ABSTRACT
Background: The severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) is a highly contagious illness associated with a hyperactivated and dysregulated host immune humoral response. In detail, there is a cytokine storm which may take to the release of interleukin IL-6 as a critical mediator for respiratory failure, shock and multiorgan dysfunction. Such dysregulation may act as a target for therapeutics and, in this view, a blockade of IL-6 function by an anti-IL-6 receptor antibody (tocilizumab) has been described to be effective for the treatment of the inflammatory process COVID-19-related. Timing of administration of therapy was reported in literature to have a critical role in benefit for patients;thus, the aim of the present study is to compare two different methods for the IL-6 assessment: the Human IL-6 ELISA Kit (Invitrogen) and the Elecsys IL-6 (Roche). Method(s): IL-6 levels of 128 COVID-19 patients, who were consequently admitted to the Emergency and Medicine Department of AOU Careggi (Hospital in Florence -Italy ) between April and May 2020, were assessed by using the two above mentioned methods and were analysed through Passing-Bablok regression fit and Bland-Altman plot. Result(s): The analyses showed that the two methods correlate, but do not agree in terms of numeric results. In particular, further investigation were performed on the Bland-Altman results, showing that the maximum number of samples for which the differences between the two methods is close to "0" (p > 0.05) (which means a good overlap between the two methods) is 49 (p=0.07), and among them, 40 samples showed a complete agreement of results (p=0.95). These results can be attributed to the different methods' linearities: 3.1-200 pg/mL for ELISA and 1.5#5000 pg/mL for ECLIA, which could be extended to 50 000 pg/mL. Conclusion(s): Although a small percentage of data overlapping in a certain range, still a high correlation among the two methods can be found;given the overall analytical performance of the ECLIA, it can be considered more adequate for different reasons: i) it is available on a fully automated platform h24, ii) it uses of a small sample volume, iii) it is low cost and no-time consuming and iiii) the different timing for measuring IL-6 is much attractive.
ABSTRACT
Introduction: Since 2020 COVID-19 pandemic has spread throughout the world and became an ongoing global health crisis due to SARS-CoV-2 virus. Elderly and pre-existing disorders including hypertension, heart problems, diabetes, cancer, autoimmune diseases and IBD are found associated with an increased risk of COVID-19. Although COVID-19 leads to mild flu-like symptoms in the majority of patients, the disease may cause severe complications and death. To date, a few clinical studies suggested that IBD and/or immunomodulation may reduce the susceptibility to COVID-19;however, the mechanisms through which this is happening is largely unknown. Aims & Methods: Aim of this study is to investigate the effects of IBD and different therapies on the risk of SARS-CoV-2 infection and COVID-19 severity through serum proteomics and metabolomics. Between April 2020 and April 2022, 238 IBD patients (N=145 Crohn disease, N=93 Ulcerative colitis) and 45 healthy controls (HC) of the North Italy area were enrolled and serum samples were collected. To evaluate the exposure to SARSCoV- 2, both clinical data were collected and seroprevalence of anti-SARSCoV- 2 Ab were analyzed by means of multiplex technology, the BioPlex 2200 Sars-Cov-2 IgG Panel (biorad, Italy). Serum samples underwent untargeted metabolomics analysis and the frequency of a serum metabolomics signature associated with protection were evaluated in IBD compared to HC and also between anti-TNF and Vedolizumab biological therapies for IBD patients. Result(s): The seroprevalence of anti-SARS-CoV-2 Ab in IBD cohort (22/238) indicates an overall lower incidence of COVID-19 in comparison with the general population of Lombardy. Our data indicated that IBD patients in treated with biologic drugs as anti-TNF (10,5%) and Vedolizumab (7,5%) have a lower incidence than IBD patients treated with conventional therapies (28,0%). Accordingly, we observed that serum metabolomics signature associated with protection was more frequent in IBD patients treated with anti-TNF (N=50, 70%), and with Vedolizumab (N=57, 85%) than healthy controls (N=45, 50%). The metabolomic protective profile is characterized by the presence of fat-soluble Tocopherols family members and Cholecalciferol and also of omega-3 and omega-6 polyunsaturated fatty acid. Conclusion(s): Our study indicates that IBD population treated with biologics has an overall lower risk to contract SARS-CoV-2 infection and a serum proteomic/metabolomic protection profile. The increased presence in IBD patients of radical scavengers such as tocopherols which are incorporated into cell membranes and protect against oxidative damage and anti-inflammatory and immunomodulating fatty acids suggest a better response to SARS-CoV-2 infection. Also increased levels of omega;-3 interfere with the entry of the virus by modulating the Lipid Rafts where ACE2 and TMPRSS2 are mainly expressed and PUFAs inhibit the attachment of SARS-CoV-2 virions to the human ACE2 receptor by interacting directly with the RBD sequence. Mechanistically understanding how this protection profile exerts its effects on COVID-19 severity might shed light on potential targets to increase resistance in higher risk subgroups of patients.
