The neutrophil/lymphocyte ratio as a prognostic factor in COVID-19 patients: a case-control study.

OBJECTIVE: SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) has been identified in China as responsible for viral pneumonia, now called COVID-19 (Coronavirus Disease 2019). Patients infected can develop common symptoms like cough and sore throat, and, in severe cases, acute respiratory syndrome and even death. To optimize the available resources, it is necessary to identify in advance the subjects that will develop a more serious illness, therefore requiring intensive care.The neutrophil / lymphocyte ratio (NLR) parameter, resulting from the blood count, could be a significant marker for the diagnosis and management of risk stratification. PATIENTS AND METHODS: A retrospective, single-center case-control observational study was conducted. The differential cell count of leukocytes, the NLR and the clinical course of patients hospitalized in intensive care with COVID-19 were analyzed, comparing them with other patients (COVID-19 and non-COVID-19) and healthy individuals selected among workers of the Teaching Hospital Policlinico Umberto I in Rome. RESULTS: 370 patients (145 cases and 225 controls) were included in the case-control study, 211 males (57%) and 159 females (43%). The average age of the population was 63 years (SD 16.35). In the group of cases, out of 145 patients, 57 deaths and 88 survivors were recorded, with a lethality rate of 39.3%. The group of cases has an NLR of 7.83 (SD = 8.07), a much higher value than the control group where an NLR of 2.58 was recorded (SD = 1.93) (p <0.001). The Neutrophils / Lymphocytes ratio may prove to be a diagnostic factor for COVID-19, an NLR> 3.68 revealed an OR 10.84 (95% CI = 6.47 - 18.13) (p <0.005). CONCLUSIONS: The value of NLR considered together with the age variable allows a risk stratification and allows the development of diagnostic and treatment protocols for patients affected by COVID-19. A high neutrophil to lymphocyte ratio suggests worse survival. Risk stratification and management help alleviate the shortage of medical resources and reduce the mortality of critically ill patients.

How are the undocumented migrants in Rome? Assessment of quality of life and its determinants among migrant population.

Aim: The aim of this study is to evaluate the level of Health-Related Quality of Life (HRQoL) and its determinants among migrants in irregular situations in Italy. Methods: This cross-sectional study was held in Rome in 2014. HRQoL was assessed through SF-12 questionnaire and physical (PCS) and mental component scores (MCS) were calculated; socio-demographic information and medical conditions were collected. Bivariate and multivariate analyses were performed to assess the impact of demographic and pathological variables on the HRQoL. Results: The median PCS among the 200 migrants enrolled was 46.5 and the median MCS was 37.9, some points below the Italian average. The multivariate analysis revealed a negative association between PCS and age (P < 0.01), respiratory (P: 0.03) and Poverty-Related Diseases (PRDs) (P < 0.01). MCS, on the other hand, resulted negatively associated with neuropsychiatric diseases (P: < 0.01) and PRDs (P < 0.01). Conclusion: Although multivariate analyses revealed that gender acts as an effect modifier the negative association between PRDs and the two dimensions of HRQoL is confirmed in both genders. This suggests a great impact of socio-economic status on the HRQoL. Public health could contribute to improve the HRQoL of migrants only taking into account social aspects of diseases and tailoring intervention on the specific needs of migrants.

In vitro effect of anti-human immunodeficiency virus CCR5 antagonist maraviroc on chemotactic activity of monocytes, macrophages and dendritic cells.

