Search

The COVID-19 pandemic has produced an extraordinary care setting where physicians played, and continue to play, a critical role in containing viral spread and treating affected patients. Frontline workers have been receiving day-to-day new information about therapeutic advances. The purpose of the study is to analyse COVID-19 drug consumption trends in both acute and intensive care settings comparing Defined Daily Doses and the release of scientific clinical data from January to December 2020.

A case of hepatitis B virus (HBV) infection in an Eritrean migrant was described to provide an epidemiological approach based on phylogenetic analysis useful in developing countries with lacking information. Migrant, positive for HBsAg and HBeAg, carried HBV at high copy number. A sequence of HBV HBsAg region was used for phylogenetic relationships and genetic variability investigation. In the phylogenetic tree, the sequence corresponded to D2 HBV genotype and the cluster root dated 7 years ago. These data compared with the date of landing in Italy, suggest that he was infected at least 7 years before his arrival. This approach by ‘mirror effect’ allows the reconstruction of HBV epidemiology in the country of origin, analyzing the migrant population in the host country.

Background Few data about predictors and outcomes associated with prolonged SARS-CoV-2 RNA shedding (VS) are available. Methods Retrospective study including all patients admitted with COVID-19 in an Italian reference hospital for infectious diseases between March 1 and July 1, 2020. Predictors of viral clearance (VC) and prolonged VS from upper respiratory tract were assessed by Poisson regression and logistic regression analyses. The causal relation between duration of VS and probability of clinical outcomes was evaluated through inverse probability weighted Cox model. Results 536 subjects were included. Median duration of VS from symptoms onset was 18 days (IQR 12-26). The estimated 30-day probability of VC was 70.2% (95%CI:65-75). At multivariable analysis, patients with comorbidities (aIRR = 0.88, p = 0.004), lymphopenia at hospital admission (aIRR = 0.75, p = 0.032) and with moderate/severe respiratory disease (aIRR = 0.42, p 1000 ng/mL at admission (aOR = 1.76, p = 0.035) independently predicted prolonged VS. The achievement of VC doubled the chance of clinical recovery (aHR = 2.17, p

Background Eritrean migrants accounted for the majority of people who drowned in the Mediterranean this year. Recently, data deficit about international migration has been reported. Methods One hundred and thirty-three Eritrean migrants, upon their arrival in Italy, were accommodated at the asylum seekers center of Castelnuovo di Porto, together with 25 sanitary workers, and received microbiological surveillance at the Clinical Pathology and Microbiology Unit of the University Campus Bio-Medico of Rome. Results Gram-negative bacteria, mostly Klebsiella pneumoniae and Klebsiella oxytoca, were frequently recovered in surveillance swabs. Gram-positive bacteria, represented by Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus dysgalactiae and Staphylococcus aureus, were also isolated in surveillance swabs. In migrants, polymicrobial swabs were recovered at high frequency, and unusual microorganisms, rarely found in our country, were isolated. Conclusion The polymicrobial colonization and the high prevalence of gram-negative bacteria represent the baseline characteristics of this migrant population from Eritrea. The presence of unusual microorganisms can be potentially pathogenic to asymptomatic carriers at risk of developing clinical disease if immunosuppressed or subjected to invasive procedures. In conclusion, active microbiological surveillance can represent an advantage for the host country in terms of data collection and by tracing unusual or resistant microorganisms by monitoring migrants' health status.

Background In recent years Europe has experienced a dramatic increase in migration flows. Nevertheless, limited data is available about the occurrence of neglected parasitic diseases among migrant population. The purpose of the present study was to evaluate the prevalence of intestinal and urinary parasites in newly arrived asylum seekers. Methods A total of 364 newly arrived migrants hosted at the Asylum Seekers Centre of Castelnuovo di Porto (Italy) were screened during 8 months period for intestinal and urinary parasites. Each enrolled subject was interviewed using a standardized questionnaire, with focus on socio-demographical data and risk factors of parasitic infections. Results Stool analysis showed a prevalence of intestinal parasites of 20.6%. The travel route did not affect the prevalence of intestinal parasites (p = 0.096), while a significant negative correlation was found between the length of travel and the prevalence of parasite infection (p = 0.019). No statistically significant correlation between gastrointestinal symptoms and the presence of intestinal parasites was detected. Conclusion The prevalence of intestinal parasitosis reported in asylum seekers does not necessarily reflect the prevalence of the parasitosis in the motherland. An anamnestic and syndromic approach may not be sufficient to highlight the problem of intestinal parasitic infestations in a screening setting.

