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Archives of Pathology & Laboratory... Aug 2013Immunoassays are commonly used for clinical diagnosis, although interferences have been well documented. The streptavidin-biotin interaction provides an efficient and...
Immunoassays are commonly used for clinical diagnosis, although interferences have been well documented. The streptavidin-biotin interaction provides an efficient and convenient method to manipulate assay components and is currently used in several immunoassay platforms. To date, there has been no report in the literature of interference from endogenous anti-streptavidin antibodies; however, such antibodies would potentially affect multiple diagnostic platforms. We report results from a patient being treated for thyroid dysfunction who demonstrated a T-uptake result of less than 0.2 and a nonlinear thyroid stimulating hormone dilution that suggested an immunoassay interference. Protein-A sepharose pretreatment corrected the nonlinear dilution and revealed an interference trend of falsely decreased results, as measured by sandwich assay, and falsely elevated results, as measured by competitive assay. The results of streptavidin-agarose adsorption were comparable to adsorption with protein-A sepharose. To our knowledge, this is the first published description of an endogenous anti-streptavidin antibody interfering with clinical laboratory assays.
Topics: Antibodies, Heterophile; Biotin; Diagnostic Errors; Humans; Hyperthyroidism; Immunoassay; Immunosorbent Techniques; Male; Middle Aged; Sepharose; Streptavidin; Thyrotropin; Thyroxine
PubMed: 23899071
DOI: 10.5858/arpa.2012-0270-CR -
Scientific Reports Jan 2024Knowledge of factors associated with semen quality may help in investigations of the aetiology and pathophysiology. We investigated the correlation between biomarkers...
Knowledge of factors associated with semen quality may help in investigations of the aetiology and pathophysiology. We investigated the correlation between biomarkers for testicular cell function (anti-müllerian hormone, AMH, Inhibin B, testosterone, free androgen-index (testosterone/sex-hormone binding globulin), insulin like peptide 3, INSL-3), alkaline phosphate (ALP), canine prostate-specific esterase (CPSE), and heterophilic antibodies with dog variables, semen quality, and fertility. Blood and semen were collected from 65 Bernese Mountain Dogs. We evaluated total sperm count, motility and morphological parameters. The semen quality ranged from poor to excellent, with an average total sperm count of 1.1 × 10 and 50% morphologically normal spermatozoa (MNS). Age and abnormal testicular consistency correlated with decreased motility and MNS. Higher ALP correlated with higher total sperm count. AMH could not be detected in seminal plasma. AMH in blood correlated with head defects and high AMH concentration correlated with a severe decline in several semen parameters. Testosterone was negatively and CPSE positively correlated with age. No correlations were found for INSL-3, inhibin B, or heterophilic antibodies. Our findings contribute to the understanding of factors associated with semen quality in dogs, particularly related to Sertoli cell function.
Topics: Male; Dogs; Animals; Semen Analysis; Semen; Anti-Mullerian Hormone; Body Fluids; Peptide Hormones; Testosterone; Antibodies, Heterophile; Esterases
PubMed: 38184699
DOI: 10.1038/s41598-024-51242-0 -
The Journal of Experimental Medicine May 1965A direct correlation between the amount of kidney-fixing antibody and the degree of associated renal injury was demonstrated for rabbit and duck nephrotoxic antibodies....
A direct correlation between the amount of kidney-fixing antibody and the degree of associated renal injury was demonstrated for rabbit and duck nephrotoxic antibodies. No evidence for a qualitative difference among nephrotoxic antibodies of a given type was obtained. It appeared that duck nephrotoxic antibody was directed against a wider spectrum of rat renal antigens than was rabbit nephrotoxic antibody. In order to produce immediate proteinuria an amount of rabbit or duck gamma-2 kidney-fixing antibody capable of occupying approximately 45 per cent or more of the capillary filtration surface was needed. For immediate proteinuria an amount of rabbit gamma-2 kidney-fixing antibody capable of reacting with more than one-half of the available antigenic sites was needed. Less than twice that amount of rabbit antibody completely saturated available antigenic sites in the kidney. Virtually all nephrotoxic antibodies in hyperimmune rabbit nephrotoxic sera were of the gamma-2 variety while nephrotoxic antibodies in comparable duck nephrotoxic sera were found in gamma-2 (with 5.8 and 7.4S sedimentation coefficients) and gamma-1M fractions. Gamma-1M duck nephrotoxic antibody was 60 times more potent a nephritogen than gamma-2 duck nephrotoxic antibody on a molecular basis. Mercaptoethanol abolished the nephrotoxicity of gamma-1M duck antibody and reduced that of gamma-2 duck antibodies but had no effect on rabbit gamma-2. In no case did mercaptoethanol treatment have an effect on the kidney-fixing properties of the antibodies involved. After injection of nephrotoxic antibodies there appeared to be a prompt fixation of a majority of the antibody to tissue antigens primarily in the kidney. However, a small amount of potentially kidney-fixing antibody remained in the circulation for a considerable period apparently reflecting a dissociation of less avid antibodies with an equilibrium between fixed and free antibody. The role of this more easily dissociable antibody in the progression of nephrotoxic nephritis is not certain but it is possible that it could play a role in the early progression of the disease.
