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Journal of Biochemistry Aug 1986The specificities of five heterophile Hanganutziu and Deicher (HD) antibody-containing sera from four different cancer patients and one other diseased patients were...
The specificities of five heterophile Hanganutziu and Deicher (HD) antibody-containing sera from four different cancer patients and one other diseased patients were compared. Three glycosphingolipids and one glycoprotein antigens and their chemically modified derivatives were used. The antibodies of all whole sera showed similar specificities. IgG and IgM antibody fractions of each serum were separated. Although antibodies of the same class showed similar specificities, differences were detected between the specificities of IgG and IgM. IgG antibody specificities were dependent on the hydrophobic (ceramide) group while IgM antibodies were directed more to the terminal sialic acid moiety of the glycosphingolipid antigens. The results suggested that a similar population of IgG-producing lymphocytes is stimulated in patients. Due to the similarities in specificities of HD antibodies, the results of this study will facilitate the future isolation of either IgG or IgM antibody-producing lymphocyte(s) from a patient with HD antibodies and the establishment of a monoclonal antibody through hybridization with a human myeloma cell line.
Topics: Antibodies; Antigens, Heterophile; Carbohydrate Sequence; Glycoproteins; Glycosphingolipids; Hemagglutination Inhibition Tests; Humans; Immunoglobulin G; Immunoglobulin M; Lymphocytes
PubMed: 3491066
DOI: 10.1093/oxfordjournals.jbchem.a121735 -
PloS One 2009Pathogenic avian influenza virus (H5N1) has the potential to cause a major global pandemic in humans. Safe and effective vaccines that induce immunologic memory and... (Randomized Controlled Trial)
Randomized Controlled Trial
BACKGROUND
Pathogenic avian influenza virus (H5N1) has the potential to cause a major global pandemic in humans. Safe and effective vaccines that induce immunologic memory and broad heterotypic response are needed.
METHODS AND FINDINGS
Healthy adults aged 18-60 and > 60 years (n = 313 and n = 173, respectively) were randomized (1:1) to receive two primary and one booster injection of 7.5 microg or 15 microg doses of a subunit MF59-adjuvanted H5N1 (A/Vietnam/1194/2004) (clade 1) vaccine. Safety was monitored until 6 months after booster. Immunogenicity was assessed by hemagglutination inhibition (HI), single radial hemolysis (SRH) and microneutralization assays (MN). Mild injection-site pain was the most common adverse reaction. No serious adverse events relating to the vaccine were reported. The humoral immune responses to 7.5 microg and 15 microg doses were comparable. The rates for seroprotection (HI>40; SRH>25 mm(2); MN > or = 40) after the primary vaccination ranged 72-87%. Six months after primary vaccination with the 7.5 microg dose, 18% and 21% of non-elderly and elderly adults were seroprotected; rates increased to 90% and 84%, respectively, after the booster vaccination. In the 15 microg group, seroprotection rates among non-elderly and elderly adults increased from 25% and 62% after primary vaccination to 92% and 88% after booster vaccination, respectively. A heterologous immune response to the H5N1/turkey/Turkey/05 strain was elicited after second and booster vaccinations.
CONCLUSIONS
Both formulations of MF59-adjuvanted influenza H5N1 vaccine were well tolerated. The European Union requirement for licensure for pre-pandemic vaccines was met by the lower dose tested. The presence of cross-reactive antibodies to a clade 2 heterologous strain demonstrates that this vaccine may be appropriate for pre-pandemic programs.
TRIAL REGISTRATION
(ClinicalTrials.gov) NCT00311480.
Topics: Adjuvants, Immunologic; Adolescent; Adult; Age Distribution; Aged; Antibodies, Heterophile; Antibody Formation; Cross Reactions; Female; Hemagglutination; Hemolysis; Humans; Immunization, Secondary; Immunologic Memory; Influenza A Virus, H5N1 Subtype; Influenza Vaccines; Male; Neutralization Tests; Polysorbates; Squalene; Vaccination
PubMed: 19197383
DOI: 10.1371/journal.pone.0004384 -
Xenotransplantation Mar 2020Engineering of α-Galactosyltransferase gene-knockout pigs circumvented hyperacute rejection of pig organs after xenotransplantation in non-human primates. Overcoming...
BACKGROUND
Engineering of α-Galactosyltransferase gene-knockout pigs circumvented hyperacute rejection of pig organs after xenotransplantation in non-human primates. Overcoming this hurdle revealed the importance of non-α-Gal carbohydrate antigens in the immunobiology of acute humoral xenograft rejection.
