dose (T2) after stimulation with a pool of HPV16 and HPV18 L1 peptide libraries. An evaluation of IFN-γ in supernatants was also carried out. Baseline CD4 T-cell count was defined at the first dose. An increase in the percentage of responding and polyfunctional T-cells was found in both HPV − and HPV + groups, with no significant differences between them at any time point. When the population was stratified according to baseline CD4 T-cell count, an increase was found only in > 500 cells/μL group for both the responding and polyfunctional T-cells. Furthermore, a higher frequency of responding CD4 T-cells at T1 (p = 0.0049), a higher frequency of responding CD8 T-cells at T1 and T2 (p = 0.05 and p = 0.02, respectively), a higher frequency of polyfunctional CD4 T-cells at T1 (p = 0.0242) and a higher frequency of polyfunctional CD8 T-cells at T2 (p = 0.02) were found in > 500 cells/μL group than in their relative counterpart. The analysis of the estimated change from T0 revealed no statistically significant differences between PWH with or without prior HPV infection, whereas significant associations between higher CD4 counts and improved immune responses were found. Our study revealed a coordinated T-cell response against HPV16 and 18 after Gardasil9® vaccine in PWH, including those with a history of HPV infection. PWH with a lower CD4 T-cell count remains a group who may not mount a fully protective immune response and, therefore, may require additional protection or ad hoc vaccination strategies. These data reinforce the key role of basal CD4 level in modulating the immune response over time, while prior HPV infection did not significantly affect the rate and quality of the response.

People living with HIV (PLWH) remain at high risk of mortality and morbidity from vaccine-preventable diseases, even though antiretroviral therapy (ART) has restored life expectancy and general well-being. When, which, and how many doses of vaccine should be administered over the lifetime of PLWH are questions that have become clinically relevant. Immune responses to most vaccines are known to be impaired in PLWH. Effective control of viremia with ART and restored CD4+ T-cell count are correlated with an improvement in responsiveness to routine vaccines. However, the presence of immune alterations, comorbidities and co-infections may alter it. In this article, we provide a comprehensive review of the literature on immune responses to different vaccines in the setting of HIV infection, emphasizing the potential effect of HIV-related factors and presence of comorbidities in modulating such responses. A better understanding of these issues will help guide vaccination and prevention strategies for PLWH.

INTRODUCTION:Skeletal tuberculosis (TB) accounts for about 10 to 35% of extrapulmonary cases and the knee is the most frequent site after the spine and hip. The diagnosis is difficult and largely clinical.CASE PRESENTATION:This is a case of a young Pakistani man with a history of joint pain for about 4 years, who was diagnosed with chronic arthritis of the right knee. Microscopy of synovial fluid and conventional diagnostic tests to identify Mycobacterium tuberculosis were negative, while a non-classical method based on intracellular cytokine flow cytometry response of CD4 T-cells in synovial fluid helped us to address the diagnosis, which was subsequently confirmed by Polymerase Chain Reaction (PCR).CONCLUSIONS:Thanks to an innovative immunological approach, supported by PCR for detection of M. tuberculosis DNA, we were able to diagnose tuberculous arthritis of the knee, which allowed prompt initiation of treatment to reduce morbidity and mortality.

Gender medicine is now an approach that can no longer be neglected and must be considered in scientific research. We investigated the systemic and mucosal immune response in a population of women living with HIV (WLWH) who were receiving successful ART and the sexual and psychological repercussions of HIV infection on the women's health. As control group, healthy women (HW) matched for age and sex distribution, without any therapy, were included. In summary, our study highlighted the persistence of immune-inflammatory activation in our population, despite virological suppression and a normal CD4 cell count. We found a hyperactivation of the systemic monocyte and an increase in inflammatory cytokine concentrations at the systemic level. The analysis carried out showed a significantly higher risk of HPV coinfection in WLWH compared to HW. Furthermore, our data revealed that WLWH have a profile compatible with sexual dysfunction and generalized anxiety disorders. Our study underlines that patients living with HIV should be evaluated by multidisciplinary teams. These findings also support the idea that more and different immunological markers, in addition to those already used in clinical practice, are needed. Further studies should be carried out to clarify which of these could represent future therapy targets.

