chemokine mixture with cxcl2  (R&D Systems)

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Single-Cell Analysis Reveals Malignant Cells Reshape the Cellular Landscape and Foster an Immunosuppressive Microenvironment of Extranodal NK/T-Cell Lymphoma.

doi: 10.1002/advs.202303913

Figure Lengend Snippet: Figure 7. Schematic diagrams of cross-talks among malignant NK cells and immune cells in the TME of NKTCL. EBV-infected malignant NK cells and immune cells together participate in the development of NKTCL. 1) Upon EBV infection, LMP1 may contribute to the malignant transformation of NK cells (Figure 2H). 2) Malignant NK cells and TAMs secret a variety of chemokines (including CCL2, CCL3, CCL4, CCL5, etc.) and thereby recruit multiple types of immune cells from peripheral blood through corresponding chemotactic interactions (Figure 3A). 3) Soluble DPP4 secreted by malignant NK cells can truncate and rapidly degrade CXCL2, CXCL9, and CXCL10 in NKTCL TME, whereby hampering the recruitment of CXCR2+CXCR3+ immune cells (Figure 3B–H). 4) Malignant NK cells (especially LMP1+ ones) expressing CD86 and PD-L1 can negatively regulate the immune response of tumor-infiltrating T cells including exhausted and regulatory T cells (CD8+ TEX, CD4+ TEX, and Treg; Figures 4A–C and 5A–D). 5) TAMs not only secrete immunosuppressive IL10 and angiogenic VEGFA, but also interact with tumor-infiltrating T cells through suppressive interactions of CD86-CTLA4 and PDL1-PD1 (Figures 4B and 5A).

Article Snippet: DPP4 protein (Biolegend; Cat #764102) and the supernatants from NKTCL cells (YT and NK-92) were incubated with DPP4 inhibitor (Linagliptin; Selleck; Cat #S3031) or DMSO (Sigma-Aldrich; Cat #D4540) as control for 1 h, which were then incubated with culture medium without chemokines (PBS) or containing 100 ng mL−1 of chemokine mixture with CXCL2 (R&D Systems, USA; Cat #276-GB-010), CXCL9 (R&D Systems; Cat #392-MG010), and CXCL10 (R&D Systems; Cat #266-IP-010) for a 6-h pretreatment and subsequently added into lower chambers.

Techniques: Infection, Transformation Assay, Expressing

Journal: Journal of Molecular Cell Biology

Article Title: Tumor cells educate mesenchymal stromal cells to release chemoprotective and immunomodulatory factors

doi: 10.1093/jmcb/mjz090

Figure Lengend Snippet: Determination of the CXCR1/2 ligands secreted by CA-MSCs and iCA-MSCs. ( A – C ) The upregulation of genes coding for CXCL1, CXCL2 and IL-8 was validated by RT-qPCR performed on RNA extracted from BM-MSCs and different types of iCA-MSCs (induced by IGROV-1 CM, SKOV-3 CM, or ascites). The data from BM-MSCs were set to 1 and the relative quantity of mRNA is shown. CXCL1 ( A ), CXCL2 ( B ), IL-8 ( C ). ( D – F ) The concentrations of CXCL1, CXCL2, and IL-8 in the CM were quantified using ELISA kits. The CM from CA-MSCs was also tested. Histograms show the mean concentrations of CXCL1 ( D ), CXCL2 ( E ), and IL-8 ( F ) from three independent experiments performed in triplicate (mean ± SEM). ( G ) The CXCL1, CXCL2, and IL-8 concentrations were determined using ELISA kits on samples of serum from patients with ovarian adenocarcinoma collected at diagnosis. ( H ) The sum of the CXCL1, CXCL2, and IL-8 concentrations was obtained by adding together the serum concentration of these three chemokines. P -values of <0.05 (*) using a Wilcoxon–Mann–Whitney test indicate a significant difference.

