Journal: The Journal of clinical endocrinology and metabolism

Article Title: Association of postpartum thyroid dysfunction with antepartum hormonal and immunological changes.

doi: 10.1210/jc.2002-021219

Figure Lengend Snippet: FIG. 1. Graphical output from the FACScan analyzer (Becton Dickinson and Co.). Before analysis lymphocytes were stained with R-phyco- erythrin-Cy5 mouse antihuman CD3 fluorescein isothiocyanate-conjugated mouse antihuman CD4 together with either phycoerythrin- conjugated rat antihuman IL-4 or phycoerythrin-conjugated mouse antihuman IFN-. The output was gated on the basis of side scatter and CD3 staining. Shown are control cells in the left panels and stimulated cells in the right panels. Lymphocytes gated for right-angle light scatter and CD3 staining (a and b), CD4 expression (c and d), IL-4 expression (c–f), and IFN- expression (e and f) are shown.

Article Snippet: The culture plate was placed in a humidified incubator at 37 C, gas phase 5% CO2 in air, for 4 h. Aliquots (350 l) from the stimulated and unstimulated wells were transferred into fluorescenceactivated cell sorter (FACS) tubes and the surface antigen (CD3 and CD4) was stained by adding 5 l conjugated antibody (R-phycoerythrin-Cy5 conjugated mouse antihuman CD3, DAKO Corp. Ltd., Ely, UK; fluorescein isothiocyanate-conjugated mouse antihuman CD4, Serotec Ltd., Oxford, UK) and incubated in the dark for 30 min.

Techniques: Staining, Control, Expressing

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