Anti‐insulin and regulatory anti‐idiotypic antibodies use the same germ‐line V<sub>H</sub>IX gene

Thomas, James W.

doi:10.1002/eji.1830220938

Cited by 7 publications

(4 citation statements)

References 22 publications

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“…For competitive inhibition with insulin, diluted sera were preincubated with insulin at 0.05–100 μg/ml for 2 h pretransfer to insulin-coated plates (25, 26). Insulin binding in the presence or absence of soluble inhibitor was measured using goat anti-mouse IgG conjugated to alkaline phosphatase at room temperature in washing buffer.…”

Section: Methodsmentioning

confidence: 99%

Therapeutic Targeting of Syk in Autoimmune Diabetes

Colonna

Catalano

Chew

et al. 2010

The Journal of Immunology

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In APCs, the protein tyrosine kinase Syk is required for signaling of several immunoreceptors, including the BCR and FcR. We show that conditional ablation of the syk gene in dendritic cells (DCs) abrogates FcγR-mediated cross priming of diabetogenic T cells in RIP-mOVA mice, a situation phenocopied in wild-type RIP-mOVA mice treated with the selective Syk inhibitor R788. In addition to blocking FcγR-mediated events, R788 also blocked BCR-mediated Ag presentation, thus broadly interrupting the humoral contributions to T cell-driven autoimmunity. Indeed, oral administration of R788 significantly delayed spontaneous diabetes onset in NOD mice and successfully delayed progression of early-established diabetes even when treatment was initiated after the development of glucose intolerance. At the DC level, R788 treatment was associated with reduced insulin-specific CD8 priming and decreased DC numbers. At the B cell level, R788 reduced total B cell numbers and total Ig concentrations. Interestingly, R788 increased the number of IL-10–producing B cells, thus inducing a tolerogenic B cell population with immunomodulatory activity. Taken together, we show by genetic and pharmacologic approaches that Syk in APCs is an attractive target in T cell-mediated autoimmune diseases such as type 1 diabetes.

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Section: Methodsmentioning

confidence: 99%

Therapeutic Targeting of Syk in Autoimmune Diabetes

Colonna

Catalano

Chew

et al. 2010

The Journal of Immunology

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“…Although five hybridomas expressed J558 V H family genes, they were encoded by distinct V H germline segments (7,(26)(27)(28)32). The sixth hybridoma, 11C6-E3, expressed a VGAM3.8 V H family gene and was encoded by the BALB/c VFM1 germline gene (30). The heavy-chain V regions of the antimyosin MAbs showed an average of 96.8% nucleotide homology to their germline counterparts, with homology ranging from 89.3 to 99.5%.…”

Section: Resultsmentioning

confidence: 99%

T-Cell-Dependent Antibody Response to the Dominant Epitope of Streptococcal Polysaccharide,N-Acetyl-Glucosamine, Is Cross-Reactive with Cardiac Myosin

Malkiel¹,

Liao²,

Cunningham

et al. 2000

Infect Immun

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Autoantibodies against myosin are associated with myocarditis and rheumatic heart disease. In this study, the antigenic cross-reactivity of myosin and N-acetyl-glucosamine (GlcNAc), the dominant epitope of Group A streptococcal polysaccharide, was examined. Six antimyosin monoclonal antibodies (MAbs) derived from mice with cardiac myosin-induced myocarditis were characterized. All MAbs cross-reacted with GlcNAc, mimicking a subset of MAbs derived from rheumatic carditis patients that bind both myosin and streptococcal polysaccharide. Variable (V) region gene usage was diverse, with five of six MAb heavy-chain V regions encoded by distinct members of the J558 family and the sixth encoded by a member of the VGAM3.8 family. Light-chain V-region segments were derived from the Vk1, Vk4/5, Vk10, and Vk21 families. These antimyosin, anti-GlcNac MAbs demonstrated several T-cell-dependent features: they were predominantly immunoglobulin G, were encoded by V-region genes expressed late in development, and displayed somatic mutation. A direct correlation between the extent of somatic mutation and the affinity for myosin was observed. Affinity for GlcNAc also increased with the frequency of mutation, demonstrating that affinity maturation can occur simultaneously for both self antigen and foreign antigen. Based on these observations, we immunized mice with GlcNAc coupled to bovine serum albumin and demonstrated that a T-cell-dependent response to GlcNAc leads to antimyosin reactivity. We speculate that the pathogenic antibody response in rheumatic carditis may reflect the conversion of a T-cell-independent response to GlcNAc to a T-cell-dependent cross-reactive response to GlcNAc and myosin.