Topics: Animals; Antibodies; Antibodies, Heterophile; Antigen-Antibody Reactions; Antigens; Autoantibodies; Glomerulonephritis; Immunoelectrophoresis; Kidney; Nephritis; Poultry; Proteinuria; Rabbits; Rats; Research
PubMed: 14278226
DOI: 10.1084/jem.121.5.697 -
Clinical and Experimental Immunology May 1972Infusion of cultured human lymphoblastoid cells into ten patients with advanced malignancies resulted in a significant rise in antibody titre to Epstein-Barr virus in...
Infusion of cultured human lymphoblastoid cells into ten patients with advanced malignancies resulted in a significant rise in antibody titre to Epstein-Barr virus in all ten. There was a significant rise in antibody titre to herpes simplex virus (HSV) in six of the ten patients. The four patients who did not respond in their antibody to HSV had no detectable HSV antibody before the cell infusion. In contrast, four patients had detectable antibody titre to cytomegalovirus (CMV) prior to the cell infusion; none of the ten showed a rise in CMV antibody after cell infusion. Observed antibody responses to EBV and HSV occurred regardless of the presence or absence of detectable herpesviruses in the lymphoblastoid cell lines infused. Six of the ten patients, negative for the heterophile antibody before cell infusion, developed a significant titre of the heterophile antibody after infusion. Except for the heterophile antibody response, the failure to detect IgM antibody to Epstein-Barr virus and to induce antibody to herpes simplex virus in the nonimmune patients suggests that the observed antibody responses are secondary responses.
Topics: Antibody Formation; Antibody-Producing Cells; Antigens, Neoplasm; Bone Marrow; Bone Marrow Cells; Breast Neoplasms; Cell Line; Cells, Cultured; Cytomegalovirus; Fluorescent Antibody Technique; Herpesviridae; Herpesvirus 4, Human; Humans; Immunoglobulins; Kidney Neoplasms; Lung Neoplasms; Lymphocytes; Melanoma; Neoplasms; Osteosarcoma; Rhabdomyosarcoma; Simplexvirus; Transplantation, Homologous
PubMed: 4338950
DOI: No ID Found -
BMC Immunology Mar 2009In certain cases, anti-idiotypic antibodies that recognize an antigen-combining site of an antibody can mimic the structure and/or function of certain nominal antigens....
BACKGROUND
In certain cases, anti-idiotypic antibodies that recognize an antigen-combining site of an antibody can mimic the structure and/or function of certain nominal antigens. This feature makes them particularly useful if conventional experimental approaches fail to fulfil expectations, especially when the molecule of interest is infectious, toxic or difficult to isolate and purify. We suggest the application of an anti-idiotype concept to the field of prion biology, with the aim of evoking a humoral immune response against the pathological isoform of the prion protein (PrPSc). Different ways to induce anti-idiotypic responses were studied in mice and chickens using various forms of V5B2, a PrPSc-specific monoclonal antibody we have described previously.
RESULTS
The preparation of anti-idiotypic monoclonal antibodies was achieved with well-defined strategies of immunization, selection and subsequent characterization. Our results demonstrate that it is possible to induce a strong anti-idiotypic immune response against the V5B2 monoclonal antibody in both xenogeneic and syngeneic experimental systems. From the competition seen between polyclonal and monoclonal anti-idiotypic antibodies and the original immunogen, the P1 peptide, and even more importantly, the ultimate target antigen, PrPSc, we conclude that selected antibodies bind to the antigen-combining site of the V5B2 monoclonal antibody and might even resemble the PrPSc-specific epitope. The involvement of both antigen-combining sites in the interaction between V5B2 and the most promising monoclonal anti-idiotypic antibody was further supported by molecular docking.
CONCLUSION
The results of the present study not only provide an example of the successful production of Ab2 monoclonal antibodies based on a well planned strategy for selection, but should also provide a new experimental approach that is applicable to the field of prion diseases.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Heterophile; Antibodies, Monoclonal; Binding, Competitive; Cerebral Cortex; Chickens; Epitope Mapping; Epitopes, B-Lymphocyte; Hybridomas; Immunization, Secondary; Immunohistochemistry; Mice; Mice, Inbred BALB C; Models, Immunological; PrPSc Proteins; Prion Diseases
PubMed: 19298674
DOI: 10.1186/1471-2172-10-16 -
Xenotransplantation Jul 2017The results of the assay for measuring anti-non-Gal antibodies (which affect pig xenograft survival) in recipients are important. Serum incubation time and concentration...
