T cell receptor ?-chain third complementarity-determining region gene usage is highly restricted among Sm-B autoantigen-specific human T cell clones derived from patients with connective tissue disease

Talken, Beth L.; Holyst, Margaret‐Mary; Lee, David R.; Hoffman, Robert W.

doi:10.1002/1529-0131(199904)42:4<703::aid-anr13>3.0.co;2-7

Cited by 23 publications

(21 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We used the purified hnRNP-MaBP-FP to stimulate peripheral blood mononuclear cells (PBMCs) obtained by apheresis, and cloned the resulting T cell blasts by limiting dilution, as previously described (10,(13)(14)(15)(16). Briefly, cells were obtained by apheresis and then subjected to density gradient centrifugation using Histopaque (Sigma).…”

Section: Methodsmentioning

confidence: 99%

“…Cells were restimulated with 5 ϫ 10 6 autologous APCs that had been irradiated with 30 Gy and hnRNP-FP in fresh medium on day 7. On day 14, T cell blasts were separated by centrifugation on a density gradient and cloned by limiting dilution in the presence of hnRNP-MaBP-FP, fresh irradiated autologous APCs, and 10 units/ml interleukin-2 (IL-2; R&D Systems, Minneapolis, MN) (10,(13)(14)(15)(16). Cells were cloned by limiting dilution and those that were positive for growth (seeded at the lowest number of cells per well) were selected for expansion.…”

Section: Methodsmentioning

confidence: 99%

“…Cells were cloned by limiting dilution and those that were positive for growth (seeded at the lowest number of cells per well) were selected for expansion. T cell clones were tested for proliferation as measured by 3 H-thymidine ( 3 H-TdR) incorporation with the hnRNP-MaBP-FP, the hnRNP peptides, and unconjugated MaBP in the presence of irradiated autologous PBMCs as APCs (10,(13)(14)(15)(16).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Human T cell clones specific for heterogeneous nuclear ribonucleoprotein A2 autoantigen from connective tissue disease patients assist in autoantibody production

Greidinger¹,

Gazitt²,

Jaimes³

et al. 2004

Arthritis & Rheumatism

Self Cite

View full text Add to dashboard Cite

Objective. To identify and characterize human T cells reactive with heterogeneous nuclear RNP A2 (hnRNP A2) antigen, and to determine the ability of hnRNP-reactive T cells to assist in the production of human autoantibodies.Methods. T cells from patients with high serum levels of anti-hnRNP IgG autoantibody were stimulated with an hnRNP recombinant fusion protein, and the cells were cloned by limiting dilution. The surface phenotype and cytokine profiles of the T cells were examined by flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively. T cell clones were cultured with highly purified autologous B cells, and the ability of T cells to enhance autoantibody production under a variety of conditions was measured by ELISA.Results. Human T cells reactive with hnRNP antigen were cloned from 2 patients with systemic lupus erythematosus (SLE) and 1 patient with mixed connective tissue disease (MCTD). The T cells were CD4؉ and had a Th1-like functional phenotype. In coculture in vitro with autologous B cells, T cell clones augmented anti-hnRNP autoantibody production and did so without the need for direct T cell-B cell contact.Conclusion. This study provides direct evidence for a role of anti-hnRNP-reactive T cells in autoantibody production in SLE and MCTD. These findings support the notion that hnRNP-reactive T cells play a role in the pathogenesis of these diseases.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Human T cell clones specific for heterogeneous nuclear ribonucleoprotein A2 autoantigen from connective tissue disease patients assist in autoantibody production

Greidinger¹,

Gazitt²,

Jaimes³

et al. 2004

Arthritis & Rheumatism

Self Cite

View full text Add to dashboard Cite

show abstract

“…The fusion proteins were produced by subcloning fulllength cDNA for snRNP polypeptide 70 kDa, for snRNP polypeptide B3 or for snRNP polypeptide D1 into the pMAL-c2 plasmid vector (New England Biolabs, Beverly, MA, USA), as reported [1]. The synthetic peptides for Sm-B [13] and U1±70 kDa [14] have previously been described. An identical strategy was used to construct the Sm-D peptides.…”

