Neutralising antibodies (NAbs) against SARS-CoV-2 in BNT162b2 vaccine recipients with a history of advanced HIV infection: cPass as a simpler alternative to gold-standard tests

Background: Established SARS-CoV-2 NAb tests are labor-intensive. We prospectively measured NAbs vs Wuhan-1 and Omicron BA.2 using the novel GenScript cPass assay and examined correlations with responses measured by gold-standard plaque reduction neutralisation test (PRNT) (Cotugno, Ruggiero et al. Cell Rep 2021) and with anti-Spike IgG quantified by Roche Elecsys. Given the paucity of data, we selected BNT162b2 vaccine recipients with a history of advanced HIV infection (prior AIDS-defining conditions and/or nadir CD4 <200 cells). Method(s): In Mar 2021-Apr 2022, 55 PWH received 2 vaccine doses median 3 weeks apart [IQR 3-3] and a 3rd dose 27 weeks later [23-31]. Plasma samples (n=147) were stored immediately before dose-1 (T0), median 4 weeks [3-5] after dose-2 (T1) and median 13 weeks [9-19] after dose-3 (T2) for batch testing. Result(s): Participants' characteristics: 74% male, 85% white, all on ART, 82% HIV-RNA <50 cps/ml;median age 55 years, ART duration 7 years, nadir CD4 83 cells [36-211], current CD4 440 cells [270-710], CD4:CD8 ratio 0.6 [0.4-1.0];73% had a history of advanced HIV infection;15% received a COVID-19 diagnosis during the study. At T0, T1 and T2, proportions with quantifiable anti-S IgG (>0.8 U/ml) were 11/49 (22%), 50/54 (93%) and 43/43 (100%), respectively;their median anti-S IgG titres were 30 [15-124], 15949 [596-3389] and 8527 [3146-17190] U/ml. Proportions showing Wuhan-1 neutralisation by cPass were 6/50 (12%), 45/53 (85%) and 40/43 (93%), with median neutralisations of 67% [47-70], 97% [91-98] and 98% [98-98] and corresponding NAb titres of 1332 [792-1436], 5354 [3529-6187] and 6242 [5765-6766] U/ml. At T2, 25/28 (89%) showed BA.2 neutralisation by cPass (median 83% [68-93];NAb titre 7836 [3172-12173] U/ml) (Fig 1A). Two participants lacking NAbs at T2 had a history of advanced HIV infection. cPass data were highly correlated with anti-S IgG titres (rho 0.82;p<0.0001) and with PRNT data for both Wuhan-1 (n=27, Fig 1B) and Omicron BA.2 (n=28, Fig 1C). Conclusion(s): cPAss offers a simple methodology for measuring SARS-CoV-2 NAbs. Despite prior advanced HIV infection, neutralising activity improved with successive vaccinations and most participants showed NAbs against both Wuhan-1 and Omicron BA.2 after 3 vaccine doses. (Figure Presented).

SARS-CoV-2 T-cell specific responses in allogeneic hematopoietic stem cell transplant recipients after second and third dose of BNT162b2 mRNA vaccine

