clone 1a8  (Novus Biologicals)

Journal: Cancer Research Communications

Article Title: Neutrophil Antigen Presentation Reprograms the Tumor Microenvironment and Elicits Durable and Broad Antitumor Immunity

doi: 10.1158/2767-9764.CRC-25-0509

Figure Lengend Snippet: αCD16-Ova–induced T-cell expansion in the spleen requires neutrophils and MHC on neutrophils, whereas Batf3 + cDC1s and splenic macrophages are dispensable. Mice were treated as in , and the role of neutrophils ( A and B ), MHC-I on neutrophils ( C and D ), and the contribution of cDC1 ( E and F ) and splenic macrophages ( G and H ) in αCD16-Ova–mediated T-cell proliferation was evaluated. A and B, Role of neutrophils. Treatment with 1A8 resulted in marked neutrophil depletion in blood but only partial depletion in the spleen ( A ), with a corresponding partial reduction in splenic CD8 OT-I and CD4 OT-II T-cell proliferation ( B ). C and D, Requirement for neutrophil MHC-I. In CD16B/γ −/− mice with neutrophil-specific MHC-I deletion (Ly6G-Cre/MHC-I fl/fl ), MHC-I expression was partially reduced on splenic neutrophils but intact on cDC1s ( C ); isotype control was used to set gates. CD8 OT-I proliferation was proportionally reduced in Ly6G-Cre/MHC-I fl/fl , consistent with incomplete MHC-I deletion in neutrophils ( D ). E and F, Requirement for Batf3 + cDC1s. In CD16B/γ −/− mice with a deletion in Batf3 (CD16B/γ −/− Batf3 −/− ), there was a marked loss of cDC1 but intact cDC2 populations ( E ). cDC1 loss did not affect αCD16-Ova induced CD8 OT-I or CD4 OT-II T-cell proliferation ( F ). G and H, Anti-CD115 antibody–mediated immunodepletion resulted in a significant but partial reduction of F4/80 + macrophages in the spleen compared with isotype control I (Ctr) treatment ( G ). CD115 antibody treatment did not alter αCD16-Ova–induced CD8 T-cell proliferation ( H ). Sp, spleen. Data are presented as mean ± SEM. Two-group comparisons were done using an unpaired two-tailed Student t test; comparisons among ≥3 groups were done using one-way ANOVA with Tukey post hoc test. Significance was defined as follows: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Ab, antibody; KO, knockout.

Article Snippet: CD16B-CD32A +/− /γ +/− male mice received 350 μg/mouse anti-Ly6G (clone 1A8, Bio X Cell, cat. #BP0075-1) for neutrophil depletion or isotype control rat IgG2a, κ (clone 2A3, Bio X Cell, cat. #BP0089) intraperitoneally on days −2 and −1 prior to OTI/OTII injection, then every 2 days thereafter.

Techniques: Expressing, Control, Immunodepletion, Two Tailed Test, Knock-Out