Journal: Frontiers in Immunology

Article Title: RIG-I and TLR-7/8 agonists as combination adjuvant shapes unique antibody and cellular vaccine responses to seasonal influenza vaccine

doi: 10.3389/fimmu.2022.974016

Figure Lengend Snippet: Source of reagents and kits used in the study.

Article Snippet: Mouse IL-4 ELISPOT kit , RnD systems , EL404.

Techniques: In Vivo, Luciferase, Enzyme-linked Immunospot, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Isolation

Journal: Scientific Reports

Article Title: Gene Set Enrichment Analysis Identifies LIF as a Negative Regulator of Human Th2 Cell Differentiation

doi: 10.1038/srep00464

Figure Lengend Snippet: Panel a shows results of IL-4 ELISPOT experiments. Culture conditions are mentioned at the bottom. Error bars represent standard error of mean and * denotes significant (p value 0.017) difference between the two sample (Two tailed paired sample t test). In panel b blue line represents IL-4 level detected in the supernatant at given time points and red line is a measure of IL-4R expression level at corresponding time points. Panel c shows a result obtained in a CFSE labeling experiment in ERK silenced cells. Red, blue and green colors represent not activated, TCR + IL-4 stimulated and TCR + IL-4 stimulated at day 1 followed by additional doses of IL-4 at day 3,4 and 5. Less fluorescence indicates more cell divisions because as the cells divide fluorescent label is diluted.

Article Snippet: ELISPOT assays for IL-4were performed with human IL-4 ELISPOT kits (R & D systems).

Techniques: Enzyme-linked Immunospot, Two Tailed Test, Expressing, Labeling, Fluorescence

Journal: Scientific Reports

Article Title: Gene Set Enrichment Analysis Identifies LIF as a Negative Regulator of Human Th2 Cell Differentiation

doi: 10.1038/srep00464

Figure Lengend Snippet: Panel a shows mRNA expression profile of STAT3 upstream molecules. Panel b shows ELISPOT results for IL-4 producing cells in differentially activated populations. Panel c shows results obtained in LIF ELISA experiments. Panel d and e are mRNA expression data adapted form microarray of LIF and IL-6 respectively. Fold change is calculated with respect to untreated sample. In all cases mean of three biological and two technical replicates are shown. Error bars represent standard deviation and * represents significance at p value of less than 0.05 (see text).

Article Snippet: ELISPOT assays for IL-4were performed with human IL-4 ELISPOT kits (R & D systems).

Techniques: Expressing, Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay, Microarray, Standard Deviation

Journal: Gene therapy

Article Title: RNA-loaded CD40-activated B cells stimulate antigen-specific T-cell responses in dogs with spontaneous lymphoma.

doi: 10.1038/gt.2008.22

Figure Lengend Snippet: Figure 3 Canine CD40-B (cCD40-B) cells loaded with canine distemper virus hemagglutinin (CDV-HA) mRNA elicit CDV-specific IFN-g responses in healthy dogs and dogs with spontaneous lymphoma. cCD40-B cells were generated from (a) three healthy dogs, (b) two dogs with lymphoma at the time of diagnosis and (c) two dogs with lymphoma during induction chemotherapy. All dogs had previously received the attenuated live CDV vaccine as a matter of routine veterinary care. cCD40-B cells harvested on day 14 of culture were electroporated with CDV-HA mRNA and co-cultured with autologous PBMC in the presence of rcIL-4, IL-7 and IL-2. On day 7, T cells were harvested and assayed for their ability to secrete IFN-g in response to autologous PHA blasts electroporated with CDV-HA mRNA by cIFN-g ELISpot assay. Experiments were performed in triplicate. Results for seven individual dogs (healthy donors A, B, C and patients A, B, C, D) are shown.

Article Snippet: ELISPOT analysis was performed using the canine IFN-g ELISPOT kit to detect antigen-specific responder cells (R&D Systems).

Techniques: Virus, Generated, Biomarker Discovery, Cell Culture, Enzyme-linked Immunospot

Journal: Cancer Immunology, Immunotherapy

Article Title: Linear doggybone DNA vaccine induces similar immunological responses to conventional plasmid DNA independently of immune recognition by TLR9 in a pre-clinical model

doi: 10.1007/s00262-017-2111-y

Figure Lengend Snippet: Evaluation of CD8+ and CD4+ T cell responses after vaccination with DB or PL DNA vaccines encoding E6E7. Mice were injected i.m. with 50 µg of the E6E7 DB or PL vaccines with or without electroporation. a Serial blood samples were analysed using an E7 49–57 tetramer after priming (left panel) and after boosting (right panel). Values are tetramer positive cells as a percentage of total CD8+ cells. The results are representative of 2 independent experiments each with n = 5 mice per group. * p < 0.05, ** p < 0.01, ns non-significant. b Evaluation of CD8 + responses by IFN-γ ELISPOT after priming and boosting using overlapping peptide pools for E6 and E7. E6 pep and E7 pep are the immunodominant peptides E6 48–57 and E7 49–57 , respectively. Graph is representative of 2 independent experiments each with n = 3 mice per group. Bars represent individual mice. c CD4+ Th responses were measured after priming and boosting using IFN-γ or IL-4 ELISPOT using the E7 43–77 peptide containing a well characterised Th epitope E7 44–60 . Plotted values have the non-specific background (media alone) subtracted. The cut-off line represents two times the response from naive control mice

Article Snippet: Induction of peptide-specific CD8+ or CD4+ T cells in individual mice was assessed ex vivo using an IFN-γ ELISPOT kit or IL-4 ELISPOT kit (CD4+) (both from BD Pharmingen).

Techniques: Injection, Electroporation, Enzyme-linked Immunospot