Journal: Cancer immunology research

Article Title: Radiofrequency ablation remodels the tumor microenvironment and promotes neutrophil-mediated abscopal immunomodulation in pancreatic cancer

doi: 10.1158/2326-6066.CIR-22-0379

Figure Lengend Snippet: Tumor size was recorded 4 days before (Initial), right before (Pre) and 4 days after (Post) Sham or RFA treatment. Proteome arrays were performed in locally ablated tumors and serum of ablated mice and compared to Sham-treated mice (Control). A, Experimental design of RFA-treated mice. B, Growth curves show Control tumors (n = 8) significantly increased in size 4 days after treatment when compared to RFA-treated (n = 8) and non-RFA treated (n = 7) tumors. C, At the time of euthanization only Sham-treated tumors (n = 8) had significantly increased in size compared with pretreatment size; no difference in size was observed in RFA-treated (n = 8) and non-RFA treated (n = 7) tumors Pre and Post RFA. D, ImageJ quantification of necrosis, which was detected by H&E staining. RFA significantly increased necrosis on the RFA- and non-RFA-treated tumors compared to control Sham-treated tumors. E, Representative composite H&E staining of control, RFA, and non-RFA treated tumors showing necrotic areas inside dashed lines. F, ImageJ quantification showing RFA increased cleaved caspase 3+ cells in the RFA-treated and non-RFA treated tumors compared to control Sham-treated control tumors, as assessed by IHC. G, Representative IHC staining for cleaved caspase 3 in control, RFA, and non-RFA treated tumors. H, ImageJ quantification revealed RFA significantly increased the number of granzyme B+ cells in the RFA-treated tumors compared to controls and non-RFA treated tumors, as assessed by IHC. I, IHC staining for granzyme B in control, RFA, and non-RFA treated tumors. J, RFA-treated tumors (n = 3) presented increased expression of C5/C5a, IL-23 and CXCL12 compared to control (n = 2) tumor content. K, CXCL10, CXCL12, CXCL13 and TIMP-1 were significantly elevated in serum from RFA-treated (n = 4) mice compared to Sham-treated (n = 3) control serum. Time x Treatment comparisons were performed using Two-way ANOVA, treatment only comparisons by One-way ANOVA and proteome arrays were analyzed by multiple t test. Bar plots showing mean with SEM were used to represent data. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; n.s., not significant. Scale bars are 50μM.

Article Snippet: CXCL13 ELISA Mouse CXCL13/BLC/BCA-1 Quantikine ELISA Kit assay was used to determine CXCL13 levels in mouse serum, splenocytes and tumors homogenates, following the manufacturer instructions (R&D Systems, cat #MCX130).

Techniques: Control, Staining, Immunohistochemistry, Expressing

Journal: Cancer immunology research

Article Title: Radiofrequency ablation remodels the tumor microenvironment and promotes neutrophil-mediated abscopal immunomodulation in pancreatic cancer

doi: 10.1158/2326-6066.CIR-22-0379

Figure Lengend Snippet: A-B, IMC analysis of tumors 4 days after Sham or RFA treatment revealed Ly6G+CD11b+CD44+ neutrophils are enriched in non-RFA treated tumors. C, Neighborhood analysis identified immune cells and markers with strong neutrophil co-localization. D, Cluster and Cell Phenotype information of Neighborhood analysis of IMC data. E, Experimental design for neutrophil depletion in vivo followed by RFA. F, RFA locally ablated tumors treated with IgG2a isotype control (VEH, n = 6) or anti-Ly6G (ND, n = 8) did not show differences in tumor size right before (Pre) and 4 days after (Post) RFA ablation. In non-RFA treated tumors, anti-Ly6G (ND, n = 8) treatment revealed an increase in tumor size Post RFA treatment when compared to IgG2a isotype control (VEH, n = 6) treated tumors. G, Neutrophil depletion (ND; anti-Ly6G treated group) did not alter αSMA staining, detected by IHC, in RFA-treated tumors when compared to RFA-treated tumors with IgG2a (VEH); on the contrary, neutrophil depletion (ND) revealed non-RFA treated tumors presented a significant increase in αSMA compared to control non-neutrophil depleted (VEH) group. H, Neutrophil depletion did not alter CD31 staining, detected by IHC, in any of the groups. I, Neutrophil-depleted RFA treated tumors presented a significant reduction in CXCL13 content compared to both VEH + RFA and non-RFA treated tumors when assayed using a cytokine array. No differences were found in non-RFA treated tumors between treatments. J, Neutrophil depletion presented a trend in reducing systemic CXCL13 levels in RFA treated mice. Tumor volume was analyzed by paired Student’s t test. Tumor chemokine levels were studied by Two-way ANOVA. IHC and serum protein expression levels were analyzed by unpaired Student’s t test. Bar plots indicate mean with SEM. *, P ≤ 0.05; **, P ≤ 0.01; ****, P ≤ 0.0001; n.s., not significant.