Section: Methodsmentioning

confidence: 99%

Structural Analysis of TCRα and β Chains from Human T‐Cell Clones Specific for Small Nuclear Ribonucleoprotein Polypeptides Sm‐D, Sm‐B and U1–70 kDa: TCR Complementarity Determining Region 3 Usage Appears Highly Conserved

et al. 2001

View full text Add to dashboard Cite

Systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD) are systemic autoimmune diseases that are characterized by the presence of autoantibodies reactive with U small nuclear RNP (snRNP) autoantigens. Both B and T cells are important in the pathogenesis of the disease, and T-and B-cell immunity against snRNP polypeptides have been shown to be linked in vivo. Currently, several alternative hypotheses for the pathogenesis of these diseases have been proposed. These include loss of tolerance, modified self-antigens, molecular mimicry and nondirected immune activation. To help distinguish between the various models of disease pathogenesis, we have characterized the T-cell receptor (TCR) CDR3 from a large panel of well-characterized human T-cell clones and lines specific for individual snRNP polypeptides. The results presented here reveal highly restricted TCR usage across patients by the snRNP-reactive T cells based on the deduced amino acid sequence of the CDR3 loop. These data support the hypothesis that T-cell responses against self antigens in SLE and MCTD are antigen driven and that there are a limited number of T-cell epitopes present on the snRNP autoantigens.

show abstract

“…The nature and specificity of the relevant autoreactive T cells have not been fully characterized. Some autoimmune T cells appear to be specific for nuclear Ags, such as histone (8,9), small nuclear ribonucleoprotein (snRNP) (10,11), heterogeneous nuclear (hnRNP) (12), Smith Ag (13), and DNA topoisomerase I (14). Although these studies have provided important information about T cell epitopes in autoimmune disease, the approach as a whole has permitted analysis of a limited number of epitopes and cannot confirm the actual epitopes present under physiological conditions.…”

Section: S Ystemic Lupus Erythematosus (Sle)mentioning

confidence: 99%

T Cell Reactivity to MHC Class II-Bound Self Peptides in Systemic Lupus Erythematosus-Prone MRL/lpr Mice

Suh

Freed

Cohen

2003

The Journal of Immunology

View full text Add to dashboard Cite

The epitopes recognized by pathogenic T cells in systemic autoimmune disease remain poorly defined. Certain MHC class II-bound self peptides from autoimmune MRL/lpr mice are not found in eluates from class II molecules of MHC-identical C3H mice. Eleven of 16 such peptides elicited lymph node cell and spleen cell T cell proliferation in both MRL/lpr (stimulation index = 2.03–5.01) and C3H mice (stimulation index = 2.03–3.75). IL-2 and IFN-γ production were detected, but not IL-4. In contrast to what was seen after immunization, four self peptides induced spleen cell proliferation of T cells from naive MRL/lpr, but not from C3H and C57BL/6.H2k, mice. These peptides were derived from RNA splicing factor SRp20, histone H2A, β2-microglobulin, and MHC class II I-Akβ. The first three peptides were isolated from I-Ek molecules and the last peptide was bound to I-Ak. T cell responses, evident as early as 1 mo of age, depended on MHC class II binding motifs and were inhibited by anti-MHC class II Abs. Thus, although immunization can evoke peripheral self-reactive T cells in normal mice, the presence in MRL/lpr mice of spontaneous T cells reactive to certain MHC-bound self peptides suggests that these T cells actively participate in systemic autoimmunity. Peptides eluted from self MHC class II molecules may yield important clues to T cell epitopes in systemic autoimmunity.

show abstract

T cell receptor ?-chain third complementarity-determining region gene usage is highly restricted among Sm-B autoantigen-specific human T cell clones derived from patients with connective tissue disease

Abstract: TCR CDR3 gene usage is highly conserved by Sm-B autoantigen-specific T cell clones, and this appears to be related to the recognition of a limited number of T cell epitopes on the Sm-B autoantigen presented in the context of HLA-DR.

Cited by 23 publications

References 28 publications

Human T cell clones specific for heterogeneous nuclear ribonucleoprotein A2 autoantigen from connective tissue disease patients assist in autoantibody production

Human T cell clones specific for heterogeneous nuclear ribonucleoprotein A2 autoantigen from connective tissue disease patients assist in autoantibody production

Structural Analysis of TCRα and β Chains from Human T‐Cell Clones Specific for Small Nuclear Ribonucleoprotein Polypeptides Sm‐D, Sm‐B and U1–70 kDa: TCR Complementarity Determining Region 3 Usage Appears Highly Conserved

T Cell Reactivity to MHC Class II-Bound Self Peptides in Systemic Lupus Erythematosus-Prone MRL/lpr Mice

Contact Info

Product

Resources

About