Compounds targeting the chemokine receptor CCR5 have recently been approved for treatment of human immunodeficiency virus (HIV) infection. Given the central role of CCR5 in inflammation and recruitment of antigen-presenting cells (APC), it is important to investigate the immunological consequences of pharmacological inhibition of CCR5. We evaluated the in vitro effect of different concentrations of CCR5 antagonist maraviroc (MVC) on cell migration of monocytes, macrophages (MO) and monocyte-derived dendritic cells (MDC) towards peptide formyl-methionyl-leucyl-phenylalanine (fMLP) and chemokines regulated upon activation normal T cell expressed and secreted (RANTES) and CCL4/macrophage inflammatory protein-1 (MIP-1ß) and CCL2/monocyte chemotactic protein-1 (MCP-1). Results of flow cytometric analysis showed that monocytes treated in vitro with MVC exhibited a significant dose-dependent reduction of chemotaxis towards MIP-1ß and MCP-1. fMLP-induced chemotactic activity decreased only at higher concentration (1 µM and 10 µM of MVC). In addition, all concentrations of MVC (0·1, 1 and 10 µM) induced in vitro a significant inhibition of chemotaxis of MO and MDC in response to all tested chemoattractants. No change in phenotype (CD1a and CD14) and CCR1, CCR4, CCR5 and formyl peptide receptor (FPR) expression was seen after in vitro treatment with MVC. These findings suggest that CCR5 antagonist MVC may have the in vitro ability of inhibiting the migration of innate immune cells by mechanism which could be independent from the pure anti-HIV effect. The drug might have a potential role in the down-regulation of HIV-associated chronic inflammation by blocking the recirculation and trafficking of MO and MDC.

Adherence and genotypic drug resistance mutations in HIV-1-infected patients failing current antiretroviral therapy.

The aim of this study was to assess the correlation between the estimated adherence of HIV-1 infected patients with antiretroviral (ARV) therapy failure and drug-resistant mutations. We studied 40 patients with virological and immunological ARV-therapy failure. In order to assess the adherence of patients we used the SERAD questionnaire. Genomic sequencing of the HIV-1 pol gene was performed. 100% adherence was reported by 27 patients (67.5%) (adherent patients). Multivariate analysis showed that only baseline and nadir CD4+ counts maintained a significant correlation with the adherence. For PR and NNRTI mutations, we did not find any difference between the two groups of patients. Baseline NRTI mutations were higher in adherent patients than in non-adherent patients (p<0.05). No differences were found between plasma mutations and PBMC mutations. The authors conclude that genotypic resistance mutations were found in the majority of patients with ARV-therapy failure despite a good self-reported adherence to therapy. Adequate adherence to therapy is not the only key factor in viral suppression.

A seven-year prospective study on spondylodiscitis: epidemiological and microbiological features.

BACKGROUND: The aim of this paper was to enlarge the available knowledge on clinical and etiological aspects of patients affected by spondylodiscitis. PATIENTS AND METHODS: All patients with spondylodiscitis admitted between January 2001 and December 2007 at the 1,300-bed University Hospital "Policlinico Umberto I" of Rome, Italy, were followed. Demographic characteristics, underlying diseases, invasive procedures, imaging studies, isolated microorganisms, treatment, complications, and outcome were recorded. RESULTS: Eighty-one patients of mean age 57.7 +/- 14.7 years with lumbosacral (72.8%), thoracic (14.8%), and cervical tract (12.3%) site of infection were included, of which 38 developed community-acquired (CA) spondylodiscitis and 43 developed hospital-acquired (HA) spondylodiscitis. Underlying disease was present in 49.4% of patients. HA spondylodiscitis was diagnosed earlier (46.8 +/- 49.7 days) than CA spondylodiscitis (65.0 +/- 55.4 days) (P < 0.05). The most frequently isolated microorganisms were Staphylococcus aureus (28 strains, 43.1%), coagulase-negative staphylococci (CNS) (eight strains, 12.3%), Pseudomonas aeruginosa (eight strains, 12.3%), and three methicillin-resistant S. aureus (MRSA) strains were isolated in CA spondylodiscitis. Fungi and yeasts, isolated in six patients, represented 9.2% of all strains but 17.6% when considering only HA spondylodiscitis. Over 85% of patients were managed by conservative treatment alone, and the treatment time depended on clinical and laboratory evidence. Poor outcome was recorded in 12 (14.8%) patients, and was associated with neurological deficit symptoms (relative risk [RR] 2.87; 95% confidence interval [CI] 1.02-8.07; P < 0.05) and the time between diagnosis and the onset of symptoms > or = 60 days (RR 2.65; 95% CI 0.92-7.59; P < 0.05). CONCLUSIONS: Infectious spondylodiscitis affects most frequently the elderly population, who are more exposed to healthcare contacts. Consequently, the infection etiology includes a growing proportion of multi-resistant bacteria and fungi.