The armed conflict in Mali caused a migration crisis since 2012. Most Malian refugees were in Italy. In Sub-Saharan Africa, the seroprevalence of anti-HBV antibodies is particularly high. Genotype E is the most prevalent throughout a crescent covering area from Angola to Senegal, including Mali. We report 16 HBV positive individual from 136 Malian asylum seekers in order to investigate the genetic diversity of HBV in this population. Sequencing and phylogenetic analysis has been used. The HBV genotype E isolates from Mali did not cluster together but were intermixed, with the other African sequences. Only three supported clade were evidenced and closely related to sequences from Burkina Faso. The estimated evolutionary rate was 9.29 × 104 . The root of the tree dated back to February 2008 in (95% HPD: 2006-2011). From this ancestor six main statistically supported clusters (pp > 0.80) were identified. The most recent Clade dated back to May 2015. The BSP showed that the effective number of infections softly increased from 2011 to the 2015. Phylogenetic analysis helped in understanding how two on sixteen individuals, have been infected in Italy, and give an important improvement in prevention campaigns and monitoring of the viral infection in migrants. J. Med. Virol. 89:639-646, 2017. © 2016 Wiley Periodicals, Inc.

Background: Parallel upregulation of several T-cell effector functions (ie, polyfunctionality) is believed to be critical for the protection against viruses but thought to decrease in large T-cell expansions, in particular at older ages. The factors determining T-cell polyfunctionality are incompletely understood. Here we revisit the question of cytomegalovirus (CMV)–specific T-cell polyfunctionality, including a wide range of T-cell target proteins, response sizes, and participant ages. Methods: Polychromatic flow cytometry was used to analyze the functional diversity (ie, CD107, CD154, interleukin 2, tumor necrosis factor, and interferon γ expression) of CD4+ and CD8+ T-cell responses to 19 CMV proteins in a large group of young and older United Kingdom participants. A group of oldest old people (age >85 years) was included to explore these parameters in exceptional survivors. Polyfunctionality was assessed for each protein-specific response subset, by subset and in aggregate, across all proteins by using the novel polyfunctionality index. Results: Polyfunctionality was not reduced in healthy older people as compared to young people. However, it was significantly related to target protein specificity. For each protein, it increased with response size. In the oldest old group, overall T-cell polyfunctionality was significantly lower. Discussion: Our results give a new perspective on T-cell polyfunctionality and raise the question of whether maintaining polyfunctionality of CMV-specific T cells at older ages is necessarily beneficial.

Summary Background Three years of civil war in Syria have caused death and increase of communicable diseases. The suffering population has been forced to migrate creating a fertile condition for epidemic spread of infection within the refugee camps. Methods Forty-eight Syrian migrants, upon their arrival in Italy, were accommodated at the asylum seekers centre of Castelnuovo di Porto. They received a physical examination and were subjected to microbiological surveillance by blood, rectal, pharyngeal and nasal swabs collection and delivering to the Clinical Pathology and Microbiology Laboratory of the University Campus Bio-Medico of Rome. Results All refugees resulted negative for HBV, HCV and HIV infections. In swabs a large number of unusual gram-negative bacteria species were isolated, such as Pseudomonas putida , Pseudomonas monteilii , Pseudomonas fulva , Pseudomonas moselii , Aeromonas veronii , Aeromonas caviae , Aeromonas hydrophila , Acinteobacter guilloviae , Acinteobacter lowffii ; Acinetobacter johnsonii ; Acinteobacter tjernbergae ; Pantoea agglomerans ; Pantoea calida . Among isolates, strains resistant to carbapenems, ESBL producers and methicillin resistant were found . Conclusions The microbiological surveillance performed represents a useful action to understand refugees health status and to trace unusual microorganisms movement even carriers of antimicrobial resistance during migrants traveling.