Background. Cell-mediated immunity (CMI) after anti-Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV)-2 vaccines has been poorly explored in recipients of allogeneic hematopoietic stem-cell transplantation (HSCT), especially with regard to the 3rd dose (booster). Aim of the study was to assess the specific T-cell responses before and after the 3rd dose of BNT162b2 mRNA vaccine in a cohort of allogeneic HSCT recipients, and compare it with healthy donors (HD). Methods. Allogenic HSCT recipients and HD were enrolled before receiving the 3rd dose of BNT162b2 mRNA vaccine. Whole blood for T-cell specific responses was collected before (T1) and 8 weeks after (T2) the booster administration. T-cell responses were assessed with an Interferon (IFN)-gamma release assay (IGRA), after overnight stimulation of heparin whole blood with pools of lyophilized peptides, covering the immunodominant sequence of the Spike (S) protein. IFN-gamma production was assessed with an enzyme linked immunosorbent assay (ELISA). Statistical analysis was performed with GraphPadPrism. Results. 14 HSCT recipients (8M, 6F) and 15 HD (7M, 8F) were enrolled (table 1). Median age was 47 [39-59] and 41 [31-48] years in the HSCT and HD groups, respectively. Time between the vaccine 2nd dose and T1 was significantly longer in HD than HSCT recipients (p< .001), while the time between T1 and T2 did not differ between the two groups. SARS-CoV-2 S specific T-cell responses at T1 were inferior in HSCT recipients compared to HD (median IFN-gamma production: 463 vs 231 ng/ml, respectively), although the difference did not reach the statistical significance. No differences were observed at T2. In a before-after analysis, SARS-CoV-2 S specific T-cell responses were significantly increased in HSCT recipients at T2 compared to T1 (median IFN-gamma production: 267 vs 881 ng/ml, p=0.02) (Figure 1). At T1, 3 HSCT recipients showed very low or no IFN-gamma production, while at T2 only 1 patient still had undetectable IFN-gamma production after S peptide stimulation. Conclusion. SARS-CoV-2 IGRA represents a useful tool to assess CMI, also in immunocompromised hosts. An additional 3rd BNT162b2 mRNA vaccine booster dose seems to enhance CMI in allogenic HSCT recipients. Further studies are needed to evaluate the duration of SARS-CoV-2 CMI in HSCT recipients compared to HD.

Oral Antivirals Against SARS-CoV-2: a New Valid Strategy Against COVID-19

Background. Since the outbreak of COVID-19 pandemic the scientific community efforts have been focused on finding a vaccine and a treatment for infected people. Several antivirals with activity against SARS-CoV-2 were investigated and since the end of 2021 orally administered antivirals have been used in positive patients. Methods. We conducted a retrospective, single-center study including data from 135 outpatients who resulted positive for SARS-CoV-2 and were selected according to AIFA criteria to receive Molnupiravir (M) or Nirmatrelvir/Ritonavir (N/R) in the InfectiousDiseaseClinic of the TorVergataHospital fromJanuary 2022 to February 2022. Results. Our cohort included 135 patients with a median age of 71 years (IQR 56,5 - 80,5), 51% were male, 91% received M and 9% received N/R. The median time of antiviral administration from symptoms onset was of 3,4 days. 75% of patients were vaccinated with booster dose, 19% were vaccinated with two doses and 7% were unvaccinated. The most frequent criteria of eligibility are summarized in Figure 1. Only 2 patients were hospitalized receiving oxygen support, 1 patient died, 84% of patients did not need hospitalization and 14% of the enrolled subjects were lost at follow up. Time of negativization at the nasopharyngeal swab (NPS) had a negative correlation to the value of anti-Spike (r=-0.29;p=0.01). The difference between cycle threshold (Ct) value ofNPS at T7 and Ct value at the first positive NPS (T0) was higher among not hospitalized (NH) vs hospitalized (H) patients, for Gene E [12.1 (SD 5.8) vs -1.7 (SD 10.8);p=0.02], Gene N [12.3 (SD 10.8) vs -0.9 (SD 11.6);p=0.01] and Gene RdRp [11.8 (SD 5.3) vs -1.5 (SD 11.1);p=0.01]. Anti-Spike were higher among NH vs H patients [1714 (SD 1044.7) vs [43 (SD 60.3);p=0.028]. Delta Ct E, delta Ct N and delta Ct RdRp were significantly higher in patients without neoplastic disease (p=0.04, p=0.02, p=0.01, respectively) and had a negative correlation with creatinine levels (r=-0.36, p< 0.001;r=-0.3, p=0.03;r=-0.32, p=0.02). Percentage of eligibility criteria of our patients The majority of patients had more than one criterion Conclusion. Oral antivirals represent a new effective treatment against COVID-19 in selected patients, reducing the time of negativization and the risk of hospitalization and death. Solid tumor or oncohematologic diseases are associated to a slower negativization of NPS.

