Identification of a HSP14‐3‐3 in Setaria cervi and its cross‐reactivity with W bancrofti ‐infected human sera

2020 ◽  
Vol 42 (11) ◽  
Author(s):  
Faiyaz Ahmad ◽  
Ranjeet Kumar ◽  
Sarika Gupta ◽  
Sushma Rathaur
Keyword(s):  
Cross Reactivity ◽  
Setaria Cervi ◽  
Human Sera

scholarly journals Peptide arrays incubated with three collections of human sera from patients infected with mosquito-borne viruses

F1000Research ◽  
2020 ◽  
Vol 8 ◽  
pp. 1875
Author(s):  
Maria del Pilar Martinez Viedma ◽  
Nurgun Kose ◽  
Leda Parham ◽  
Angel Balmaseda ◽  
Guillermina Kuan ◽  
...  
Keyword(s):  
Vaccine Development ◽  
Cell Epitope ◽  
Cross Reactivity ◽  
Human Pathogens ◽  
Peptide Arrays ◽  
Serological Tests ◽  
Array Data ◽  
Prevalence Studies ◽  
Diagnostic Potential ◽  
Human Sera

Background: Global outbreaks caused by emerging or re-emerging arthropod-borne viruses (arboviruses) are becoming increasingly more common. These pathogens include the mosquito-borne viruses belonging to the Flavivirus and Alphavirus genera. These viruses often cause non-specific or asymptomatic infection, which can confound viral prevalence studies. In addition, many acute phase diagnostic tests rely on the detection of viral components such as RNA or antigen. Standard serological tests are often not reliable for diagnosis after seroconversion and convalescence due to cross-reactivity among flaviviruses. Methods: In order to contribute to development efforts for mosquito-borne serodiagnostics, we incubated 137 human sera on individual custom peptide arrays that consisted of over 866 unique peptides in quadruplicate. Our bioinformatics workflow to analyze these data incorporated machine learning, statistics, and B-cell epitope prediction. Results: Here we report the results of our peptide array data analysis, which revealed sets of peptides that have diagnostic potential for detecting past exposure to a subset of the tested human pathogens including Zika virus. These peptides were then confirmed using the well-established ELISA method. Conclusions: These array data, and the resulting peptides can be useful in diverse efforts including the development of new pan-flavivirus antibodies, more accurate epitope mapping, and vaccine development against these viral pathogens.

Concentrations of 2-hydroxyoestrogens in human sera measured by a heterologous immunoassay with an 125I-labelled ligand

1982 ◽  
Vol 100 (1) ◽  
pp. 154-160 ◽  
Author(s):  
D. Berg ◽  
F. Thaler ◽  
E. Kuss
Keyword(s):  
Standard Deviation ◽  
Binding Sites ◽  
Luteal Phase ◽  
Healthy Subjects ◽  
Cross Reactivity ◽  
Follicular Phase ◽  
Underlying Assumption ◽  
Human Sera ◽  
Tube Accuracy ◽  
Serum Components

Abstract. A heterologous immunoassay for 2-hydroxyoestrogens1 has been established in which antibodies raised against 2-hydroxyoestradiol-17-succinyl-BSA serve as binding protein and 2-hydroxyoestrone-17-cmo-[125I]iodohistamine as radioligand. Lipophilic serum components competing for binding sites in this system were defined as 'total 2-hydroxyoestrogens'. The underlying assumption of specificity was supported by the pattern of cross-reactivity evaluated with structural related steroids and o-diphenols and by the fact, that an additional chromatography of the serum extracts preceding the competing reaction had little if any effect. Sensitivity: 2.8 ± 1 pg/tube; accuracy: Y = 0.91x + 2.2; r = 0.989; precision: 5.8% intra-assay; 6.5% inter-assay. The following concentrations ( ± standard deviation) were found in the sera of healthy subjects. Young men: 29 ± 5 pg/ml (n = 11); women follicular phase: 32 ± 8 pg/ml (n = 25); luteal phase: 53 ± 13 pg/ml (n = 23); postmenopausal women: 13 ± 4 pg/ml (n = 10); pregnant women 11th–20th week: 70 ± 16 mg/ml (n = 64); 36th–40th week: 240 ± 23 pg/ml (n = 40); newborn cord blood: 604 ± 43 pg/ml (n = 48).

scholarly journals Antibodies in human sera to oncorna virus-like proteins from normal or leukemia marrow cell cultures.

