Problems encountered in conventional HIV 1/2 Algorithms: lack of necessity for immunoblot assays to confirm repeated ELISA reactive results

Background: The use of conventional (serologically based) HIV 1/2 diagnostic algorithms has become controversial in recent years.Objectives: Sera from patients who underwent verification tests were evaluated because repeated ELISA-reactive results demonstrated a HIV1+HIV2 positive band pattern.Methods: The line immunoassay (LIA) test was used for repeated HIV enzyme immunoassays (EIA)-reactive sera in patients at three centers. The Bio-Rad Geenius™ HIV 1/2 and the HIV-1 RNA tests were used. HIV-1 and RNA HIV-2 were investigated using PCR.Results: LIA was used to evaluate 3,224 out of 10,591 samples with repeated ELISA reactivity (30%). We found that 32 (1%) of the sera, along with HIV1 bands and HIV2 gp36 bands, were positive. Only 28 of the 32 verified serum samples with gp36 bands were repeated, and no gp36 band positivity was detected using the Bio-Rad Geenius™ HIV-1/2 confirmatory assay in these serum samples. The HIV-2 proviral DNAs were also negative. Therefore, we excluded the possibility of HIV1+2 co-infection. All samples from the 32 patients were positive for HIV-1 RNA.Conclusion: Our findings highlight the need to exclude confirmatory tests like the LIA test from the current diagnostic HIV algorithm and replace it with rapid HIV-1 and HIV-2 confirmatory immunochromotographic tests.Keywords: HIV, AIDS, HIV-2

BNT162b2 COVID-19 vaccination elicited protective robust immune responses in pediatric patients with inborn errors of metabolism

IntroductionSARS-CoV-2 infection can lead to a life-threatening acute metabolic decompensation in children with inborn errors of metabolism (IEM), so vaccination is mandatory. However, IEMs can also impair innate or adaptive immunity, and the impact of these immune system alterations on immunogenicity and vaccine efficacy is still unknown. Here, we investigated humoral immune responses to the BNT162b2 mRNA COVID-19 vaccine and clinical outcomes in pediatric IEM patients.MethodsFifteen patients between 12-18 years of age with a confirmed diagnosis of IEM, and received BNT162b2 were enrolled to the study. Patients with an anti-SARS-CoV-2 IgG concentration >50 AU/mL before vaccination were defined as “COVID-19 recovered” whereas patients with undetectable anti-SARS-CoV-2 IgG concentration were defined as “COVID-19 naïve”. Anti-SARS-CoV-2 Immunoglobulin G (IgG) and SARS-CoV-2 neutralizing antibody (nAb) titers were measured to assess humoral immune response.ResultsAnti-SARS-CoV-2 IgG titers and nAb IH% increased significantly after the first dose. The increase in antibody titers after first and second vaccination remained significant in COVID-19 naïve patients. Complete anti-SARS-CoV-2 IgG seropositivity and nAb IH% positivity was observed in all patients after the second dose. Vaccination appears to be clinically effective in IEM patients, as none of the patients had COVID-19 infection within six months of the last vaccination.DiscussionHumoral immune response after two doses of BNT162b2 in pediatric IEM patients was adequate and the immune response was not different from that of healthy individuals

Problems encountered in conventional HIV 1/2 Algorithms: lack of necessity for immunoblot assays to confirm repeated ELISA reactive results

Background: The use of conventional (serologically based) HIV 1/2 diagnostic algorithms has become controversial in recent years. Objectives: Sera from patients who underwent verification tests were evaluated because repeated ELISA-reactive results demonstrated a HIV1+HIV2 positive band pattern. Methods: The line immunoassay (LIA) test was used for repeated HIV enzyme immunoassays (EIA)-reactive sera in patients at three centers. The Bio-Rad Geenius\u2122 HIV 1/2 and the HIV-1 RNA tests were used. HIV-1 and RNA HIV-2 were investigated using PCR. Results: LIA was used to evaluate 3,224 out of 10,591 samples with repeated ELISA reactivity (30%). We found that 32 (1%) of the sera, along with HIV1 bands and HIV2 gp36 bands, were positive. Only 28 of the 32 verified serum samples with gp36 bands were repeated, and no gp36 band positivity was detected using the Bio-Rad Geenius\u2122 HIV-1/2 confirmatory assay in these serum samples. The HIV-2 proviral DNAs were also negative. Therefore, we excluded the possibility of HIV1+2 co-infection. All samples from the 32 patients were positive for HIV-1 RNA. Conclusion: Our findings highlight the need to exclude confirmatory tests like the LIA test from the current diagnostic HIV algorithm and replace it with rapid HIV-1 and HIV-2 confirmatory immunochromotographic tests

Antibody Response to inactive SARS-CoV-2 vaccination in a cohort of elderly patients living with obesity

Introduction Obesity and aging negatively affect the immune system and host defense mechanisms, increasing vulnerability to, worsening prognosis of infectious diseases and leading to vaccine failure. Our aim is to investigate the antibody response against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) spike antigens, and the risk factors affecting antibody levels in elderly people living with obesity (PwO) after inactive SARS-CoV-2 vaccine (CoronaVac) administration. Methods One hundred twenty-three consecutive elderly patients with obesity (age>65 years, Body Mass Index (BMI)>30kg/m2) and 47 adults with obesity (age 18-64 years, BMI>30kg/m2) admitted between August and November 2021 were enrolled. Seventy five non-obese elderly people (age >65 years, BMI 18.5-29.9 kg/m2) and 105 non-obese adults (age 18-64 years, BMI 18.5-29.9 kg/m2) were recruited from subjects who visited Vaccination Unit. SARS-CoV-2 spike-protein antibody titers were measured in patients with obesity and non-obese controls who received two doses of CoronaVac. Results SARS-CoV-2 levels of patients with obesity were found to be significantly lower than those of non-obese elderly individuals who had non-prior infection.There was no difference in SARS-CoV-2 levels between patients with obesity and non-obese individuals with prior infection. Age and SARS-CoV-2 level were found to be highly correlated in the correlation analysis in the group of elderly individuals (r:-0.184). In multivariate regression analysis, when SARS-CoV-2 IgG was regressed on age, sex, BMI, Type 2 Diabetes Mellitus (T2DM) and Hypertension (HT), HT was found to be an independant factor on SARS-CoV-2 level (β:-2730). Conclusion In non-prior infection group, elderly patients with obesity generated significantly reduced antibody titers against SARS-CoV-2 spike antigen after CoronaVac vaccine compared to non-obese people. It is anticipated that the results obtained will provide invaluable information about SARS-CoV-2 vaccination strategies in this vulnerable population. Antibody titers may be measured and booster doses should be delivered accordingly in elderly PwO for optimal protection

Risk factors influencing seroconversion after inactive SARS-CoV-2 vaccination in people living with obesity

Introductio

Comparison of SARS-CoV-2 Antibody Levels after a Third Heterologous and Homologous BNT162b2 Booster Dose

This study aimed to determine the anti-S (receptor binding protein) RBD IgG antibody titers formed against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2) and the neutralizing antibody inhibition percentages (nAb IH%) in blood samples taken after two doses of inactive or mRNA-based vaccine and a booster dose. Volunteers with two doses of inactivated CoronaVac (heterologous group; n = 75) and BioNTech (BNT)162b2 mRNA vaccine (homologous group; n = 75) were included in this study. All participants preferred the BNT162b2 vaccine as a booster dose. First, peripheral blood samples were taken 3 months after the second vaccine dose. Second, peripheral blood samples were taken 1 month after the booster dose. Anti-S-RBD IgG titers were determined by CMIA (SARS-CoV-2 IgG II Quant). Neutralizing antibodies were detected by a surrogate neutralization assay (SARS-CoV-2 NeutraLISA, Euroimmun, Lübeck, Germany). The median age of the volunteers was 40 (IQR 29–47) years old. After the heterologous booster dose, anti-S-RBD IgG levels and neutralizing antibodies increased approximately 50-fold and 9-fold, respectively. Anti-S-RBD IgG titers increased by 9 and 57 times, respectively, while nAb IH% increased by 1.5 and 16 times, respectively, among those with heterologous reminder doses and those with and without a prior history of coronavirus disease (COVID-19). This study showed that after the administration of a heterologous booster dose with BNT162b2 to those whose primary vaccination was with inactivated CoronaVac, the binding and neutralizing antibody levels were similar to those who received a homologous BNT162b2 booster dose. It was observed that the administration of heterologous and homologous booster doses resulted in the development of similar levels of neutralizing antibodies, independently from a prior history of COVID-19

Waning of SARS-CoV-2 Vaccine-Induced Immune Response over 6 Months in Peritoneal Dialysis Patients and the Role of a Booster Dose in Maintaining Seropositivity

Introduction: Although lower than general population, newly developed SARS-CoV-2 vaccines generate immune responses in end-stage kidney disease patients. However, the persistence of immune responses in the long term is not known yet. This study aimed to evaluate humoral immune responses in peritoneal dialysis (PD) patients over 6 months and to analyze the effects of the booster dose. Methods: Humoral immune responses of PD patients were measured after initial SARS-CoV-2 vaccinations and after 6 months following initial vaccinations. Immune responses were compared between patients who received and did not receive booster doses. PD patients were compared with 41 hemodialysis (HD) patients and 61 healthy controls. Humoral immune responses were measured by a commercial test that detects antibodies toward the receptor-binding domain of the spike protein of SARS-CoV-2. Results: Twenty PD patients were evaluated over 6 months. The initial seropositivity rate was 90.9% with inactivated vaccine and 100% with mRNA vaccine. Seropositivity decreased to 44.4% after 6 months, and a booster dose helped in maintaining the 100% of seropositivity (p = 0.005). Magnitude of humoral response at the 6th month was also higher in patients who received the third dose (1,132.8 +/- 769.6 AU/mL vs. 400.0 +/- 294.6 AU/mL; p = 0.015). Among patients who did not receive the third dose, those who got mRNA vaccine could maintain higher seropositivity than others who got inactivated vaccine (75% vs. 40% for PD, 81.8% vs. 50% for HD). Seropositivity and antibody levels were similar for PD and HD patients after 6 months (p = 0.24 and 0.56) but lower than healthy controls (p = 0.0013). Conclusion: SARS-CoV-2 vaccine-induced antibody levels and seropositivity of PD patients significantly fall after 6 months. A booster dose after around 3 months following initial immunization might help in maintaining seropositivity