Longitudinal Evaluation of the QuantiFERON-TB Gold Plus Assay in Hospitalized COVID-19 Patients with a First Indeterminate Result: Resolution of Inflammation and Restoration T-lymphocyte Counts and Interferon-gamma Production

Background. Several studies reported an increased rate of indeterminate QuantiFERON-TB Gold Plus (QFT-P) assay results in patients with severe Coronavirus Disease (COVID)-19. Methods. Aim of the study was to longitudinally evaluate QFT-P responses in patients who survived COVID-19, with a previous indeterminate result. Results. We observed 223 patients with an indeterminate QFT-P assay among 949 patients hospitalized because of COVID-19 (23,5%) during 2020 and 2021. 36 patients among those with an indeterminate QFT-P assay were enrolled for reassessing the test. In 12 patients peripheral blood lymphocyte subsets were also reassessed. Considering disease severity, 30 were classified as severe and 6 as non-severe. Median age was 57,5 (interquartile range [IQR]: 49,5-63,8), with a prevalence of male sex (M/F: 24/12);median Charlson Comorbidity Index was 2 (IQR: 1-3). The second QFT-P assay was performed after at least 1 month from the first assay (median time 7 months, IQR: 5-12 months). All QFT-P assays gave a determined result: 2 positive (5.5%) and 34 negatives (94,4%). A statistically significant difference was observed after comparing the laboratory parameters at the time of the first and the second QFT-P assay: the absolute counts of total lymphocyte, total CD3+, CD4+ and CD8+ T-lymphocytes were significantly increased (p< 0.001) while neutrophil absolute counts, neutrophil to lymphocyte (N/L) ratio, D-dimer,fibrinogen, ferritin, C-reactive protein (CRP) were significantly reduced (p< 0.0001). Concerning the QFT-P assay, interferon gamma (INF-gamma) production in the Mitogen-Nil, TB1-Nil and TB2-Nil conditions were significantly increased (p< 0.0001;p=0.0019;p=0.0205, respectively) (Table 1 and Figure 1). Conclusion. Once the acute phase of COVID-19 is resolved, inflammatory markers and peripheral blood leucocyte counts tend to normalize with an effective INF-gamma production after specific and nonspecific stimulation. All the 36 QFT-P showed a determinate result. Moreover, we observed 2 positive QFT-P assay, supporting the importance of retesting patients with indeterminate result to identify latent tuberculosis infection and monitor patients for possible reactivation because of the immunesuppression associated with COVID-19.

Monoclonal Antibodies Against SARS-CoV-2 in Fragile Outpatients: Clinical and Laboratory Risk Factors and Protective Role of Vaccination

Background. To cope with the SARS-CoV-2 pandemic, several treatments were studied and out of these, monoclonal antibodies (MAbs) have shown efficacy to prevent the development of pneumonia after the infection Methods. We conducted a retrospective, single-center study including patients with SARS-CoV-2 infection, treated with MAbs (bamlanivimab/etesevimab (B/E), casirivimab/ imdevimab (C/I) or sotrovimab (S)) from March 2021 to February 2022 Results. We included 504 patients with a median age of 62 years (IQR 49-72), 51% were males and 66% had completed the vaccination schedule according to the current Italian regulations. The most frequent eligibility criteria are summarized in figure 1. As for MAbs combination, patients were treated with B/E (54%), followed by C/I (30%) and S (16%). Outcomes are shown in Table 1. Nasopharyngeal swab (NPS) negativization time had a positive correlation with patients' age (r=0.16;p=0.001), C-reactive protein (CRP) (r=0.26;p< 0.001) and creatinine values (r=0.22;p< 0.001) assessed at baseline (infusion day). Time to NPS negativization was 6.9 (95% C.I. [4.5-9.2]) days shorter for vaccinated compared to unvaccinated patients (p< 0.001). Patients treated with C/I had a negative NPS on average 4.5 (95% C.I.= [1.8-7.3] days earlier than patients treated with B/E;patients who received S reached negativization 6.0 (95% C.I.= [2.2, 9.9]) days earlier than those treated with B/E (p=0.004). Patients with positive outcome had a negative NPS on average 14.3 (95% C.I.= [6.8, 23.1)], 25.5 (95% C.I.= [18.9, 33.4] and 68.3 (95% C.I.= [47.7, 90.2]) days earlier than patients who needed hospitalization and patients who died (p< 0.001, p< 0.001, respectively). Unvaccinated patients had a higher rate of oxygen support need compared to vaccinated ones (p=0.006). Patients with worse outcomes were significantly older and had higher values of CRP and creatinine at baseline (p=0.04, p< 0.001, p< 0.001, respectively) Conclusion. MAbs reduce the risk of hospitalization in fragile patients. Vaccinated patients had shorter time of NPS negativization and lower probability of hospitalization. Older age, higher CRP and creatinine values assessed at baseline, correlated with worse outcomes. S was the most effective treatment amongst MAbs used in our study.

