Journal: Vaccines

Article Title: Immunogenicity of the Xcl1 -SARS-CoV-2 Spike Fusion DNA Vaccine for COVID-19

doi: 10.3390/vaccines10030407

Figure Lengend Snippet: Xcl1-S vaccination elicited a significant increase in cellular immune responses. ( A , B ) The number of IFN-γ-expressing cells ( A ) and IL-4-expressing cells ( B ) from splenocytes were measured by ELISpot. Data are mean ± SEM of the spleens from n = 5 mice per group. ( C , D ) Splenocytes were stimulated with a combined mixture of S peptide pools for 18 h. The frequency of S-specific IFN-γ CD4 + and CD8 + T cells was determined using a flow cytometry analysis. Summary graphs showing the frequencies of IFN-γ CD4 + and CD8 + T cells after vaccination. Data shown in the graphs represent the average of five mice in each group, and error bars represent SEM. ★, p < 0.05, ★★, p < 0.01.

Article Snippet: Splenocytes were counted and 2 × 10 5 splenocytes per well were plated into anti-IFN-γ or anti-IL-4 plates and restimulated with the N terminal domain (NTD), receptor binding domain (RBD) and S1 peptides pools with 5 µg/mL concentration for 18 h at 37 °C 5% CO 2 (Sino Biological, cat. PP001, PP002 and PP003-A, Beijing, China).

Techniques: Expressing, Enzyme-linked Immunospot, Flow Cytometry

Journal: Mucosal immunology

Article Title: IL-13-mediated immunological control of enterochromaffin cell hyperplasia and serotonin production in the gut.

doi: 10.1038/mi.2012.58

Figure Lengend Snippet: Figure 7 Phorbol ester (phorbol-12-myristate-13-acetate (PMA)) induces the release of 5-hydroxytryptamine (5-HT) from BON cells after 1 h in a dose-dependent manner, but recombinant human interleukin- 13 (rhIL-13) at three different concentrations does not. BON cells were treated with serum-free media (Control) containing either PMA or rhIL- 13 for 1 h at 37 ° C. Media were collected and 5-HT concentration was determined by enzyme-linked immunosorbent assay (ELISA). Each bar represents mean ± s.e.m. of six biological replicates (wells). a, b and c represent statistical significance. a is significantly different from b and c, b is significantly different than a and c, and all the conditions with c are not significantly different from each other.

Article Snippet: Colon samples were analyzed using a mouse IL-13 and IL-4 ELISA kit (R & D Systems, Minneapolis, MN) according to the manufacturer ’ s instructions.

Techniques: Recombinant, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Live attenuated Salmonella displaying HIV-1 10E8 epitope on fimbriae: systemic and mucosal immune responses in BALB/c mice by mucosal administration

doi: 10.1038/srep29556

Figure Lengend Snippet: ( a ) Serum total IgG levels at day 14 after the final 10E8 peptide boost were measured by quantitative ELISA. The correlations between the serum total IgG concentration and total IgG ASC in BM ( b ), and the specific antibody titer ( c ) were shown. ( d ) IgG subclasses at day 14 after the final 10E8 peptide boost were analyzed by quantitative ELISA. ( e ) The Th1/Th2 bias of immunity was shown as the ratio of IgG2a to IgG1 in concentration. ( f ) The levels of Th1 cytokines (IL-10, IFN-γ, TNF-α) and Th2 cytokines (IL-4, IL-6, IL-21) in sera of immunized mice were detected by quantitative ELISA. R, correlation coefficient. Data shown as mean ± SD of 5 mice in each group. The asterisk indicates a significant difference versus PBS control (* P < 0.05).

Article Snippet: Secretory levels of cytokines IL-4, IL-6, IL-10, IL-21, IFN-γ and TNF-α in the mouse sera were quantitatively determined using the ELISA kits (BOSTER, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control

