Journal: Diagnostics

Article Title: Expression of IL-21 and IL-33 in Intestinal Mucosa of Inflammatory Bowel Disease: An Immunohistochemical Study

doi: 10.3390/diagnostics13132185

Figure Lengend Snippet: Pre-treatment IL-21 and IL-33 expression in IBD patients and controls.

Article Snippet: The antibodies used were the IL 21 rabbit polyclonal (ab154767, ABCAM Ltd., Cambridge, UK) and the IL 33 monoclonal (clone 12B3C4, NOVUS Biologicals Ltd., Oxon, UK).

Techniques: Expressing, Staining

Journal: Diagnostics

Article Title: Expression of IL-21 and IL-33 in Intestinal Mucosa of Inflammatory Bowel Disease: An Immunohistochemical Study

doi: 10.3390/diagnostics13132185

Figure Lengend Snippet: Post-treatment IL-21 and IL-33 expression in IBD patients.

Article Snippet: The antibodies used were the IL 21 rabbit polyclonal (ab154767, ABCAM Ltd., Cambridge, UK) and the IL 33 monoclonal (clone 12B3C4, NOVUS Biologicals Ltd., Oxon, UK).

Techniques: Expressing, Staining

Journal: Diagnostics

Article Title: Expression of IL-21 and IL-33 in Intestinal Mucosa of Inflammatory Bowel Disease: An Immunohistochemical Study

doi: 10.3390/diagnostics13132185

Figure Lengend Snippet: ( a , b ) Colonic tissues from UC patient pre- ( a ) and post- ( b ) biologic treatment. Weak (+) tissue staining for IL-21(arrows). ×200.

Article Snippet: The antibodies used were the IL 21 rabbit polyclonal (ab154767, ABCAM Ltd., Cambridge, UK) and the IL 33 monoclonal (clone 12B3C4, NOVUS Biologicals Ltd., Oxon, UK).

Techniques: Staining

Journal: Diagnostics

Article Title: Expression of IL-21 and IL-33 in Intestinal Mucosa of Inflammatory Bowel Disease: An Immunohistochemical Study

doi: 10.3390/diagnostics13132185

Figure Lengend Snippet: ( a , b ) Colonic tissues from CD patient post- ( a ) and pre- ( b ) biologic treatment. Weak (+) tissue staining for IL-21. ×200.

Article Snippet: The antibodies used were the IL 21 rabbit polyclonal (ab154767, ABCAM Ltd., Cambridge, UK) and the IL 33 monoclonal (clone 12B3C4, NOVUS Biologicals Ltd., Oxon, UK).

Techniques: Staining

Journal: Diagnostics

Article Title: Expression of IL-21 and IL-33 in Intestinal Mucosa of Inflammatory Bowel Disease: An Immunohistochemical Study

doi: 10.3390/diagnostics13132185

Figure Lengend Snippet: Synopsis of study results.

Article Snippet: The antibodies used were the IL 21 rabbit polyclonal (ab154767, ABCAM Ltd., Cambridge, UK) and the IL 33 monoclonal (clone 12B3C4, NOVUS Biologicals Ltd., Oxon, UK).

Techniques: Activity Assay, Expressing

Journal: Cell Death & Disease

Article Title: Single-cell profiling of immune cells after neoadjuvant pembrolizumab and chemotherapy in IIIA non-small cell lung cancer (NSCLC)

