immunospot mouse ifn γ elispot kit  (Cellular Technology Ltd)

Journal: Journal of Biomedicine and Biotechnology

Article Title: Immunogenicity of a Recombinant Influenza Virus Bearing Both the CD4+ and CD8+ T Cell Epitopes of Ovalbumin

doi: 10.1155/2011/497364

Figure Lengend Snippet: T cell responses in WSN-OVA I/II virus-infected mice. C57BL/6J mice were immunized i.n. with 5 × 10 2 PFU of WSN virus or 3 × 10 5 PFU of WSN-OVA I/II virus, and sacrificed on day 9 for measuring epitope-specific CD8+ T cell responses in mediastinal lymph nodes (MLN) and spleens (a) and CD4+ T cell responses in MLN (b), respectively, by use of an IFN- γ ELISPOT assay with the indicated peptides. The values presented are averages of triplicate wells; the error bars indicate the standard deviation. The data are representative of three experiments. * P < 0.05 comparing WSN- versus WSN-OVA I/II -infected mice.

Article Snippet: Nine days later, five mice from each group were sacrificed to obtain spleens and mediastinal lymph nodes (MLN), and epitope-specific T cell responses were determined by using a mouse IFN- γ enzyme-linked immunospot assay (ELISPOT) kit (Pharmingen, BD).

Techniques: Infection, Enzyme-linked Immunospot, Standard Deviation

Journal: Journal of Biomedicine and Biotechnology

Article Title: Immunogenicity of a Recombinant Influenza Virus Bearing Both the CD4+ and CD8+ T Cell Epitopes of Ovalbumin

doi: 10.1155/2011/497364

Figure Lengend Snippet: Protective and antigen-specific immunity to melanoma B16 cells expressing OVA. C57BL/6J mice were immunized once with either the parental WSN virus or the recombinant WSN-OVA I/II virus. Three weeks after immunization, mice were challenged i.v. with 3 × 10 5 B16-OVA cells or with PBS. The number of pulmonary metastases (a) and the frequency of SIINFEKL-specific IFN- γ -producing T cells, as measured by using the ELISPOT assay on spleen-derived lymphocytes (b) was determined 12 days after the tumor cell inoculation. All of the experiments included 8 mice per group and were repeated three times with similar results. * P < 0.001 and ** P < 0.0001, comparing WSN-OVA I/II -versus WSN-infected and naïve mice.

Article Snippet: Nine days later, five mice from each group were sacrificed to obtain spleens and mediastinal lymph nodes (MLN), and epitope-specific T cell responses were determined by using a mouse IFN- γ enzyme-linked immunospot assay (ELISPOT) kit (Pharmingen, BD).

Techniques: Expressing, Recombinant, Enzyme-linked Immunospot, Derivative Assay, Infection

Journal: PLoS ONE

Article Title: A Whole Virus Pandemic Influenza H1N1 Vaccine Is Highly Immunogenic and Protective in Active Immunization and Passive Protection Mouse Models

doi: 10.1371/journal.pone.0009349

Figure Lengend Snippet: Balb/c mice were immunized with pandemic H1N1 (H1N1 A/California/7/2009), seasonal H1N1 (H1N1 A/Brisbane/59/2007), and pandemic H5N1 (H5N1 A/Vietnam/1203/2004) vaccines. Spleen cells were collected 7 days after the first, or 21 days after the booster immunization (i.e. 42 days after the first), and stimulated with various seasonal or pandemic influenza virus antigens, before determination of cells responding by secretion of either IFN-g or IL-4 by an ELISPOT assay. Anti-HA IgG subclass responses were analyzed by ELISA using sera collected on day 42.

Article Snippet: The frequency of IFN-γ- or Interleukin-4 (IL-4)-secreting cells was analyzed using mouse IFN-γ and IL-4 enzyme-linked immunospot (ELISPOT) kits (Mabtech AB, Nacka, Sweden), as previously described , using vaccine antigen at a concentration of 0.3 µg HA/ml.

Techniques: Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay

Journal: Cancer immunology research

Article Title: Pathogen-boosted adoptive cell transfer therapy induces endogenous antitumor immunity through antigen spreading

doi: 10.1158/2326-6066.CIR-19-0251

Figure Lengend Snippet: (A-B) One month after ReACT-mediated tumor eradication, CD8+ T cells were isolated from draining lymph nodes, stimulated ex vivo with irradiated B16-F10 cells, and assessed for IFNγ secretion using an ELISPOT assay. IFNγ-producing cells are shown in (A) representative images along with (B) quantification. (C-D) Naïve and ReACT memory mice (two months after tumor clearance) were intravenously injected with 1 × 105 B16-F10 cells and sacrificed 15 days later. (C) Representative images of lungs and (D) quantification of tumor foci in lungs isolated 15 days after tumor challenge. Data represents cumulative results from two independent experiments with n=6/group for (A-B) and n=10/group for (C-D). The significance was determined by t-test. Mean±SEM is shown. **P<0.01, ***P<0.001.

Article Snippet: Enzyme-linked immunospot assay (ELISPOT) For ELISPOT assays, an Immunospot mouse IFNγ ELISPOT Kit from CTL (Cleveland, OH) was used.

