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: Plant Direct

Article Title: The maize premature senesence2 encodes for PHYTOCHROME‐DEPENDENT LATE‐FLOWERING and its expression modulation improves agronomic traits under abiotic stresses

doi: 10.1002/pld3.295

Figure Lengend Snippet: Cloning and characterization of the Pre2 candidate gene. (a) Gene structure of the pre2 candidate gene, Zm00001d0053300. Exons and introns are represented by filled rectangles and thin lines, respectively. Insertions of a Mutator ( Mu ) in intron1 and a retrotransposon (RTE) in intron8 in the pre2‐1 and pre2‐2 mutant alleles are represented by triangles, respectively. The gene‐specific primers (GSPs) used in the PCR‐fingerprinting and RT‐PCR expression analyses are shown above the exons and arrows are pointing in their directions. (b) Full‐length cDNA of Pre2 amplified by RT‐PCR using 3′‐RACE. Lanes 1 and 2 are both represent the FL‐cDNAs amplified by using PHN137957 and PHN137958 GSPs from the 5′‐UTR of the pre2 candidate gene. (c) RT‐PCR of the pre2‐1 allele and its wild type sib using PHN137983 and PHN137985 GSPs from the exon1 and exon2 of the pre1 candidate gene, respectively. Three additional mature transcripts of ~301 bp, ~339 bp, and ~552 bp sizes with variable intensities were detected in the pre2‐1 allele as compared to only one functional transcript of ~179 bp in its WT sib. The blue and green regions of bars on the right side of Figure represent the predicted polypeptides of the differentially spliced mature transcripts resulted in frame shift (FS) and/or early termination (T) as compared to the functional predicted polypeptide represented by the black bars. Total RNA from ten days old seedlings was used in the RT‐PCR analysis. (d) Southern blot (SB) analysis of a segregating population detecting an RFLP of ~8.0 Kb/ EcoRI associated with the pre2‐2 mutant allele phenotype. The FL‐cDNA was used as a DNA probe for hybridization in SB analysis. (E) PCR‐fingerprinting of 7 F1 plants (lanes 2–8) of pre2‐1 and pre2‐2 allelic cross using PHN137983 in combination with Mu ‐TIR (upper lane in Figure ), GSP‐Exon8‐F2 with RTE‐R1 primer (middle lane in Figure ), and PHN137983 with PHN137985 (lower lane in Figure ). (f) The RT‐PCR expression analysis of the pre2‐2 mutant allele and its WT sib using GSP‐Exon8‐F2 and GSP‐Exon9‐R1 primers of the pre2 candidate gene. Two additional mature transcripts were detected in the pre2‐2 mutant allele as compared to a functional transcript of ~700 bp size in its WT sib. The colored bars on the right side of Figure are representing the predicted polypeptides as described in Figure . Amplification of the ZmActin1 was used as a control in RT‐PCR analysis

Article Snippet: Since only a few partial ESTs, representing 3′ end of the Zm00001d053300 gene, were found in both the public and Corteva Agriscience maize databases, we amplified and cloned ~4.0 kb full‐length cDNA (FL‐cDNA) by RT‐PCR using two GSPs from the 5′UTR of the pre2 candidate gene in 3′‐RACE (Figure ).

Techniques: Cloning, Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Expressing, Amplification, Functional Assay, Southern Blot, Hybridization, Control