To evaluate ex vivo the efficacy of an amino acid buffered hypochlorite solution supplemented to surface debridement with air-powder abrasion in removing bacterial biofilm following open-flap decontamination of implants failed due to peri-implantitis. This study was an ex vivo, single-blind, randomized, intra-subject investigation. Study population consisted of 20 subjects with at least three implants failed for peri-implantitis (in function for > 12 months and progressive bone loss exceeding 50

We investigated specific humoral and T-cell responses in people living with HIV (PLWH) before (T0), after two (T1) and after six months (T2) from the third dose of the BNT162b2 vaccine. Healthy donors (HD) were enrolled. The specific humoral response was present in most PLWH already after the second dose, but the third dose increased both the rate of response and its magnitude. Collectively, no significant differences were found in the percentage of responding T-cells between PLWH and HD. At T0, stratifying PLWH according to CD4 cell count, a lower percentage of responding T-cells in <200 cells/µL subgroup compared to >200 cells/µL one was observed. At T1, this parameter was comparable between the two subgroups, and the same result was found at T2. However, the pattern of co-expression of IFNγ, IL2 and TNFα in PLWH was characterized by a higher expression of TNFα, independently of CD4 cell count, indicating a persistent immunological signature despite successful ART. mRNA vaccination elicited a specific response in most PLWH, although the cellular one seems qualitatively inferior compared to HD. Therefore, an understanding of the T-cell quality dynamic is needed to determine the best vaccination strategy and, in general, the capability of immune response in ART-treated PLWH.

Background CD163, a haptoglobin-hemoglobin scavenger receptor mostly expressed by monocytes and macrophages, is involved in the regulation of inflammatory processes. Following proteolytic cleavage after pro-inflammatory stimulation, CD163 is shed from the cell surface and its soluble form in plasma, sCD163, is a biomarker of monocyte/macrophage lineage activation. The assessment of sCD163 plasmatic levels in an early stage of the disease could have clinical utility in predicting the severity of COVID-19 pneumonia. The use of tocilizumab (monoclonal antibody anti-IL-6 receptor) in COVID-19 patients reduces lethality rate at 30 days. The aim of the study was to investigate the effect of tocilizumab on sCD163 plasmatic levels in a cohort of COVID-19 patients. Methods In COVID-19 patients, on hospital admission (T0), after 7 days from hospitalization (T7) and after 45 days from discharge (T45) sCD163 plasmatic levels were evaluated, along with other laboratory parameters. COVID-19 patients were stratified into tocilizumab (TCZ) and non-tocilizumab (non-TCZ) groups. TCZ group was further divided into responder (R) and non-responder (NR) groups. Patients who died or required mechanical ventilation were defined as NR. As control group, healthy donors (HD) were enrolled. Results Seventy COVID-19 patients and 47 HD were enrolled. At T0, sCD163 plasmatic levels were higher in COVID-19 patients compared to HD (p<0.0001) and the longitudinal evaluation showed a reduction in sCD163 plasmatic levels at T7 compared to T0 (p=0.0211). At T0, both TCZ and non-TCZ groups showed higher sCD163 plasmatic levels compared to HD (p<0.0001 and p=0.0147, respectively). At T7, the longitudinal evaluation showed a significant reduction in sCD163 plasmatic levels (p=0.0030) only in the TCZ group, reaching levels comparable to those of HD. Conversely, not statistically significance in non-TCZ group was observed and, at T7, a statistically significance was found comparing non-TCZ group to HD (p=0.0019). At T0, R and NR groups showed not statistically significance in sCD163 plasmatic levels and both groups showed higher levels compared to HD (p=0.0001 and p=0.0340, respectively). The longitudinal evaluation showed significant reductions in both groups (R: p=0.0356; NR: p=0.0273) independently of the outcome. After 45 days of follow-up sCD163 plasmatic levels remain stable. Conclusion sCD163 plasmatic levels are increased in COVID-19 pneumonia and is efficiently down-regulated by tocilizumab treatment regardless of the clinical outcome.

In the direct-acting antiviral (DAA) era, it is important to understand the immunological changes after HCV eradication in HCV monoinfected (mHCV) and in HIV/HCV coinfected (HIV/HCV) patients. In this study, we analyzed sub-populations of monocytes, dendritic cells (DCs), T-lymphocytes and inflammatory biomarkers following initiation of DAA in 15 mHCV and 16 HIV/HCV patients on effective antiretroviral therapy at baseline and after sustained virological response at 12 weeks (SVR12). Fifteen age- and sex-matched healthy donors (HD) were enrolled as a control group. Activated CD4+ and CD8+ T-lymphocytes, mDCs, pDCs, MDC8 and classical, non-classical and intermediate monocytes were detected using flow cytometry. IP-10, sCD163 and sCD14 were assessed by ELISA while matrix metalloproteinase-2 (MMP-2) was measured by zymography. At baseline, increased levels of IP-10, sCD163 and MMP-2 were found in both HIV/HCV and mHCV patients compared to HD, whereas sCD14 increased only in HIV/HCV patients. After therapy, IP-10, sCD163 and sCD14 decreased, whereas MMP-2 persistently elevated. At baseline, activated CD8+ T-cells were high in HIV/HCV and mHCV patients compared to HD, with a decrease at SVR12 only in HIV/HCV patients. Activated CD4+ T-cells were higher in HIV/HCV patients without modification after DAAs therapy. These results suggest complex interactions between both viruses and the immune system, which are only partially reversed by DAA treatment.

Between November 2018 and October 2019, carbapenem-resistant Enterobacterales carrying New Delhi Metallo-beta-lactamase (NDM) caused one of the largest and persistent outbreaks occurred in Italy and intensified surveillance measures have been taken in all Italian hospitals. In this study we analyzed NDM-5-producing Escherichia coli identified in 2 hospitals of the Lazio region in Italy. Epidemiological and microbiological data demonstrated that in 2018-2019 the NDM-5-producing high-risk E. coli ST167 clone circulated in patients from both hospitals. In 2019, another NDM-5-producing E. coli clone, identified by MLST as ST617 was introduced in one of the 2 hospitals and caused an outbreak. This study describes an application of genomics as a useful method to discern endemic and outbreak clones when applied to strains of the same species (E. coli) with the same resistance determinant (NDM-5) and the relevance of screening patients admitted in critical units for carbapenemase producers to prevent outbreaks. (C) 2021 Elsevier Inc. All rights reserved.

BACKGROUND:NK cells seem to be mainly involved in COVID-19 pneumonia. Little is known about NKT cells which represent a bridge between innate and adaptive immunity.METHODS:We characterized peripheral blood T, NK and NKT cells in 45 patients with COVID-19 pneumonia (COVID-19 subjects) and 19 healthy donors (HDs). According to the severity of the disease, we stratified COVID-19 subjects into severe and non-severe groups.RESULTS:Compared to HDs, COVID-19 subjects showed higher percentages of NK CD57+ and CD56dim NK cells and lower percentages of NKT and CD56bright cells. In the severe group we found a significantly lower percentage of NKT cells. In a multiple logistic regression analysis, NKT cell was independently associated with the severity of the disease.CONCLUSIONS:The low percentage of NKT cells in peripheral blood of COVID-19 subjects and the independent association with the severity of the disease suggests a potential role of this subset.

In this study, we investigated VIM-1-producing Escherichia coli , Klebsiella oxytoca , Klebsiella pneumoniae , Citrobacter freundii , and Enterobacter cloacae strains, isolated in 2019 during a period of active surveillance of carbapenem-resistant Enterobacterales in a large university hospital in Italy. VIM-1-producing strains colonized the gut of patients, with up to three different VIM-1-positive bacterial species isolated from a single rectal swab, but also caused bloodstream infection in one colonized patient.

Objectives To compare, using an ex vivo model, the biofilm removal of three surface decontamination methods following surgical exposure of implants failed for severe peri-implantitis. Materials and methods The study design was a single-blind, randomized, controlled, ex vivo investigation with intra-subject control. Study participants were 20 consecutive patients with at least 4 hopeless implants, in function for >12 months and with progressive bone loss exceeding 50%, which had to be explanted. Implants of each patient were randomly assigned to the untreated control group or one of the three decontamination procedures: mechanical debridement with air-powder abrasion, chemical decontamination with hydrogen peroxide and chlorhexidine gluconate, or combined mechanical-chemical decontamination. Following surgical exposure, implants selected as control were retrieved, and afterwards, test implants were decontaminated according to allocation and carefully explanted with a removal kit. Microbiological analysis was expressed in colony-forming-units (CFU/ml). Results A statistically significant difference ( p < 0.001) in the concentrations of CFU/ml was found between implants treated with mechanical debridement (531.58 ± 372.07) or combined mechanical-chemical decontamination (954.05 ± 2219.31) and implants untreated (37,800.00 ± 46,837.05) or treated with chemical decontamination alone (29,650.00 ± 42,596.20). No statistically significant difference (p = 1.000) was found between mechanical debridement used alone or supplemented with chemical decontamination. Microbiological analyses identified 21 microbial species, without significant differences between control and treatment groups. Conclusions Bacterial biofilm removal from infected implant surfaces was significantly superior for mechanical debridement than chemical decontamination. Clinical relevance The present is the only ex vivo study based on decontamination methods for removing actual and mature biofilm from infected implant surfaces in patients with peri-implantitis.