Background HIV infection in women in disproportionate ratios as compared to men has been a grave concern over the years. It is in proportion to reproductive and hormonal differences making women more vulnerable. It elicits an Immune response which can be monitored by analysing various factors such as mucosal immunity, sexual behaviour, biomarkers in the plasma, serum and vaginal lavage and vaginal infections. Aim Evaluating and comparing the systemic and mucosal immuno-inflammatory status, the female sexual function (FSF) and generalized anxiety in HIV+ women on successful HAART with healthy women (HW). Methods We enrolled 53 subjects (23 HIV+ women on successful HAART and 30 Healthy women (HW)) with no statistical differences in age. The figure (named: methodology) below explains the methods applied: Cytometry and Kit ELISA were used to estimate lymphocytes and all cytokines. Women were also tested for co-morbidities such as diabetes, blood pressure, HCV, cervical cancer etc. Statistical analysis was performed using PRISM 8.0. Methodology Results Higher CD4 and CD8 cell count was observed in HW compared to HIV+ women (p=0.02,p=0.004).Plasma levels of sCD 163, CXCL-10, IL-1, IL-6 and IL-8 were significantly higher in HIV women as compared to HW(p< 0.001), while IL-6 and IL8 were lower in the VL of HIV women. An ASCUS in HW was found for PAP Test. CXCL-10 was correlated to estradiol levels (r=0.8, p=0.02). 57% reported FSD and 43% had a FSFI score≤10. A significant difference between the two groups in the FSFI score (p=0.007) was found, particularly in sexual desire, arousal and pain. A positive correlation between level of testosterone and FSFI score was found only in HIV+ women (p=0.02; r= 0.74). 17% of women presented an anxiety disorder. Z-index was associated with orgasm domains (p=0.01; r=-0.4) and CD4+ T cells (p=0.02; r=-0.45). Conclusion Higher plasma levels of the cytokines despite successful antiretroviral therapy were observed. At the mucosal level evaluating the balance within pro anti-inflammatory cytokines and micro-biome will be interesting to study. FSD is detected in more than half of HIV infected women and seems to be related to testosterone levels. The comparison with uninfected women underlying a persistent gap in quality of life of young HIV women should be bridged. Disclosures All Authors: No reported disclosures

Chikungunya fever is caused by Chikungunya virus (CHIKV) and is generally considered a self-limiting disease. However, severe clinical presentations with a high mortality rate have been reported in association with underlying medical conditions. This study reports the molecular characterization of the virus and an abnormal pattern of circulating cytokines in a unique lethal CHIKV case during the 2017 outbreak in Italy, which involved an elderly patient with underlying cardiac disease. Analysis of inflammatory cytokines revealed a strong increase of interferon (IFN)-α and IFN-β, as well as interleukin-6, suggesting a possible role of type-I IFN in the cytokine storm, which may be correlated with unfavorable prognosis of CHIKV infection.

Objectives The management of infectious outbreaks in closed settings represents an important public health issue. An outbreak of acute febrile syndrome affecting 22 refugees resident at the Asylum Seekers Centre of Castelnuovo di Porto in Rome has been reported, and the preventive and control measures adopted have been described as an example of public health safety. Methods Pharyngeal swab and whole-blood samples were collected from 22 cases observed and analyzed for standard bacterial cultures and respiratory and herpesviruses by qualitative CLART PneumoVir2 and Entherpex microarray. Results A possible respiratory-transmitted etiology and a concomitant reactivation of multiple herpesviruses have been evidenced. The epidemiological investigation showed that the spread of the epidemic was promoted because patients were hosted in neighboring rooms or in the same room, facilitating the rapid spread of infectious disease. Conclusions The potential way of transmission was supposed, and preventive measures for infection control were adopted. The measures adopted are an example of best practice for outbreak management, and the microbiological surveillance is recommended for public health improvement., Highlights • Outbreak of acute febrile syndrome affecting refugees resident at an Italian Asylum Seeker Center. • Respiratory-transmitted etiology and a concomitant reactivation of multiple herpesviruses. • Spread of epidemic promoted by patients hosted in neighboring rooms or in the same room. • Preventive measures for infection control adopted to limit the outbreak. • Example of best practice for outbreak management.

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.

