Journal: Cell

Article Title: Loss of Bcl-6-Expressing T Follicular Helper Cells and Germinal Centers in COVID-19

doi: 10.1016/j.cell.2020.08.025

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Article Snippet: These specimens were incubated with the following antibodies: anti-CD3 (clone: A045229-2; DAKO), anti-CD4 (clone: EPR6855;Abcam), anti-CD19 (clone: SKU310; Biocare Medical), anti-Bcl6 (clone: LN22; Biocare Medical), anti-AID (clone: ZA001; Invitrogen), anti-T-bet (clone: ab150440; Abcam), GATA3 (clone: CM405A; Biocare), ICOS (clone: 89601; Cell Signaling Technology), Rorc (clone: ab212496; Abcam), CXCR5 (clone: MAB190; R&D Systems), Foxp3 (clone: 98377; Cell Signaling Technology), anti-CD8 (clone: ab85792; Abcam), anti-IgD (clone: AA093; DAKO), anti-CD27 (clone: ab131254; Abcam), anti-IgG (clone: ab109489; Abcam), anti-TNF-α (clone: ab6671; Abcam), and anti-CD35 (clone: ab25; Abcam) followed by incubation with a secondary antibody using an Opal Multiplex Kit (Perkin Elmer).

Techniques: Recombinant, Multiplex Assay, Blocking Assay, Staining, Expressing, Saline, Software

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Tight co-twin similarity of monozygotic twins for hTERT protein level of T cell subsets, for telomere length and mitochondrial DNA copy number, but not for telomerase activity

doi: 10.1007/s00018-017-2738-z

Figure Lengend Snippet: Reagents for immunophenotyping analysis of lymphocyte subpopulations

Article Snippet: Table 1 Antibody Reactivity Host Dilutions Manufacturer Clone/isotype hTERT-FITC Human Rabbit 1:100 Novus Biologicals Polyclonal FoxP3-Pe Human Mouse/IgG1, kappa 1:50 eBioscience 236A/E7 CD8-Pe Human Mouse/IgG1 1:50 cytognos 143-44 CD4-PerCP-Cy5.5 Human Mouse/IgG2b, kappa 1:50 eBioscience OKT4 CD3-APC Human Mouse/IgG2a 1:50 cytognos 33-2A3 Open in a separate window Reagents for immunophenotyping analysis of lymphocyte subpopulations.

Techniques:

Journal: The Journal of Clinical Investigation

Article Title: Inhibition of cyclooxygenase-2 in hematopoietic cells results in salt-sensitive hypertension

doi: 10.1172/JCI81550

Figure Lengend Snippet: (A) Immunoblotting demonstrated that high-salt diet–treated Cox2–/–-WT BMT mouse kidneys had increased protein levels of F4/80 (a marker of macrophages/dendritic cells), CD3 (a marker of T cells), and iNOS (a marker of M1 phenotype), but decreased protein levels of a Treg marker, FOXP3. (B-D) High-salt diet–treated Cox2–/–-WT BMT mouse kidneys had decreased mRNA levels (B) and decreased immunoreactivity (C) of MR as well as decreased protein levels of arginase-1, markers of an M2 phenotypic macrophages/dendritic cells (D). ***P < 0.001 vs. high-salt diet–treated WT-WT BMT mice. n = 4 in each group. Original magnification, ×250. (E) High-salt diet–treated Cox2–/–-WT BMT mouse kidneys had increased mRNA levels of M1/Th1 markers/cytokines Inos, Ccl3, Tnfa, Il1a, and Il1b. *P < 0.05; ***P < 0.001 vs. high-salt diet–treated WT-WT BMT mouse kidneys. n = 4 in each group. All values are shown as mean ± SEM. All P values were calculated by Student’s t test.

Article Snippet: Rabbit anti-murine COX-2 (160106) was purchased from Cayman Chemical; rat anti-mouse F4/80 (MCA497R), Ly-6G (MCA2387), CD3 (MCA1477), and CD8α (MCA2694) were purchased from AbD Serotec; rabbit anti–TNF-α (ab6671) was from R&D Systems; FOXP3 (NB100-39002) was from Novus Biologicals; rabbit anti-iNOS (ab3523) and MR (CD206, ab64693) were from Abcam; goat VEGF-C (sc-1881) and podoplanin (sc-23564), rat anti-lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1) (sc-80170), and mouse anti-SGK1 (sc28338) were from Santa Cruz Biotechnology Inc. Sheep anti–p-NCC (at Thr45, Thr50, and Thr55) was obtained from Hillary McLauchlan/James Hastie (University of Dundee, Dundee, United Kingdom).

