Journal: Journal for immunotherapy of cancer

Article Title: PARP inhibitors enhance antitumor immune responses by triggering pyroptosis via TNF-caspase 8-GSDMD/E axis in ovarian cancer.

doi: 10.1136/jitc-2024-009032

Figure Lengend Snippet: Figure 5 Deficiency of GSDME blunts the immune response triggered by PAPR inhibitor in vivo. (A–L) Immunocompetent C57BL/6 mice were transplanted with wild type (WT) or Gsdme-deficient ID8 cells and challenged with niraparib for about 4 weeks. Tumors were harvested and subjected to bulk T-cell receptor (TCR) sequencing and evaluation of immune status. The frequency of T-cell clonotypes in the top 25% of TCR repertoires was shown using pie charts ((A) n=4) and quantified (B). The clonal expansion was evaluated using clonality (C). The TCR diversity was calculated by normalized Shannon diversity entropy (D). The proportion of dendritic cell differentiation (CD11c+MHCIIHigh) and maturation (CD11c+MHCIIHighCD86High) in tumors (E), lymph nodes (F), and spleens (G) was determined by flow cytometry. The proportion of CD3+ T cells and NK cells among CD45+ immune cells in tumors was determined by flow cytometry (H). The production of IFN-γ in tumor-infiltrated CD4/8+ T and NK cells was examined by flow cytometry (I). The expression of granzyme B in tumor-infiltrated CD8+ T and NK cells was evaluated by mean fluorescence intensity (J). Representative images of GSDME, CD4, CD8, and GZMB IHC staining in tumor sections (K) and numbers of indicated immune cells in a ×20 field of a microscope (L). Scale bars: 200 µm. WT or Gsdme-deficient ID8 cells were intrabursally transplanted into C57BL/6 mice and received niraparib and/or anti-PD-1 treatment. Representative bioluminescent images of mice bearing WT and Gsdme-KO ID8 tumors at the endpoint (M). The tumor weight was quantified in each mouse treated with niraparib and/or anti-PD-1 antibodies ((N) n=7). Mean values±SEM. *P<0.05, **p<0.01, and ***p<0.001 by Student’s t-test in (B–J, L, and N).

Article Snippet: Subsequently, these isolated T cells were stimulated with anti- CD3 (BioXCell, BE0002) and anti- CD28 antibodies (BioXCell, BE0328) in the presence or absence of olaparib/niraparib.

Techniques: In Vivo, Sequencing, Cell Differentiation, Flow Cytometry, Expressing, Fluorescence, Immunohistochemistry, Microscopy

Journal: Frontiers in Immunology

Article Title: Desmoglein-4 Deficiency Exacerbates Psoriasiform Dermatitis in Rats While Psoriasis Patients Displayed a Decreased Gene Expression of DSG4

doi: 10.3389/fimmu.2021.625617

Figure Lengend Snippet: Dsg-4 deficiency induces high mRNA expression of inflammatory genes in response to IMQ. RNA from skin was extracted, and subsequently real-time PCR and cDNA synthesis were performed to evaluate mRNA abundance of cytokines: IL-1β (A) , IL-8 (B) , IL-17 (C) , IL-10 (D) , and TGF-β (E) performing Kruskal–Wallis tests analysis. Additionally, TGF-β/IL-17 mRNA abundance means ratio was calculated (F) . Similarly, chemokine receptors mRNA levels were measured: CCR1 (G) , CCR2 (H) , CCR3 (I) , CCR5 (J) , and CXCR5 (K) . Kruskal–Wallis and ANOVA tests were used; data represent the median with values range; * P < 0.05, ** P < 0.01, *** P < 0.001. # P < 0.05, ## P < 0.01 t -test between SD and IMQ-SD datasets. (L) Pseudocolor dot plots represent counts of infiltrating CD45 + cells and CD3 + cells from total skin cells analyzed by flow cytometry derived from SD and OFA rats. (M,N) Graphs represent the percentage and absolute cell counts of CD45 + CD3 + cells from total skin cells. ( n rats /group = 6; two independent experiments). (O,P) Spleen and inguinal lymph node weights. FACS and lymphatic tissue weight data are represented as the mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001 with ANOVA and Bonferroni post hoc test was used. # P < 0.05, ## P < 0.01 t -test between SD and IMQ-SD datasets.

Article Snippet: The cells were then resuspended in PBS/2% FBS and stained with PE-Cy5-conjugated anti-rat CD45 (clone OX-1; isotype mouse IgG1 k, BD Biosciences, San Jose, USA) and FITC-conjugated anti-rat CD3 (clone IF4; isotype mouse IgM, Cedarlane Laboratories, US).

Techniques: Expressing, Real-time Polymerase Chain Reaction, cDNA Synthesis, Flow Cytometry, Derivative Assay

Journal: Heliyon

Article Title: Erzhi pills reverse PD-L1-mediated immunosuppression in melanoma microenvironment

doi: 10.1016/j.heliyon.2024.e24988

Figure Lengend Snippet: EZP increased the percentage of CD4 + T cells and enhanced the function of CD8 + T in vivo . (A–D) T cells were analyzed by flow cytometry. (A) The gate strategy for CD4 + and CD8 + T cells. Flow cytometry plots (B) and bar graphs present percentages of CD3 + CD4 + T cells (C) and CD3 + CD8 + T cells (D) in the spleen from each group mice. n = 5. (E–H) IHC staining for the tumor tissues. IHC images (E) and quantifications of CD4 + T cells (F), CD8 + T cells (G) and GZMB + cells (H) in tumor tissues. Scale bar, 100 μm. n = 3. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01 and *** P < 0.001 vs DMSO. IHC, immunohistochemistry; GZMB, granzyme B.

Article Snippet: PerCP/Cyanine5.5 anti-mouse CD3 antibody (E-AB-F1013J), FITC anti-mouse CD8a antibody (E-AB-F1104C) and PE anti-mouse CD4 antibody (E-AB-F1097D) were obtained from Elabscience (Wuhan, China).

Techniques: In Vivo, Flow Cytometry, Immunohistochemistry

Journal: STAR Protocols

Article Title: Protocol to assess the tolerogenic properties of adoptively transferred dendritic cells during murine experimental autoimmune encephalomyelitis

doi: 10.1016/j.xpro.2022.101653

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD3e-redFluor710 (Clone 17A2) , Tonbo Biosciences , Cat# 80-0032, RRID: AB_2621971.

Techniques: Recombinant, Adjuvant, Staining, Software

Journal: STAR Protocols

Article Title: Protocol to assess the tolerogenic properties of adoptively transferred dendritic cells during murine experimental autoimmune encephalomyelitis

doi: 10.1016/j.xpro.2022.101653

Figure Lengend Snippet: FACS Antibody Staining mix

Article Snippet: Anti-mouse CD3e-redFluor710 (Clone 17A2) , Tonbo Biosciences , Cat# 80-0032, RRID: AB_2621971.

Techniques: Staining, Control