Journal:

Article Title: Proanthocyanidins Inhibit UV-Induced Immunosuppression Through IL-12-Dependent Stimulation of CD8 + Effector T Cells and Inactivation of CD4 + T Cells

doi: 10.1158/1940-6207.CAPR-10-0224

Figure Lengend Snippet: Prevention of UVB-induced suppression of the CHS response by GSPs requires IL-12. Dietary GSPs prevent UVB-induced suppression of the CHS response in wild-type mice but not in IL-12 KO mice. Wild-type (C3H/HeN) (Panel A) or IL-12 KO C3H/HeN (Panel B) mice that received either a standard diet or a diet supplemented with GSPs (0.5% or 1.0%, w/w) and exposed to UVB radiation (150 mJ/cm2) on four-consecutive days, sensitized to DNFB and the CHS response to application of DNFB on ear skin (challenge) assessed by measurement of ear swelling response 24 h later, as described in the Materials and Methods. Treatment group number 1 in each panel indicates that mice were not sensitized with DNFB but only challenged with DNFB in ear skin. One group of wild-type mice were injected with anti-IL-12 antibody (Panel A, 8th bar), and one group of IL-12 KO mice (Panel B, 8th and 9th bar) were injected with 1,000 ng of IL-12, 3 h before DNFB sensitization. The change in ear thickness is reported in millimeter (mm ×10-2) as the mean ± SD, with n=5 per group. The experiment was repeated once with similar results. *Significant sensitization versus UVB exposure in the absence of GSPs treatment (5th bar), P<0.001; ¶Significant inhibition versus the positive control of sensitization in the absence of UVB irradiation or GSPs treatment (2nd bar), P<0.005.

Article Snippet: Grape seed proanthocyanidins (GSPs) and dietary supplementation The purified GSPs were obtained from the Kikkoman Corporation (Tokyo, Japan) and the chemical composition has been described earlier ( 17 , 18 ).

Techniques: Injection, Inhibition, Positive Control, Irradiation

Journal:

Article Title: Proanthocyanidins Inhibit UV-Induced Immunosuppression Through IL-12-Dependent Stimulation of CD8 + Effector T Cells and Inactivation of CD4 + T Cells

doi: 10.1158/1940-6207.CAPR-10-0224

Figure Lengend Snippet: GSPs protect against UVB-induced immunosuppression and does so even after cessation of treatment. Mice (C3H/HeN) that received a standard diet or a GSPs-supplemented diet were UVB-irradiated, sensitized through the UVB-irradiated skin, challenged by application of DNFB on the right ear (primary challenge), and the ear skin swelling measured 24 h later. The CHS response after primary challenge of the UVB-exposed mice that did not receive GSPs was significantly lower than the CHS response of the mice that were not UVB-irradiated, whereas mice that received GSPs in the diet before and during the CHS protocol mounted a CHS response to the primary DNFB challenge that was comparable to the response in the mice that were not UVB-irradiated. When the mice were rested for 4 weeks after primary challenge, then further challenged with DNFB (secondary challenge) on the left ear similar results were obtained. The change in ear swelling response in each group is reported as mean ± SD, n=5 per group. The experiment was repeated once with similar observations. *Significant increase versus UVB exposure in the absence of GSPs treatment, P<0.001; ¶Significant increase versus UVB in the absence of GSPs treatment, P<0.005; †Significant inhibition versus the positive control (unirradiated mice) (2nd bar from the top), P<0.001.

Article Snippet: Grape seed proanthocyanidins (GSPs) and dietary supplementation The purified GSPs were obtained from the Kikkoman Corporation (Tokyo, Japan) and the chemical composition has been described earlier ( 17 , 18 ).

Techniques: Irradiation, Inhibition, Positive Control

Journal:

Article Title: Proanthocyanidins Inhibit UV-Induced Immunosuppression Through IL-12-Dependent Stimulation of CD8 + Effector T Cells and Inactivation of CD4 + T Cells

doi: 10.1158/1940-6207.CAPR-10-0224

Figure Lengend Snippet: GSPs prevent transferable UVB-induced suppression through activation of T cells. A, Donor mice (C3H/HeN) that received either a standard diet or a diet supplemented with GSPs were sensitized, UVB-irradiated, sacrificed 5 days later and single-cell suspensions prepared from the regional lymph nodes and spleens, as detailed in the Materials and Methods. Syngeneic Recipient mice were injected i.v. with 5× 107 spleen and lymph node cells obtained from syngeneic donor mice. Recipient mice were sensitized with DNFB 24 h after transfer, ear challenge was performed 5 days later, and ear skin thickness was measured before and 24 h after challenge. B, The donor mice were treated as described in Panel A, except that CD8+ T cells were positively selected from the spleen and lymph nodes cell preparations. The CD8+ T cells (8× 106) were injected i.v. into naïve mice, the recipient mice were challenged immediately and the ear swelling response was measured 24 h later. In one group of mice, the donor mice were administered recombinant IL-12 (1000 ng/mouse) i.p. 3 h before sensitization. In another group, donor mice received an i.p. injection of anti-IL-12 (500 ng/mouse) 24 and 3 h before DNFB sensitization. Control mice received rat IgG1 (isotype control of anti-IL-12). The change in ear thickness is reported as the mean of millimeters (mm × 10-2) ±SD, n=5 per group. *Significantly greater CHS response versus UVB irradiation in the absence of GSPs treatment, P<0.001; ¶Significantly greater CHS response versus recipient of T cells from UVB+ DNFB treated mice, P<0.01; †Significantly lower CHS response versus the positive control (DNFB-sensitized) group, P<0.001