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“…5), up to the CDR3 region, was almost identical to that of an anti-idiotypic antibody to an anti-insulin antibody, except for three amino acids in framework regions at positions 10, 28, and 39 (27). Its CDR3 sequence, however, was markedly different: VFYDGYYAWFPY vs. EYAPYFDV in 82H.…”

Section: Isolationmentioning

confidence: 93%

Idiotypic mimicry and the assembly of a supramolecular structure: an anti-idiotypic antibody that mimics taxol in its tubulin-microtubule interactions.

Leu

Chen²,

Diamanduros³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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Taxol, rii y extrced fro the bark of the western yew, Taxus brenfolia, is reportedly the first of a new class of anti-cancer agent. It Taxol, reportedly the first of a new class of anti-cancer agents, was originally extracted from the bark of the western yew, Taxus brevifolia (1). In vivo, it prevents the disassembly of microtubules, causing the arrest of cells at the G2/M step of the cell cycle (2). Its mechanism differs from that of other anti-cancer agents that have tubulin as their target, such as colchicine, podophyllotoxin, or vinblastine, which inhibit tubulin polymerization (3). The site of action of taxol on tubulin is being investigated in several laboratories. Photoaffinity-labeling experiments implicate the N-terminal region of (3-tubulin as the site of attachment (4, 5).With the aim of using immunological techniques to investigate the action of taxol, we sought to prepare an anti-idiotypic antibody that mimics taxol, using an auto-anti-idiotypic strategy (6, 7). We have succeeded in obtaining an antibody, 82H, that, as intact antibody or Fab fiagment, mimics taxol in its tubulinmicrotubule interactions to the extent that it can cause tubulin to assemble into microtubules. MATERIALS AND METHODSPreparation of and Screening for Anti-Idiotypic Antibodies. The object was to screen for monoclonal antibodies that showed taxol-inhibitable binding to Fab fragments ofaffinitypurified rabbit anti-taxol antibody R585 (8).Preparation of Fab fragments of R585. Rabbit anti-taxol antiserum R585 (8) was affinity-purified as follows. Two affinity columns were prepared. Either 10 mg of bovine serum albumin (BSA) or 10 mg of taxol covalently linked to rabbit serum albumin (taxol-RSA) (8) was dissolved in 1 ml of 0.1 M 2-(N-morpholino)-ethanesulfonic acid (Mes) buffer (pH 4.8) and added to 2 ml of Affi-Gel 10 (Bio-Rad). The mixtures were rocked gently overnight at 40C. The gels were then washed with several column volumes of phosphatebuffered saline (PBS, pH 7.3) and with 0.2 M glycine hydrochloride buffer (pH 2.5) and then equilibrated with PBS. Six milliliters of R585 antiserum was first recirculated through the taxol-RSA-Affi-Gel 10 column overnight at 40C at a rate of0.2 ml/min. The column was then extensively washed with PBS at a rate of 1.0 ml/min to remove all nonspecifically bound proteins. The antibodies were then eluted with 0.2 M glycine hydrochloride (pH 2.5) at a rate of 0.5 ml/min. The eluted antibodies were immediately neutralized with 2 M Tris base and dialyzed against PBS at 40C overnight. On the following day, the eluted antibodies were recycled through the BSA-Affi-Gel 10 column at 40C overnight to remove BSA-binding antibodies. The effluent, which contained antitaxol antibodies in PBS, was collected and concentrated by a Centriprep-10 concentrator (Amicon) to a volume of 1-2 ml.Fab fragments were prepared from the affinity-purified antibodies as described (9).Screening for anti-idiotypic antibodies. A competitive ELISA was used to assay for taxol-inhibitable binding to the Fab fiagments of ...

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Anti‐insulin and regulatory anti‐idiotypic antibodies use the same germ‐line V_HIX gene

Cited by 7 publications

References 22 publications

Therapeutic Targeting of Syk in Autoimmune Diabetes

Therapeutic Targeting of Syk in Autoimmune Diabetes

T-Cell-Dependent Antibody Response to the Dominant Epitope of Streptococcal Polysaccharide,N-Acetyl-Glucosamine, Is Cross-Reactive with Cardiac Myosin

Idiotypic mimicry and the assembly of a supramolecular structure: an anti-idiotypic antibody that mimics taxol in its tubulin-microtubule interactions.

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Anti‐insulin and regulatory anti‐idiotypic antibodies use the same germ‐line VHIX gene

Cited by 7 publications

References 22 publications

Therapeutic Targeting of Syk in Autoimmune Diabetes

Therapeutic Targeting of Syk in Autoimmune Diabetes

T-Cell-Dependent Antibody Response to the Dominant Epitope of Streptococcal Polysaccharide,N-Acetyl-Glucosamine, Is Cross-Reactive with Cardiac Myosin

Idiotypic mimicry and the assembly of a supramolecular structure: an anti-idiotypic antibody that mimics taxol in its tubulin-microtubule interactions.

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Anti‐insulin and regulatory anti‐idiotypic antibodies use the same germ‐line V_HIX gene