doi: 10.1038/s41419-022-05057-4

Figure Lengend Snippet: A The heatmap showing GSEA enrichment of the CXCL13, Tfh cell, and 12-chemokine signatures in treatment-naive, non-MPR, and MPR tumor tissues. P adj > 0.05 in the enrichment pathway was adjusted to 0 to draw the heat map. B The higher TLS density in neoadjuvant tumor lesions verified by mIHC staining in a single-cell sequencing cohort. C Higher intratumoral C2-CD4-IL7R and C4-CD4-TCF7 cluster proportions in neoadjuvant tumor lesions in scRNA profiles. D IL21 expression in different tissue types tested with Wilcoxon in scRNA profiles showing that IL21 was most prevalent in tumor tissues. **** p < 0.0001. E Go biological function analysis of the IL21 expression cells by clusterprofiler in scRNA profiles. F Flow cytometry of IL-21 in CD4 + T cells in tumor tissues. Data presented as mean ± SEM. Data were summarized from n = 8 treatment-naive, n = 5 non-MPR and n = 15 MPR tumor samples. P values were determined by ordinary one-way ANOVA. * p < 0.05, ** p < 0.01, ns: not significant. G Comparison of paired plasma IL-21 before and after neoadjuvant chemoimmunotherapy. P values were determined by Wilcoxon matched-pairs signed-rank test, two-tailed. * p < 0.05. H , I MIHC staining showing CD4 + IL21 + and IgG-positive cells were much abundant in MPR tumor tissues (image under 10× and 20×). Data presented as mean ± SEM. Data were summarized from n = 6 treatment-naive, n = 9 non-MPR and n = 18 MPR tumor samples. P values were determined by ordinary one-way ANOVA. * p < 0.05, ns: not significant.

Article Snippet: The primary antibodies and IHC metrics were: rabbit monoclonal anti-human CD4 antibody (Abcam, Cat# ab133616, diluted at 1:1000), rabbit monoclonal anti-human CD20 antibody (Abcam, Cat# ab78237, diluted at 1:2000), mouse monoclonal anti-human IgG antibody (Abcam, Cat# ab200699, diluted at 1:800), rabbit polyclonal anti-human IL-21 antibody (Invitrogen, Cat# PA5-34801, diluted at 1:2000), mouse monoclonal anti-human CD21 antibody (Invitrogen, Cat# MA5-11417, diluted at 1:200), mouse monoclonal anti-human FoxP3 antibody (Abcam, Cat# ab20034, diluted at 1:500), rabbit monoclonal anti-human IgG1 antibody (Abcam, Cat# ab108969, diluted at 1:1000), rabbit monoclonal anti-human IgG3 antibody (Abcam, Cat# ab193172, diluted at 1:800), recombinant rabbit monoclonal anti-human IgA antibody (Invitrogen, Cat# MA5-32575, diluted at 1:800).

Techniques: Staining, Sequencing, Expressing, Flow Cytometry, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Single-cell profiling of immune cells after neoadjuvant pembrolizumab and chemotherapy in IIIA non-small cell lung cancer (NSCLC)

doi: 10.1038/s41419-022-05057-4

Figure Lengend Snippet: A Validation cohort: Surgical resected tumor tissues from 26 treatment-naive IIIA/IIIB NSCLC and 30 resectable IIIA/IIIB NSCLC who have received two cycles of neoadjuvant pembrolizumab and chemotherapy before surgery. B Neoadjuvant chemoimmunotherapy promoted more TLSs formation in non-MPR and MPR tumor lesions in our validation cohort. Data presented as mean ± SEM. Data were summarized from n = 26 treatment-naive, n = 10 non-MPR and n = 20 MPR tumor samples. P values were determined by ordinary one-way ANOVA. * p < 0.05, *** p < 0.001, ns: not significant. C , D MIHC staining of B cell isotypes, CD4 and IL-21 showing that CD20, IgG1, IgG3, CD4, and IL-21 were significantly higher in MPR tumor lesions, whereas IgA was much lower in MPR tumor lesions. Data presented as mean ± SEM. Data were summarized from n = 26 treatment-naive, n = 10 non-MPR and n = 20 MPR tumor samples. P values were determined by ordinary one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant. E Pearson correlation coefficient analysis found that IgG1 and IgG3 were positively correlated with IL-21 within tumor lesions. F Plasma IL-21 was significantly higher in MPR patients than that in treatment-naive and non-MPR patients in our validation cohort. Data presented as mean ± SEM. Data were summarized from n = 18 treatment-naive, n = 5 non-MPR and n = 10 MPR tumor samples. P values were determined by ordinary one-way ANOVA. * p < 0.05, ns: not significant.