1976 ◽  
Vol 144 (5) ◽  
pp. 1243-1253 ◽  
Author(s):  
S Louie ◽  
J E Curtis ◽  
J E Till ◽  
E A McCulloch
Keyword(s):  
Lupus Erythematosus ◽  
Competitive Inhibition ◽  
Marrow Cell ◽  
Leukemia Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cross Reactivity ◽  
Antibody Activity ◽  
Human Sera ◽  
Blood Specimens ◽  
Radioimmunoprecipitation Assay

Some human marrows in culture release particles with oncornavirus-like properties. This study was designed to examine the immunological properties of similar particles in human marrow culture supernates. Leukemic and nonleukemic marrows were cultured for 5-7 days in the presence of [14C]uridine and [3H]leucine or [3H]glucosamine. Labeled supernatant components banding in sucrose gradient densities of 1.20-1.24 g/ml were used as antigen in a double antibody immunoprecipitation assay. The assay was validated by end point titrations and competition with unlabeled antigen; purified myeloma proteins were used as negative controls. Cross-reactivity with mammalian oncornaviruses, as judged by competitive inhibition of precipitation by these viruses, was slight and at the border of the sensitivity of the method. Precipitated antigens analyzed by SDS polyacrylamide gel electrophoresis contained three distinct polypeptides of about 70,000, 45,000 and 30,000 mol wt; these comigrated with the gp 70, pg 45, and p 30 of a murine leukemia virus. Similar polypeptides were obtained from both leukemic and nonleukemic marrow culture supernates. As determined by the radioimmunoprecipitation assay, 32 of 45 leukemic sera (71%), 36 of 45 normal sera (80%), 15 of 19 sera from family contacts of leukemic patients (79%), 14 of 21 cord blood specimens (67%), and 21 of 23 sera (91%) from patients with systemic lupus erythematosus had detectable antibody activity.

Measurement of serum bone gla-protein (BGP) in humans with an ovine BGP-based radioimmunoassay

1990 ◽  
Vol 36 (9) ◽  
pp. 1620-1624 ◽  
Author(s):  
P Pastoureau ◽  
P D Delmas
Keyword(s):  
Amino Acids ◽  
Primary Hyperparathyroidism ◽  
Renal Osteodystrophy ◽  
Clinical Investigation ◽  
Gel Filtration ◽  
Normal Subjects ◽  
Conformational Epitope ◽  
Cross Reactivity ◽  
Bone Gla Protein ◽  
Human Sera

Abstract Most RIAs of serum bone gla-protein (BGP; also called osteocalcin) used for clinical investigation are based on bovine BGP for standard, tracer, and immunogen because of the homology between bovine and human BGP. However, ovine BGP differs from human BGP by only five amino acids, being identical from residues 11 to 49, as compared with homology at residues 20-49 between bovine and human BGP. In screening various anti-ovine BGP polyclonal anti-sera we selected one (R310) that exhibits apparently complete cross-reactivity with human BGP, as assessed by dilutions of 13 human sera from normal subjects and from patients with bone disease. This RIA gave a 42% binding at a 10,000-fold final dilution, with intra- and interassay variations less than 7% and 11%, respectively. Gel-filtration chromatography of human serum showed a single immunoreactive peak. Synthetic fragments of human BGP 1-10, 7-19, 25-37, and 37-49 were not recognized by R310, suggesting that either a mid-molecule region or a conformational epitope was its target. Using this RIA, we determined that serum BGP increased with age in women (P less than 0.02), by a mean of 90% from ages 30 to 70 years. Serum BGP was also increased in patients with primary hyperparathyroidism, renal osteodystrophy, and Paget's disease. In contrast with the "normal" concentrations of BGP detected with an anti-bovine BGP antiserum (R102), serum BGP was increased in patients with postmenopausal osteoporosis as measured with the R310 ovine assay, suggesting a greater sensitivity for the latter assay.

scholarly journals Two-Way Antigenic Cross-Reactivity between Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Group 1 Animal CoVs Is Mediated through an Antigenic Site in the N-Terminal Region of the SARS-CoV Nucleoprotein

2007 ◽  
Vol 81 (24) ◽  
pp. 13365-13377 ◽  
Author(s):  
Anastasia N. Vlasova ◽  
Xinsheng Zhang ◽  
Mustafa Hasoksuz ◽  
Hadya S. Nagesha ◽  
Lia M. Haynes ◽  
...  
Keyword(s):  
Amino Acid ◽  
Severe Acute Respiratory Syndrome ◽  
Antigenic Site ◽  
Amino Acid Identity ◽  
Cross Reactivity ◽  
Acid Identity ◽  
N Protein ◽  
Human Sera ◽  
The Cross ◽  
Group 1