Prevalence of hepatitis C virus estimates of undiagnosed individuals in different Italian regions: a mathematical modelling approach by route of transmission and fibrosis progression with results up to January 2021

This study provides an update on hepatitis C virus (HCV) estimates across Italy up to January 2021. A mathematical probabilistic modelling approach, including a Markov chain for liver disease progression, was used to estimate current HCV viraemic burden. Prevalence was defined by geographic area using an estimated annual historical HCV incidence by age, treatment, and migration rate from the Italian National database (ISTAT). Viraemic infection was estimated for the main HCV transmission routes by stages F0-F3 (patients without liver cirrhosis, i.e., potentially asymptomatic liver disease) and F4 (patients with liver cirrhosis, i.e., potentially symptomatic liver disease). By January 2021, we estimated that there were 398,610 individuals in Italy with active HCV infection (prevalence of 0.66%;95% CI: 0.66-0.67), of which 287,730 (0.48%;95% CI: 0.46-0.59%) were stage F0-F3. Prevalence values for all individuals with active HCV infection were: North 0.54% (95% CI: 0.53-0.54%), Central 0.88% (95% CI: 0.87-0.89%), South 0.72% (95% CI: 0.71-0.73%), and the Isles 0.67% (95% CI: 0.66-0.68%). The population at risk for previous/current drug injection accounted for 48.6% of all individuals with active HCV infection. A modelling approach such as this to estimate and update the prevalence of active HCV infection could be a useful methodology for the evaluation of healthcare policies related to HCV elimination plans.

HOW HIV MODULATES the SAFETY and IMMUNOGENICITY of the BNT162b2 COVID-19 VACCINE

Background: The pivotal BNT162b2 trials included only ∼60 vaccine recipients, all with well controlled HIV, and there is a need to gather more information on vaccine safety and immunogenicity in diverse populations. This prospective study evaluated solicited and unsolicited adverse events (AEs) and anti-S and anti-NC serological profiles in a diverse cohort of people with HIV undergoing BNT162b2 vaccination (2 doses 3 weeks apart). Methods: Participants completed structured questionnaires modelled on the BNT162b2 trials (FDA submission, Nov 2020) to report solicited and unsolicited AEs in the 7 days after each vaccine dose, indicating severity and duration. Serum samples collected prior to dose-1 (T0) and 3-6 weeks after dose-2 (T1) underwent qualitative anti-NC and quantitative anti-S testing by Elecsys®. Factors associated with T1 anti-S titres were explored in linear regression models including all available parameters. Results: Overall, 259 adults received dose-1 (26% female, 77% white, 44% MSM, 44% history of advanced disease, 31% ≥1 comorbidity, 10% HIV RNA >50 cps/ml [median 122 cps], 7% prior COVID-19 diagnosis, 15% anti-NC positive;median age 48 years, ART duration 7 years, nadir/current CD4 count 225/708 cells/mm3, CD4:CD8 ratio 0.8);257 received dose-2. Local AEs were more common after dose-1 than dose-2 (70% vs. 62%, p=0.015), whereas systemic AEs increased with dose-2 (50% vs 60%;p=0.006) (Fig 1a-c);22% experienced moderate-severe systemic AEs after dose-2. Unsolicited AEs (mainly nausea and light-headedness) were reported by 7% after dose-1 and 9% after dose-2. Among 206 participants with T1 samples, 205 (99%) had measurable anti-S (>0.8 U/ml). Anti-S levels were significantly lower at CD4 counts <200 cells/mm3 (Fig 1d). In adjusted regression analyses, factors associated with anti-S titres comprised anti-NC positivity (fold-change 7.39;95% CI 3.92-13.91;p<0.01), HIV viraemia (FC 0.24;0.11-0.50;p<0.01), reporting moderate-severe systemic AEs after dose-2 (FC 1.77;1.03-3.04;p=0.04) and either the CD4 count (FC 1.01;1.00-1.01;p=0.04) or CD4:CD8 ratio (FC 1.05;1.00-1.10;p=0.05). Conclusion: In this cohort with HIV, AE patterns after vaccination were similar to those seen in the pivotal BNT162b2 trials and most AEs were mild and short-lived. Whilst prior exposure to SARS-CoV-2 predicted higher anti-S responses, CD4 counts <200 cells/mm3 and low-level viraemia predicted reduced anti-S responses, thus identifying a subset potentially vulnerable to reduced vaccine efficacy.

