Generation and functional characterization of anti-clonotype antibodies to human T-cell receptors

Steenbakkers, Peter G. A.; Boots, Annemieke M. H.; Rijnders, A.W.M.

doi:10.1016/s0022-1759(97)00176-2

Cited by 4 publications

(9 citation statements)

References 33 publications

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“…The technique we used, was used recently for the successful generation of antibodies against human TCRs [23]. Although this technique included several selection steps, most hybridomas we obtained were not specific for the A2b T cells.…”

Section: Discussionmentioning

confidence: 99%

“…Nuclear and cellular debris was removed by centrifugation. To load beads with the TCR/CD3 complexes, goat anti mouse (GAM) paramagnetic beads (Dynal 11006 Oslo, Norway) were first incubated with R73 (mouse antiratTCR) as described previously [23]. After incubation, the beads were washed and the TCR/CD3 complexes of A2b or Z1a T cells were added.…”

Section: Loading Beads With Tcr/r73mentioning

confidence: 99%

“…To preclear spleen cells for B cells reactive to the constant region of the TCR chains or to rat CD3, cells were subjected to two rounds of negative selection [23]. Therefore, spleen cells were incubated with beads coupled to TCR/CD3 complexes of Z1a T cells (AV2 and BV8S2 [17]).…”

Section: Selection Of A2b Specific B Cellsmentioning

confidence: 99%

“…For this assay, 50 l supernatant of B cell cultures or hybridoma cultures was mixed with 2×10 5 paramagnetic GAM-beads and 5×10 4 A2b T cells or Z1a T cells [23]. After 3 h incubation at 37°C, agglutination was examined under a light microscope.…”

Section: T-cell Agglutination Assaymentioning

confidence: 99%

“…To increase the likelihood of finding a clonotype specific antibody, we used an earlier described method [23,24]. B cells derived from the A2b T cell immunized mice were selected for the production of Ig molecules specific for these T cells.…”

Section: Generation Of Three Hybridomas Specific For the A2b T Cell Cmentioning

confidence: 99%

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Generation and Characterization of a Clonotypic Antibody Specific for the T Cell Receptor of an Arthritogenic T Cell Clone—Studies in Adjuvant Arthritis

Tienhoven

Steenbakkers²,

Veenstra

et al. 2000

Journal of Autoimmunity

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Section: Discussionmentioning

confidence: 99%

Section: Loading Beads With Tcr/r73mentioning

confidence: 99%

Section: Selection Of A2b Specific B Cellsmentioning

confidence: 99%

Section: T-cell Agglutination Assaymentioning

confidence: 99%

Section: Generation Of Three Hybridomas Specific For the A2b T Cell Cmentioning

confidence: 99%

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Generation and Characterization of a Clonotypic Antibody Specific for the T Cell Receptor of an Arthritogenic T Cell Clone—Studies in Adjuvant Arthritis

Tienhoven

Steenbakkers²,

Veenstra

et al. 2000

Journal of Autoimmunity

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Methods for Construction of Yeast Display Libraries of Four-Domain T-Cell Receptors

Sádio

Stadlmayr

Eibensteiner

et al. 2019

Methods in Molecular Biology

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T‐cell anergy induced by clonotype‐specific antibodies: modulation of an autoreactive human T‐cell clone in vitro

Steenbakkers¹,

Boots²,

Rijnders³

1999

Immunology

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SUMMARYMonoclonal antibodies (mAb) specific for the clonotype of an autoreactive T cell may be useful reagents in the modulation of autoimmune disease. We have previously reported the generation of a set of mAb specific for the clonotypic structure of a human T-cell clone recognizing an epitope of human cartilage gp-39. This glycoprotein was recently identified as a candidate autoantigen in rheumatoid arthritis. Here, we demonstrate for the first time that small amounts of immobilized anticlonotype mAb can induce anergy in the autoreactive clone. Following the anergic stimulus, T cells failed to proliferate upon restimulation as a result of a lack of interleukin-2 (IL-2) gene transcription. In addition, a diminished interferon-c (IFN-c) production was found. Our data indicate that anergy was not a result of T-cell receptor (TCR) downmodulation or the absence of free TCR. The anergic state was induced independent of costimulation or the presence of IL-2 and no protein synthesis was required for the induction of anergy. Anticlonotype mAbinduced anergy was prevented by cyclosporin A, suggesting that active signalling via the calcium/ calcineurin pathway was required for the induction of anergy. In coculture experiments, anergic T cells were found to suppress the response of reactive cells from the same clone. This bystander suppression led to 90% inhibition of peptide-induced proliferation. Together, these findings suggest that mAb to the clonotypic structure of autoreactive T cells may be suitable reagents for the functional inactivation of these T cells in autoimmune diseases.

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Generation and functional characterization of anti-clonotype antibodies to human T-cell receptors

Cited by 4 publications

References 33 publications

Generation and Characterization of a Clonotypic Antibody Specific for the T Cell Receptor of an Arthritogenic T Cell Clone—Studies in Adjuvant Arthritis

Generation and Characterization of a Clonotypic Antibody Specific for the T Cell Receptor of an Arthritogenic T Cell Clone—Studies in Adjuvant Arthritis

Methods for Construction of Yeast Display Libraries of Four-Domain T-Cell Receptors

T‐cell anergy induced by clonotype‐specific antibodies: modulation of an autoreactive human T‐cell clone in vitro

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