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Article Title: Major peptide autoepitopes for nucleosome-specific T cells of human lupus

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Figure Lengend Snippet: IL-2 production by anti-DNA autoantibody–inducing T-cell clone DD2 in response to nucleosomal histone peptides (10 μM each) presented by autologous EBV B-cell APCs. Bars represent mean of triplicate values. Values higher than 3 SD (horizontal line) above the mean of background (DD2 + APCs without added peptide produced 132 pg/mL IL-2) were considered as stimulatory. IL-2 production by DD2 in response to anti-CD3 stimulation was 554 pg/mL.

Article Snippet: Purified mAb’s for ELISA, phycoerythrin-conjugated (PE-conjugated) mAb to human IL-10, and FITC-conjugated mAb to human IL-2 were from PharMingen (San Diego, California, USA).

Techniques: Produced

Journal:

Article Title: Major peptide autoepitopes for nucleosome-specific T cells of human lupus

doi:

Figure Lengend Snippet: (a) IL-2 production by a representative short-term CD4 T-cell line L-EB, with different concentrations of histone H449–63 peptide-pulsed autologous EBV B-cell APCs. Stimulation with 0.01–100 μM peptide showed significant differences (*P < 0.05 at 0.01 μM, Student’s t test) from background (T cells plus APCs cultured without peptide). (b) An example of MHC class II–dependent recognition of nucleosomal peptides. Presentation of H449–63 peptide by autologous EBV B cells to short-term T-cell line L-EB was inhibited mainly by anti-DR mAb.

Article Snippet: Purified mAb’s for ELISA, phycoerythrin-conjugated (PE-conjugated) mAb to human IL-10, and FITC-conjugated mAb to human IL-2 were from PharMingen (San Diego, California, USA).

Techniques: Cell Culture

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Article Title: Major peptide autoepitopes for nucleosome-specific T cells of human lupus

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Figure Lengend Snippet: Example of a pepscan of L-SC, a short-term CD4 T-cell line from a patient with lupus, with histone peptides presented by autologous EBV B-cell APCs. The baseline for y axis is set at 3 SD above the mean of background values obtained with T cells cultured with APCs alone. IL-2 production responses to peptides above that value are shown.

Article Snippet: Purified mAb’s for ELISA, phycoerythrin-conjugated (PE-conjugated) mAb to human IL-10, and FITC-conjugated mAb to human IL-2 were from PharMingen (San Diego, California, USA).

Techniques: Cell Culture

Journal:

Article Title: Major peptide autoepitopes for nucleosome-specific T cells of human lupus

doi:

Figure Lengend Snippet: IL-2 production by N-JV, a short-term CD4 T-cell line from a representative normal donor to histone peptides using autologous EBV B-cell APCs. The horizontal line in each panel demarcates 3 SD above the mean of background values. None of the normal T-cell lines showed a response to any of the peptides that was 3 SD above their respective background values.

Article Snippet: Purified mAb’s for ELISA, phycoerythrin-conjugated (PE-conjugated) mAb to human IL-10, and FITC-conjugated mAb to human IL-2 were from PharMingen (San Diego, California, USA).

Techniques:

Journal:

Article Title: Major peptide autoepitopes for nucleosome-specific T cells of human lupus

doi:

Figure Lengend Snippet: Summary of IL-2 production responses of short-term T-cell lines to nucleosomal histone peptides in 10 patients with lupus. The results are expressed as percentages of maximal responses of the T-cell lines stimulated by anti-CD3 mAb. Bars represent mean values from 10 lupus T-cell lines, with error bars in SEM. Values of T cells cultured with autologous EBV B-cell APCs alone were considered as background. The baseline for y axis is set at 22%, which is 3 SD above the mean of background values (14.5%). A peptide was considered stimulatory when it elicited a positive response above this baseline. For instance, the average of responses of all 10 T-cell lines to H416–30 peptide was 23 ± 2.6. All the stimulatory peptides in the panels elicited positive responses in all 10 lupus T-cell lines to levels greater than 3 SD above their respective background values. The extreme right bar in the bottom right (H4) panel was the mean value of responses of the 10 lines to whole nucleosome preparation.

Article Snippet: Purified mAb’s for ELISA, phycoerythrin-conjugated (PE-conjugated) mAb to human IL-10, and FITC-conjugated mAb to human IL-2 were from PharMingen (San Diego, California, USA).

Techniques: Cell Culture

Journal:

Article Title: Major peptide autoepitopes for nucleosome-specific T cells of human lupus

doi:

Figure Lengend Snippet: Representative 2-color intracellular cytokine staining of gated CD4+ T cells freshly obtained from 2 patients with lupus who were in remission (R-WG and R-SC). IFN-γ and IL-10 response (a) and IL-2 and IL-4 production (b) in response to stimulation by anti-CD3, nucleosomes, or the histone peptide epitopes. Demarcation of the quadrants was set based on background staining of the T cells cultured in medium alone. Percentage values for the respective cytokine stainings can be found in Figure ​Figure88.

Article Snippet: Purified mAb’s for ELISA, phycoerythrin-conjugated (PE-conjugated) mAb to human IL-10, and FITC-conjugated mAb to human IL-2 were from PharMingen (San Diego, California, USA).

Techniques: Staining, Cell Culture