Background and aims Measles is an infectious disease that represents a serious public health problem worldwide, being associated with increased susceptibility to secondary infections, especially in the respiratory and gastrointestinal tracts. The aim of this study was to evaluate sCD163 and sCD14 levels in measles virus (MV) infected patients, as markers of immune activation, in order to better understand their role in the pathogenesis of the disease. TNF-α plasma levels were also evaluated. Methods sCD163, sCD14 and TNF-α were measured by ELISA in plasma samples of 27 MV infected patients and 27 healthy donors (HD) included as controls. Results At the time of hospital admission, sCD163 and sCD14 levels were significantly higher in MV infected patients than in HD, while a decrease in TNF-α levels were found even if without statistical significance. sCD163 and sCD14 levels were significantly decreased after two months from acute infection compared to hospital admission although they remained significantly higher compared to HD. TNF-α levels increased significantly during the follow-up period. Considering clinical parameters, sCD163 levels positively correlated with aspartate aminotransferase, white blood cell count and neutrophils rate, while negatively correlated with the lymphocyte percentage. sCD14 levels positively correlated with the neutrophil and lymphocyte percentages. Conclusions These results indicate that, despite the resolution of symptoms, an important macrophage/monocyte activation persists in measles patients, even after two months from infection.

Human immunodeficiency virus type 2 (HIV-2) infection prevalence is increasing in some European countries. The increasing migratory flow from countries where HIV-2 is endemic has facilitated the spread of the virus into Europe and other regions. We describe a case of HIV-2 infection in a migrant individual in the Asylum Seekers Centre (ASC) in Italy. The patient's virus was sequenced and found to be a typical HIV-2 genotype A virus. Bayesian evolutionary analysis revealed that the HIV-2 sequence from migrant dated back to 1986 in a subcluster, including sequences from Guinea Bissau. This was coherent with the history of the migrant who lived in Guinea Bissau from his birth until 1998 when he was 13 years old. Monitoring for HIV-2 infection in migrants from western Africa is necessary using adequate molecular tools to improve the diagnosis and understand the real origin of infection.

Background Despite a large measles outbreak is taking place in WHO European region, currently no data are available on measles immunization coverage in the asylum seeker and migrants hosted in this area. Methods Two hundred and fifty-six migrants upon their arrival in Italy on March, April and May 2016 were screened for measles virus IgG antibodies by chemiluminescence immunoassay (Liaison XL analyzer, Diasorin, Italy). The virus susceptibility in this cohort, the differences between the official country reported and the observed measles immunization coverage and the impact of current measles outbreak on the asylum seekers hosted in the largest Asylum Seeker centres of Italy, were evaluated. Results The prevalence of subjects with positive result for measles IgG antibodies ranged between 79.9% and 100%. In Senegal, Mali, Nigeria, Pakistan and Bangladesh, the measles IgG seroprevalence observed was greater than the vaccinal coverage reported by WHO after I dose of vaccine. Based on data regarding the II dose coverage, the ASs population presented a seroprevalence greater to that expected. Conclusion On the basis of the results obtained, extraordinary screening and vaccination campaigns in the migrant population, especially in the course of large outbreaks, could represent a resource to reach an adequate measles immunization coverage and to control this infectious disease.

Data on diseases' determinants and health status of asylum seekers (ASs) are limited.We performed a cross-sectional retrospective study in a large ASs centre in Italy. Data were collected during a 1-year period. Descriptive statistics were calculated. A χ(2) test was used to assess the association between socio-demographics characteristics of ASs and screening test results. A multiple logistic regression analysis was performed to identify diseases' predictors by using ICD-10 diagnoses classification as outcome variable, socio-demographic characteristics as independent variable and visits' number as confounding variable.Overall, data on 792 ASs (mean age 27 years, 80% males, 58% from Africa) were assessed, 43% underwent voluntary infectious diseases screening and 2843 diagnoses were recorded. The most frequent diagnoses were: respiratory diseases, symptoms/signs not elsewhere classified, digestive diseases and infectious diseases. Gender was the most frequent predictor of ICD-10 diagnoses, while African origin, civil status and education were, respectively, predictive of cardiovascular and infectious diseases, genitourinary diseases and pregnancy-related disorders. Higher mean age was associated with syphilis, HIV and HCV infection and African origin with HIV infection.Communicable diseases were not prevalent in the ASs population we analysed. A stronger cultural mediation support is needed to facilitate prevention, access and continuity of care for ASs.

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus belonging to the Togaviridae family [1]. Chikungunya is a Makonde word (Bantu language) meaning The one which bends up. The term is re...