Article Snippet: The primary antibodies and IHC metrics were: rabbit monoclonal anti-human CD4 antibody (Abcam, Cat# ab133616, diluted at 1:1000), rabbit monoclonal anti-human CD20 antibody (Abcam, Cat# ab78237, diluted at 1:2000), mouse monoclonal anti-human IgG antibody (Abcam, Cat# ab200699, diluted at 1:800), rabbit polyclonal anti-human IL-21 antibody (Invitrogen, Cat# PA5-34801, diluted at 1:2000), mouse monoclonal anti-human CD21 antibody (Invitrogen, Cat# MA5-11417, diluted at 1:200), mouse monoclonal anti-human FoxP3 antibody (Abcam, Cat# ab20034, diluted at 1:500), rabbit monoclonal anti-human IgG1 antibody (Abcam, Cat# ab108969, diluted at 1:1000), rabbit monoclonal anti-human IgG3 antibody (Abcam, Cat# ab193172, diluted at 1:800), recombinant rabbit monoclonal anti-human IgA antibody (Invitrogen, Cat# MA5-32575, diluted at 1:800).

Techniques: Staining

Journal: Cell Death & Disease

Article Title: Single-cell profiling of immune cells after neoadjuvant pembrolizumab and chemotherapy in IIIA non-small cell lung cancer (NSCLC)

doi: 10.1038/s41419-022-05057-4

Figure Lengend Snippet: Neoadjuvant chemoimmunotherapy promotes LAMP3 + DCs aggregation in tumor lesions, and the increased LAMP3 + DCs acquire tumor antigen, migrate and interact with lymphocytes, and participate in the activation, recruitment, and regulation of T/B lymphocytes through L-R interactions. Anti-PD-1 therapy reinvigorates the expansion of intratumoral CD4 + T cells and peripheral cytotoxic CD8 + T clones and promoted CD8 + T cells migration to tumor tissues to exert an anti-tumor effect. Neoadjuvant chemoimmunotherapy promotes more TLSs formation in NSCLC tumor tissues. In tumor-associated TLSs, IL-21 secreted by Tfh cells promotes B cells class switching to IgG1 and IgG3 but not IgA to mediate anti-tumor response. IgG1 and IgG3 antibodies bind to Fcγ receptor (FcγR) and trigger ADCC and antibody-mediated phagocytosis and mediate complement-based cytotoxicity. In addition, the decrease of immunosuppressive IgA + plasma cells and TNFRSF4 + Tregs lead to the repression of immunosuppression during chemoimmunotherapy.

Article Snippet: The primary antibodies and IHC metrics were: rabbit monoclonal anti-human CD4 antibody (Abcam, Cat# ab133616, diluted at 1:1000), rabbit monoclonal anti-human CD20 antibody (Abcam, Cat# ab78237, diluted at 1:2000), mouse monoclonal anti-human IgG antibody (Abcam, Cat# ab200699, diluted at 1:800), rabbit polyclonal anti-human IL-21 antibody (Invitrogen, Cat# PA5-34801, diluted at 1:2000), mouse monoclonal anti-human CD21 antibody (Invitrogen, Cat# MA5-11417, diluted at 1:200), mouse monoclonal anti-human FoxP3 antibody (Abcam, Cat# ab20034, diluted at 1:500), rabbit monoclonal anti-human IgG1 antibody (Abcam, Cat# ab108969, diluted at 1:1000), rabbit monoclonal anti-human IgG3 antibody (Abcam, Cat# ab193172, diluted at 1:800), recombinant rabbit monoclonal anti-human IgA antibody (Invitrogen, Cat# MA5-32575, diluted at 1:800).

Techniques: Activation Assay, Clone Assay, Migration

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Induction of Interferon‐γ and Tissue Inflammation by Overexpression of Eosinophil Cationic Protein in T Cells and Exosomes

doi: 10.1002/art.41920

Figure Lengend Snippet: Induction of tumor necrosis factor (TNF) signaling and interferon‐γ (IFNγ) production by overexpression of ECP in T cells. A , Distribution and classification of T cells from Lck‐ECP–transgenic and WT mice. Data were visualized using Uniform Manifold Approximation and Projection (UMAP). Values listed for each cluster are the percentage of the cell subset among all T cells. B , Volcano plot showing the selected differentially expressed genes (DEGs) in Lck‐ECP–transgenic versus WT mouse T cells. The q values were determined by Fisher's exact test. C , Violin plots showing the expression of selected DEGs in WT and Lck‐ECP–transgenic mouse T cells. P values were determined by Wilcoxon's rank sum test. D , Violin plots showing the expression of follicular helper T (Tfh) cell markers (interleukin‐21 [IL‐21], CXCR5, inducible costimulator [ICOS], and programmed death 1 [PD‐1]) in WT and Lck‐ECP–transgenic mouse T cells. P values were determined by Wilcoxon's rank sum test. ab = antibody; GH = growth hormone; TNFSF8 = tumor necrosis factor superfamily 8 (see Figure for other definitions).

