Journal: Cancers

Article Title: Bimodal Effect of NKG2A Blockade on Intratumoral and Systemic CD8 T Cell Response Induced by Cancer Vaccine

doi: 10.3390/cancers16112036

Figure Lengend Snippet: NKG2A blockade reduces peripheral CD8 T cell immune response induced by KISIMA—VSV-GP-TAg vaccination. C57BL/6J mice were injected subcutaneously with 10 5 TC-1 cells at D0, vaccinated with KISIMA vaccine at D6 and with VSV-GP-TAg at D13. Mice received the first injection of aNKG2A at D14 followed by a second injection 3 days later. One week post VSV-GP-TAg boost, blood was collected, and CD8 T cell response was analyzed by flow cytometry. ( A ) Frequency and number of E7-specific CD8 T cells measured in the blood one week post VSV-GP-TAg boost. ( B ) Frequency of E7-specific CD8 T cells expressing activation and exhaustion markers analyzed in the blood. One representative of two independent experiments. ( n = 7). ( C ) Representative FACS plot of E7-specific CD8 T cells in the blood. ( D ) Frequency and number of E7-specific CD8 T cells in the spleen. ( E ) Cytokine production by antigen specific CD8 T cells in the spleen was measured by intracellular staining after ex vivo restimulation with E7 peptide. Mann-Whitney test was used except for figure E where Two-way ANOVA test with Sidak’s multiple comparisons was used * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. One representative of two independent experiments ( n = 5–7).

Article Snippet: Anti-mouse NKG2A (InVivoMab BioXCell Cat BE0339) was diluted to 2 mg/mL in 1X phosphate buffer saline (PBS, Gibco) prior to injection.

Techniques: Injection, Flow Cytometry, Expressing, Activation Assay, Staining, Ex Vivo, MANN-WHITNEY

Journal: Cancers

Article Title: Bimodal Effect of NKG2A Blockade on Intratumoral and Systemic CD8 T Cell Response Induced by Cancer Vaccine

doi: 10.3390/cancers16112036

Figure Lengend Snippet: NKG2A blockade dampens long-term protection elicited by KISIMA—VSV-GP-TAg vaccination. TC-1 tumor-bearing mice were vaccinated with KISIMA-Mad25 vaccine at D6 post tumor implantation followed by a boost with VSV-GP-HPV at D13 and a second boost with KISIMA at D27. Mice were treated with anti-NKG2A starting 4 days post VSV-GP-HPV injection for a total of five injections. Mice were sacrificed at D45, tumor, spleen and bone marrow were harvested, processed and CD8 T cell immune response was analyzed by flow cytometry. ( A ) Frequency of E7-specific among CD8 T cells in blood, spleen, and bone marrow. ( B ) Cytokine production by antigen specific CD8 T cells in the spleen was measured by intracellular staining after ex-vivo restimulation with E7 peptide in the presence of Golgi inhibitor. ( C ) Memory phenotype of E7-specific CD8 T cells analyzed by flow cytometry in blood, spleen, and bone marrow at D45 post tumor implantation. ( D ) Complete responders were rechallenged at D75 with 10 5 TC-1 cells and tumor growth was followed. Two-way ANOVA with Sidak’s multiple comparisons were used (* p < 0.05, ** p < 0.01, **** p < 0.0001) except in fig. A where Mann–Whitney test was used (* p < 0.05, ** p < 0.01). One representative of two independent experiments ( n = 6–7).

Article Snippet: Anti-mouse NKG2A (InVivoMab BioXCell Cat BE0339) was diluted to 2 mg/mL in 1X phosphate buffer saline (PBS, Gibco) prior to injection.

Techniques: Tumor Implantation, Injection, Flow Cytometry, Staining, Ex Vivo, MANN-WHITNEY