Background and aims Increased levels of chemokine interferon-gamma (IFN-γ)-inducible protein-10 (CXCL10), soluble CD163 (sCD163) and soluble CD14 (sCD14) have been reported in HCV infection. The aim of this study was to compare, sCD163 and sCD14 levels in HCV-infected patients undergoing direct acting antiviral (DAA)-containing regimens with or without interferon (IFN). Methods sCD163, sCD14 and CXCL10 were longitudinally measured by ELISA in 159 plasma samples from 25 HCV-infected patients undergoing IFN-based treatment plus telaprevir or boceprevir and 28 HCV infected subjects treated with DAA IFN-free regimens. Twenty-five healthy donors (HD) were included as controls. Results At baseline CXCL10, sCD163 and sCD14 levels were higher in HCV-infected patients than in HD. CXCL10 and sCD163 levels were significantly decreased in responder (R) patients who achieved sustained virological response (SVR), with both IFN-based and IFN-free regimens, while they were persistently elevated in non-responders (NR) patients who stopped IFN-based treatments because of failure or adverse events. Conversely, sCD14 levels were apparently unchanged during therapy, but at the end of treatment the levels reached normal ranges. Comparing the two regimens, the extent of CXCL10 reduction was more pronounced in patients undergoing DAA IFN-free therapies, whereas sCD163 and sCD14 reduction was similar in the two groups. Interestingly, only in IFN-based regimens baseline sCD163 levels were significantly higher in NR than in R patients, while in the IFN-free treatment group also patients with high sCD163 plasma levels obtained SVR. At the end of therapy, even if the biomarkers were largely decreased, their levels remained significantly higher compared to HD. Only in the early fibrosis stages, sCD163 values tended to normalize. Conclusions These results indicate that IFN-free regimens including newer DAA induce an early and marked decrease in circulating inflammatory biomarkers. However, the full normalization of biomarkers was not obtained, especially in patients with advanced fibrosis, thus underlying the need for a treatment in the early stages of HCV infection.

Infection with Cytomegalovirus is associated with accelerated immunosenescence. Expansions of CMV-specific T cell responses have previously been demonstrated to affect the ability of T cells to respond to other infections. Most people above 60 years of age display M. tuberculosis-specific immunity because of vaccination, exposure, or both. T-cell responses can be assessed by measuring intracellular IFN-γ in vitro after tuberculin stimulation. Here we investigated tuberculin-specific CD4 T-cell responses in independently living healthy older people in the South of England using flow-cytometry. Individuals were investigated for tuberculin and CMV-specific T-cell immunity using in vitro antigen stimulation followed by intracellular staining for IFN-γ, TNF-α, IL2, as well as degranulation and CD154 upregulation. We also examined a control group of younger individuals (20–35 years of age). There was no significant difference between older and young people in regards to tuberculin responsiveness of CD4 T-cells; however, older people seemed to show more outliers. Increased responsiveness to tuberculin was significantly correlated to CMV responsiveness but not age. In older donors, the memory phenotype of tuberculin-induced T-cells was significantly skewed towards a more terminal differentiation phenotype in CMV-infected compared to uninfected individuals and the degree of skewing correlated quantitatively with the size of the CMV-specific CD4 T-cell response. This is a fundamental advance over previous reports of changes of the tuberculin-specific CD4 T-cell response with CMV serostatus. Our results show that how the immune system responds to CMV has a fundamental impact on the phenotype and function of the immune response to mycobacterial antigens in older life., Highlights • We examine the CD4 T-cell response to tuberculosis antigens in older people. • The CD4 T-cell response to Cytomegalovirus is explored in parallel. • CMV infection changes the profile of the tuberculin-specific-response. • The size of the CMV T-cell response is linked to these changes in a quantitative way. • The way we respond to CMV (‘mode’) affects our T-cell immunity to other pathogens.

Background. Cytomegalovirus (CMV) infection directly targets vascular endothelium and smooth muscle and at older ages is associated with accelerated vascular pathology and mortality. CMV-specific cellular immunity might directly contribute to this process. Methods. Conventional ex vivo activation–induced T-cell responses to 19 dominant CMV antigens, along with CMV-specific inducible regulatory-type CD4+ T cells (iTregs), were measured in healthy older people, using a novel protocol that included classic Treg markers alongside the activation marker CD134. Measurements were correlated with diastolic, systolic, and mean arterial blood pressure, a surrogate marker for arterial stiffness. Results. CMV-specific iTregs recognized the same antigens as conventional CD4+ T cells and were significantly more frequent at older ages. They suppressed antigen-specific and nonspecific proliferation and in large part expressed Foxp3. Frequencies of CMV-specific iTregs and CD8+ T cells (summated response) were significantly associated with diastolic and mean arterial pressures. Confounders, including age, body mass index, smoking, antihypertensive medication use, or C-reactive protein levels, did not explain these observations. Conclusions. A novel CMV-induced regulatory-type CD4+ T-cell subset is readily detectable in CMV-infected people and, like the aggregate CD8+ T-cell response to the most dominant CMV antigens, is quantitatively associated with arterial stiffness in older life. Whereas CD8+ effector T cells might directly cause vascular injury, iTregs may attenuate this response.