Article Snippet: CXCL13 ELISA Mouse CXCL13/BLC/BCA-1 Quantikine ELISA Kit assay was used to determine CXCL13 levels in mouse serum, splenocytes and tumors homogenates, following the manufacturer instructions (R&D Systems, cat #MCX130).

Techniques: In Vivo, Control, Staining, Expressing

Journal: Science Advances

Article Title: A prokaryotic-eukaryotic hybrid viral vector for delivery of large cargos of genes and proteins into human cells

doi: 10.1126/sciadv.aax0064

Figure Lengend Snippet: ( A ) Formulations used for mice vaccinations. ( B ) Prime-boost immunization scheme. The days of prebleeding (pre.), bleeding, prime, boost, and challenge with Y. pestis are indicated. ( C and D ) Sera from the PBS, T4, T4(HA4900), and T4(HA4900)-AAV(VRCO1) groups were assessed for specific anti-HA IgG/G1/G2a antibody titers by ELISA on day 14 (prime) (C) and day 35 (boost) (D). ( E ) Measurement of anti-HA IgG2a antibody titers in sera from groups T4, T4(HA4900), and T4(HA4900)-AAV(VRCO1) at serial time points (on prebleeding day and days 14, 35, 60, 120, and 180). ( F ) Serum CXCL13 concentration (ng/ml) in mice immunized with T4(HA4900) or T4(HA4900)-AAV(VRCO1) at prebleeding day and days 14, 35, 60, and 180 after immunization. ( G ) Comparison of anti-HA antibody titers in sera from groups T4, T4(HA4900), T4(HA4900)-AAV(VRCO1), and F1mutV-T4(HA4900)-AAV(VRCO1) on day 35. ( H and I ) Anti-F1mutV–specific antibody (IgG/G1/G2a) titers were determined by ELISA using sera from mice vaccinated with T4(HA4900)-AAV(VRCO1) and F1mutV-T4(HA4900)-AAV(VRCO1) on days 14 (H) and 35 (I). ( J ) Comparison of anti-F1mutV IgG antibody titers in sera from groups T4, AAV, F1mutV-T4(HA4900), and F1mutV-T4(HA4900)-AAV on day 35. ( K ) Kaplan-Meier survival curves of mice against intranasal pneumonic plague challenge with 295 LD 50 (median lethal dose) of Y. pestis CO92 (1 LD 50 = 100 colony-forming units in BALB/c mice). ( L ) Anti-HA–specific antibody titers in immunized sera from the F1mutV-T4-AAV(HA4900) group at various time points. ( M ) Comparison of anti-F1mutV antibody titers in sera from groups T4(HA4900)-AAV, T4-AAV(HA4900), F1mutV-T4(HA4900)-AAV, and F1mutV-T4-AAV(HA4900) on day 35. The values represent means ± SD of three independent replicates. ** P < 0.01 compared with the T4(HA4900) vaccination. * P < 0.05 compared with the F1mutV-T4 vaccination. Statistical significance was determined using two-tailed Student’s t tests. N.D., not detected. See Materials and Methods for additional details.

Article Snippet: For CXCL13 quantification in sera, the Mouse CXCL13 ELISA Kit (Boster Biological Technology, CA) was used by following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Comparison, Two Tailed Test