scholarly journals Cross-Reactivity between Chemical Antibodies Formed to Serum Proteins and Thyroid Axis Target Sites

2020 ◽  
Vol 21 (19) ◽  
pp. 7324
Author(s):  
Datis Kharrazian ◽  
Martha Herbert ◽  
Aristo Vojdani
Keyword(s):  
Conformational Change ◽  
Protein Misfolding ◽  
Serum Proteins ◽  
Cross Reactivity ◽  
Surface Topology ◽  
Antigenic Structure ◽  
Thyroid Peroxidase ◽  
Immune Reactivity ◽  
Target Sites ◽  
Thyroid Axis

In some instances, when chemicals bind to proteins, they have the potential to induce a conformational change in the macromolecule that may misfold in such a way that makes it similar to the various target sites or act as a neoantigen without conformational change. Cross-reactivity then can occur if epitopes of the protein share surface topology to similar binding sites. Alteration of peptides that share topological equivalence with alternating side chains can lead to the formation of binding surfaces that may mimic the antigenic structure of a variant peptide or protein. We investigated how antibodies made against thyroid target sites may bind to various chemical–albumin compounds where binding of the chemical has induced human serum albumin (HSA) misfolding. We found that specific monoclonal or polyclonal antibodies developed against thyroid-stimulating hormone (TSH) receptor, 5′-deiodinase, thyroid peroxidase, thyroglobulin, thyroxine-binding globulin (TBG), thyroxine (T4), and triiodothyronine (T3) bound to various chemical HSA compounds. Our study identified a new mechanism through which chemicals bound to circulating serum proteins lead to structural protein misfolding that creates neoantigens, resulting in the development of antibodies that bind to key target proteins of the thyroid axis through protein misfolding. For demonstration of specificity of thyroid antibody binding to various haptenic chemicals bound to HSA, both serial dilution and inhibition studies were performed and proportioned to the dilution. A significant decline in these reactions was observed. This laboratory analysis of immune reactivity between thyroid target sites and chemicals bound to HSA antibodies identifies a new mechanism by which chemicals can disrupt thyroid function.

scholarly journals Immunological Reactivity Using Monoclonal and Polyclonal Antibodies of Autoimmune Thyroid Target Sites with Dietary Proteins

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
Datis Kharrazian ◽  
Martha Herbert ◽  
Aristo Vojdani
Keyword(s):  
Thyroid Hormones ◽  
Dietary Protein ◽  
Polyclonal Antibodies ◽  
Thyroid Peroxidase ◽  
Immune Reactivity ◽  
Dietary Proteins ◽  
Thyroxine Binding Globulin ◽  
Autoimmune Thyroid ◽  
Target Sites ◽  
Thyroid Axis

Many hypothyroid and autoimmune thyroid patients experience reactions with specific foods. Additionally, food interactions may play a role in a subset of individuals who have difficulty finding a suitable thyroid hormone dosage. Our study was designed to investigate the potential role of dietary protein immune reactivity with thyroid hormones and thyroid axis target sites. We identified immune reactivity between dietary proteins and target sites on the thyroid axis that includes thyroid hormones, thyroid receptors, enzymes, and transport proteins. We also measured immune reactivity of either target specific monoclonal or polyclonal antibodies for thyroid-stimulating hormone (TSH) receptor, 5′deiodinase, thyroid peroxidase, thyroglobulin, thyroxine-binding globulin, thyroxine, and triiodothyronine against 204 purified dietary proteins commonly consumed in cooked and raw forms. Dietary protein determinants included unmodified (raw) and modified (cooked and roasted) foods, herbs, spices, food gums, brewed beverages, and additives. There were no dietary protein immune reactions with TSH receptor, thyroid peroxidase, and thyroxine-binding globulin. However, specific antigen-antibody immune reactivity was identified with several purified food proteins with triiodothyronine, thyroxine, thyroglobulin, and 5′deiodinase. Laboratory analysis of immunological cross-reactivity between thyroid target sites and dietary proteins is the initial step necessary in determining whether dietary proteins may play a potential immunoreactive role in autoimmune thyroid disease.

scholarly journals Antibodies to Thyroid Peroxidase Arise Spontaneously with Age in NOD.H-2h4 Mice and Appear after Thyroglobulin Antibodies

