Journal: EMBO Molecular Medicine
Article Title: Fibrolytic vaccination against ADAM12 reduces desmoplasia in preclinical pancreatic adenocarcinomas
doi: 10.1038/s44321-024-00157-4
Figure Lengend Snippet: ( A ) Quantitative analysis by flow cytometry of absolute count of CD4 + T cells (left) and CD8 + T cells (middle) in tumor samples from subcutaneous PDAC model treated with control vaccine (v-CTRL) and ADAM12 vaccine (v-A12), along with corresponding gating strategy (right). ( n = 12 mice in v-CTRL and v-A12 respectively. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test.). ( B ) Quantitative analysis of CD4 + T cell population at the edge of PDAC tumor tissue (left, upper) and interior of PDAC tissue (left, bottom), along with corresponding representative images. ( n = 5 mice in v-CTRL and n = 4 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test. Scale bar 50 μm). ( C ) Quantitative analysis of CD8 + T cell population at the edge of PDAC tumor tissue (left, upper) and interior of PDAC tissue (left, bottom), along with corresponding representative images. ( n = 5 mice in v-CTRL and n = 6 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test. Scale bar 50 μm). ( D ) Representative flow cytometry gating (upper) of CD62L - CD4 + effector T cells between control-vaccinated (v-CTRL) mice and ADAM12-vaccinated (v-A12) mice; corresponding quantitative analysis of naïve CD4 + T cells (bottom, left) and CD62L - CD4 + effector T cells (bottom, right) within splenic CD4 + T cells from PDAC tumor-bearing mice treated with control vaccine (v-CTRL) and ADAM12 vaccine (v-A12). ( n = 12 mice in v-CTRL and v-A12 respectively. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test.). ( E ) Representative flow cytometry gating of CD62L - CD8 + effector T cells (upper) between control-vaccinated mice and ADAM12-vaccinated mice; corresponding quantitative result of naïve CD8 + T cells (bottom, left) and CD62L - CD8 + effector T cells (bottom, right) within splenic CD8 + T cells from PDAC tumor-bearing mice treated with control vaccine (v-CTRL) and ADAM12 vaccine (v-A12). ( n = 12 mice in v-CTRL and v-A12 respectively. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test.). ( F ) Spot-forming cells (SFC) in IFN-γ ELISpot of splenic CD4 + T cell and CD8 + T cell populations purified from tumor-bearing animals vaccinated with v-CTRL or v-A12 and restimulated with MutuDC1940 cells expressing ADAM12-derived epitopes (ADAM12 target) or with corresponding CTRL target (TurboGFP alone). ( n = 6 independent samples in v-CTRL and v-A12 respectively. Data were presented as mean ± SEM. Statistical test: two-way ANOVA.). ( G ) Gene expression analysis of Adam12 in YAC-1, “KPPC”-derived KP2 PDAC cells (KP2) and ADAM12-expressing NIH 3T3 cells (NIH 3T3 A12). ( H ) Spontaneous baseline killing of YAC-1 cells by purified splenic CD8 + T cells from PDAC tumor-bearing mice vaccinated with v-CTRL or v-A12. ( n = 3 independent samples in v-CTRL and v-A12, respectively. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test.). ( I ) Cytotoxic assay of purified splenic CD8 + T cells from PDAC tumor-bearing mice vaccinated with v-CTRL or v-A12 with target cells (KP2, NIH 3T3 A12). Data represented as fold change to v-CTRL. ( n = 3 samples in v-CTRL and v-A12, respectively. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test.). ( J ) Spot-forming cells (SFC) in IFN-γ ELISpot of purified splenic CD8 + T cells from PDAC tumor-bearing mice vaccinated with control vaccine (v-CTRL) or ADAM12 vaccine (v-A12) co-cultured with target cells (KP2, NIH 3T3 A12). ( n = 3 samples in v-CTRL and v-A12, respectively. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t -test). .
Article Snippet: Murine IFN-γ ELISpot assay , Diaclone , Cat# 862.031.010S.
Techniques: Flow Cytometry, Control, Two Tailed Test, Enzyme-linked Immunospot, Purification, Expressing, Derivative Assay, Gene Expression, Cell Culture