International audience; Cytomegalovirus (CMV) infection sometimes causes large expansions of CMV-specific T cells, particularly in older people. This is believed to undermine immunity to other pathogens and to accelerate immunosenescence. While multiple different CMV proteins are recognized, most publications on age-related T-cell expansions have focused on dominant target proteins UL83 or UL123, and the T-cell activation marker interferon-γ (IFN-γ). We were concerned that this narrow approach might have skewed our understanding of CMV-specific immunity at older ages. We have, therefore, widened the scope of analysis to include in vitro-induced T-cell responses to 19 frequently recognized CMV proteins in "young" and "older" healthy volunteers and a group of "oldest old" long-term survivors (>85 years of age). Polychromatic flow cytometry was used to analyze T-cell activation markers (CD107, CD154, interleukin-2 [IL-2], tumor necrosis factor [TNF], and IFN-γ) and memory phenotypes (CD27, CD45RA). The older group had, on average, larger T-cell responses than the young, but, interestingly, response size differences were relatively smaller when all activation markers were considered rather than IFN-γ or TNF alone. The oldest old group recognized more proteins on average than the other groups, and had even bigger T-cell responses than the older group with a significantly larger central memory CD4 T-cell component.

Summary Background and objective Interferon-gamma (IFN-γ)-inducible protein-10 (IP-10), soluble (s) CD163 and sCD14 play an important role in the pathogenesis of HCV and HIV infection and are involved in inflammation and liver fibrosis. The aim of the present study was to evaluate at a single time point, plasma soluble biomarkers and inflammatory monocytes subsets in different groups of subjects: (i) HIV monoinfected patients on suppressive ART; (ii) HIV/HCV coinfected patients on ART, with undetectable HIV viremia (including either subjects who had active HCV replication or those who cleared HCV); (iii) HCV monoinfected individual with active viral replication. Methods Hundred and twenty-nine plasma samples were analyzed including HCV and HIV monoinfected patients, HIV/HCV coinfected patients, with active HCV infection (AHI) or with HCV viral clearance (VHC) and healthy donors (HD). Levels of IP-10, sCD163 and sCD14 were measured by ELISA. Absolute cell counts of monocyte subpopulations were enumerated in whole blood by using flow cytometric analyses. Results IP-10 and sCD163 plasma levels were higher in HCV monoinfected and in AHI coinfected pts compared to HIV monoinfected and HD, whereas sCD14 levels were higher only in HIV monoinfected patients. Considering the degree of fibrosis, sCD163 and sCD14 levels positively correlated with kPa values (as assessed by fibroscan) and FIB-4 in HCV monoinfected group. On the other hand, IP-10 did not correlate with the fibrosis stage and it was found increased also in patients with low fibrosis. Moreover, we found an increase of the inflammatory NCM subset, in non-cirrhotic HCV subjects, while no alterations were observed in HIV, AHI and VHC. Conclusions Our study suggests a scenario in which active HCV infection is associated with a strong pro-inflammatory state, even in the initial stage of liver fibrosis, regardless the presence of HIV coinfection, thus underlying the need of an early anti-HCV treatment.