2010 ◽  
Vol 31 (4) ◽  
pp. 600-600
Author(s):  
Chun-Rong Chen ◽  
Sepehr Hamidi ◽  
Helen Braley-Mullen ◽  
Yuji Nagayama ◽  
Catherine Bresee ◽  
...  
Keyword(s):  
Hashimoto’S Thyroiditis ◽  
Time Course ◽  
Thyroid Autoimmunity ◽  
Cross Reactivity ◽  
Eukaryotic Cells ◽  
Human Thyroid ◽  
Hashimoto's Thyroiditis ◽  
Thyroid Peroxidase ◽  
Thyroglobulin Antibodies ◽  
Self Tolerance

Abstract Hashimoto’s thyroiditis, a common autoimmune disease, is associated with autoantibodies to thyroglobulin (Tg) and thyroid peroxidase (TPO). TPO, unlike abundant and easily purified Tg, is rarely investigated as an autoantigen in animals. We asked whether antibodies (Abs) develop to both TPO and Tg in thyroiditis in mice that is induced (C57BL/6 and DBA/1 strains) or arises spontaneously (NOD.H-2h4). Screening for TPOAbs was performed by flow cytometry using mouse TPO-expressing eukaryotic cells. Sera were also tested for binding to purified mouse Tg and human TPO. The antibody data were compared with the extent of thyroiditis. Immunization with mouse TPO adenovirus broke self-tolerance to this protein in C57BL/6 mice, but thyroiditis was minimal and TgAbs were absent. In DBA/1 mice with extensive granulomatous thyroiditis induced by Tg immunization, TPOAbs were virtually absent despite high levels of TgAbs. In contrast, antibodies to mouse TPO, with minimal cross-reactivity with human TPO, arose spontaneously in older (7–12 months) NOD.H-2h4 mice. Unexpectedly, TgAbs preceded TPOAbs, a time course paralleled in relatives of probands with juvenile Hashimoto’s thyroiditis. These findings demonstrate a novel aspect of murine and human thyroid autoimmunity, namely breaking B cell self-tolerance occurs first for Tg and subsequently for TPO.

scholarly journals Inhibition of Na,K-ATPase by oleandrin and oleandrigenin, and their detection by digoxin immunoassays

1996 ◽  
Vol 42 (10) ◽  
pp. 1654-1658 ◽  
Author(s):  
S A Jortani ◽  
R A Helm ◽  
R Valdes
Keyword(s):  
Sodium Pump ◽  
Serum Proteins ◽  
Cross Reactivity ◽  
Pump Activity ◽  
Ic50 Values ◽  
On Line ◽  
Atpase Inhibition ◽  
Free Serum ◽  
Derivatives Of ◽  
Rule Out

Abstract Ingestion of oleander plant, containing the cardiac glycoside oleandrin, has been reported to induce fatal poisonings. Derivatives of oleandrin are structurally similar to digoxin. We investigated the cross-reactivities of oleandrin and its aglycone metabolite, oleandrigenin, in several commercially available digoxin immunoassays; assessed their ability to inhibit Na,K-ATPase catalytic activity; and measured their binding to proteins in serum. As assayed with ACS:180, Stratus, RIA, On-Line, and TDx digoxin assays, oleandrin at 100 micromol/L in digoxin-free serum gave apparent digoxin values of 0, 0.83, 2.24, 2.37, and 5.34 nmol/L, respectively, whereas oleandrigenin at that concentration gave results of 0, 0.52, 0.77, 4.94, and 1.40 nmol/L. Study of Na,K-ATPase inhibition showed IC50 values (micromol/L) of 0.22 for ouabain, 0.62 for oleandrin, 1.23 for oleandrigenin, and 2.69 for digoxin. At 25 degrees C, 96% of oleandrin and 48% of oleandrigenin were bound to serum proteins. Because detection of oleandrin and oleandrigenin by digoxin immunoassays is variable between assays as well as between congeners, assessment of cross-reactivity is warranted for each assay. The inhibition of Na,K-ATPase by oleandrin and oleandrigenin confirms that they likely exert their toxic effects through inhibition of sodium pump activity. In cases of digitalis-like poisoning with suspicion of oleander ingestion, a combination of digoxin immunoassays may be useful to effectively rule out the presence of oleander.

Highly sensitive assays of autoantibodies to thyroglobulin and to thyroid peroxidase.

1989 ◽  
Vol 35 (9) ◽  
pp. 1949-1954 ◽  
Author(s):  
K Beever ◽  
J Bradbury ◽  
D Phillips ◽  
S M McLachlan ◽  
C Pegg ◽  
...  
Keyword(s):  
Solid Phase ◽  
Protein A ◽  
Cross Reactivity ◽  
Thyroid Peroxidase ◽  
Scatchard Analysis ◽  
Thyroid Autoantibodies ◽  
Serum Samples ◽  
Highly Sensitive ◽  
Phase Protein ◽  
High Concentrations

Abstract These highly sensitive assays are based on the interaction between thyroid autoantibodies and 125I-labeled autoantigens. Serum samples are incubated with labeled thyroid peroxidase (TPO) or thyroglobulin (Tg) to allow the formation of antibody-labeled antigen complexes. The complexes are then precipitated by addition of solid-phase Protein A. In the presence of high concentrations of TPO antibody or Tg antibody, more than 50% of the respective labeled antigen was precipitated, whereas only 1-2% was precipitated in the absence of autoantibody. Interassay CVs were 3.2% and 5.7%, respectively, for the anti-TPO and anti-Tg assays. There was no cross-reactivity between Tg antibody and TPO antibody. Results correlated highly significantly with results from other assay systems based on antigen-coated cells or plastic supports, but the assays described here were considerably more sensitive. Scatchard analysis of the assay data provided information on the affinity and serum concentration of TPO autoantibodies (ka approximately 10(9) L/mol and concentrations up to 1 g/L) and Tg autoantibodies (ka approximately 4 x 10(10) L/mol and concentrations up to 1 g/L). Overall, these assays provide a sensitive, precise, and convenient system for measuring and investigating the properties of thyroid autoantibodies.

scholarly journals Detection of Islet Cell Immune Reactivity with Low Glycemic Index Foods: Is This a Concern for Type 1 Diabetes?

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Datis Kharrazian ◽  
Martha Herbert ◽  
Aristo Vojdani
Keyword(s):  
Insulin Receptor ◽  
Glycemic Index ◽  
Autoimmune Diabetes ◽  
Acid Decarboxylase ◽  
Immune Reactivity ◽  
Dietary Proteins ◽  
Receptor Alpha ◽  
Target Sites ◽  
Gad 65 ◽  
Receptor Beta

Dietary management of autoimmune diabetes includes low glycemic foods classified from the glycemic index, but it does not consider the role that immunoreactive foods may play with the immunological etiology of the disease. We measured the reactivity of either monoclonal or polyclonal affinity-purified antibodies to insulin, insulin receptor alpha, insulin receptor beta, zinc transporter 8 (ZnT8), tyrosine phosphatase-based islet antigen 2 (IA2), and glutamic acid decarboxylase (GAD) 65 and 67 against 204 dietary proteins that are commonly consumed. Dietary protein determinants included unmodified (raw) and modified (cooked and roasted) foods, herbs, spices, food gums, brewed beverages, and additives. There was no immune reactivity between insulin or insulin receptor beta and dietary proteins. However, we identified strong to moderate immunological reactivity with antibodies against insulin receptor alpha, ZnT8, IA2, GAD-65, and GAD-67 with several dietary proteins. We also identified 49 dietary proteins found in foods classified as low glycemic foods with immune reactivity to autoimmune target sites. Laboratory analysis of immunological cross-reactivity between pancreas target sites and dietary proteins is the initial step necessary in determining whether dietary proteins may play a potential immunoreactive role in autoimmune diabetes.

scholarly journals STUDY OF THE STATE OF REPRODUCTIVE HEALTH OF WOMEN IN THE CONDITIONS OF PRODUCTION EXPOSURE TO ACRYLONITRILE

2018 ◽  
Vol 97 (1) ◽  
pp. 59-64
Author(s):  
Irina V. Leshkova ◽  
E. M. Vlasova ◽  
V. E. Belitskaya
Keyword(s):  
Menstrual Cycle ◽  
Reproductive Health ◽  
Working Conditions ◽  
Inflammatory Responses ◽  
Thyroid Peroxidase ◽  
Immune Reactivity ◽  
Endocrine Regulation ◽  
Menstrual Dysfunction ◽  
Female Workers ◽  
The Impact

At workplaces in the observation group, working conditions are characterized by a combined effect of the industrial noise and a chemical factor (at 13.4% of working places the class of working conditions is 3.1, at 63.3% of workplaces the class of working conditions is 3.2). In the comparison group, working conditions are characterized by the impact of electromagnetic radiation from a personal computer, the class of working conditions corresponds to an allowable 2nd class. The executed research showed that under working conditions with an acrylonitrile concentration of 0.01 ± 0.003 mg/m3 (MPC of 0.5 mg/m3), working conditions do not provide safety for the reproductive health of female workers. An analysis of the results of the studies showed that female workers have menstrual dysfunction (menstrual irregularities in the form of hypomenorrhea, algomenorrhea, and irregular menstrual cycle were more often presented); absence of pregnancy in the first 5 years of the family life; hormonal disorders, there is also an activation of inflammatory responses, an increase in the immune reactivity; specific sensitization to acrylonitrile. Under conditions of production exposure to acrylonitrile, in workers there is observed an activation of autoimmune processes (increase in the level of antibodies to thyroid peroxidase (TPO), antibodies to phosphatidylserine), promoting lowering of immunological tolerance (elevation of antisperm antibodies), which has the impact on the formation of reproductive disorders (on fertilization and fetal development in the early stages of pregnancy), a high risk of abnormal pregnancy. The development of pathological processes of the cervix and breast is also associated with a deterioration of immunological reactivity and endocrine regulation. There was established a very high degree of the occupational conditionality of dyshormonal disorders (RR = 3.28, 95% CI = 1.22, EF = 69.48%); disorders of the menstrual cycle (RR = 4.63, 95% CI = 2.4-2.7, EF = 76.7%), the average degree of production conditionality of the pregnancy pathology (RR = 1.9, 95% CI = 1, 1-1,7, EF = 47,4%), pathological processes of the cervix (RR = 1.6, 95% CI = 1.3-1.7, EF = 37.5%), mastopathy (RR = 2.2, 95%, CI = 1, 9-2.7, EF = 50%).

scholarly journals Development of Vasoinhibin-Specific Monoclonal Antibodies

2021 ◽  
Vol 12 ◽  
Author(s):  
Nils Müller ◽  
Juan Pablo Robles ◽  
Magdalena Zamora ◽  
Johannes Ebnet ◽  
Hülya Markl-Hahn ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Serum Proteins ◽  
Limit Of Detection ◽  
Sandwich Elisa ◽  
Vascular Diseases ◽  
High Specificity ◽  
Cross Reactivity ◽  
Dimensional Structure ◽  
Serum Samples ◽  
Limit Of Quantitation

Vasoinhibin is a protein hormone with antiangiogenic, antivasodilatatory, and antivasopermeability effects generated by the proteolytic cleavage of prolactin. The discovery of its role in diabetic retinopathy and peripartum cardiomyopathy led to the evaluation of new pharmacological treatments in clinical interventional trials. However, the quantitative evaluation of vasoinhibin in biological samples from patients has not been possible due to the lack of vasoinhibin-specific antibodies. Recently, loop 1 of vasoinhibin was identified to have a different three-dimensional structure compared to PRL, and thus to contain vasoinhibin-specific epitopes. Here, we report the development of two sets of vasoinhibin-specific monoclonal antibodies against two neighboring regions of the vasoinhibin loop 1. An experimental sandwich ELISA with two monoclonal anti-vasoinhibin antibodies was developed, which had no cross-reactivity to recombinant human full-length prolactin. The ELISA had a quantitation limit of 100 ng/ml, and intra-assay- and inter-assay coefficients of variation of 12.5% and 14%, respectively. The evaluation of 15 human serum samples demonstrated concentrations of below limit of detection (n=3), below limit of quantitation (n=1) and between 0.23 µg/ml (230 ng/ml) to 605 µg/ml (n=12) in the quantifiable range. Despite the high specificity of the monoclonal-monoclonal antibody sandwiches which discriminate vasoinhibin from PRL, there might be cross-reactivities by serum proteins other than vasoinhibin. A fully established vasoinhibin ELISA may support diagnostic and therapeutic measures in vascular diseases.

scholarly journals Reaction of Human Monoclonal Antibodies to SARS-CoV-2 Proteins With Tissue Antigens: Implications for Autoimmune Diseases

2021 ◽  
Vol 11 ◽  
Author(s):  
Aristo Vojdani ◽  
Elroy Vojdani ◽  
Datis Kharrazian
Keyword(s):  
Monoclonal Antibodies ◽  
Autoimmune Diseases ◽  
Human Tissue ◽  
Molecular Mimicry ◽  
Disease Process ◽  
Cross Reactivity ◽  
Immune Reactivity ◽  
Human Tissues ◽  
Human Monoclonal Antibodies ◽  
Tissue Antigens