Ensuring the implementation of healthcare policies able to eliminate hepatitis C at both central and regional levels during the COVID-19 pandemic

Viral hepatitis C is an important public health problem and its elimination by 2030, defined by the World Health Organization, is an ambitious goal. The chance of free screening for HCV infection represents an important achievement that requires a successful State-Regions coordination and an effective regional organisation, that guarantees an interdisciplinary course between local and specialized healthcare. A structured communication program to increase the sensitivity of target populations as well as health professionals is the key for success. The implementation of the proactive screening, defined by the Milleproroghe Law, is crucial because it will define the tracks for the whole HCV costeffective screening strategies (1948-1988 birth cohorts) required to achieve the HCV elimination targets in Italy by 2030.

Low testosterone levels and high estradiol to testosterone ratio are associated with hyperinflammatory state and mortality in hospitalized men with COVID-19.

OBJECTIVE: Evidence supports a sex disparity in clinical outcomes of COVID-19 patients, with men exhibiting higher mortality rates compared to women. We aimed to test the correlation between serum levels of sex hormones [total testosterone, estradiol (E2), estradiol to testosterone (E2/T) ratio, progesterone), prolactin and 25-hydroxyvitamin D [25(OH)D] and markers of inflammation, coagulation and sepsis at admission in hospitalized men with COVID-19. PATIENTS AND METHODS: We conducted an exploratory retrospective study including symptomatic men with confirmed SARS-CoV-2 infection who were consecutively admitted to our Institution between April 1 and May 31, 2020. RESULTS: Patients were divided into survivors (n=20) and non-survivors (n=39). As compared to survivors, non-survivors showed significantly higher median neutrophil-to-lymphocyte ratio (NLR) values, D-dimer and procalcitonin (PCT) levels, along with significantly lower median 25(OH)D levels and total testosterone levels. Non-survivors exhibited significantly higher median values of E2/T ratio (a marker of aromatase activity). Spearman's correlation analysis revealed that total testosterone levels were significantly and inversely correlated with NLR, high-sensitivity C-reactive protein (hsCRP), interleukin-6, D-dimer and PCT. Conversely, E2/T ratio values were significantly and positively correlated with the aforementioned markers and with white blood cell (WBC) count. In a multivariate analysis performed by a logistic regression model after adjusting for major confounders (age, body mass index, hypertension and cardiovascular disease, diabetes mellitus and malignancy), total testosterone levels were significantly and inversely associated with risk of COVID-19-related in-hospital mortality. CONCLUSIONS: Low total testosterone levels and elevated E2/T ratio values at admission are associated with hyperinflammatory state in hospitalized men with COVID-19. Low total testosterone levels at admission represent an independent risk factor for in-hospital mortality in such patients. Therefore, total testosterone and E2/T ratio may serve as prognostic markers of disease severity in this population.

Milestones to reach Hepatitis C Virus (HCV) elimination in Italy: From free-of-charge screening to regional roadmaps for an HCV-free nation

Although Italy has been on track for Hepatitis C Virus (HCV) elimination since 2019, it fell off track due to the decrease in the number of treated patients. HCV elimination in Italy will be possible if immediate action is taken. A health policy was implemented beginning in 2021, consisting of screening among key populations and birth cohorts (1969-1989), estimated to have a high prevalence of undiagnosed individuals. The active screening requires regional governance that manages the processes' complexity integrating a well-organized network between territory assistance and hospital to achieve an effective HCV care cascade. This document aims to support the regional decision-making process by defining paths for screening and linkage-to-care. Implementing active screening strategies beyond a risk-based approach is required as a General Practitioners' task. Simplified paths must be drawn for the key populations screening. The infrastructure built for COVID-19 vaccination could be used also for HCV screening. According to a multidisciplinary care delivery, screening should be supplemented with rapid linkage-to-care and treatment of newly diagnosed patients. The realization of the proactive screening during the first two years is vital because it will define the tracks for the whole HCV cost-effective screening of 1948-1988 birth cohorts in Italy.