METHODS
This study analyzed serum from seven naïve cynomolgus monkeys (blood type O/B/AB = 3/2/2) for the intensity of natural IgM and IgG signals using carbohydrate antigen microarray, which included historically reported α-Gal and non-α-Gal carbohydrate antigens with various modifications.
RESULTS
The median (range) of IgM and IgG signals were 12.71 (7.23-16.38) and 9.05 (7.23-15.90), respectively. The highest IgM and IgG signals with narrowest distribution were from mono- and disaccharides, followed by modified structures. Natural anti-α-Gal antibody signals were medium to high in IgM (11.2-15.9) and medium in IgG (8.5-11.6) spectra, and was highest with Lac core structure (Galα1-3Galβ1-4Glc, iGb3) and lowest with LacNAc core structure (Galα1-3Galβ1-4GlcNAc). Similar signal intensities (up to 15.8 in IgM and up to 11.8 in IgG) were observed for historically detected natural non-α-Gal antigens, which included Tn antigen, T antigen, GM2 glycolipid, and Sd antigen. The hierarchical clustering analysis revealed the presence of clusters of anti-A antibodies and was capable of distinguishing between the blood group B and AB non-human primates.
CONCLUSIONS
The results presented here provide the most comprehensive evaluation of natural antibodies present in cynomolgus monkeys.
Topics: Animals; Antibodies; Antigens, Heterophile; Disaccharides; Galactosyltransferases; Graft Rejection; Heterografts; Macaca fascicularis; Primates; Transplantation, Heterologous
PubMed: 31762117
DOI: 10.1111/xen.12567 -
Archives of Endocrinology and Metabolism Nov 2021Falsely elevated estradiol is rare, may result from heterophile antibody interference, and can result in unnecessary investigation and intervention. We present the case...
Falsely elevated estradiol is rare, may result from heterophile antibody interference, and can result in unnecessary investigation and intervention. We present the case of a 56-year-old female with falsely elevated estradiol levels inconsistent with her overall clinical picture, which ultimately led to an unnecessary surgical procedure. With the use of alternative analytical platforms and a heterophile antibody blocking agent, we determined the false elevation was due to heterophile antibody interference. Clinicians must suspect and investigate for laboratory error when the clinical picture contradicts laboratory results.
Topics: Antibodies, Heterophile; Estradiol; False Positive Reactions; Female; Humans; Immunoassay; Middle Aged
PubMed: 33587834
DOI: 10.20945/2359-3997000000324 -
Australian Family Physician Jun 2014Health professionals rely on the accuracy and validity of laboratory investigations in managing patients. Occasionally the results of laboratory investigations do not...
BACKGROUND
Health professionals rely on the accuracy and validity of laboratory investigations in managing patients. Occasionally the results of laboratory investigations do not correlate with the clinical scenario. Incorrect pathology results may lead to unnecessary further investigation, inappropriate therapeutic interventions, and considerable anxiety for the patient and doctor. Heterophile antibodies are endogenous antibodies in human serum/plasma that may interfere with immunoassays resulting in false elevation, or rarely false depression of measured values.
OBJECTIVE
To alert health professionals to clinical situations in which heterophile antibodies may result in misleading results and potentially compromise patient care.
DISCUSSION
Heterophile antibodies may interfere with a number of immunoassays commonly used in clinical practice. Awareness of the possibility of interference by heterophile antibodies is important to prevent inappropriate management on the basis of erroneous laboratory results.
Topics: Adult; Antibodies, Heterophile; Child; Diagnostic Errors; False Positive Reactions; Humans; Immunoassay; Male; Middle Aged; Myocardial Infarction; Prostate-Specific Antigen; Prostatic Neoplasms; Thyroid Diseases; Thyrotropin; Troponin I
PubMed: 24897990
DOI: No ID Found -
Indian Journal of Endocrinology and... 2024Considering the inherent vulnerability of immunoassays for heterophilic interference and the potential of Rheumatoid Factor (RF) to act as a heterophile-like antibody,...
INTRODUCTION
Considering the inherent vulnerability of immunoassays for heterophilic interference and the potential of Rheumatoid Factor (RF) to act as a heterophile-like antibody, we conducted this study to investigate if RF leads to any such heterophilic interference in seropositive rheumatoid arthritis (RA) patients. The study was done on the TSH assay as it is a noncompetitive, double antibody sandwich assay, which is known to be vulnerable to heterophilic interference.
METHODS
In this cross-sectional observational study, eighty-four consecutive newly diagnosed RF-positive RA patients underwent TSH, Free T4, and anti-TPO estimation using the chemiluminescence technique (CLIA) on Siemens Immulite 1000 platform. The samples were screened for TSH interference using four methods: 1) analysis on a different platform, 2) assessment of linearity using doubling dilutions, 3) polyethylene glycol (PEG) precipitation, and 4) addition of a commercial blocker.