Phytochemical profiling of Salsola tetragona Delile by LC-HR/MS and investigation of the antioxidant, anti-inflammatory, cytotoxic, antibacterial and anti-SARS-CoV-2 activities

This study aimed to investigate the phytochemical composition and biological activity of Salsola tetragona Delile. (Amaranthaceae), a medicinal plant. The study evaluated the antioxidant potential of the crude extract and five fractions of S. tetragona using DPPH•, ABTS•+, CUPRAC, and metal chelating assays. The anti-inflammatory activity was determined using a protein denaturation assay, and the antibacterial activity was determined by the Minimum inhibitory concentrations (MICs) for the growth of Escherichia coli and Staphylococcus aureus strains. The MTT test and an in vitro scratch assay evaluated the effects on cell viability and cell migration. The potential anti-SARS-CoV-2 activity was assessed by analyzing the effects on the interaction between ACE2 and Spike protein. The bioactive compounds present in the plant were identified using LC-HR/MS analysis. The crude hydromethanolic extract (STM) and five fractions of S. tetragona, n-hexane (STH), dichloromethane (STD), ethyl acetate (STE), n-butanol (STB), and aqueous (STW) showed significant antioxidant activity in four different tests. In the anti-inflammatory assay, the ethyl acetate fraction exhibited significantly higher activity than Aspirin® (IC50 = 13 ± 5 µg/mL). The crude extract and its fractions showed positive antibacterial activity with similar MICs. In the cytotoxicity assay against the breast cancer cell line MCF7, the dichloromethane fractions (STD) were very effective and demonstrated superiority over the other fractions (IC50 = 98 µg/mL). Moreover, the potential of the extract and fractions as anti-SARS-CoV-2, the ethyl acetate, and dichloromethane fractions demonstrated important activity in this test. LC-HR/MS analysis identified 16 different phenolic compounds, Eleven of which had not been previously reported in the genus Salsola. The results suggest that the extracts of S. tetragona have the potential to become new sources for developing plant-based therapies for managing a range of diseases

Neopterin and soluble CD14 (sCD14) levels as immunoactivation markers in aHBc-only cases

Aim: Isolated hepatitis B core antibodies (aHBc)-only pattern complicates the diagnosis of HBV infections. We evaluated neopterin and sCD14 levels in HBV infections. Methods: aHBc-only (n: 102), healthy control (healthy control group [HCG], n: 100), and chronic hepatitis (CHB) groups (n: 70) were investigated. Competitive and sandwich ELISA were used. Results: The mean neopterin levels were significantly lower in the aHBc-only group than those in the CHB group (p = 0.0001). No significant difference was found between the aHBc-only group and the HCG (p = 0.854). The mean sCD14 levels were lower in the aHBc-only group than those in the CHB group (p = 0.0001), but no significant difference was found between the aHBc-only group and HCG (p = 0.402). No significant difference was detected between aHBc-only and HCG for mean sCD14 (p = 0.402) and neopterin levels (p = 0.854). Conclusion: These two biomarkers are not useful for diagnosing the aHBc only pattern

Helicobacter pylori-miRNA interaction in gastric cancer tissues: First prospective study from Turkey

Helicobacter pylori (H. pylori) is involved in the etiology of gastric cancer (GC). miRNAs are short RNAs that regulate gene expression by marking mRNAs for degradation. miRNAs are involved in tumorigenesis, metastasis, and cell proliferation. We aimed to investigate the miRNA expression profiles of tissues from H. pylori Wand (-) GC patients. Forty GC patients, 20 H. pylori Wand 20 H. pylori (-), and a healthy control group were included. The miRNA expression levels were investigated by microarrays and quantitative RT-PCR. We detected 9 upregulated and 4 downregulated miRNAs by microarray. We selected 5 upregulated and 5 downregulated miRNAs for the quantitative RT-PCR assay. The relative fold changes of miRNAs in the cancerous tissue and non-tumor mucosa specimens of H. pylori (+) GC patients for hsa-miR-194 were 4.24- and 3.83-fold higher, respectively, whereas the hsa-miR-145 expression levels were downregulated 0.33-fold and 0.43-fold, respectively, in the same group. The presence of H. pylori significantly upregulated hsa-miR-194 and downregulated hsa-miR-145 expression levels in H. pylofi(+)GC cases, compared to H. pylori (-) GC cases. Regional differences in the virulence of H. pylori strains may also be involved in the up- or downregulation of miRNA expression levels