Circulating dendritic cells and interferon-alpha production in patients with tuberculosis: correlation with clinical outcome and treatment response.

Dendritic cells (DC) have been characterized recently as having an important role in the initiation and control of immunological response to Mycobacterium tuberculosis infection. Blood DC have been subdivided into myeloid (mDC) and plasmacytoid (pDC) subsets, on the basis of differences in phenotype markers and function. Little is known about the enumeration and functional evaluation of circulating DC in patients with tuberculosis and their correlation with clinical outcome during the course of anti-tuberculous treatment. We assessed circulating mDC and pDC counts measured by a newly developed single-platform flow cytometric assay based on TruCOUNT, as well as the production of interferon (IFN)-alpha after in vitro stimulation by herpes simplex virus (HSV-1) in 24 patients with active tuberculosis (TB) and 37 healthy donors. Absolute numbers of both DC subsets were decreased significantly in patients with active TB compared to controls. Similarly, the production of IFN-alpha was highly impaired. In 13 patients these parameters were assessed longitudinally, before and after the specific anti-microbial treatment. Most interestingly, in all nine patients with successful anti-tuberculous therapy there was a significant and marked increase of pDC counts and IFN-alpha production. In contrast, no significant longitudinal variations in DC counts and IFN-alpha production were observed in four patients with lack of response to specific treatment. In conclusion, active TB is associated with a defect in blood DC numbers and IFN-alpha production that is restored after bacterial clearance and clinical improvement, as a result of effective anti-tuberculous treatment.

Ex vivo and in vitro effect of human immunodeficiency virus protease inhibitors on neutrophil apoptosis.

Polymorphonuclear leukocytes (PMNL) from human immunodeficiency virus (HIV)-infected patients exhibit accelerated apoptosis and impaired functional activity. HIV protease inhibitor-based therapy produces improvements in both acquired and innate immune responses. Ex vivo and in vitro effects of HIV protease inhibitors on apoptosis and chemotaxis of PMNL were evaluated. After therapy, there was a rapid and significant decrease of PMNL apoptosis, which correlated with increased chemotactic function. These findings were found both in patients with immunological and virological response and in control subjects who showed an increase in CD4(+) T cell counts but no concomitant decline in HIV load. After in vitro treatment with ritonavir or indinavir, apoptosis of both HIV-infected and -uninfected PMNL markedly decreased and correlated with significant enhancement of chemotaxis. These results suggest that HIV protease inhibitors may improve the PMNL function by reducing the apoptosis rate and that this effect may, at least in part, be independent of their antiviral activity.

Interleukin-15 enhances neutrophil functional activity in patients with human immunodeficiency virus infection.

Polymorphonuclear leukocyte (PMN) dysfunction has been reported in human immunodeficiency virus (HIV)-infected patients. Interleukin (IL)-15 is a recently discovered cytokine that potentiates antimicrobial functions of normal PMNs. We evaluated the in vitro effect of IL-15 on chemotaxis and fungicidal activity of PMNs from 9 patients with untreated advanced HIV infection, 8 patients with viral suppression after 52 to 130 weeks of highly active antiretroviral therapy (HAART), and 12 patients with treatment failure. We also studied oxidative burst and apoptosis of PMNs in 5 patients with untreated advanced HIV infection. Twelve healthy donors were included as controls. Chemotaxis and fungicidal activity of unprimed PMNs was significantly lower in patients with untreated HIV infection compared with controls. After incubation with IL-15, a significant increase in PMN chemotaxis and fungicidal activity was found; moreover, IL-15 induced a significant reduction in the number of apoptotic HIV(+) PMNs. IL-15 did not modulate oxidative burst of HIV(+) PMNs as measured by chemiluminescence production. The in vitro priming of PMNs with IL-15 determined a complete reversal of defective chemotaxis and killing in all HAART-treated patients with long-term HIV suppression. IL-15 significantly enhanced chemotaxis and fungicidal activity also in patients with HAART failure. In conclusion, IL-15 is an important cytokine in the activation of the functional properties of HIV(+) PMNs, by delaying apoptosis and enhancing chemotaxis and fungicidal activity. The potent stimulant effect of IL-15 on PMN function was observed in antiretroviral naive patients as well as in individuals who were receiving HAART, including those with treatment failure. (Blood. 2000;96:1979-1984)

Changes in circulating levels of soluble cell adhesion molecules following highly active antiretroviral treatment of HIV-1-infected patients.