The new QuantiFERON-TB Gold Plus employs modified peptides optimized to elicit an IFNγ response from CD8 + cytotoxic T lymphocytes in addition to CD4 + T cells. With a view to improve the difficult identification of TB cases, we assessed the combination of two specific immunological markers comprising IFNγ secretion and T cells co-expression of CD25 and CD134 in response to Mycobacterium tuberculosis -specific antigens. A total of 34 subjects with suspected TB and 10 age-matched HD were prospectively enrolled. Assessing the performance of QFT-Plus in terms of the TB1 and TB2 results, we found that in TB patients, the quantitative IFNγ value in TB2 was similar to that in TB1, and we did not find any differences irrespective of the disease (pulmonary or extra-pulmonary). The flow cytometric CD25/CD134 assay, allowed a more accurate differentiation between M. tuberculosis -infected and uninfected patients, with a better combination of sensitivity and specificity, especially by evaluation of CD4 + T-cell subset. All individuals with negative QFT-Plus results displayed a positive CD25/CD134 response. Overall, a positive correlation was found between T cells co-expressing CD25/CD134 and IFNγ levels in response to both QFT-Plus TB antigen tubes, as well as between the QFT-Plus TB1 and TB2 tubes. We demonstrated that both TB1 and TB2 induce a higher expression of CD25 + CD134 + markers on CD4 + T cells among infected TB subjects, compared to the lower degree of CD8 + T cells, mainly induced to TB2 stimulation. We suggest that a combined use of classic QFT-Plus and specific CD25/CD134 response may be a useful means in the diagnostic workup for active TB.

HIV infection is characterized by a state of chronic activation of the immune system, which is not completely reversed by antiretroviral treatment (ART). The aim of this study was to assess myeloid and lymphoid activation markers during HIV infection, before and one year after ART initiation, in AIDS and non-AIDS presenters. Treatment naïve HIV positive patients were enrolled in this study. Myeloid dendritic cell (mDC), plasmacytoid dendritic cell (pDC), slanDC, monocyte and T-lymphocyte cell counts and activation status, were assessed by flow cytometry in peripheral blood samples. Soluble (s)CD14 and sCD163 were assessed in plasma samples using ELISA assays. Statistical analyses were performed using GraphPad Prism and Minitab Express. Thirty-four ART naïve HIV-1 infected subjects were enrolled in this study (22 non-AIDS and 12 AIDS presenters). Seventeen healthy donors (HD) were included as control group. Although circulating mDC levels resulted unchanged, HLA-DR expression was decreased on mDCs of HIV positive subjects compared to HD (p < 0,0001). AIDS presenters showed the lowest level of expression of HLA-DR on mDCs. Circulating levels of pDCs were decreased in HIV patients compared to HD (p < 0,001), without any changes in HLA-DR expression. SlanDC cell counts were extremely reduced in AIDS presenters, compared to non-AIDS presenters and HD (p < 0,01 and p < 0,0001, respectively) and showed higher HLA-DR expression in HIV patients compared to HD (p < 0,01). Intermediate monocyte (IM) cell counts were increased in AIDS and non-AIDS presenters compared to HD (p < 0,001 and p < 0,001 respectively). Furthermore, IM expansion was directly correlated to HIV viral load (p = 0,036) and independent from CD4 cell counts and activation levels. Plasma concentrations of sCD14 and sCD163 resulted increased in HIV infected subjects compared to HD (p < 0,0001 and p < 0,001), with the highest levels observed in AIDS presenters. After 1 year, ART was able to increase pDC and decrease IM absolute cell counts and modify HLA-DR expression on mDCs and slanDCs, approaching the levels observed in HD. ART reduced also CD4 and CD8 activation levels. In conclusion, in untreated HIV infected subjects circulating dendritic cells resulted altered either in numbers or in HLA-DR expression, especially in AIDS presenters. IM absolute counts were equally increased in AIDS and non-AIDS presenters. ART was able to reduce myeloid and lymphoid inflammation in both advanced and non-advanced HIV patients, confirming the role of ART in hampering disease progression and immune activation associated non-AIDS events.