Journal: Oncoimmunology

Article Title: Vaccine-induced but not tumor-derived Interleukin-10 dictates the efficacy of Interleukin-10 blockade in therapeutic vaccination

doi: 10.1080/2162402X.2015.1075113

Figure Lengend Snippet: IL-10 blockade enhances vaccine-induced antitumor innate and adaptive responses in tumor-bearing mice. C57BL/6 mice (n = 4–6/group) bearing 5 mm B16-OVA tumors received a single vaccination with topical Imiquimod administration plus i.t. OVA injection, combined with i.p. injection of neutralizing anti-IL-10R antibodies or an isotype control. (A) They were sacrificed 2 d after vaccination and CD86 and intracellular IL-12 were determined by flow cytometry in DC from tumor-draining lymph nodes. (B) In the same animals as in A, lymphocytes were cultured with the NK-sensitive YAC-1 cells line and IFN-γ production measured by ELISPOT. (C) Equivalent groups were sacrificed one week after vaccination and cells from tumor-draining lymph nodes were stimulated with OVA(257–264) peptide or B16-OVA tumor cells to measure IFN-γ-producing cells by ELISPOT. (D) OVA(257–264)-specific CD8+ T cells from mice shown in C were analyzed by flow cytometry measuring the percentage of IFN-γ+TNF-α+ and IFNγ+TNF- α +IL-2+ CD8+ T cells after 5 h of stimulation with OVA(257–264). (E) Mice with B16-OVA tumors (n=10/group) were depleted of CD8+, CD4+ or NK cells and vaccinated with OVA + Imiquimod plus IL-10 blockade. As controls mice were left untreated or vaccinated in the absence of any cell depletion. Tumor volume was measured in these animals. Results are representative of two independent experiments.

Article Snippet: CD8 + epitope peptides OVA(257–264) and HPV16 E7(49–57), with a purity >95%, were obtained from NeoMPS.

Techniques: Injection, Control, Flow Cytometry, Cell Culture, Enzyme-linked Immunospot

Journal: Oncoimmunology

Article Title: Vaccine-induced but not tumor-derived Interleukin-10 dictates the efficacy of Interleukin-10 blockade in therapeutic vaccination

doi: 10.1080/2162402X.2015.1075113

Figure Lengend Snippet: IL-10 blockade enhances (T)cell responses in an adjuvant-dependent manner. (A) C57BL/6 mice (n = 4 /group) immunized with OVA s.c. and topical Imiquimod cream application at the immunization site, combined with i.p. injection of isotype or anti-IL-10R antibodies. One week later their splenocytes were stimulated with CD8 epitope OVA(257–264) or OVA protein and IFNγ-producing cells evaluated by ELISPOT. (B) Mice were immunized with Imiquimod plus EDA-HPVE7 protein as in A with or without IL-10 blockade and responses against HPV E7(49–57) peptide were measured by ELISPOT. (C) C57BL/6 mice (n = 4) were immunized with OVA s.c. plus adjuvants CpG, poly(I:C) or anti-CD40 agonistic antibodies or with EDA-OVA plus control or anti-IL-10R blocking antibodies. One week later mice were sacrificed and their splenocytes were stimulated with CD8+ epitope OVA(257–264) and IFNγ-producing cells evaluated by ELISPOT. Results show the difference between peptide-stimulated minus unstimulated cells and are representative of two-three independent experiments.

Article Snippet: CD8 + epitope peptides OVA(257–264) and HPV16 E7(49–57), with a purity >95%, were obtained from NeoMPS.

Techniques: Adjuvant, Cream, Injection, Enzyme-linked Immunospot, Control, Blocking Assay

Journal: Oncoimmunology

Article Title: Vaccine-induced but not tumor-derived Interleukin-10 dictates the efficacy of Interleukin-10 blockade in therapeutic vaccination

doi: 10.1080/2162402X.2015.1075113

Figure Lengend Snippet: Inhibition of IL-10 when using a MAC-based therapeutic vaccine enhances antitumor responses resulting in complete tumor rejection. (A) C57BL/6 mice (n = 4) were immunized with EDA-OVA + MAC plus control or anti-IL-10R blocking antibodies. One week later mice were sacrificed and their splenocytes were stimulated with CD8 epitope OVA(257–264) or OVA protein and IFN-γ-producing cells evaluated by ELISPOT. (B) C57BL/6 mice (n = 8/group) bearing 5 mm B16-OVA tumors received three weekly cycles of vaccination with EDA-OVA + MAC, combined with i.p. injection of neutralizing anti-IL-10R antibodies or an isotype control. Tumor growth and animal survival was monitored twice per week. Results are representative of two independent experiments.

Article Snippet: CD8 + epitope peptides OVA(257–264) and HPV16 E7(49–57), with a purity >95%, were obtained from NeoMPS.

Techniques: Inhibition, Control, Blocking Assay, Enzyme-linked Immunospot, Injection