ABSTRACT
Background: The current novel coronavirus (SARS-CoV-2) pandemic is an ongoing global health crisis, which represents an important challenge for the whole society and mankind. Patients with inflammatory bowel disease (IBD) are treated with immunosuppressive drugs that are usually associated with more severe viral infections. However, the effects of the different therapies on the risk of SARS-CoV-2 infection and Covid-19 severity in IBD patients are still under investigation. Methods: Between April 2020 and April 2021, 238 IBD patients (N=145 with Crohn disease and N=93 with Ulcerative colitis) of the North Italy area have been enrolled. Both serum samples (N=238 IBD patients and N=45 healthy donors) and colon biopsies from inflamed and non-inflamed mucosa (N=88 IBD patients N=20 non-IBD control) have been collected. To evaluate the exposure to SARS-CoV-2, both clinical data and seroprevalence of anti-SARS-CoV-2 Ab have been analyzed. Serum samples were analyzed by untargeted metabolomics analysis and the frequency of a serum metabolomics signature associated with protection were evaluated in IBD versus healthy donors. Moreover, gene expression analysis of key proteins for virus entry (ACE2, TMPRSS2, TMPRSS4, ADAM17) were analyzed by qPCR in the gut mucosa biopsies of IBD patients. Results: The seroprevalence of anti-SARS-CoV-2 Ab in our cohort of IBD patients (10/238) indicates an overall lower incidence of COVID-19 in comparison with the general population of Lombardy. Accordingly, we observed that the serum metabolomics signature associated with protection was more frequent in IBD patients treated with anti-TNF (N=50, 70%), than healthy controls (N=45, 50%). Gene expression analysis of the proteins involved in SARS-CoV-2 entry also indicated that IBD patients treated with anti-TNF (N=14) had a lower mucosal level of SARS-CoV-2 receptor ACE2 and its sheddase ADAM17 than non-IBD subjects along with higher expression of the proteases TMPRSS2 and TMPRSS4. Moreover, vedolizumab-treated patients (N=7) showed a significant lower expression of ACE2, TMPRSS2 and TMPRSS4 than controls, whereas ADAM17 levels were similar. Conclusion: Our study indicates that IBD population treated with biologics has an overall lower risk to contract SARS-CoV-2 infection. Future studies to gather the mechanisms underpinning the effects of biologics on the expression of the proteins involved in SARS-CoV-2 viral entry in association with the specific metabolomics signature of viral susceptibility might shed light on potential targets to increase resistance in higher risk subgroups of patients.
ABSTRACT
Background: Homocysteine (Hcy) has been reported as a potential predictive biomarker for CoViD-19 infection severity in many studies. Hyper-homocysteinemia is related to many virus infection outcomes, including HEV, HPV and HIV. Recent data confirmed the value of Hcy in predicting the risk of severe pneumonia. Materials and Methods: Our retrospective cohort study, including 313 CoViD-19 hospitalized patients (female 34.8%;mean age 62 years), also included a broad panel of clinical laboratory data collected. Of the enrolled patients, 10.9% died during hospitalization (3% were transferred to other hospitals and were lost to follow-up). Results: Hcy was found to be the strongest predictor of CoViD- 19 critical-progression leading to death. Univariate analysis demonstrated that age (OR 1.04), Hcy (OR 1.06), and neutrophil/lymphocyte count ratio (OR 1.03) were significant predictors of critical progression leading to death and RBC (OR 0.68) and lymphocytes count (OR 0.23) with benign outcome. ROC analysis indicated Hcy cut off of 16 μmol/L for predicting CoViD- 19 infection outcome (sensitivity 40% and specificity 84%);patients with Hcy levels >16 μmol/L had significantly increased risk of in-hospital mortality (p=0.002) both as a continuous and dichotomic value. Conclusions: Our results demonstrate that Hcy is an effective predictive biomarker for hospitalized CoViD-19 patients' outcome. Hcy may be a valuable biomarker to help clinicians to identify patients who are at higher risk for severe CoViD-19 infection.
ABSTRACT
The immune system plays a pivotal role in COVID-19 infection, being responsible for the clinical manifestations and prognosis of the disease. Hyperactivation of the immune response against SARS-CoV-2 virus may result, in some cases, in the development of autoimmune disorders. In fact, a robust immune response is involved in the pathogenesis of both disease conditions. The molecular mimicry, the presence of circulating autoantibodies and the overlapping of autoimmune diseases in some patients show the potential trigger effect of SARS-CoV-2 virus on human immunity in developing autoimmune disorders. A variety of systemic or organ-specific manifestations have been reported to be associated with COVID-19, particularly: antiphospholipid antibody syndrome, Kawasaki Syndrome, Guillain-Barrè Syndrome, Miller Fisher Syndrome, idiopathic thrombocytopenic purpura, and autoimmune hemolytic anemia. However, the existing evidence, which differs according to the specific disease, is poor mainly because it is based on case reports and case series without a long-term follow-up. Since it is still unknown whether these autoimmune conditions may represent a transient phenomenon post-infection, more data are needed to thoroughly understand the relationship between COVID-19 and autoimmunity.