Increased levels of soluble cell adhesion molecules (sCAM) have been reported in HIV-1 infection and may possibly contribute to altering the adhesion mechanisms of phagocytic cells. We evaluated the effect of highly active antiretroviral therapy (HAART) on plasma levels of sL-selectin, sE-selectin, intercellular cell adhesion molecule-1 (sICAM-1), sICAM-3, and vascular cell adhesion molecule-1 (sVCAM-1). Study participants included 22 HIV-1-infected patients with a CD4+ T-cell count/microl below 500 who were started on a HAART regimen and followed up for 9 months. After the initiation of therapy, plasma sL-selectin concentrations progressively decreased to normal ranges in the majority of our patients (P < 0.001), while no changes in sE-selectin were found. In all patients sICAM-1 remained relatively constant at significantly elevated concentrations during the 9 months of therapy. A significant reduction in plasma concentrations of both sICAM-3 and sVCAM-1 was found; however, the levels of these sCAM were not normalized by HAART and remained significantly elevated throughout the study (P < 0.001). The reduced release of sL-selectin could improve the ability of phagocitic cells to migrate in response to chemotactic stimuli after starting HAART. On the other hand, the persistent elevation of sICAM-1, sICAM-3, and sVCAM-1 could reflect continuous HIV-1-mediated immune activation, despite adequate control of plasma HIV-1 replication by therapy.

Chlamydia pneumoniae seroprevalence among HIV-1-infected and uninfected people with known HIV risk factor.

OBJECTIVE: To evaluate the prevalence of Chlamydia pneumoniae antibodies in an Italian population of HIV-infected and uninfected individuals in relation to the presence of HIV risk factors. DESIGN AND METHODS: A prospective evaluation of C. pneumoniae microimmunofluorescence immunoglobulin (Ig) G and IgM titres, in relation to sex, age, HIV clinical stage, and the presence of different HIV acquisition risk factors. SETTING: The Department of Infectious and Tropical Diseases, a secondary and tertiary care institution in the 'La Sapienza' University of Rome, during 1994 and 1995. PARTICIPANTS: HIV-infected and uninfected subjects (n = 322), all of them without respiratory symptoms. RESULTS: A statistically significant higher C. pneumoniae seroprevalence was found to be related, by multivariate analysis, to sex, age, and presence of HIV risk factor, but not to the presence of HIV infection itself. Among HIV-positive subjects, C. pneumoniae seroprevalence appeared to decrease with absolute CD4+ cell count and was low in Centers for Disease Control and Prevention (CDC) stage C of HIV infection. Furthermore, high C. pneumoniae IgG titres (> or = 1:512) were not found in subjects with CDC stage C disease or in those with low CD4+ cell count (< 200 x 10(6)/l). CONCLUSION: C. pneumoniae seroprevalence is higher in injecting drug users and in subjects with promiscuous heterosexual activity. A previous report of a higher C. pneumoniae seroprevalence among HIV-1-infected subjects (in relation to the normal population) was probably due to the presence of HIV risk factor and not to the HIV infection itself. HIV-infected subjects seem to have progressively lost their C. pneumoniae IgG antibodies in middle and advanced stages of HIV infection. High C. pneumoniae IgG titres are rarely found in advanced stage HIV-infected patients.

Overexpression of monosialoganglioside GM3 on lymphocyte plasma membrane in patients with HIV infection.

