Antibody and memory B cell responses to the dengue virus NS1 antigen in individuals with varying severity of past infection

To further understand the role of NS1-specific antibodies (Abs) in disease pathogenesis, we compared neutralizing antibody levels (Nabs), NS1-Ab levels, IgG antibody subclass profiles and NS1-specific memory B-cell responses (Bmems) in individuals, with varying severity of past dengue. Nabs (Neut50 titres) were assessed using the Foci Reduction Neutralization Test (FRNT) and in-house ELISAs were used to assess NS1-Abs and NS1-Ab subclasses for all four DENV serotypes in individuals with past DF (n = 22), those with past DHF (n = 14) and seronegative (SN) individuals (n = 7). B-cell ELISpot assays were used to assess NS1-specific Bmem responses. 15/22 (68.18%) individuals with past DF and 9/14 (64.29%) individuals with past DHF had heterotypic infections. Neut50 titres were found to be significantly higher for DENV1 than DENV2 (p = 0.0006) and DENV4 (p = 0.0127), in those with past DHF, whereas there was no significant difference seen in titres for different DENV serotypes in those with past DF. Overall NS1-Ab to all serotypes and NS1-specific IgG1 responses for DENV1, 2 and 4 serotypes were significantly higher in those with past DHF than individuals with past DF. Those with past DHF also had higher IgG1 than IgG3 for DENV1 and DENV3, whereas no differences were seen in those with past DF. Over 50% of those with past DF or DHF had NS1-specific Bmem responses to >2 DENV serotypes. There was no difference in the frequency of Bmem responses to any of the DENV serotypes between individuals with past DF and DHF. Although the frequency of Bmem responses to DENV1 correlated with DENV1-specific NS1-Abs levels (Spearman r = 0.35, p = 0.02), there was no correlation with other DENV serotypes. We found that those with past DF had broadly cross-reactive Nabs, while those with past DHF had higher NS1-Ab responses possibly with a different functionality profile than those with past DF. Therefore, it would be important to further evaluate the functionality of NS1-specific antibody and Bmem responses to find out the type of antibody repertoire that is associated with protection against severe disease.     

Heparin-induced thrombocytopenia: evaluation of IgG and IgGAM ELISA assays

Background: Heparin‐induced thrombocytopenia (HIT) is a rare complication of heparin therapy resulting from antibody production to platelet factor 4 and heparin complexes (H‐PF4). Methods: We have evaluated four enzyme‐linked immunosorbent assay (ELISA)‐based screening tests to identify the best assay(s) with the highest specificity but without underdiagnosis of HIT. As functional assays are difficult to perform, ELISAs are useful to provide clinicians with a timely answer. Over a 10‐month period, all samples (N = 107) referred to our laboratory were tested for HIT antibodies using four commercially available ELISA kits, two detecting IgG/A/M anti‐H‐PF4 antibodies and the other two IgG specific. Results: Twenty‐eight samples were positive by at least one assay; IgGAM ELISAs were found to be more sensitive with 24 samples positive by Asserachrom IgGAM and 23 by Zymutest IgGAM. Only 18 samples were positive by GTI‐PF4‐IgG and Zymutest IgG. The gold standard serotonin release assay (SRA) was used as a confirmation assay, and 11/28 samples tested positive. All these SRA‐positive samples were positive by all four assays. None of the IgGAM‐only‐positive samples was found to be positive by SRA suggesting a better specificity for the IgG‐only assays. Conclusion: Our data strongly support the use of IgG‐only assays for the detection of HIT antibodies.     

Self-Selection Bias in an Epidemiological Study of Respiratory Health of a Rural Population

ABSTRACT

A common source of bias in epidemiological studies is self-selection or volunteer bias. The self-selection into cohorts for further investigation or for participation in a substudy may be an important source of selection bias. The objectives of this paper were to identify differences in selected demographic characteristics, individual and contextual factors, and variables related to respiratory health between study participants who were willing to participate in subsequent clinical and allergy testing and those who were not willing to participate. One hundred and fourteen households (207 individuals) from a small town and 54 households (99 individuals) from a nearby rural municipality participated in this study. One key informant from each household provided information about each adult in that household. The question being studied was: “We wish to find out more about respiratory health of rural people. We would like to invite you to perform breathing and allergy tests. Would you be willing to be contacted for breathing and allergy tests in a nearby location?” One hundred and four participants said “Yes” they would participate in the clinical studies, 144 said “No” they would not participate in the clinical studies, and 52 said “Would like more information” about the clinical studies. More than one half (53.8%) of male participants and 46.2% of female participants indicated that they would like to participate in breathing and allergy tests. A higher proportion of study participants (26.1% males, 30.8% females) in the lowest income category requested more information compared to those answering either “Yes” (15.7% for males, 20.5% for females) or “No” (18.5% males, 23.3% females) to the question being studied. Study participants who were willing to participate in further breathing and allergy tests had a higher proportion of self-reported chronic phlegm and ever had allergic reaction to things eaten than those who either said “No” or “Would like more information.” Among male study participants who said “Yes” to further participation, a higher proportion was exposed to one of the occupational exposures of interest compared to those who said either “No” or “Would like more information.” This pattern was not observed for females.     

Finite Element Analysis of Steel Plates with Rectangular Openings Subjected to Axial Stress

Steel plates with openings are among the important ship structural components used in the ship’s hull to withstand the hydrostatic forces of the ocean, which cause sagging and hogging moments at the ship’s bottom. The existence of openings on plates can cause structural rupture, stress concentration and a decrease in ultimate strength. This research is aimed at investigating the influence of selected parameters on the ultimate capacity of steel plates with rectangular holes subjected to axial stress, using ANSYS finite element analysis (FEA) under its non-linear static structural programme. The main parameters investigated in this paper are the plate thickness, opening aspect ratio, number of openings, position of openings, and the boundary condition of the plate. The influence of these parameters on the stress of plates and their deformation was evaluated. The comparison of the numerical simulation with the well-established analytical method using the Navier solution and Roark’s Formulas showed a good agreement.     

Identification of Novel Candidate CD8+ T Cell Epitopes of the SARS-CoV2 with Homology to Other Seasonal Coronaviruses

Cross-reactive T cell immunity to seasonal coronaviruses (HCoVs) may lead to immunopathology or protection during SARS-CoV2 infection. To understand the influence of cross-reactive T cell responses, we used IEDB (Immune epitope database) and NetMHCpan (ver. 4.1) to identify candidate CD8⁺ T cell epitopes, restricted through HLA-A and B alleles. Conservation analysis was carried out for these epitopes with HCoVs, OC43, HKU1, and NL63. 12/18 the candidate CD8⁺ T cell epitopes (binding score of ≥0.90), which had a high degree of homology (>75%) with the other three HCoVs were within the NSP12 and NSP13 proteins. They were predicted to be restricted through HLA-A*2402, HLA-A*201, HLA-A*206, and HLA-B alleles B*3501. Thirty-one candidate CD8⁺ T cell epitopes that were specific to SARS-CoV2 virus (<25% homology with other HCoVs) were predominantly identified within the structural proteins (spike, envelop, membrane, and nucleocapsid) and the NSP1, NSP2, and NSP3. They were predominantly restricted through HLA-B*3501 (6/31), HLA-B*4001 (6/31), HLA-B*4403 (7/31), and HLA-A*2402 (8/31). It would be crucial to understand T cell responses that associate with protection, and the differences in the functionality and phenotype of epitope specific T cell responses, presented through different HLA alleles common in different geographical groups, to understand disease pathogenesis.