RESULTS
Ten samples had a loss of linearity on serial dilution, indicating potential interference. After heterophile blocker treatment, five cases exhibited interference. One patient had diagnostic interpretation discordance on the second platform. No sample on PEG precipitation suggested the influence of antibodies. It is worth noting that even in cases where interference was suspected, the clinical interpretation was largely unaffected by the correction of TSH values based on mean dilution or measurement after heterophile blocker treatment.
CONCLUSION
RF can cause heterophilic interference in TSH immunoassays used commercially. However, in most cases, this interference does not affect clinical decision-making.
PubMed: 38533277
DOI: 10.4103/ijem.ijem_99_23 -
Xenotransplantation May 2017Corneal xenotransplantation is an effective solution for the shortage of human donor corneas, and the porcine cornea may be a suitable candidate for the donor cornea...
BACKGROUND
Corneal xenotransplantation is an effective solution for the shortage of human donor corneas, and the porcine cornea may be a suitable candidate for the donor cornea because of its optical similarity with humans. However, it is necessary to administer additional immunosuppressants to overcome antigenic differences. We aimed to investigate the feasibility of porcine corneas with anti-CD40 antibody-mediated costimulation blockade in a clinically applicable pig-to-non-human primate corneal xenotransplantation model.
METHODS
Five Chinese rhesus macaques underwent deep-lamellar corneal transplantation using clinically acceptable sized (7.5 mm diameter) porcine corneal grafts. The anti-CD40 antibody was intravenously administered on a programmed schedule. Graft survival, central corneal thickness, and intraocular pressure were evaluated. Changes in effector and memory T and B cell subsets and anti-αGal and donor-specific antibodies were investigated in the blood, and the changes in complement levels in the aqueous humor and blood were evaluated. Memory cell profiles in the anti-CD40 antibody-treated group were compared with those from the anti-CD154 antibody-treated group or rejected controls presented in our previous report. The changes in anti-αGal, non-αGal, and donor-specific antibodies after 6 months were compared with baseline values.
RESULTS
Anti-CD40 antibody-mediated costimulation blockade resulted in the successful survival of xenocorneal grafts (>389, >382, >236, >201, and >61 days), with 80% reaching 6 months of survival. Injection of anti-CD40 antibody considerably reduced the infiltration of inflammatory cells into the grafts and significantly blocked the complement response in the aqueous humor (P=.0159, Mann-Whitney U test). Systemic expansion of central or effector memory T cells was abrogated in the anti-CD40 antibody-treated primates compared with those in the rejected controls (P<.05, Mann-Whitney U test) or those in the anti-CD154 antibody-treated primates (P>.05, Mann-Whitney U test). The levels of anti-αGal, non-αGal, and donor-specific antibodies at 6 months were not significantly increased compared with baseline levels (P>.05, Wilcoxon signed rank test).
CONCLUSIONS
An anti-CD40 antibody-mediated blockade appears to be effective immunosuppressive approach for porcine corneal deep-lamellar xenotransplantation in primates.
Topics: Animals; Antibodies, Blocking; Antibodies, Heterophile; Antibodies, Monoclonal; CD40 Antigens; Complement C3a; Corneal Transplantation; Disaccharides; Epitopes; Graft Survival; Heterografts; Humans; Immunologic Memory; Macaca mulatta; Swine; Swine, Miniature; T-Lymphocyte Subsets; Tissue Donors; Transplantation, Heterologous
PubMed: 28393447
DOI: 10.1111/xen.12298 -
Current Opinion in Organ Transplantation Apr 2010Anti-nonGal xenoantibodies are a major barrier to the survival of genetically modified porcine xenografts. This review summarizes the contribution of anti-nonGal... (Review)
Review
PURPOSE OF REVIEW
Anti-nonGal xenoantibodies are a major barrier to the survival of genetically modified porcine xenografts. This review summarizes the contribution of anti-nonGal xenoantibodies to the activation of porcine endothelial cells and graft rejection, and further provides an update on recent advancements in defining the unique features of anti-nonGal xenoantibody structure.
RECENT FINDINGS
Anti-nonGal xenoantibodies pre-exist at low levels in humans and nonhuman primates, and are notably absent in neonates. Exposure of nonhuman primates to alpha1,3-galactosyltransferase gene knockout endothelial cells initiates an induced xenoantibody response that is restricted and encoded by the germline immunoglobulin heavy chain gene IGHV3-21. The target xenoantigen remains undetermined, but several candidate targets have been proposed, including carbohydrate xenoantigens. New advancements in molecular modeling provide insight on the mechanism by which xenoantibodies bind to structurally related carbohydrates.
