Journal: PLoS ONE

Article Title: Development, Alteration and Real Time Dynamics of Conjunctiva-Associated Lymphoid Tissue

doi: 10.1371/journal.pone.0082355

Figure Lengend Snippet: Antibody staining panel.

Article Snippet: Anti-MHC II , ER-TR2 , Novus Biologicals, UK.

Techniques: Staining

Journal: Oncoimmunology

Article Title: Tumor cell-derived IL-10 promotes cell-autonomous growth and immune escape in diffuse large B-cell lymphoma

doi: 10.1080/2162402X.2021.2003533

Figure Lengend Snippet: Neutralization of IL-10R signaling reduces the lymphoma burden in a syngeneic transplantation model and reduces the intratumoral frequencies of regulatory T-cells. a-e , C57BL/6 mice were intravenously injected with 1 Mio >90% pure lymphoma cells isolated from the spleens of Eµ-Myc-transgenic donors, and examined at the study endpoint (10–15 days post injection) with respect to their lymph node weights and the frequencies and surface marker expression of various lymph node leukocyte populations by flow cytometry. An initial dose of 500 µg anti-IL-10RA neutralizing or control IgG antibody was i.p. injected shortly before tumor cell inoculation, followed by twice weekly injections of 300 µg antibody. Control mice did not receive tumor cells. (a) Combined axillary and inguinal lymph node weights of anti-IL-10RA- or control IgG antibody-treated mice. (b) Tumor B-cell frequencies in axillary and inguinal lymph node preparations as determined by staining for CD19 and CD45, of the mice shown in A, alongside representative FACS plots of one mouse per group. The Ki67 signal of the tumor B-cells is shown as well. See suppl. Figure 5a for the complete gating strategy. (c,d) Frequencies of TCRβ + T-cells, of CD4 + T-cells and of CD8 + T-cells among all leukocytes, as determined by flow cytometry. See suppl. Figure 5c for the gating strategy. (e) Frequencies of FoxP3 + Tregs among all CD4 + T-cells, as determined by intracellular staining for FoxP3; representative FACS plots of one mouse per group are shown alongside the summary plot for all mice. f,g , Frequencies of tumor B-cells and of CD4 + T-cells in axillary and inguinal lymph node preparations as determined by staining for CD19, CD4 and CD45, of anti-IL-10R- or control IgG antibody-treated WT and Foxp3 -iDTR mice. Mice received twice weekly i.p. doses of 20 ng/g diphtheria toxin (DT) to deplete Tregs, starting from one day before tumor cell injection. Treg depletion efficiency was >90%. Four mice were each injected i.v. with 100ʹ000 Tregs that had been sorted from the tumors of Foxp3 -iDTR mice based on their GFP expression, on day four post tumor cell transplantation. h , MHCII expression as determined by flow cytometry, of a subset of the mice shown in a-e. A representative histogram of the MHCII signal of two representative mice is shown alongside the summary plot of all mice. i-k , C57BL/6 mice were intravenously injected with 1 Mio >90% pure lymphoma cells and examined at the study endpoint (10–15 days post injection) with respect to their lymph node weights (i), tumor B-cell frequencies (j) and CD4 + /CD8 + T-cell frequencies (k). An initial dose of 500 µg anti-IL-10RA neutralizing and/or MHCII-blocking or control IgG antibody was i.p. injected shortly before tumor cell inoculation, followed by twice weekly injections of 300 µg antibody. Control mice did not receive tumor cells. Data in a-e are pooled from four independent experiments; data in f,g are pooled from one to six experiments, Treg transfer was only performed once. Data in hare from three experiments and data in i-k are from two experiments; note that several mice were lost to follow-up by flow cytometry. Horizontal lines indicate medians. Statistical comparisons were performed either by one-way ANOVA (in the case of normal data distribution) or by non-parametric ANOVA (Kruskal–Wallis test, in the case of non-normal data distribution) with Tukey’s multiple comparisons correction. ns, not significant, *p < .05, **p < .01 ***p < .005, ****p < .001

Article Snippet: Anti-mouse IL-10RA (1B1.3A), anti-mouse MHCII (M5/114), anti-mouse PD-L1 (10 F.9G2), and isotype control antibodies were purchased from BioXCell.

Techniques: Neutralization, Transplantation Assay, Injection, Isolation, Transgenic Assay, Marker, Expressing, Flow Cytometry, Control, Staining, Blocking Assay