The results of the assay for measuring anti-non-Gal antibodies (which affect pig xenograft survival) in recipients are important. Serum incubation time and concentration may be important factors in the extent of antibody binding to the graft. The aim of this in vitro study was to determine the optimal incubation time and serum concentration for measuring anti-non-Gal antibody binding to porcine aortic endothelial cells (pAECs). Pooled human, naive, and sensitized baboon sera were incubated with wild-type, α1,3-galactosyltransferase gene-knockout (GTKO), and GTKO/human CD55 pAECs. IgM/IgG binding to pAECs after varying serum incubation times (0.5, 1, 2, and 3 hour) and concentrations (5, 10, 20, and 40 μL) was determined by flow cytometry. An increase in incubation time from 30 minutes to 2 hour was associated with increases in anti-non-Gal IgM/IgG binding to GTKO and GTKO/hCD55 pAECs of pooled human, naive and sensitized baboon sera (P<.05). Pooled human serum showed a significant increase in anti-non-Gal IgM (1.5 times) and a minimal increase in anti-non-Gal IgG antibody binding. IgM/IgG binding of sensitized baboon serum to GTKO pAECs after 2-hour incubation was 1.5 times and 2 times greater than after 30-minutes incubation, respectively, whereas naïve baboon sera showed minimal (non-significant) increase in anti-non-Gal IgM/IgG antibody binding. With 2-hour incubation, increasing the serum concentration from 5 μL to 20 μL significantly increased antibody binding to non-Gal antigens in pooled human and sensitized baboon serum. With naïve baboon serum, only IgG was significantly increased. Increasing the serum incubation time contributed to improve the sensitivity of detecting anti-non-Gal antibodies, without affecting cell viability in vitro.
Topics: Animals; Animals, Genetically Modified; Antibodies, Heterophile; Endothelial Cells; Gene Knockout Techniques; Graft Rejection; Graft Survival; Heterografts; Humans; Immunoglobulin G; Immunoglobulin M; Swine; Time Factors; Transplantation, Heterologous
PubMed: 28547819
DOI: 10.1111/xen.12312 -
Indian Journal of Clinical Biochemistry... Apr 2019The practice of medicine depends on the accuracy of biochemical assays. The high prevalence of incidental masses on imaging necessitates a correct biochemical diagnosis...
The practice of medicine depends on the accuracy of biochemical assays. The high prevalence of incidental masses on imaging necessitates a correct biochemical diagnosis before proceeding to radiological studies. Hormonal assays, tumour markers, and markers of cardiac injury are particularly susceptible to heterophile antibody interference which may lead to inaccurate and misleading results, inappropriate investigation and/or treatment, patient concern and potential harm. A case of heterophile antibody interference in the measurement of ACTH in a patient with Cushing's syndrome resulting in unnecessary invasive investigation is presented. Close collaboration and communication between laboratory and clinical staff is essential where laboratory results and the clinical picture are not congruent.
PubMed: 31093000
DOI: 10.1007/s12291-018-0770-x -
Avian Pathology : Journal of the W.V.P.A Apr 2019Avian pathogenic E. coli (APEC) cause severe respiratory and systemic disease. To address the genetic and immunological basis of resistance, inbred chicken lines were...
Avian pathogenic E. coli (APEC) cause severe respiratory and systemic disease. To address the genetic and immunological basis of resistance, inbred chicken lines were used to establish a model of differential resistance to APEC, using strain O1 of serotype O1:K1:H7. Inbred lines 7, 15I and C.B12 and the outbred line Novogen Brown were inoculated via the airsac with a high dose (10 colony-forming units, CFU) or low dose (10 CFU) of APEC O1. Clinical signs, colibacillosis lesion score and bacterial colonization of tissues after high dose challenge were significantly higher in line 15I and C.B12 birds. The majority of the 15I and C.B12 birds succumbed to the infection by 14 h post-infection, whilst none of the line 7 and the Novogen Brown birds developed clinical signs. No difference was observed after low dose challenge. In a repeat study, inbred lines 7 and 15I were inoculated with low, intermediate or high doses of APEC O1 ranging from 10 to 10 CFU. The colonization of lung was highest in line 15I after high dose challenge and birds developed clinical signs; however, colonization of blood and spleen, clinical signs and lesion score were not different between lines. No difference was observed after intermediate or low dose challenge. Ex vivo, the phagocytic and bactericidal activity of lung leukocytes from line 7 and 15I birds did not differ. Our data suggest that although differential resistance of inbred lines 7, 15I and C.B12 to APEC O1 challenge is apparent, it is dependent on the infectious dose. Research Highlights Lines 15I and C.B12 are more susceptible than line 7 to a high dose of APEC O1. Differential resistance is dose-dependent in lines 15I and 7. Phagocytic and bactericidal activity is similar and dose independent.
Topics: Air Sacs; Animals; Animals, Inbred Strains; Antibodies, Heterophile; Bacterial Load; Chickens; Disease Resistance; Dose-Response Relationship, Immunologic; Escherichia coli; Escherichia coli Infections; Female; Immunity, Innate; Macrophages; Male; Poultry Diseases; Specific Pathogen-Free Organisms
PubMed: 30570345
DOI: 10.1080/03079457.2018.1562154 -
Annals of Surgery Oct 2018Xenotransplantation using pig organs could end the donor organ shortage for transplantation, but humans have xenoreactive antibodies that cause early graft rejection....
OBJECTIVE
Xenotransplantation using pig organs could end the donor organ shortage for transplantation, but humans have xenoreactive antibodies that cause early graft rejection. Genome editing can eliminate xenoantigens in donor pigs to minimize the impact of these xenoantibodies. Here we determine whether an improved cross-match and chemical immunosuppression could result in prolonged kidney xenograft survival in a pig-to-rhesus preclinical model.
METHODS
Double xenoantigen (Gal and Sda) knockout (DKO) pigs were created using CRISPR/Cas. Serum from rhesus monkeys (n = 43) was cross-matched with cells from the DKO pigs. Kidneys from the DKO pigs were transplanted into rhesus monkeys (n = 6) that had the least reactive cross-matches. The rhesus recipients were immunosuppressed with anti-CD4 and anti-CD8 T-cell depletion, anti-CD154, mycophenolic acid, and steroids.
RESULTS
Rhesus antibody binding to DKO cells is reduced, but all still have positive CDC and flow cross-match. Three grafts were rejected early at 5, 6, and 6 days. Longer survival was achieved in recipients with survival to 35, 100, and 435 days. Each of the 3 early graft losses was secondary to IgM antibody-mediated rejection. The 435-day graft loss occurred secondary to IgG antibody-mediated rejection.
CONCLUSIONS
Reducing xenoantigens in donor pigs and chemical immunosuppression can be used to achieve prolonged renal xenograft survival in a preclinical model, suggesting that if a negative cross-match can be obtained for humans then prolonged survival could be achieved.
Topics: Animals; Animals, Genetically Modified; Antigens, Heterophile; Disease Models, Animal; Drug Therapy, Combination; Graft Survival; Immunoglobulin M; Immunosuppression Therapy; Immunosuppressive Agents; Kidney Transplantation; Macaca mulatta; Swine; Transplantation, Heterologous
PubMed: 30048323
DOI: 10.1097/SLA.0000000000002977 -
Anales de Pediatria Jun 2019The aim of this study is to assess epidemiological, clinical and laboratory characteristics of primary infection by Epstein-Barr virus (EBV) in children without previous...
INTRODUCTION
The aim of this study is to assess epidemiological, clinical and laboratory characteristics of primary infection by Epstein-Barr virus (EBV) in children without previous diagnosis of any immune disease and its relationship with clinical presentation.
PATIENTS AND METHODS
A retrospective study was conducted on all children from 0 to 15 years with IgM against viral capsid of EBV positive or indeterminate during a 22 month period. Epidemiological, clinical and laboratory data were analysed and compared between typical (mononucleosis syndrome) and non-typical clinical symptoms.
RESULTS
The study included a total of 103 children, with a median age of 7 years (3-12.5 years). Almost two-thirds (63%) of patients had typical clinical signs, with a mononucleosis syndrome, and 37% had a non-typical presentation. The non-typical clinical group had a lower age (P=.03) and took less antibiotic than the typical clinical group (P=.015). From laboratory studies, there were no differences between the groups, except in RCP, which was higher in typical clinical group (P=.04). Heterophile antibodies were positive in 33% of patients. An indeterminate IgM against viral capsid was present in 20% of the patients, and most of them had an oligosymptomatic or atypical presentation. An IgM positive for other viruses was found in 21%, and 3 of them were suspicious of false positive for EBV.
CONCLUSIONS
In the studied population, a primary infection due to EBV is common in younger ages, and they have usually an oligosymptomatic clinical presentation. A very low percentage of positive heterophile antibodies were found. Cases with indeterminate IgM against viral capsid are more frequent in the non-typical clinical group. Co-infection with other viruses is common.
Topics: Adolescent; Child; Child, Preschool; Epstein-Barr Virus Infections; Female; Humans; Infant; Male; Retrospective Studies
PubMed: 30529045
DOI: 10.1016/j.anpedi.2018.09.003