To the Editor—We read with interest the comments of Manfredi. The aim of our study was to determine the seroprevalence of cytomegalovirus (CMV) antibodies among human immunodeficiency virus (HIV)–positive subjects in the ICONA cohort and to define the impact of CMV serostatus on the risk of AIDS and severe non-AIDS events/death. Although we agree that our analysis has several limitations, mentioned in the discussion section of the article [1], we believe that we have contributed useful new data to the debate about whether CMV infection might be associated with the risk of clinical progression in HIV-infected individuals. CMV is a herpes virus characterized by persistence in a latent state, resulting in lifelong infection [2]. Chronic CMV infection is diagnosed on the basis of detection of persistently positive CMV immunoglobulin G (IgG) antibodies, which can be detected in immunodeficient individuals and elderly individuals, except those with common variable immunodeficiency [3]. There is incontrovertible evidence that CMV infection has an important role in human immunity [4]. As shown by many studies, there is a significant correlation between the increase in CD57+ and CD28− T cells and CMV IgG antibody positivity [5–7]. These cells, characterized by short telomeres, loss of CD28 expression, and/or gain of CD57 expression, play a significant role in various conditions associated with chronic immune activation, such as cancer and autoimmune diseases [8, 9]. Because of these findings, CMV serostatus (and not, for example, T-cell–specific CMV response or level of CMV viremia) was included in the immune risk phenotype markers panel, a cluster of immune markers predictive of increased mortality in elderly individuals. For these reasons, we believe that it is appropriate to use CMV serostatus as a marker of CMV chronic infection, assessed by a standard test for anti-CMV IgG used in clinical practice. Manfredi also pointed out that a limitation of our study is the cumulated time elapsed from the CMV-positive serologic test until the occurrence of end-point events. Our analysis was, indeed, performed using the data from a large cohort of HIV-positive, disease-free Italian subjects monitored for almost 6 years, on average (interquartile range, 2–10 years), until they developed AIDS or a severe non–AIDS-defining event/death. It is also true that a single measurement of CMV antibodies was used in the analysis, which, in most cases, was performed at enrollment (baseline) and that, therefore, potential new incident CMV infections occurring after baseline were ignored. However, previous observations showed that the CMV seroconversion rate over time is low [10], so current exposure might have been potentially misclassified only for a minority of our population. Moreover, although it is true that clinical progression can be determined by many events over follow-up, it was originally hypothesized that would take several years for chronic CMV infection to influence the analyzed clinical outcomes. Indeed, CMV serostatus was evaluated at baseline with the aim of assessing whether it was a predictor of the risk of clinical progression independently of other classical factors (eg, nadir CD4+ T-cell count, Centers for Disease Control and Prevention HIV disease stage, and age). To reduce bias due to these potential confounding factors, in particular for the risk of cardiovascular and cerebrovascular disease, we conducted a multivariable analysis. Moreover, we also explored the potential confounding role of smoking status. In contrast, the interpretation of the association between CMV serostatus, fitted as a time-dependent covariate, and the risk of progression is not free of caveats and potentially requires sophisticated modeling able to correctly control for potential time-dependent confounding factors affected by CMV seroconversion, which was beyond the scope of our analysis. Finally, Manfredi was surprised by the absence of an association between positive anti-CMV serostatus and AIDS events/death. Most people included in the analysis achieved viral suppression during antiretroviral therapy (ART) and showed a good recovery in the CD4+ T-cell count. CMV reactivation with clinical manifestations are typical of severe immunodeficiency, which is uncommon among ART recipients. In fact, the natural history of CMV infection has changed in terms of viral replication and reactivation as a result of suppression of HIV replication and improved patient immunity. In conclusion, we think that our analysis contributes to existing knowledge regarding the role of CMV in HIV disease progression. Indeed, it is the first large cohort study in which CMV/HIV-coinfected individuals were compared to HIV-monoinfected individuals in terms of their risk of future morbidity and mortality. The analysis provided strong evidence that coinfected persons are at higher risk for severe non–AIDS-related events/death than those with HIV monoinfection. Additional studies are needed to investigate the possible mechanisms through which CMV might increase this risk and whether CMV coinfection should be considered for inclusion in scores for the prediction of severe non–AIDS-related events.

Mono- and multifunctional specific CD4+and CD8+T-cell responses were evaluated to improve the immune-based detection of active tuberculosis (TB) and latent infection (LTBI). We applied flow cytometry to investigate cytokines profile (IFN-γ, TNF-α, and IL-2) of T cells after stimulation with TB antigens in 28 TB-infected subjects (18 active TB and 10 LTBI) and 10 uninfected controls. Cytokines production by CD4+T cells at single-cell levels was higher in TB-infected subjects than uninfected controlsP<0.0001. Assigning to activated CD4+T cells, producing any of the three cytokines, a cut-off >0.45%, it was possible to differentiate TB-infected (>0.45%) by uninfected subjects (+T cells, simultaneously producing all 3 cytokines, are lower in active TB than LTBI subjectsP=0.003. Thus, assigning to triple-positive CD4+T cells a cut-off 0.182%). The magnitude of CD8+T-cell responses showed no differences between active TB and LTBI. Multifunctional CD4+T-cell responses could have the potential to identify at single time point subjects without TB infection and patients having active or latent TB.

