Journal: SAGE Open Medicine

Article Title: A novel prostate cancer immunotherapy using prostate-specific antigen peptides and Candida skin test reagent as an adjuvant

doi: 10.1177/2050312118800202

Figure Lengend Snippet: IFN-γ ELISPOT assay results assessing the immunogenicity of the PSA peptides in prostate cancer patients (n = 10). The results of patients with positive response to at least one PSA peptide or peptide pool are shown (n = 4). The error bars represent standard error of means, and the boxed numbers represent positivity indices, which are calculated by dividing the mean SFUs in peptide wells with the mean SFUs in media only wells.

Article Snippet: The spots formed by IFN-γ-secreting T-cells were counted with an automated enzyme-linked immunospot (ELISPOT) analyzer (AID ELISPOT Classic Reader; AID Autoimmun Diagnostika GmbH, Strassberg, Germany).

Techniques: Enzyme-linked Immunospot

Journal: Applied Microbiology and Biotechnology

Article Title: T cell immunity rather than antibody mediates cross-protection against Zika virus infection conferred by a live attenuated Japanese encephalitis SA14-14-2 vaccine

doi: 10.1007/s00253-020-10710-z

Figure Lengend Snippet: Cross-reactive cytokine responses in mouse splenocytes. a Schedule. Three weeks after the final immunization, splenocytes from adult C57BL/6 mice were collected. b The cytokines IL-2, IL-4, and IFN-γ secreted by splenocytes were determined using ELISPOT ( n = 7). The numbers of cytokine-positive cells are reported as the mean SFU/3 × 10 5 splenocytes + SD. Asterisk indicates P < 0.05; double asterisk indicates P < 0.01; triple asterisk indicates P < 0.001

Article Snippet: Three weeks after the final immunization, the cytokines IL-2, IL-4, and IFN-γ secreted by the splenocytes of C57BL/6 mice were determined using enzyme-linked immunospot (ELISPOT) kits (BD, USA) according to the manufacturer’s instructions and the previous protocol (Wang et al. , ).

Techniques: Enzyme-linked Immunospot

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Potent anti-myeloma efficacy of dendritic cell therapy in combination with pomalidomide and programmed death-ligand 1 blockade in a preclinical model of multiple myeloma

doi: 10.1007/s00262-020-02654-0

Figure Lengend Snippet: Activation of cytotoxic T lymphocytes (CTLs) and cytokine production induced by treatment with DC vaccination combined with pomalidomide/dexamethasone and PD-L1 blockade. The number of interferon (IFN)-γ-secreting lymphocytes in the spleens of mice treated with PBS, DCs + pomalidomide with dexamethasone, pomalidomide with dexamethasone + PD-L1 blockade, and DCs + pomalidomide with dexamethasone + PD-L1 blockade was counted using IFN-γ enzyme-linked immunospot (ELISPOT) assays. Treatment with pomalidomide with dexamethasone + PD-L1 blockade partially increased the number of IFN-γ-secreting splenocytes against MOPC-315 and YAC-1 cells compared to the PBS control, whereas treatment with a combination of DCs + pomalidomide with dexamethasone + PD-L1 blockade increased the number of IFN-γ-secreting lymphocytes targeting MOPC-315 and YAC-1 cells compared to all other groups at days 31 a and 34 b after treatment (*, P < 0.05; (***, P < 0.001). These results indicated that the tumor-inhibiting effects of DCs + pomalidomide with dexamethasone + PD-L1 blockade resulted from the CTL-mediated cytotoxicity response (represented by the number of IFN-γ-secreting splenocytes against MOPC-315 cells) and natural killer (NK) cell-mediated cytotoxicity response (represented by the number of IFN-γ-secreting splenocytes against YAC-1 cells). The supernatants from CTL cultures from each group of treated mice were collected for enzyme-linked immunosorbent (ELISA) assays. The combination of DCs + pomalidomide with dexamethasone + PD-L1 blockade led to the production of higher levels of IFN-γ compared to the PBS control, DCs + pomalidomide with dexamethasone, and pomalidomide with dexamethasone + PD-L1 blockade groups at days 31 c and 34 d after treatment (*, P < 0.05; ***, P < 0.001). Moreover, the IFN-γ/interleukin (IL)-10 ratio was significantly higher in the group treated with DCs + pomalidomide with dexamethasone + PD-L1 blockade compared to the other groups (e, f; **, P < 0.05; ***, P < 0.001). In contrast, transforming growth factor-beta (TGF-β) production under DCs + pomalidomide with dexamethasone + PD-L1 blockade treatment was lower compared to all other groups at days 31 g and 34 h after treatment (*, P < 0.05; **, P < 0.01; ***, P < 0.001). These results suggested that the combination of DCs + pomalidomide with dexamethasone + PD-L1 blockade enhanced Th1 immune responses in addition to CTL responses. Data are shown as mean (pg/mL) ± standard deviation (SD) of triplicate cultures from three independent experiments

Article Snippet: After restimulation, the splenocytes were assessed for tumor antigen-specific CTLs using a mouse interferon (IFN)-γ enzyme-linked immunospot (ELISPOT) assay (BD Bioscience, Franklin Lakes, NJ, USA).

Techniques: Activation Assay, Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay, Standard Deviation