We sought to determine whether immune reactivity occurs between anti-SARS-CoV-2 protein antibodies and human tissue antigens, and whether molecular mimicry between COVID-19 viral proteins and human tissues could be the cause. We applied both human monoclonal anti-SARS-Cov-2 antibodies (spike protein, nucleoprotein) and rabbit polyclonal anti-SARS-Cov-2 antibodies (envelope protein, membrane protein) to 55 different tissue antigens. We found that SARS-CoV-2 antibodies had reactions with 28 out of 55 tissue antigens, representing a diversity of tissue groups that included barrier proteins, gastrointestinal, thyroid and neural tissues, and more. We also did selective epitope mapping using BLAST and showed similarities and homology between spike, nucleoprotein, and many other SARS-CoV-2 proteins with the human tissue antigens mitochondria M2, F-actin and TPO. This extensive immune cross-reactivity between SARS-CoV-2 antibodies and different antigen groups may play a role in the multi-system disease process of COVID-19, influence the severity of the disease, precipitate the onset of autoimmunity in susceptible subgroups, and potentially exacerbate autoimmunity in subjects that have pre-existing autoimmune diseases. Very recently, human monoclonal antibodies were approved for use on patients with COVID-19. The human monoclonal antibodies used in this study are almost identical with these approved antibodies. Thus, our results can establish the potential risk for autoimmunity and multi-system disorders with COVID-19 that may come from cross-reactivity between our own human tissues and this dreaded virus, and thus ensure that the badly-needed vaccines and treatments being developed for it are truly safe to use against this disease.

Recombinant thyroid antigens: preparation and application in the diagnosis and treatment of autoimmune diseases

2021 ◽  
pp. 48-53
Author(s):  
Alexander Vladimirovich Zubkov
Keyword(s):  
Autoimmune Diseases ◽  
Hormone Receptor ◽  
Extracellular Domain ◽  
Thyroid Stimulating Hormone ◽  
Human Thyroid ◽  
Antigenic Structure ◽  
Thyroid Peroxidase ◽  
Α Subunit ◽  
Thyroid Autoantigens ◽  
Stimulating Hormone

New opportunities are opening up in the study of the relationship between the antigenic structure of thyroid autoantigens and autoantibodies in the pathogenesis of such autoimmune diseases as Graves’ disease (GD) and autoimmune thyroiditis (AIT) using synthesized recombinant proteins of the thyroid-stimulating hormone receptor (TSHR) and human thyroid peroxidase (TPO). In the present work, the results of cloning of the fragments of the TPO extracellular domain and fragments of the RNA sequence of the α-subunit of TSHR, which do not contain nucleotide substitutions, are demonstrated. Recombinant vectors for the expression of proteins of the α-subunit of thyroid-stimulating hormone receptor and fragments of the TPO extracellular domain have been obtained.

Radioimmunoassay for Serum Triiodothyronine: Evaluation of Simple Techniques to Control Interference from Binding Proteins

1974 ◽  
Vol 20 (9) ◽  
pp. 1150-1154 ◽  
Author(s):  
Victor S Fang ◽  
Samuel Refetoff
Keyword(s):  
Binding Sites ◽  
Binding Proteins ◽  
Serum Proteins ◽  
Cross Reactivity ◽  
Heat Inactivation ◽  
Nonspecific Binding ◽  
High Concentration ◽  
Assay Sensitivity ◽  
Thyroxine Binding Globulin ◽  
Fixed Concentration

Abstract Simple techniques for controlling interference from binding proteins in serum, such as thyroxine-binding globulin, in radioimmunoassay for triiodothyronine (T3) have been evaluated for their efficacy, and their effect on assay sensitivity and on recovery of added T3. Ethanol precipitation of serum proteins decreased the assay sensitivity, nonspecific binding was increased, and recoveries of added T3 were inconsistent. Heat-inactivation of thyroxine-binding globulin or use of 8-anilino-1naphthalene sulfonic acid (ANS) to displace T3 from thyroxine-binding globulin produced comparable recovery rates. The heat-inactivation method slightly decreased the sensitivity of the assay and prolonged the procedure, whereas use of ANS is simple, and the assay sensitivity is maintained. When sera contain a high concentration of thyroxine-binding globulin, a fixed concentration of ANS (175 µg/100 µl of serum) might be too low to displace T3 from all its binding sites, but a concentration of ANS greater than 200 µg/100 µl of serum interferes with T3 quantitation by competitively binding to the antibodies. The cross-reactivity of thyroxine to T3-antibodies varies with the antiserum. Thyroxine-binding globulin appears to be the only protein in serum that competes with the antibody for T3 binding.

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