Low prevalence of SARS-CoV-2 infection in inflammatory bowel disease.

OBJECTIVE: Treatments used in Inflammatory Bowel Disease (IBD) have been associated with enhanced risk of viral infections and viral reactivation, however, it remains unclear whether IBD patients have increased risk of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection. The aim of the study was to examine the prevalence of SARS-CoV-2 IgG positivity in IBD patients followed at our referral center. The role of treatments for IBD and risk factors for infection were also evaluated. PATIENTS AND METHODS: In a prospective study, all IBD patients followed at our referral centre between May 27th and July 21st, 2020 and fulfilling the inclusion criteria were tested for SARS-CoV-2 IgG. Specific IgG antibodies were evaluated by a commercial ELISA kit and SARS-CoV-2 nasopharyngeal swab was performed in seropositive patients. RESULTS: Two-hundred and eighteen patients, 128 Crohn's disease (CD) and 90 Ulcerative colitis (UC) [age 44, (19-77) years; ongoing biologics in 115 (52.7%)] were enrolled. No patient had major SARS-CoV-2-related symptoms. SARS-CoV-2 IgG were detected in 3 out of 218 (1.37%) patients with IBD (2 CD and 1 UC), all on biologics (2.6%). In all of the 3 seropositive patients, the nasopharyngeal swab was negative. There was no relationship between SARS-CoV-2 seroprevalence and the demographic/clinical characteristics of IBD patients. In contrast, history of recent travel was more frequent in the SARS-CoV-2 seropositive patients (2/3; 66.6%) than in SARS-CoV-2 seronegative patients [7/215 (3.25%); p<0.0001]. CONCLUSIONS: The prevalence of SARS-CoV-2 IgG seropositivity in IBD patients appears to be comparable to the non-IBD population and not influenced by ongoing treatments. Risk factors for infection common to the general non-IBD population should be considered when managing patients with IBD.

Subjective neurological symptoms frequently occur in patients with SARS-CoV2 infection. (Special Issue: Immunopsychiatry of COVID-19 pandemic.)

Objective: Coronavirus disease 2019 (COVID-19) represents a novel pneumonia leading to severe acute respiratory syndrome (SARS). Recent studies documented that SARS-Coronavirus2 (SARS-CoV2), responsible for COVID-19, can affect the nervous system. The aim of the present observational study was to prospectively assess subjective neurological symptoms (sNS) in patients with SARS-CoV2 infection.

High body mass index and night shift work are associated with COVID-19 in health care workers.

OBJECTIVE: To assess the magnitude of COVID-19 spread and the associated risk factors among health care workers (HCWs), we conducted an in-hospital survey in a central Italian COVID Hospital. METHODS: Participants underwent nasopharyngeal swab and/or serum collection for SARS-CoV-2 IgG examination. We divided participants according to working status, into rotating-night shift workers (r-NSW) and day-workers. RESULTS: We found 30 cases of COVID-19 infection in a total of 1180 HCWs (2.5%). Most COVID-19-positive hospital employees were r-NSWs with significantly higher BMI than that of individuals who tested negative. After adjustment for covariates, night work and BMI > 30 were associated with a markedly greater risk of COVID-19 diagnosis (OR 3.049 [95%CI 1.260-7.380] and OR 7.15 [95%CI 2.91-17.51], respectively). CONCLUSIONS: Our results describe a low prevalence of COVID-19 infection among HCWs at a central Italian COVID Hospital. COVID-19 infection risk appears to be associated with obesity and night shift work, thus supporting the need for careful health surveillance among frontline HCWs exposed to COVID-19.