Techniques: Isolation, Ex Vivo, Irradiation, Enzyme-linked Immunospot, Injection

Journal: Journal of Virology

Article Title: Expanding Regulatory T Cells Alleviates Chikungunya Virus-Induced Pathology in Mice

doi: 10.1128/JVI.00998-15

Figure Lengend Snippet: Pretreatment with IL-2 Ab Cx reduces production of proinflammatory cytokines in joint footpads of CHIKV-infected mice. WT mice ( n = 5 per group) were infected s.c. with 10 6 PFU CHIKV-SGP011 after treatment with PBS, IL-2 only, JES6-1 only, or IL-2 Ab Cx. Cells were stimulated with either IL-2 only or IL-2 plus CHIKV. ELISpot was performed in quadruplicate to detect for CHIKV-specific IFN-γ-producing cells. (A and B) Representative images of ELISpot wells depicting the number of IFN-γ-producing cells in total cells (A) and enriched CD4 + T cells (B) from joint footpads. Bar charts show average numbers of IFN-γ-producing cells per infected joint footpad after subtraction of the background of the IL-2-only stimulation control. (C) mRNA was extracted from joint footpads, and qRT-PCR was performed to detect the expression of IL-6, IL-10, ENTPD1, IFN-γ, STAT1, and CXCL10. Statistical analysis was done using one-way ANOVA across all CHIKV-infected groups, followed by Dunnett's posttest comparing to the PBS-CHIKV group. Data are means and SD from three independent experiments. *, P = 0.0127 (IL-2 Ab Cx IL-6) or 0.0113 (IL-2 Ab Cx IL-10); **, P = 0.0037 (IL-2 Ab Cx ENTPD1), 0.0036 (IL-2 Ab Cx IFN-γ), 0.0032 (IL-2 Ab Cx STAT1), or 0.0067 (IL-2 Ab Cx CXCL10); ** *, P = 0.0008 (IL-2 Ab Cx IFN-γ-producing cells per joint footpad) or 0.0126 (IL-2 Ab Cx IFN-γ-producing CD4 + T cells per joint footpad).

Article Snippet: Spot detection was done using a mouse IFN-γ enzyme-linked immunospot (ELISpot) kit with alkaline phosphatase (ALP) (Mabtech) following the manufacturer's instructions.

Techniques: Infection, Enzyme-linked Immunospot, Quantitative RT-PCR, Expressing

Journal: Journal of Virology

Article Title: Expanding Regulatory T Cells Alleviates Chikungunya Virus-Induced Pathology in Mice

doi: 10.1128/JVI.00998-15

Figure Lengend Snippet: Pretreatment with IL-2 Ab Cx reduces the frequency of CHIKV-specific responses in pLN. WT mice ( n = 5 per group) were infected s.c. with 10 6 PFU CHIKV-SGP011 after treatment with PBS, IL-2 only, JES6-1 only, or IL-2 Ab Cx. Cells from draining pLN were isolated at 6 dpi and analyzed using flow cytometry. (A through D) The bar charts show CD45 + T cells (A), CD4 + T cells (B), CD4 + Teff cells (C), and CD4 + Tregs (D) per pLN. Cells were stimulated with either IL-2 only or IL-2 plus CHIKV. ELISpot was performed in quadruplicate to detect for CHIKV-specific IFN-γ-producing cells. (E and F) Representative images of ELISpot wells depicting the number of IFN-γ-producing cells in total cells (E) and enriched CD4 + T cells (F) from pLN. The bar charts show the average numbers of IFN-γ-producing cells per pLN after subtraction of the background of the IL-2-only stimulation control. Statistical analysis was done using one-way ANOVA across all CHIKV-infected groups, followed by Dunnett's posttest comparing to the PBS-CHIKV group. Data are means and SD from three independent experiments. ***, P < 0.0001 (IL-2 Ab Cx Tregs per pLN), 0.0008 (IL-2 Ab Cx IFN-γ-producing cells per pLN), or 0.0007 (IL-2 Ab Cx IFN-γ-producing CD4 + T cells per pLN).

Article Snippet: Spot detection was done using a mouse IFN-γ enzyme-linked immunospot (ELISpot) kit with alkaline phosphatase (ALP) (Mabtech) following the manufacturer's instructions.

Techniques: Infection, Isolation, Flow Cytometry, Enzyme-linked Immunospot

Journal: PLoS ONE

Article Title: Immunogenicity and Protective Efficacy of a Vero Cell Culture-Derived Whole-Virus H7N9 Vaccine in Mice and Guinea Pigs

doi: 10.1371/journal.pone.0113963

Figure Lengend Snippet: DBA/2J mice were immunized with whole-virus (WV) H7N9 vaccine or buffer as control on days 0 and 21. Spleen cells were collected 7 days after the first, or 21 days after the booster immunization (i.e. 42 days after the first), and stimulated with whole-virus (WV) H7N9 vaccine or recombinant H7 HA (rHA) for the determination of cells responding to the antigens by secretion of either IFN-γ (A) or IL-4 (B) using an ELISPOT assay. Statistically significant differences of comparisons between T cell responses in mice receiving H7N9 vaccine or buffer are shown. H7 HA-specific IgG subclass responses were analyzed by ELISA using sera collected on day 42 (C). IgG subclass responses on day 42 were compared by ANOVA. ***, p<0.0001; ns, not significant.

Article Snippet: The frequency of H7N9-specific IFN-γ- or Interleukin-4 (IL-4)-secreting cells was analyzed using mouse IFN-γ and IL-4 enzyme-linked immunospot (ELISPOT) kits (Mabtech AB, Nacka, Sweden), as previously described [ ], using vaccine antigen at a concentration of 0.3 μg HA/ml.

Techniques: Recombinant, Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay