An enzyme-linked immunoassay for thyroid microsomal antibodies

Schardt, Christof; McLachlan, Sandra M.; Matheson, J. G.; Smith, Bernard Rees

doi:10.1016/0022-1759(82)90028-x

Cited by 84 publications

(28 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Sera from 11 patients with autoimmune thyroid disease were studied. All sera contained high levels of TPO autoantibodies (detectable by ELISA [ 31] Competition studies between Fab fragments and serum TPO antibodies ofdifferent IgG subclasses. An ELISA was used for this purpose, similar to that previously described for TPO autoantibodies measured using thyroid microsomal antigen (32), with some modifications.…”

Section: Methodsmentioning

confidence: 99%

Recognition by recombinant autoimmune thyroid disease-derived Fab fragments of a dominant conformational epitope on human thyroid peroxidase.

Portolano

Chazenbalk²,

Seto³

et al. 1992

J. Clin. Invest.

View full text Add to dashboard Cite

To characterize the nature ofthyroid peroxidase (TPO) autoantibodies present in the sera of patients with autoimmune thyroid disease, we cloned three IgGl /kappa Fab fragments which bind 125I-TPO. This was accomplished by the molecular cloning and expression in bacteria of IgG gene fragments from B cells infiltrating the thyroid of a patient with Graves' disease. The three Fab fragments (SP2, SP4, and SP5) are coded for by a common heavy chain (VH1, D, JH3) and three related, but different, light chains (VK1, JK2). The SP Fab fragments bind specifically to TPO with high affinities (6 x 10-l"-2 X 10`' M) comparable to those of serum TPO autoantibodies. TPO autoantibodies represented by the SP Fab fragments are present in all 11 patients studied, constitute a high proportion (36-72%) of serum TPO autoantibodies in individual patients and interact with a conformational epitope on TPO. (J. Clin. Invest. 1992. 90:720-726.)

show abstract

Section: Methodsmentioning

confidence: 99%

Recognition by recombinant autoimmune thyroid disease-derived Fab fragments of a dominant conformational epitope on human thyroid peroxidase.

Portolano

Chazenbalk²,

Seto³

et al. 1992

J. Clin. Invest.

View full text Add to dashboard Cite

show abstract

“…Human Tg was puri®ed from thyroid glands, obtained at operations, applying a well-established methodology (28). AChE from Electrophorus electricus (approx.…”

Section: Reagentsmentioning

confidence: 99%

B-cell autoepitopes on the acetylcholinesterase-homologous region of human thyroglobulin: association with Graves' disease and thyroid eye disease

Thrasyvoulides

Sakarellos‐Daitsiotis²,

Philippou³

et al. 2001

European Journal of Endocrinology

View full text Add to dashboard Cite

Objective: Thyroglobulin (Tg) is a large autoantigen involved in autoimmune thyroid diseases. Tg epitopes have, so far, been identi®ed within large peptides. In the present study, we used small synthetic peptides to ®nely map serological epitopes on the highly immunogenic C-terminal region of Tg. Homology of this region to acetylcholinesterase (AChE) has been implicated in the pathogenesis of thyroid eye disease (TED) through cross-reactive antibodies. Methods: We tested total IgG puri®ed from four pilot Graves' disease (GD) sera reactive with both Tg and AChE and from three healthy controls, for reactivity against overlapping 20mer peptides (pin synthesis) covering the sequence 2171±2748 of human Tg. Antibody-reactive peptides were subsequently synthesized by a solid-phase technique for con®rmation with a large number of sera: 99 GD, 32 Hashimoto's thyroiditis (HT) and 45 healthy controls. Results: Peptides TgP15, TgP26 and TgP41 (amino acids 2339±2358, 2471±2490 and 2651±2670 respectively) were found to be targets of autoantibodies on intact Tg, recognized by a statistically signi®cant proportion of GD sera (22.2%, 35.4% and 30.3% respectively), compared with HT (0%, 15.6% and 6.3% respectively) and healthy controls (0%, 4.4% and 4.4% respectively). The majority of GD sera (56.6%) were positive for at least one of the three peptides. In GD, TgP26 reactivity was found to be associated with TED (48.6% with TED versus 25.5% without TED, P , 0X05X Conclusion: Some epitopes on the C-terminal region of Tg are associated with GD. A subset of Tgreactive autoantibodies, directed to this region, is associated with TED and may be involved in the development of the disease.

show abstract

“…Sera from 10 patients with autoimmune thyroid disease were studied. All sera contained high levels of TPO autoantibodies (detectable by ELISA [24] (10,11,26) were compared to the closest, presently known germline genes. It must be emphasized that, even with the recent expansion of VH germline sequence information (27), the germline assignments are putative and cannot be regarded as definitive without characterization of the respective genes in these patients.…”

Section: Family Crosspriming (--C A---a-c-c ------------) Vkla (16) (mentioning

confidence: 99%

Human organ-specific autoimmune disease. Molecular cloning and expression of an autoantibody gene repertoire for a major autoantigen reveals an antigenic immunodominant region and restricted immunoglobulin gene usage in the target organ.

Chazenbalk

Portolano²,

Russo³

et al. 1993

J. Clin. Invest.

135

115

View full text Add to dashboard Cite

The most common organ-specific autoimmune disease in humans involves the thyroid. Autoantibodies against thyroid peroxidase (TPO) are present in the sera of virtually all patients with active disease. We report the molecular cloning of the genes for 30 high-affinity, IgG-class human autoantibodies to TPO from thyroid-infiltrating B cells. Analysis of the putative germline genes used for the TPO human autoantibodies suggests the use of only five different H and L chain combinations involving four H chains and three L chains. In addition, the same combination of H and L chains was found in multiple patients. The F(ab) proteins expressed by these genes define two major, closely associated domains (A and B) in an immunodominant region on TPO. These A and B domains contain the binding sites of 80% of IgG-class TPO autoantibodies in the sera of patients with autoimmune thyroid disease. The present information permits analysis, not previously possible, of the relationship between autoantibody H and L chain genes and the antigenic domains on an autoantigen. Our data, obtained using target organ-derived autoantibodies, indicate that there is restriction in H and L chain usage in relation to the interaction with specific antigenic domains in human, organ-specific autoimmune disease. (J. Clin. Invest. 1993. 92:62-74.)

show abstract

An enzyme-linked immunoassay for thyroid microsomal antibodies

Cited by 84 publications

References 15 publications

Recognition by recombinant autoimmune thyroid disease-derived Fab fragments of a dominant conformational epitope on human thyroid peroxidase.

Recognition by recombinant autoimmune thyroid disease-derived Fab fragments of a dominant conformational epitope on human thyroid peroxidase.

B-cell autoepitopes on the acetylcholinesterase-homologous region of human thyroglobulin: association with Graves' disease and thyroid eye disease

Human organ-specific autoimmune disease. Molecular cloning and expression of an autoantibody gene repertoire for a major autoantigen reveals an antigenic immunodominant region and restricted immunoglobulin gene usage in the target organ.

Contact Info

Product

Resources

About