Article Snippet: The concentrations of murine and human CXCL1, murine and human CXCL2, and human IL-8 were determined by ELISA using the respective ELISA kits, EK0722, EK0723, EK0452 (Boster Biological Technology), ARG80185 (Arigo biolaboratories), EK0413 (Boster Biological Technology), and the IL-8 DuoSet® ELISA Development System (R&D Systems), according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Concentration Assay, MANN-WHITNEY

Journal: Journal of Molecular Cell Biology

Article Title: Tumor cells educate mesenchymal stromal cells to release chemoprotective and immunomodulatory factors

doi: 10.1093/jmcb/mjz090

Figure Lengend Snippet: Levels of pro-inflammatory cytokines in the mouse peritonea. Levels of human and mouse chemokines CXCL1, CXCL2, and IL-8 in the mice peritonea were evaluated by ELISA (8 mice/group). P -values of <0.05 (*) using a Wilcoxon–Mann–Whitney test indicate a significant difference.

Article Snippet: The concentrations of murine and human CXCL1, murine and human CXCL2, and human IL-8 were determined by ELISA using the respective ELISA kits, EK0722, EK0723, EK0452 (Boster Biological Technology), ARG80185 (Arigo biolaboratories), EK0413 (Boster Biological Technology), and the IL-8 DuoSet® ELISA Development System (R&D Systems), according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: The Journal of Clinical Investigation

Article Title: Targeting VLA4 integrin and CXCR2 mobilizes serially repopulating hematopoietic stem cells

doi: 10.1172/JCI124738

Figure Lengend Snippet: (A–B) DBA2/J mice were treated with the VLA4 inhibitor firategrast (100 mg/kg, i.v.), the CXCR2 ligand tGro-β (2.5 mg/kg, s.c.) or both agents immediately after each other. Blood was analyzed for WBCs (A) and CFU-Cs (B). Data are mean ± SEM, n = 5. ***P < 0.001, **P < 0.01, compared with firategrast alone/compared with tGro-β alone. (C) Molecular structures. (D) G2-ALL cells were treated in duplicate with the VLA4 inhibitors shown in C. Percent inhibition of VCAM1 binding as compared with untreated samples. Data are mean ± SEM of a single experiment representative of 3 experiments. (E) DBA2/J mice were injected with tGro-β (2.5 mg/kg, s.c.), a VLA4 antagonist (3 mg/kg, i.v., for BIO5192, CWHM-823, and -842; 100 mg/kg, i.v., for firategrast), or their combination. Controls received vehicle only. Numbers of circulating CFU-Cs and LSK cells were analyzed 0.5 hours after the injection(s). Data are mean ± SEM, n = 8–10. ***P < 0.001, **P < 0.01, *P < 0.01, compared with tGro-β alone/VLA4 antagonist alone. (F) HSPC mobilization in CXCR2-KO mice using the CXCR2 ligands CXCL1, CXCL2 (tGro-β), and CXCL8 and the VLA4 antagonist CWHM-823 as well as their combinations was compared with that in WT BALB/cJ. Blood CFU-C numbers were analyzed at baseline, 15 minutes after injection of CXCR2 ligands (s.c., 1 mg/kg CXCL1 and CXCL8, 2 mg/kg tGro-β), 1 hour after injection of CWHM-823 (s.c., 3 mg/kg), and 30 minutes after the combined treatment (s.c. injection of each ligand together with CWHM-823 at same doses as single treatments). Data are mean ± SEM, n = 4–26 in mobilized groups, n = 51–78 in baseline groups. ***P < 0.001, compared with CXCR2 agonist alone/compared with CWHM-823 alone. Statistical comparisons were made using linear mixed models in A and B and ANOVA in all others, followed by step-down Bonferroni’s adjustment for multiple comparisons.

Article Snippet: Human recombinant truncated Gro-β peptide (tGro-β, CXCL2, SB-251353 from GlaxoSmithKline) was used for all in vivo experiments.

Techniques: Inhibition, Binding Assay, Injection