Article Snippet: Grape seed proanthocyanidins (GSPs) and dietary supplementation The purified GSPs were obtained from the Kikkoman Corporation (Tokyo, Japan) and the chemical composition has been described earlier ( 17 , 18 ).

Techniques: Activation Assay, Irradiation, Injection, Recombinant, Control, Positive Control

Journal:

Article Title: Proanthocyanidins Inhibit UV-Induced Immunosuppression Through IL-12-Dependent Stimulation of CD8 + Effector T Cells and Inactivation of CD4 + T Cells

doi: 10.1158/1940-6207.CAPR-10-0224

Figure Lengend Snippet: Treatment of mice with GSPs or rIL-12 enhances the levels of production of IL-2 and IFNγ by CD8+ T cells. Mice were treated and CD8+ T-cells isolated as described under Figure 3. The CD8+ T-cells were then co-cultured with DNSB-labeled BMDC for 48 h, as detailed in the Materials and Methods. The concentrations of cytokines in the cell supernatants were estimated by ELISA and are presented as the mean± SD in terms of pg or ng per 2 million cells. n=5/group. *Significant increase versus UVB+DNFB group, P<0.001

Article Snippet: Grape seed proanthocyanidins (GSPs) and dietary supplementation The purified GSPs were obtained from the Kikkoman Corporation (Tokyo, Japan) and the chemical composition has been described earlier ( 17 , 18 ).

Techniques: Isolation, Cell Culture, Labeling, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Proanthocyanidins Inhibit UV-Induced Immunosuppression Through IL-12-Dependent Stimulation of CD8 + Effector T Cells and Inactivation of CD4 + T Cells

doi: 10.1158/1940-6207.CAPR-10-0224

Figure Lengend Snippet: (A) GSPs prevent transferable suppression by UVB through the inactivation of UVB-induced CD4+ suppresser T cells. Mice were treated as described in Figure 3 and CD4+ T cells purified from the splenocytes and lymphocytes by positive selection. Twenty-four hours after i.v. injection of CD4+ T cells into naïve mice, the mice were sensitized with DNFB, and ear challenged 5 days after sensitization, as detailed in the Materials and Methods. Those naïve mice that received CD4+ T cells from UVB-exposed donor mice that were GSPs treated showed a greater CHS response than UVB-exposed mice that were not GSPs treated. Those naïve mice that received CD4+ T cells from UVB-exposed rIL-12-injected donor mice showed a significantly higher CHS response than UVB-exposed mice that were not injected with rIL-12. The change in the ear swelling response in each group is reported as mean ± SD (n=5 per group). Significantly greater CHS response versus UVB irradiation in the absence of GSPS or rIL-12 treatment, †P<0.001; Significantly lower CHS response versus positive control group (2nd bar), ¶P<0.01. (B) Treatment of mice with GSPs or rIL-12 decreases the production of IL-4 and IL-10 by CD4+ T cells. Mice were sensitized to DNFB after UVB-irradiation described in Figure 3, the CD4+ T cells isolated by positive selection and then co-cultured with DNSB-labeled BMDC for 48 h, as detailed in the Materials and Methods. The concentrations of cytokines in the cell supernatants were estimated by ELISA and are presented as the mean ±SD in terms of pg/2 million cells. Experiment was repeated once, n=5. Significant decrease versus UVB+DNFB group, †P<0.001; Significant decrease versus positive control (DNFB alone), ¶P<0.001; Significant increase versus positive control (DNFB alone), *P<0.001

Article Snippet: Grape seed proanthocyanidins (GSPs) and dietary supplementation The purified GSPs were obtained from the Kikkoman Corporation (Tokyo, Japan) and the chemical composition has been described earlier ( 17 , 18 ).

Techniques: Purification, Selection, Injection, Irradiation, Positive Control, Isolation, Cell Culture, Labeling, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Acetyl-Phosphate Is Not a Global Regulatory Bridge between Virulence and Central Metabolism in Borrelia burgdorferi

doi: 10.1371/journal.pone.0144472

Figure Lengend Snippet: Strains, vectors, and primers used in this study.

Article Snippet: Tailed cDNA was subsequently used in PCR reactions with an internal GSP primer ( ackA -GSP2 or pta -GSP2— ) and an abridged anchor primer provided by Invitrogen.

Techniques: Sequencing