ABSTRACT In 2002, severe acute respiratory syndrome-associated coronavirus (SARS-CoV) emerged in humans, causing a global epidemic. By phylogenetic analysis, SARS-CoV is distinct from known CoVs and most closely related to group 2 CoVs. However, no antigenic cross-reactivity between SARS-CoV and known CoVs was conclusively and consistently demonstrated except for group 1 animal CoVs. We analyzed this cross-reactivity by an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using specific antisera to animal CoVs and SARS-CoV and SARS patient convalescent-phase or negative sera. Moderate two-way cross-reactivity between SARS-CoV and porcine CoVs (transmissible gastroenteritis CoV [TGEV] and porcine respiratory CoV [PRCV]) was mediated through the N but not the spike protein, whereas weaker cross-reactivity occurred with feline (feline infectious peritonitis virus) and canine CoVs. Using Escherichia coli-expressed recombinant SARS-CoV N protein and fragments, the cross-reactive region was localized between amino acids (aa) 120 to 208. The N-protein fragments comprising aa 360 to 412 and aa 1 to 213 reacted specifically with SARS convalescent-phase sera but not with negative human sera in ELISA; the fragment comprising aa 1 to 213 cross-reacted with antisera to animal CoVs, whereas the fragment comprising aa 360 to 412 did not cross-react and could be a potential candidate for SARS diagnosis. Particularly noteworthy, a single substitution at aa 120 of PRCV N protein diminished the cross-reactivity. We also demonstrated that the cross-reactivity is not universal for all group 1 CoVs, because HCoV-NL63 did not cross-react with SARS-CoV. One-way cross-reactivity of HCoV-NL63 with group 1 CoVs was localized to aa 1 to 39 and at least one other antigenic site in the N-protein C terminus, differing from the cross-reactive region identified in SARS-CoV N protein. The observed cross-reactivity is not a consequence of a higher level of amino acid identity between SARS-CoV and porcine CoV nucleoproteins, because sequence comparisons indicated that SARS-CoV N protein has amino acid identity similar to that of infectious bronchitis virus N protein and shares a higher level of identity with bovine CoV N protein within the cross-reactive region. The TGEV and SARS-CoV N proteins are RNA chaperons with long disordered regions. We speculate that during natural infection, antibodies target similar short antigenic sites within the N proteins of SARS-CoV and porcine group 1 CoVs that are exposed to an immune response. Identification of the cross-reactive and non-cross-reactive N-protein regions allows development of SARS-CoV-specific antibody assays for screening animal and human sera.

scholarly journals Peptide arrays of three collections of human sera from patients infected with mosquito-borne viruses

F1000Research ◽  
2020 ◽  
Vol 8 ◽  
pp. 1875
Author(s):  
Maria del Pilar Martinez Viedma ◽  
Nurgun Kose ◽  
Leda Parham ◽  
Angel Balmaseda ◽  
Guillermina Kuan ◽  
...  
Keyword(s):  
Vaccine Development ◽  
Cell Epitope ◽  
Cross Reactivity ◽  
Human Pathogens ◽  
Peptide Arrays ◽  
Serological Tests ◽  
Array Data ◽  
Prevalence Studies ◽  
Diagnostic Potential ◽  
Human Sera

Background: Global outbreaks caused by emerging or re-emerging arthropod-borne viruses (arboviruses) are becoming increasingly more common. These pathogens include the mosquito-borne viruses belonging to the Flavivirus and Alphavirus genera. These viruses often cause non-specific or asymptomatic infection, which can confound viral prevalence studies. In addition, many acute phase diagnostic tests rely on the detection of viral components such as RNA or antigen. Standard serological tests are often not reliable for diagnosis after seroconversion and convalescence due to cross-reactivity among flaviviruses. Methods: In order to contribute to development efforts for mosquito-borne serodiagnostics, we incubated 137 human sera on individual custom peptide arrays that consisted of over 866 unique peptides in quadruplicate. Our bioinformatics workflow to analyze these data incorporated machine learning, statistics, and B-cell epitope prediction. Results: Here we report the results of our peptide array data analysis, which revealed sets of peptides that have diagnostic potential for detecting past exposure to a subset of the tested human pathogens including Zika virus. These peptides were then confirmed using the well-established ELISA method. Conclusions: These array data, and the resulting peptides can be useful in diverse efforts including the development of new pan-flavivirus antibodies, more accurate epitope mapping, and vaccine development against these viral pathogens.

