Journal: Cancer Science

Article Title: Chemokine C‐C motif ligand 21 synergized with programmed death‐ligand 1 blockade restrains tumor growth

doi: 10.1111/cas.15110

Figure Lengend Snippet: Chemokine C‐C motif ligand 21 (CCL21) was positively associated with CD8+ T cell infiltration, but not survival. A, The landscape of immune infiltration in 128 tumor tissues arranged by CCL21 expression from low to high in The Cancer Genome Atlas (TCGA) cohort. B, Analysis of differential immune cells between the low and high CCL21 expression group in TCGA cohort. C, Correlation analysis of CCL21 and CD8 T cells based on TCGA data. Kaplan‐Meier survival curves of CCL21 in (D) TCGA and (E) International Cancer Genome Consortium (ICGC) cohorts

Article Snippet: The CCL21 concentration was measured by the Mouse CCL21 ELISA Kit (Boster Biological Technology).

Techniques: Expressing

Journal: Cancer Science

Article Title: Chemokine C‐C motif ligand 21 synergized with programmed death‐ligand 1 blockade restrains tumor growth

doi: 10.1111/cas.15110

Figure Lengend Snippet: Chemokine C‐C motif ligand 21 (CCL21) promoted T cell infiltration as well as increased programmed death‐ligand 1 (PD‐L1) expression in vivo. A‐C, (A) Western blot analysis, (B) quantitative real‐time PCR analysis, and (C) ELISA analysis for CCL21 in CCL21 overexpressing (OE) and relative control Panc02 (CON) cells. D, Images (left), volumes (middle), and weights (right) of the CCL21 OE vs control Panc02 tumors in C57BL/6 mice. E, Immunohistochemical images of CCL21 and PD‐L1 expression levels and tumor‐infiltrating lymphocyte densities, and immunofluorescent images of TUNEL staining in CCL21 OE vs control Panc02 tumors in C57BL/6 mice. Representative images are shown. Scale bars, 100 μm. F, Quantification of immunohistochemistry and immunofluorescence results of CD4 + T cells, CD8 + T cells, and TUNEL + cells in CCL21 OE vs control Panc02 tumors in C57BL/6 mice. Each group contained six mice. Statistical significance was calculated by unpaired two‐tailed Student’s t tests. ** P < .01, *** P < .001

Article Snippet: The CCL21 concentration was measured by the Mouse CCL21 ELISA Kit (Boster Biological Technology).

Techniques: Expressing, In Vivo, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control, Immunohistochemical staining, TUNEL Assay, Staining, Immunohistochemistry, Immunofluorescence, Two Tailed Test

Journal: Cancer Science

Article Title: Chemokine C‐C motif ligand 21 synergized with programmed death‐ligand 1 blockade restrains tumor growth

doi: 10.1111/cas.15110

Figure Lengend Snippet: Chemokine C‐C motif ligand 21 (CCL21) upregulated programmed death‐ligand 1 (PD‐L1) expression to promote immune escape in vitro. A, Western blot analysis of PD‐L1 abundance in MIA‐PaCa‐2 and SW1990 cells treated with CCL21 at the concentration of 0, 12, 25, 50, or 100 ng/mL for 48 h. B, Western blot analysis of PD‐L1 abundance in MIA‐PaCa‐2 and SW1990 cells treated with CCL21 at a concentration of 50 ng/mL for 0, 12, 24, 36, and 48 h after 24 h starvation. C, Flow cytometry analysis of PD‐L1 expression in MIA‐PaCa‐2 and SW1990 cells treated with 50 ng/mL CCL21 for 48 h. D, Quantitative real‐time PCR analysis of PD‐L1 mRNA level when MIA‐PaCa‐2 and SW1990 cells were treated with CCL21 (50 ng/mL) or PBS for 12 h. E, T cell‐mediated tumor cell killing assay analysis of different groups of cancer cell survival after coculture with or without activated T cells or anti‐PD‐L1 Ab for 4 d. F, Modified T cell‐mediated tumor cell killing assay using the Transwell system. Pancreatic cancer cells were seeded in the lower chamber prior to addition of the corresponding tumor cell line‐specific T cells in the upper chamber. Surviving tumor cells were visualized by crystal violet staining

Article Snippet: The CCL21 concentration was measured by the Mouse CCL21 ELISA Kit (Boster Biological Technology).