SUMMARY
Genetic manipulation of porcine donors has significantly prolonged the survival of grafts placed into nonhuman primate recipients, but anti-nonGal xenoantibodies and thrombosis limit the ability of these grafts to function on a long-term basis. Recent developments defining pre-existing anti-nonGal xenoantibody levels, the restriction in the anti-nonGal xenoantibody response and the identification of key sites defining xenoantibody-carbohydrate interactions now provide the information necessary to develop new approaches to preventing xenoantibody-mediated rejection.
Topics: Amino Acid Sequence; Animals; Animals, Genetically Modified; Antibodies, Heterophile; Antigens, Heterophile; Endothelial Cells; Galactosyltransferases; Gene Knockout Techniques; Genes, Immunoglobulin Heavy Chain; Graft Rejection; Graft Survival; Immunity, Humoral; Molecular Sequence Data; Species Specificity; Swine; Transplantation Tolerance; Transplantation, Heterologous; Trisaccharides
PubMed: 20075731
DOI: 10.1097/MOT.0b013e328336b854 -
Xenotransplantation 2008Experience with non-antigenic galactose alpha1,3 galactose (alphaGal) polymers and development of alphaGal deficient pigs has reduced or eliminated the significance of...
BACKGROUND
Experience with non-antigenic galactose alpha1,3 galactose (alphaGal) polymers and development of alphaGal deficient pigs has reduced or eliminated the significance of this antigen in xenograft rejection. Despite these advances, delayed xenograft rejection (DXR) continues to occur most likely due to antibody responses to non-Gal endothelial cell (EC) antigens.
METHODS
To gauge the diversity of the non-Gal antibody response we used antibody derived from CD46 transgenic heterotopic cardiac xenografts performed without T-cell immunosuppression, Group A (n = 4) and Gal knockout (GT-KO) heart transplants under tacrolimus and sirolimus immunosuppression, Group B (n = 8). Non-Gal antibody was measured by flow cytometry and by western blots using GT-KO EC membrane antigens. A nanoLC/MS/MS analysis of proteins recovered from 2D gels was used to identify target antigens.
RESULTS
Group A recipients exhibited a mixed cellular and humoral rejection. Group B recipients mainly exhibited classical DXR. Western blot analysis showed a non-Gal antibody response induced by GT+ and GT-KO hearts to an overlapping set of pig aortic EC membrane antigens. Proteomic analysis identified 14 potential target antigens but failed to define several immunodominant targets.
CONCLUSIONS
These experiments indicate that the non-Gal antibody response is directed to a number of stress response and inflammation related pig EC antigens and a few undefined targets. Further analysis of these antibody specificities using alternative methods is required to more fully define the repertoire of non-Gal antibody responses.
Topics: Animals; Animals, Genetically Modified; Antibodies, Heterophile; Antibody Specificity; Disaccharides; Graft Rejection; Heart Transplantation; Humans; Papio anubis; Proteomics; Swine; Transplantation, Heterologous
PubMed: 18957049
DOI: 10.1111/j.1399-3089.2008.00480.x -
Scandinavian Journal of Immunology Feb 2010IgG4 has been implicated in a diverse set of complex pathologies - e.g. autoimmune pancreatitis (AIP), idiopathic membranous nephropathy - and carries unique features...
IgG4 has been implicated in a diverse set of complex pathologies - e.g. autoimmune pancreatitis (AIP), idiopathic membranous nephropathy - and carries unique features including lack of activation of the classical complement pathway and a dynamic Fab-arm exchange. We recently showed that the rheumatoid factor (RF)-like activity of IgG4 is achieved through a hitherto unknown, Fc-Fc (and not Fab-Fc as is the case in classical RF; CRF) interaction; hence the name, novel RF (NRF). Here, we further explore the resemblance/difference between CRF and NRF. As heterophilic interactions of human IgM RF (CRF) are well known, we checked whether this is the case for IgG4. Human IgG4 showed variable reactivity to animal IgGs: reacting intensely with rabbit and mouse IgGs, but weakly with others. The binding to rabbit IgG was not through the Fab (as in CRF) but via the Fc piece, as was recently shown for human IgG (NRF). This binding correlates with the IgG4 concentration per se and could therefore be of diagnostic usage and incidentally explain some observed interferences in biological assays. In conclusion, here is defined a novel heterophilic antibody interaction and is established the universality of the unique Fc-Fc binding, both involving IgG4.
Topics: Adult; Aged; Animals; Antibodies, Heterophile; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin Fc Fragments; Immunoglobulin G; Male; Mice; Middle Aged; Pancreatitis, Chronic; Protein Binding; Rabbits
PubMed: 20384862
DOI: 10.1111/j.1365-3083.2009.02353.x