Techniques: Western Blot, Marker

Journal: Cell Reports Medicine

Article Title: LIFUS-driven engineered bacteria reprogram immunosuppressive niches via mechano-NOTCH signaling

doi: 10.1016/j.xcrm.2026.102658

Figure Lengend Snippet: Mechanical immunotherapy effect of LIFUS combined with VNP /ARG-GV for tumors (A) Schematic illustrating the five-cycle LIFUS + VNP /ARG-GVs treatment regimen over 21 days. (B and C) Repeated LIFUS activation of intratumoral VNP /ARG-GVs resulted in marked suppression of tumor growth compared with all control and monotherapy groups (∗∗∗∗ p < 0.0001; n = 6) (B). (D) Kaplan-Meier survival analysis demonstrating significantly prolonged survival in the combination group (median 47.5 days), with several mice surviving beyond 60 days. All control and monotherapy cohorts succumbed by days 30–40 (control and LIFUS alone: median 20 days, VNP /ARG: 27.5 days, VNP /ARG-GVs: 25 days, n = 10). (E and G) Flow cytometry analysis (E) showing differential immune remodeling across groups. The details of the flow cytometry gating strategy are shown in . LIFUS alone did not alter CD4 + /CD8 + infiltration. VNP /ARG monotherapy induced about 1.5-fold increase in CD8 + T cells. Combination treatment produced a pronounced expansion of cytotoxic CD8 + T cells (63.96% ± 4.90% vs. 14.31% ± 2.0% in control; ∗∗∗∗ p < 0.0001; n = 4) (G). (F and H) The combination group showed the strongest suppression of FOXP3 + Tregs (7.76% ± 1.60%), significantly lower than controls (28.38% ± 2.59%; ∗∗∗ p < 0.005; n = 4) (H). (I and J) ELISA quantification of effector cytokines. IFN-γ levels were highest in the combination group (37.4 ± 6.88 ng/mL), followed by LIFUS alone (12.1 ± 2.3 ng/mL) and control (5.6 ± 1.4 ng/mL) ( n = 4) (I). TNF-α expression followed a similar trend (1.37 ± 0.41 ng/mL vs. 0.42 ± 0.15 ng/mL and 0.18 ± 0.06 ng/mL) ( n = 4) (J). (K and L) Representative Ki67 and hematoxylin and eosin staining showing reduced proliferation and increased apoptosis across groups, with the strongest effects observed in the LIFUS + VNP /ARG-GVs cohort. Scale bars, 100 μm. Data are representative of three (B, D, I, and J) or two (G and H) independent experiments. Data are mean ± SD or chi-square (D). Statistical analysis was performed using one-way ANOVA (B and H–K), two-way ANOVA (G), or log rank test (D). See also .

Article Snippet: Anti-FOXP3 antibody , Proteintech , Cat# 22228-1-AP; RRID: AB_11182376.

Techniques: Activation Assay, Control, Flow Cytometry, Produced, Enzyme-linked Immunosorbent Assay, Expressing, Staining

Journal: Frontiers in Pharmacology

Article Title: Protective effect of Huanglian Pingwei San on DSS-induced ulcerative colitis in mice through amelioration of the inflammatory response and oxidative stress

doi: 10.3389/fphar.2024.1484532

Figure Lengend Snippet: HLPWS restored the immune balance. Effects of HLPWS on the levels of IL-17 (A) , IL-22 (B) , and IL-23 (C) in colon homogenates, as detected via ELISA. Representative photographs of CD4 + CD25 + Foxp3 + (Treg) cells in the spleen analysed by flow cytometry (D) and quantitative analysis of the percentage of Treg cells (E) . Representative photographs of CD4+IL-17A (Th17) cells in the spleen analysed by flow cytometry (F) and quantitative analysis of the percentage of Th17 cells (G) . The results are expressed as the mean ± SEM ( n = 3). ## p < 0.01 vs. the control group; *** p < 0.001, ** p < 0.01, * p < 0.05 vs. the DSS group.

Article Snippet: The APC-conjugated anti-mouse CD4 antibody (E-AB-F1097UE), PE-conjugated anti-mouse Foxp3 antibody (E-AB-F1238D), and FITC-conjugated anti-mouse CD25 antibody (E-AB-F1102UC) were obtained from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Control