In HIV-positive patients (pts), CMV co-infection has been proposed as a key factor in sustaining immune activation, which in turn could play a role in determining immune senescence. We evaluated the prevalence and predictors of CMV co-infection in a cohort of HIV+ pts and assessed the impact of CMV co-infection on the risk of AIDS and non-AIDS events. We included pts in the ICONA study with

Large cytomegalovirus (CMV)-specific CD8 T-cell responses are observed in both young and, somewhat more often, old people. Frequent CMV reactivation is thought to exhaust these cells and render them dysfunctional so that larger numbers of them are needed to control CMV. Expansions of CMV-specific CD4 T cells are also seen but are less well studied. In this study, we examined the T-cell response to the dominant CMV pp65 and IE-1 antigens in healthy CMV-infected people across a wide age range (20 to 84 years) by using multicolor flow cytometry. CMV-specific T cells were characterized by the activation markers CD40 ligand (CD40L), interleukin-2 (IL-2), tumor necrosis factor alpha (TNF-α), and gamma interferon (IFN-γ) and the memory markers CD27 and CD45RA. The proportions of effector memory T cells increased in large responses, as did the proportions of polyfunctional CD8 (IFN-γ + IL-2 +/− TNF-α + ) and CD4 (CD40L +/− IFN-γ + IL-2 + TNF-α + ) T-cell subsets, while the proportion of naïve T cells decreased. The bigger the CD4 or CD8 T-cell response to pp65, the larger was the proportion of T cells with an advanced memory phenotype in the entire (including non-CMV-specific) T-cell compartment. In addition, the number of activation markers per cell correlated with the degree of T-cell receptor downregulation, suggesting increased antigen sensitivity in polyfunctional cells. In summary, our findings show that polyfunctional CMV-specific T cells were not superseded by dysfunctional cells, even in very large responses. At the same time, however, the memory subset composition of the entire T-cell compartment correlated with the size of the T-cell response to CMV pp65, confirming a strong effect of CMV infection on the immune systems of some, but not all, infected people.

Background Pediatric tuberculous meningitis is a highly morbid, often fatal disease. Its prompt diagnosis and treatment saves lives, in fact delays in the initiation of therapy have been associated with high mortality rates. Case presentation This is a case of an Italian child who was diagnosed with tuberculous meningitis after a history of a month of headache, fatigue and weight loss. Cerebrospinal fluid analysis revealed a lymphocytic pleocytosis with predominance and decreased glucose concentration. Microscopy and conventional diagnostic tests to identify Mycobacterium tuberculosis were negative, while a non classical method based on intracellular cytokine flow cytometry response of CD4 cells in cerebral spinal fluid helped us to address the diagnosis, that was subsequently confirmed by a nested polymerase chain reaction amplifying a 123 base pair fragment of the M. tuberculosis DNA. Conclusions We diagnosed tuberculous meningitis at an early stage through an innovative immunological approach, supported by a nested polymerase chain reaction for detection of M. tuberculosis DNA. An early diagnosis is required in order to promptly initiate a therapy and to increase the patient’s survival. Electronic supplementary material The online version of this article (doi:10.1186/s13104-017-2444-9) contains supplementary material, which is available to authorized users.

Myeloid dendritic cells (mDCs) play a complex role in HIV infection regardless of viral replication. The aim of our study was to analyse mDCs in long term antiretroviral therapy (ART)- suppressed HIV-infected patients. We evaluated the numbers of mDCs and slanDC in the context of different degree of CD4+ T cell recovery, persistent T cell activation (as HLA-DR+/CD3+ expression) and monocyte-macrophage activation assessed in terms of circulating levels of both sCD14 and sCD163. We enrolled 72 aviremic patients under effective ART and 34 healthy donors (HD). Patients were divided into two groups on the bases of ΔCD4, indicating the difference between the value of CD4 at the time of sampling and CD4 nadir. Higher levels of mDCs and slanDC were found in patients with ΔCD4>200/mmc in comparison to HD. In those patients also an increased level of sCD14 was found, whereas sCD163 seemed to be at normal levels. An augmentation of activated CD4 T lymphocytes was found, although less pronounced in patients with ΔCD4