scholarly journals Multi-peptide ELISAs overcome cross-reactivity and inadequate sensitivity of conventional Chlamydia pneumoniae serology

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Kh Shamsur Rahman ◽  
Bernhard Kaltenboeck
Keyword(s):  
Chlamydia Pneumoniae ◽  
High Specificity ◽  
Cross Reactivity ◽  
Peptide Antigens ◽  
Specificity And Sensitivity ◽  
Peptide Antibody ◽  
Antibody Assays ◽  
Human Sera ◽  
Composite Reference Standard ◽  
Individual Peptide

Abstract Cross-reactivity of classical chlamydial antigens compromises Chlamydia (C.) pneumoniae serology. By testing with 185 human antisera, we expanded 18 previously discovered C. pneumoniae-specific B-cell epitopes to 48 peptide antigens from 12 C. pneumoniae immunodominant proteins. For specific detection of antibodies against C. pneumoniae, we developed novel ELISAs with strongly reactive individual peptide antigens and mixtures of these peptides. By comparison to a composite reference standard (CRS) for anti-C. pneumoniae antibody status of human sera, the top-performing CpnMixF12 peptide assay showed 91% sensitivity at 95% specificity, significantly higher than 4 commercial anti-C. pneumoniae IgG ELISAs (36-12% sensitivity at 95% specificity). Human C. pneumoniae (Cpn) and C. trachomatis (Ctr) seroreactivity was 54% biased towards co-positivity in commercial Cpn and Ctr ELISAs, but unbiased in Cpn and Ctr peptide antibody assays, suggesting severe cross-reactivity of commercial ELISAs. Using hyperimmune mouse sera against each of 11 Chlamydia spp., we confirm that commercial Cpn and Ctr ELISA antigens are cross-reactive among all Chlamydia spp., but Cpn and Ctr peptide antigens react only with antisera against the cognate chlamydial species. With simultaneously high specificity and sensitivity, and convenient use for non-specialized laboratories, these ELISAs have the potential to improve serodiagnosis of C. pneumoniae infection.

scholarly journals AN ALTERNATIVE MECHANISM FOR THE PROPERDIN SYSTEM

1958 ◽  
Vol 108 (4) ◽  
pp. 515-535 ◽  
Author(s):  
Robert A. Nelson
Keyword(s):  
N Uptake ◽  
Chelating Agent ◽  
Wide Distribution ◽  
Cross Reactivity ◽  
Normal Serum ◽  
Natural Resistance ◽  
Alternative Mechanism ◽  
Ancillary Data ◽  
Undiluted Serum ◽  
Human Sera

Evidence is presented that phenomena ascribed to the properdin system may be explained in terms of classical antibody (Ab) in combination with three of the components of complement (C'), i.e., C'1, C'4, and C'2 respectively. The results suggest that the complex of properdin (P) with zymosan (Z) represents a mixture of Z·Ab, Z·Ab·C'1, 4, Z·Ab-C'1, 4, 2, and Z·Ab·C'1, 4, 2 in a decayed state. The purported preferential reactivity of Z with the third component of C' (C'3) is not supported by the present experiments in which Z was reacted with C' in both guinea pig and human sera. Approximately the same number of reactive units of C'1, 4 and of C'3 were inactivated by Z, as well as by D. pneumoniae and a washed specific precipitate of bovine albumin-anti-albumin. The latter evidence that small amounts of antigen-antibody complex fix significant amounts of C'3 stands in contrast with the classical concept that C'3 is not fixed in ordinary complement fixation reaction. The observed reactivity of C'3 is explained on the basis of the present use of essentially undiluted serum as a source of C'3 and of 37° as the temperature for fixation. Ancillary data indicate that purified properdin contains Ab. In the presence of a chelating agent and at 0° properdin agglutinated Z granules. Measurements of nitrogen (N) uptake by Z suggest that 0.25 µg. N were contributed by solutions stated to contain 1 unit of properdin. The broad spectrum of reactivity or cross-reactivity of the Ab in normal serum is likely due to the wide distribution in nature of closely related polysaccharides. Further immunochemical studies are necessary to establish definitively the origin and mode of action in "natural resistance" of antibodies reactive with these polysaccharides.

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