ABSTRACT
Background: Inadequate knowledge about extent of coronavirus disease 2019 (COVID19) epidemic challenges health response and planning. COVID19 mortality among cancer patients (pts) is higher than in general population. The identification of asymptomatic COVID19 cancer pts is important from both a personal and a health system point of view as immunosuppression increases COVID19 disease severity. Screening for asymptomatic carriers is being tested in some categories. The best method for screening and monitoring is yet unknown. Material and methods: Azienda Usl Toscana Centro Oncology department tested pts receiving chemo-or radio-therapy. Aims were checking the prevalence of asymptomatic COVID19 cancer pts and evaluating the need and the best method for subsequent monitoring. Pts were offered both a quantitative serologic IgM and IgG test (Qt-ST) and a RT-PCR test for SARS-CoV-2 in nasopharyngeal and oropharyngeal swabs (NOS). As the Qt-ST is costly and requires hours for response, in S.M. Annunziata Hospital (SMA), pts were also tested with a qualitative serologic IgM and IgG method (Ql-ST) to collect information on different serologic assays. Results: Between May 11th and 17th, 1148 pts receiving an active antineoplastic treatment signed a written informed consent and were screened with RT-PCR for SARS-CoV-2 in NOS and Qt-ST. 317 pts in SMA were also tested with Ql-ST. 16 pts refused RT-PCR, 3 Qt-ST and none Ql-ST. 89 pts with haematological malignancy didn't receive ST. Only 0.44% (5/1132) of asymptomatic cancer pts had a positive RT-PCR. 1 of them had COVID19 disease and was declared healed 14 days before screening. All pts with a positive RT-PCR in NOS had a positive Qt-ST for IgM. 1/5 had also IgG positivity. Qt-ST was positive in 26/1145 pts (2.3%): 3 were IgM+/ IgG+, 10 IgM+/IgG-and 13 IgM-/IgG+. 19.2% of pts with a positive Qt-ST (5/26) were asymptomatic carriers of COVID19 disease. In SMA, 6/317 (1.9%) had a positive Qt-ST, 21 (6.6%) a positive Ql-ST and all were negative for RT-PCR in NOS. Conclusions: With a positivity of 0.44%, RT-PCR for SARS-CoV-2 in NOS may not be cost-effective for screening in asymptomatic cancer pts under antineoplastic treatment from a patient point of view. However, RT-PCR may improve both the compliance and the safety's sense of pts and operators in hospital and is mandatory in case of ST positivity. Qt-ST is more accurate than Ql-ST. Both Qt-ST and Ql-ST were safe and may be a proper option for monitoring cancer pts based on local organization.
ABSTRACT
Diagnostic tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, named also COVID-19) infection are always more widespread and constantly evolving. The WHO provides the reverse-transcription polymerase chain reaction (RT-PCR) as laboratory testing strategy recommendation for COVID-19 performed in upper and lower respiratory specimens.SARSCoV-2 infection can also be detected indirectly through serological testing;however, systematic data on the diagnostic performance and the predictive value of these tests are still lacking. The aim of this study was to evaluate the role of serological tests and their predicitive values, in combination with RT-PCR, to support diagnosis of COVID-19. We conducted a retrospective study on 637 patients admitted to the Emergency Department and Internal Medicine Ward of S.Giuseppe Hospital (Empoli,Italy) for suspected COVID-19. All sera were first tested by rapid test for qualitative detection of anti-SARSCoV-2 IgG/IgM. Sera positive for IgG and/or IgM were subsequently tested for the quantitative detection using a chemiluminescent method. Of the 637 patients, 59 had a COVID-19 diagnosis;104 were positive for anti-SARSCoV-2 IgG/IgM antibodies by a qualitative test and 57 for the viral RNA by RT-PCR, with a concordance rate between the two tests of 24% (25/104). Moreover, 32 out of the 57 patients with positive RT-PCR were negative for anti-SARS-CoV-2 IgG/IgM antibodies by qualitative test with a concordance rate of 56.1% and, on the contrary, two COVID-19 patients were negative for RT-PCR but positive for anti-SARS-CoV-2 antibodies. Double positive tests (qualitative and quantitative) over the total number of positive results ranged from 27% to 37%, according to the single isotype or combination of antibodies used. The best odd ratio (OR) for associations of anti-SARS-CoV-2 positive tests with COVID-19 was reached by combining 3 or 4 tests (OR:12.9-22.7). Since our results showed a weaker association between SARS-CoV-2 infection and the positive single qualitative test (31-37%), compared to the double positive qualitative test or the double, triple, and quadruple qualitative and quantitative tests (63-90%), the significance of the qualitative test should be evaluated according to the isotype detected and to its combination with the quantitative test.