Journal: PLoS ONE

Article Title: NKT Cells Stimulated by Long Fatty Acyl Chain Sulfatides Significantly Reduces the Incidence of Type 1 Diabetes in Nonobese Diabetic Mice

doi: 10.1371/journal.pone.0037771

Figure Lengend Snippet: ( A , B ) C16:0 and C24:0 sulfatide stimulate different CD4 + T cell cytokine secretion profiles. NOD spleen CD4 + T cells were co-cultured for 72 h with mitomycin treated CD4 – cells at a CD4 + :CD4 – ratio of 100∶1 in the presence of control vehicle or sulfatide (C16:0 or C24:0, 50 mg/ml), as described in Fig. 3A. The concentration of IFN-g (A) and IL-10 (B) secreted into cell supernatants were analyzed by ELISA. IL-2 and IL-4 were not detected in these supernatants. Data shown were obtained from one of three representative and reproducible experiments. ( C , D ) Treatment with sulfatide inhibits the induced anti-islet diabetogenic T cell cytokine responses. Female NOD mice (4 week-old, 4 mice/group) were injected i.p. with sulfatide (20 mg/mouse) or vehicle/PBS. One week later, the treated mice were immunized with 100 mg of either the insulin p9–23, GAD206-220, GAD524-543 or hsp277 peptide. Ten days following antigenic challenge, PLN lymphocytes were assayed for their proliferative ([ 3 H]-thymidine incorporation) and cytokine (IFN-g, IL-4) secretion responses (Elispot assay). The average frequencies of IFN-γ ( C ) or IL-4 ( D ) secreting cells reactive to different islet antigens are shown. In comparison to control vehicle values, statistically significant reductions in the frequencies of cytokine-secreting islet antigen-reactive T cells were found for the insulin and GAD peptides but not Hsp peptide, as follows. Insulin 9–23 (P = 0.0001 for IFN-g, P = 0.0078 for IL-4); GAD 206-220 (P = 0.012 for IFN-g, P = 0.0018 for IL-4); GAD 524-543 (P<0.0001 for IFN-g, P = 0.0328 for IL-4); and Hsp 277 (Not significant, P = 0.0542 for IFN-g; Not significant, P = 0.0952 for IL-4).

Article Snippet: The wells were then incubated with biotin-conjugated anti-IFN-g or anti-IL-4 mAbs followed by incubation with avidin peroxidase (Vector Laboratories).

Techniques: Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay, Injection, Enzyme-linked Immunospot

Journal: The Journal of Clinical Investigation

Article Title: Gasdermin C sensitizes tumor cells to PARP inhibitor therapy in cancer models

doi: 10.1172/JCI166841

Figure Lengend Snippet: ( A ) GZMB-mediated GSDMC cleavage by caspase-6 in MDA-MB-157 and Hs578t cells. Caspase-6i, caspase-6 inhibitor; caspase-8i, caspase-8 inhibitor. ( B ) MDA-MB-436 cells expressing WT GSDMC ( GSDMC -WT) or the D365A mutant ( GSDMC -mut) were treated with GZMB or inhibitors of caspase-6 or caspase-8. Immunoblotting of GSDMC cleavage. ( C ) Same as B , except that cells were cocultured with T cells instead of GZMB treatment. ( D ) Cell death measured by LDH release (LDH-released cell death) induced by GZMB in MDA-MB-157 and Hs578t cells ( n = 3). ( E ) LDH-released cell death induced by cytotoxic T cells in MDA-MB-157 and Hs578t cells treated with caspase-6 siRNA (6si) and/or caspase-8 siRNA (8si) ( n = 3). ( F ) LDH-released cell death induced by cytotoxic T cell in MDA-MB-436 cells with expression of vector, GSDMC -WT, and GSDMC -mut ( n = 3). ( G ) Quantification of cytokine levels by ELISA in tumors of Figure 2F. ( H ) GSDMC induction by cytokines indicated in BT549 and HCC38 cells. ( I ) IFN-γ enhanced LDH-released cell death induced by olaparib at indicated concentration in BT549 and HCC38 cells ( n = 3). ( J and K ) IFN-γ enhanced LDH-released cell death in BT549 and HCC38 cells treated with cytosolic delivery of GZMB ( J ) or cocultured with cytotoxic T cells ( K ) ( n = 3). Data represent mean ± SD. 1-way ANOVA was used for D – F . 2-way ANOVA was used for I . Unpaired 2-tailed t test was used for G , J , and K . * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ELISA kits for detection of TGF-β (ab119557), IL-1α (ab199076), IL-1β (ab100705), and IL-4 (ab100710) were purchased from Abcam.

Techniques: Expressing, Mutagenesis, Western Blot, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Concentration Assay