Article Snippet: Alexa 488–conjugated anti‐mouse IL‐21 (bs‐2621R‐A488) antibody was purchased from Bioss.

Techniques: Over Expression, Transgenic Assay, Expressing

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Induction of Interferon‐γ and Tissue Inflammation by Overexpression of Eosinophil Cationic Protein in T Cells and Exosomes

doi: 10.1002/art.41920

Figure Lengend Snippet: Increase in follicular helper T (Tfh) cell and plasma B cells numbers, and autoantibody levels, in Lck‐ECP–transgenic mice. A , Flow cytometric analysis of Tfh (CD3+CD4+IL‐21+ or CD3+CD4+CXCR5+) cells in the spleens (Sp) of 8‐week‐old and 16‐week‐old WT and Lck‐ECP–transgenic mice. B , Flow cytometric analysis of B220+ B cells (B220+CD21+/mature B or B220+CD23+/naive B cells) in the spleens of 8‐week‐old and 16‐week‐old WT and Lck‐ECP–transgenic mice. C , Flow cytometric analysis of plasma B (B220+CD138+) cells in the bone marrow (BM) of 8‐week‐old and 16‐week‐old WT and Lck‐ECP–transgenic mice. Values next to the outlined areas in A–C are the number of positive cells. D , Serum antinuclear antibody (ANA) and rheumatoid factor (RF) levels in 8‐week‐old and 16‐week‐old WT mice (blue) and Lck‐ECP–transgenic mice (pink), determined by enzyme‐linked immunosorbent assay. Each symbol represents an individual mouse; bars show the mean ± SEM. * = P < 0.05; ** = P < 0.01, by Student's 2‐tailed t ‐test. See Figure for other definitions.

Article Snippet: Alexa 488–conjugated anti‐mouse IL‐21 (bs‐2621R‐A488) antibody was purchased from Bioss.

Techniques: Transgenic Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Disruption of Tfh:B Cell Interactions Prevents Antibody-Mediated Rejection in a Kidney Transplant Model in Rats: Impact of Calcineurin Inhibitor Dose

doi: 10.3389/fimmu.2021.657894

Figure Lengend Snippet: Markers associated with activation and function of T follicular helper cells. (A) splenic mRNA expression of PD-1 and ICOS, (B) ICOS ligand (C) CD40 ligand and (D) IL-21 and IL-21 receptor. From controls (n=5), hiCNI (n=6) and loCNI (n=8), (E) representative immunofluorescent staining of IL-21 within splenic follicles. (F) Serum IL-21 concentrations of hiCNI and loCNI at d0 (hiCNI n=3, loCNI n=3), d14 (hiCNI n=2, loCNI n=8) and d28 (hiCNI n=6, loCNI n=8) post-Ktx. Splenic mRNA expression of IL-6 (G) and TACI (H) from controls (n=5), hiCNI (n=6) and loCNI (n=8). Expression of mRNA was normalized to house-keeping gene HPRT and expressed as delta CT (AU). Data is shown as mean and individual data points; statistical significance is denoted as *p≤ 0.05, **p<0.01 and ***p<0.001. In (F) , # p≤0.05 for loCNI, *p≤0.05 for hiCNI.

Article Snippet: Primary antibodies used were anti-rat IgG-Alexa Fluor 647 (ThermoFisher, A21472) for plasma cells, anti-IL-21 (Bioss bs-2621R-a350, Boston, USA), anti-CD20 (Santacruz sc-393894, Dallas, USA) for B cells, anti-Ki67-Fitc (ThermoFisher 11-5698) for proliferating GC cells, and anti-CD3 (Abcam 5690, Cambridge, UK) for Tfh, which were counted within Ki67 + GC (activated Tfh) and the MZ (resting Tfh) regions as demonstrated in .

Techniques: Activation Assay, Expressing, Staining