Techniques: Expressing, In Vitro, Western Blot, Concentration Assay, Flow Cytometry, Real-time Polymerase Chain Reaction, Modification, Staining

Journal: Cancer Science

Article Title: Chemokine C‐C motif ligand 21 synergized with programmed death‐ligand 1 blockade restrains tumor growth

doi: 10.1111/cas.15110

Figure Lengend Snippet: Chemokine C‐C motif ligand 21 (CCL21) stabilized programmed death‐ligand 1 (PD‐L1) through the AKT‐glycogen synthase kinase‐3β (GSK‐3β) signaling pathway. A, Western blot analysis of PD‐L1 abundance in MIA‐PaCa‐2 and SW1990 cells treated with 80 μΜ cycloheximide (CHX) for 0, 4, 8, 12, or 16 h after CCL21 stimulation (right) or not (left). B, Western blot analysis of AKT‐GSK‐3β signaling pathway changes of starved cells treated with serum‐free medium contained CCL21 (50 ng/mL) for 0, 15, 30, 60, and 120 min. C, Western blot analysis of the AKT‐GSK‐3β signaling pathway changes of starved cells pretreated with DMSO, AKT inhibitor (LY294002, 20 μmol/L) or anti‐CC‐chemokine receptor 7 (CCR7, 10 µg/mL) for 2 h, and then treated with CCL21 (50 ng/mL) or PBS for an additional 2 h. D, Western blot analysis of PD‐L1 expression in starved cells pretreated with DMSO, LY29400,2 or anti‐CCR7 for 2 h, and then treated with CCL21 (50 ng/mL) or PBS for an additional 48 h. E, Coimmunoprecipitation assays in MIA‐PaCa‐2 and SW1990 cells transfected with indicated plasmids

Article Snippet: The CCL21 concentration was measured by the Mouse CCL21 ELISA Kit (Boster Biological Technology).

Techniques: Western Blot, Expressing, Transfection

Journal: Cancer Science

Article Title: Chemokine C‐C motif ligand 21 synergized with programmed death‐ligand 1 blockade restrains tumor growth

doi: 10.1111/cas.15110

Figure Lengend Snippet: Chemokine C‐C motif ligand 21 (CCL21) combined with programmed death‐ligand 1 (PD‐L1) blockade enhanced therapeutic efficacy for pancreatic cancer. A‐C, (A) Tumor images, (B) volumes, and (C) weights (means ± SD) of control Panc02 or CCL21‐overexpressing (OE) Panc02 tumor subcutaneously inoculated in C57BL/6 mice treated with IgG control Ab or anti‐PD‐L1 Ab as indicated. D, Immunohistochemistry analysis of CD4, CD8, and granzyme B, and immunofluorescent image of TUNEL staining in tumors from mice that received the treatments described above. Representative images are shown. Scale bars, 100 μm. E, Quantification of immunohistochemistry and immunofluorescence results above. Each group contained six mice. Statistical significance was calculated by unpaired two‐tailed Student’s t tests. ** P < .01, *** P < .001

Article Snippet: The CCL21 concentration was measured by the Mouse CCL21 ELISA Kit (Boster Biological Technology).

Techniques: Drug discovery, Control, Immunohistochemistry, TUNEL Assay, Staining, Immunofluorescence, Two Tailed Test

Journal: Cancer Science

Article Title: Chemokine C‐C motif ligand 21 synergized with programmed death‐ligand 1 blockade restrains tumor growth

doi: 10.1111/cas.15110

Figure Lengend Snippet: Schematic drawing for the mechanism of chemokine C‐C motif ligand 21 (CCL21) synergistic with programmed death‐ligand 1 (PD‐L1) blockade. CCL21 in the tumor microenvironment can promote infiltration of T cells and enhance their antitumor immune response. However, CCL21 also stabilizes and upregulates PD‐L1 expression on pancreatic cancer cells through the AKT‐glycogen synthase kinase‐3β (GSK‐3β) signaling pathway at the same time, which could partly impair the local immune response. Thus, PD‐L1 blockade was undertaken in combination with CCL21, which showed a powerful synergism and promising therapeutic efficacy. CCR7, CC‐chemokine receptor 7

Article Snippet: The CCL21 concentration was measured by the Mouse CCL21 ELISA Kit (Boster Biological Technology).

Techniques: Expressing, Drug discovery

Journal: Immunity

Article Title: Visualization of T Cell Migration in the Spleen Reveals a Network of Perivascular Pathways that Guide Entry into T Zones

doi: 10.1016/j.immuni.2020.03.010

Figure Lengend Snippet:

Article Snippet: QuantiTect Primers Ccl21a , QIAGEN , QT00284753.

Techniques: Software, Imaging, Light Microscopy