Molecular cloning and characterization of human thyroid peroxidase autoantibodies of lambda light chain type

Portolano, Stefano; Prummel, Mark F.; Rapoport, Basil; McLachlan, Sandra M.

doi:10.1016/0161-5890(95)00060-7

Cited by 26 publications

(26 citation statements)

References 44 publications

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“…Several TPO Fabs have been produced using either bacteriophage vector (7,9) or phage display vector pComb3 (10,11,16,35,36). Despite the fact that individual H and L chain genes of the TPO Fab already described are clearly those used in vivo, it has yet to be proved whether separate amplification of heavy and light chains and their random recombination produce the same H and L chains pairing as in vivo.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the germline gene V 1-51 (Humlv117) that encodes the light chain of the other two TPO scFvs (ICA1 and ICB7) is also used in autoimmune anti-DNA Abs (46) and in acetylcholine receptor autoantibodies (47). Only four anti-TPO Fab/ light chain combinations have been described (16,48). One of them (TR1.41) used the VH1-69 germline gene that is used by one of our TPO scFvs (ICA5).…”

Section: Discussionmentioning

confidence: 99%

“…Despite the large number of applications of combinatorial library methods to the study of human disease-associated autoantibodies and although the high affinity and the epitope specificity of TPO recombinant Abs are similar to autoantibodies in the serum, one major question raised by many investigators remains: Does the V H and V L chain pairing of these antibodies from random combinatorial libraries reflect the in vivo pairing (13)(14)(15)(16)? The production of human mAbs by cell fusion or by transformation of B cells with EBV could provide answers to this question.…”

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confidence: 99%

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Human Anti-Thyroid Peroxidase Single-Chain Fragment Variable of Ig Isolated from a Combinatorial Library Assembled In-Cell: Insights into the In Vivo Situation

Chapal¹,

Péraldi-Roux²,

Bresson³

et al. 2000

The Journal of Immunology

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In an attempt to explore the natural variable heavy and light chain (VH/VL) pairing of autoantibodies involved in Graves’ disease, we constructed a phage-displayed Ab library obtained by in-cell PCR of thyroid-infiltrating cells. We report here the molecular cloning and characterization of human single-chain fragment variable regions (scFv) specific for thyroid peroxidase (TPO) generated from this library. On the basis of the nucleotide sequences, three different scFvs were obtained (ICA1, ICB7, and ICA5). All were encoded by genes derived from the VH1 and Vλ1 gene families. Using BIACORE for epitope mapping and kinetic analysis, we showed that these scFvs exhibited high affinity (Kd = 1 nM) for TPO and recognized three different epitopes. The biological relevance of these scFvs as compared with serum anti-TPO autoantibodies was assessed by competition studies. Sera from all the 29 Graves’ disease patients tested were able to strongly inhibit (60–100%) the binding of the 3 scFvs to TPO. These data demonstrate that the in-cell PCR library generated human anti-TPO scFvs that retained the VH/VL pairing found in vivo and that the different epitope specificities defined by these scFvs overlapped with those found in the sera of patients with autoimmune thyroid disease.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Human Anti-Thyroid Peroxidase Single-Chain Fragment Variable of Ig Isolated from a Combinatorial Library Assembled In-Cell: Insights into the In Vivo Situation

Chapal¹,

Péraldi-Roux²,

Bresson³

et al. 2000

The Journal of Immunology

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“…SS-A/Ro 60-kDa peptide Sjögren's syndrome [37] 3 l DPL 16/ 3 dsDNA SLE [32] VL3.1 5 × neutrophil cytoplasmic antigen (ANCA) Healthy, nonimmunized [38] (IGLV3S1) Bloodgroup H1 (self antigen) Healthy, nonimmunized [39] Bloodgroup D (self antigen) Healthy, nonimmunized [39] Bloodgroup panagglutinin Healthy, nonimmunized [39] 2 a2 2a2.27A12/ 3 (2) a) dsDNA SLE [32] DPL 11 Thyroid peroxidase Autoimmune thyroid disease [35] 2 × neutrophil cytoplasmic antigen (ANCA) Healthy, nonimmunized [38] Bloodgroup E Healthy, nonimmunized [39] Bloodgroup Kp (self antigen) Healthy, nonimmunized [39] a) Number in parentheses indicates number of different clones count. Four weeks later, he again became thrombocytopenic and was treated with prednisone (100 mg/day), danazol, and vincristine (2 mg/week for 3 weeks).…”

Section: Patientsmentioning

confidence: 99%

“…Although they were derived from the same germ-line genes, many of these autoantibodies derived from autoimmune patients were highly specific for the respective autoantigens. These included dsDNA [32], the acetylcholine receptor [33], thyroid peroxidase [34,35], the pyruvate dehydrogenase E2 subunit [36], and SS-A/ Ro 60 [37]. In addition, autoantibodies from healthy, nonimmunized individuals [38,39] and a synthetic library [40] were also derived from those gene loci (Table 3).…”

Section: Genetic Origin Of Ivig-reactive Light and Heavy Chainsmentioning

confidence: 99%

Genetic origin of IgG antibodies cloned by phage display and anti-idiotypic panning from three patients with autoimmune thrombocytopenia

et al. 1998

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The beneficial use of intravenous immunoglobulins (IVIG) in certain groups of patients with autoimmune thrombocytopenic purpura (AITP) has been proven. AITP is a severe disease in children with a still unknown etiology. It is not clear how IVIG functions in this and other autoimmune diseases. To analyze and compare patient-derived monoclonal IgG antibodies that are bound by IVIG in an anti-idiotypic manner, the combinatorial antibody phage display system was applied. From three different patients with AITP, a large number of clones specifically reacting with IVIG molecules were enriched. The heavy and light chain variable regions were sequenced and compared with each other and with databases. Many variable regions showed extensive replacement mutations within the complementarity-determining regions, while two were identical to germ-line genes. Our data show that the most frequently used germ-line gene loci of these IVIG binders are identical to those observed for many other autoantibodies. This implicates a specific interaction of IVIG particularly with autoantibodies and B cell receptors derived from germ-line genes that are often used for the generation of autoantibodies.

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The Use of Phage Display in Neurobiology

Bradbury

2010

CP Neuroscience

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Phage display has been extensively used to study protein-protein interactions, receptor- and antibody-binding sites, and immune responses, to modify protein properties, and to select antibodies against a wide range of different antigens. In the format most often used, a polypeptide is displayed on the surface of a filamentous phage by genetic fusion to one of the coat proteins, creating a chimeric coat protein, and coupling phenotype (the protein) to genotype (the gene within). As the gene encoding the chimeric coat protein is packaged within the phage, selection of the phage on the basis of the binding properties of the polypeptide displayed on the surface simultaneously results in the isolation of the gene encoding the polypeptide. This unit describes the background to the technique, and illustrates how it has been applied to a number of different problems, each of which has its neurobiological counterparts. Although this overview concentrates on the use of filamentous phage, which is the most popular platform, other systems are also described.

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Molecular cloning and characterization of human thyroid peroxidase autoantibodies of lambda light chain type

Cited by 26 publications

References 44 publications

Human Anti-Thyroid Peroxidase Single-Chain Fragment Variable of Ig Isolated from a Combinatorial Library Assembled In-Cell: Insights into the In Vivo Situation

Human Anti-Thyroid Peroxidase Single-Chain Fragment Variable of Ig Isolated from a Combinatorial Library Assembled In-Cell: Insights into the In Vivo Situation

Genetic origin of IgG antibodies cloned by phage display and anti-idiotypic panning from three patients with autoimmune thrombocytopenia

The Use of Phage Display in Neurobiology

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