Efficient generation of monoclonal antibodies from preselected antigenspecific B cells

Steenbakkers, Peter G. A.; Hubers, Henk A. J. M.; Rijnders, A.W.M.

doi:10.1007/bf00997158

Cited by 17 publications

(14 citation statements)

References 18 publications

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“…Sections (5 m thick) were deparaffinized and used for immunohistochemistry analysis. An anti-MIA monoclonal antibody (mAb) was produced by immunization of BALB/c mice with recombinant MIA according to routine procedures (6,31,32). Clone 14A produced a high-affinity anti-MIA mAb of the IgG1 isotype, which was further tested for monospecificity by Western blotting of cell line and tissue extracts (results not shown).…”

Section: Methodsmentioning

confidence: 99%

Endogenous HLA–DR–restricted presentation of the cartilage antigens human cartilage gp‐39 and melanoma inhibitory activity in the inflamed rheumatoid joint

Lierop¹,

Hoed²,

Houbiers³

et al. 2007

Arthritis & Rheumatism

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Objective. The cartilage proteins melanoma inhibitory activity (MIA) and human cartilage gp-39 (HC gp-39) are candidate autoantigens in rheumatoid arthritis (RA). The present study was undertaken to investigate the endogenous HLA-DR4-restricted presentation of these self proteins, in order to seek in vivo evidence in support of their potential immunologic role. The mechanisms that cause and sustain rheumatoid arthritis (RA) remain poorly elucidated. The association of the disease with certain class II major histocompatibility complex (MHC) haplotypes, however, strongly suggests that specific antigens are presented by antigen-presenting cells (APCs) to CD4ϩ T lymphocytes (1,2). Previous studies indicate that cartilage could be the source of some of these antigens in RA (3-7). In the present study we investigated the local presence and presentation of 2 cartilage-derived proteins that have distinct properties: human cartilage gp-39 (HC gp-39) and a protein called melanoma inhibitory activity (MIA).

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Section: Methodsmentioning

confidence: 99%

Endogenous HLA–DR–restricted presentation of the cartilage antigens human cartilage gp‐39 and melanoma inhibitory activity in the inflamed rheumatoid joint

Lierop¹,

Hoed²,

Houbiers³

et al. 2007

Arthritis & Rheumatism

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“…In a previous study, we used the EL4.B5 system to culture B cells of anti-HLA Ab seropositive, multiparous females in limiting dilution format. This enabled us to determine B cell precursor frequencies (BCPF) 3 toward HLA class I Ags (11). In the present study, we used tetrameric HLA-A2 as an affinity ligand to identify and isolate alloantigen-specific B cells and confirmed their HLA-A2 specificity by induction of specific Ab secretion in culture.…”

Section: T He Characterization Of B Cells That Participate In a Specificmentioning

confidence: 97%

“…Beside their low efficiency, these approaches do not provide a genuine representation of the entire B cell response as immortalization is confined to certain subpopulations of B cells (1,2). Ag-guided selection of B cells provides the yields that are necessary to establish specific Ab-producing B cell lines without the need for immortalization (3).…”

Section: T He Characterization Of B Cells That Participate In a Specificmentioning

confidence: 99%

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

Mulder

Eijsink

Kardol

et al. 2003

The Journal of Immunology

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Characterizing the individual B cells that participate in the production of anti-HLA Abs requires isolation and culture of these cells and a suitable assay for detection of Abs produced in these B cell cultures. We previously showed that B cell precursors, programmed for anti-HLA Ab secretion, are present at measurable frequencies in peripheral blood of women immunized by pregnancy. In this study, we show that tetrameric HLA-A2, although designed for characterization of CTLs, provides a suitable affinity ligand for isolation of allospecific B cells, which subsequently can be induced to produce HLA-A2 Ab in a CD40-driven culture system. The validity of this concept was established by assaying human hybridomas, producing anti-HLA Abs, for specific tetrameric HLA-A2 binding. The availability of anti-HLA Ab-producing B cell cultures that are established without immortalization will be of value when T-B cell interaction is studied at an alloantigen-specific level.

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“…Eight days after expansion of the selected B cells, the cells were tested for specificity in the agglutination assay (see below). The B cell cultures specific for A2b T cells were then submitted to a mini-electrofusion procedure with NS-1 myeloma cells as described previously [24]. After expansion for 1 week, cultures were examined for hybridoma growth and screened in a T-cell agglutination assay.…”

Section: Selection Of A2b Specific B Cellsmentioning

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%

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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Efficient generation of monoclonal antibodies from preselected antigenspecific B cells

Cited by 17 publications

References 18 publications

Endogenous HLA–DR–restricted presentation of the cartilage antigens human cartilage gp‐39 and melanoma inhibitory activity in the inflamed rheumatoid joint

Endogenous HLA–DR–restricted presentation of the cartilage antigens human cartilage gp‐39 and melanoma inhibitory activity in the inflamed rheumatoid joint

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

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

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