SUMMARY: This study was undertaken to analyze both the GM3 expression on peripheral blood lymphocytes of HIV-infected patients and the relationship between ganglioside content and anti-GM3 reactivity. GM3 expression was determined as a percentage of lipid-bound sialic acid and by cytofluorimetric analysis in 25 AIDS patients, 20 anti-HIV+ asymptomatic subjects, 25 patients with different viral disease, and 25 healthy donors. GM3 distribution was analyzed by immunofluorescence and immunoelectron microscopy. A follow-up study to detect anti-lymphocytic GM3 antibodies was performed in progressive and nonprogressive anti-HIV+ subjects. Lymphocytes from HIV-infected patients showed a significant increase of plasma membrane GM3 content; no difference was found between CD4+ and CD8+ cells. Immunofluorescence and immunoelectron microscopic analysis showed that GM3 was distributed in large clusters over the cell plasma membrane. The follow-up study revealed that the occurrence of anti-lymphocytic GM3 antibodies was significantly higher in patients with progressive disease, compared with asymptomatic non-progressive subjects. These findings revealed that (1) the increased GM3 content in HIV-infected patients is detected at the plasma membrane level, (2) GM3 overexpression is able to induce an increased reactivity with anti-GM3 antibodies, and (3) the appearance of anti-lymphocytic GM3 antibodies in asymptomatic anti-HIV+ subjects could have prognostic relevance for the risk of developing AIDS.

Myelin degrading activity in the CSF of HIV-1-infected patients with neurological diseases.

Degradation of purified myelin basic protein (MBP) was studied by SDS gel electrophoresis after addition of CSF samples obtained from HIV-1-infected patients. An increase in MBP degradation was detected in patients with neurological complications, such as AIDS dementia complex (ADC) or progressive multifocal leukoencephalopathy (PML), when compared with patients with no neurological symptoms (NA) or with other neurological opportunistic infections (OI). In the ADC and PML patients, in addition to CSF proteolytic activity, an increase in CSF-MBP levels and presence of white matter lesions were also observed by neuroimaging (MRI). In other opportunistic infections of the brain, MBP levels but not anti-MBP proteolytic activity increased. Results suggest the involvement of proteases in the virus-induced demyelination.

Autoantibodies against ganglioside GM3 represent a portion of anti-lymphocyte antibodies in AIDS patients.

In this study we analysed the relationship between anti-lymphocytic ganglioside antibodies and anti-lymphocyte antibodies in AIDS patients. Anti-lymphocytic ganglioside antibodies were detected by thin layer chromatography (TLC) immunostaining; three colour flow cytometry was used to analyse circulating antibodies against different lymphocyte subsets. Anti-lymphocytic ganglioside antibodies were detected in 23 out of 49 AIDS patients sera (46.9%). All positive sera reacted selectively with the GM3 comigrating band from AIDS lymphocytes. Twenty two out of the 23 anti-lymphocytic GM3 positive sera also had antibodies against CD4+T cells, versus 17/26 anti-GM3 negative. Furthermore, patients with lymphocytic GM3 antibodies showed a significantly higher antibody reactivity against CD4+ T cells than patients in which these antibodies were not detected. The absorption tests revealed that preincubation of positive sera with GM3 was followed by a decrease in the reaction with target lymphocytes. These findings suggest that anti-GM3 antibodies are a portion, but not the majority, of antibodies reacting with CD4+ T cells.

Tumour Necrosis Factor-alpha Production and Immune Cell Activation in Tuberculous Meningitis.

The local production of tumour necrosis factor-alpha (TNFalpha) was evaluated in the cerebrospinal fluid (CSF) from ten patients with tuberculous meningitis (TBM). The degree of intrathecal immune activation was also studied by assessing the CSF levels of beta(2)-microglobulin (beta(2)-M) and adenosine deaminase activity (ADA). Results indicate that elevated CSF concentrations of TNFalpha, beta(2)-M and ADA were found in all TBM patients. Moreover, TNFalpha is produced and selectively concentrated for a long period of time, while beta(2)-M and ADA values progressively decline during the course of TBM. Our findings suggest that in TBM patients, after an early activation of immune cells, there is an enhanced and continuous production of TNFalpha at the site of infection.