rabbit polyclonal anti ccl5 antibody  (R&D Systems)

Journal: Cancer letters

Article Title: PARP inhibitor plus radiotherapy reshape the immune suppressive microenvironment and potentiate the efficacy of immune checkpoint inhibitors in tumors with IDH1 mutation.

doi: 10.1016/j.canlet.2024.216676

Figure Lengend Snippet: Fig. 3. IDH1 mutation downregulates the interferon pathway. (A) GSEA was performed to analyze different IFN-related signaling pathways. (B) WB was conducted to measure the expression of STAT1 in GL261 and MC38 cells overexpressing mutant IDH1 and cells overexpressing an empty vector. (C) and (D) qRT-PCR was used to assess the expression of Ifnb, Cxcl10, and Ccl5 in cells and mouse tumor tissues overexpressing mutant IDH1 and cells with an empty vector. (E) Multifactorial analysis of CXCL10 and CCL5 secretion in tumor tissues from GL261 and MC38 cells overexpressing mutant IDH1 and cells with an empty vector. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data in the bar graphs are presented as mean ± SD. Statistical analysis was performed using unpaired t tests.

Article Snippet: Subsequently, slides were incubated overnight at 4 ◦C with primary antibodies specific for CD8 (1:500, abcam, AB199016, UK), CCL5 (1:300, Origene, AA01152, USA), and CXCL10 (1:300, Proteintech, 10937-1-AP, China).

Techniques: Mutagenesis, Protein-Protein interactions, Expressing, Plasmid Preparation, Quantitative RT-PCR

Journal: Cancer letters

Article Title: PARP inhibitor plus radiotherapy reshape the immune suppressive microenvironment and potentiate the efficacy of immune checkpoint inhibitors in tumors with IDH1 mutation.

doi: 10.1016/j.canlet.2024.216676

Figure Lengend Snippet: Fig. 6. RT + PARPi enhances the activation of cGAS-STING pathway. (A) and (C) depict the detection of γ-H2AX using immunofluorescence. (B) and (D) show the detection of γ-H2AX using WB. (E) and (J) show WB analysis of the activation of the cGAS-STING pathway and the IFNβ-JAK-STAT1 pathway in GL261-IDH1mut and MC38-IDH1mut cells following RT + PARPi. (F) and (K) represent RT-qPCR analysis of the expression of downstream chemokines IFNβ, CXCL10, and CCL5 in GL261- IDH1mut and MC38-IDH1mut cells related to the cGAS-STING pathway. (H) and (M) show RT-qPCR analysis of the expression of downstream chemokines IFNβ, CXCL10, and CCL5 in tumor tissues of GL261-IDH1mut and MC38-IDH1mut. (G) and (L) demonstrate the secretion of CXCL10 and CCL5 in the cell culture supernatant of GL261-IDH1mut and MC38-IDH1mut cells using a multiplex assay. (I) and (N) display the secretion of CXCL10 and CCL5 in tumor tissues of GL261-IDH1mut and MC38-IDH1mut using a multiplex assay. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Subsequently, slides were incubated overnight at 4 ◦C with primary antibodies specific for CD8 (1:500, abcam, AB199016, UK), CCL5 (1:300, Origene, AA01152, USA), and CXCL10 (1:300, Proteintech, 10937-1-AP, China).

Techniques: Activation Assay, Immunofluorescence, Quantitative RT-PCR, Expressing, Cell Culture, Multiplex Assay

Journal: Cancer letters

Article Title: PARP inhibitor plus radiotherapy reshape the immune suppressive microenvironment and potentiate the efficacy of immune checkpoint inhibitors in tumors with IDH1 mutation.

doi: 10.1016/j.canlet.2024.216676

Figure Lengend Snippet: Fig. 7. cGAS knockout abolishes CD8+T cell chemotaxis and anti-tumor effects of RT + PARPi. (A) and (C) show the WB detection of cGAS expression in GL261- IDH1mut and MC38-IDH1mut cells after cGAS KO. (B) and (D) depict WB analysis of the activation of the JAK-STAT1 pathway by IFNβ in the four groups, with the combination group without cGAS KO serving as a control. (E) and (F) present qRT-PCR analysis of the expression of IFNβ, CXCL10, and CCL5 in GL261-IDH1mut and MC38-IDH1mut cells before and after cGAS KO. (G) and (I) count the number of CD8+ T cells passing through the pores per minute using flow cytometry in each group. (H) and (J) show multiplex assay measurement of CXCL10 and CCL5 in the cell culture supernatant of GL261-IDH1mut and MC38-IDH1mut cells in the control, RT + PARPi, and RT + PARPi cGAS KO groups. (K) and (L) display the survival curves and growth curves of mouse tumors, comparing the survival and tumor growth of mice in the combination group with and without cGAS KO in GL261-IDH1mut in situ tumor model and MC38-IDH1mut subcutaneous tumor model. Tumor growth was analyzed using two-way ANOVA. Data are presented as mean ± SD. Survival curves were plotted using Kaplan-Meier analysis. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Subsequently, slides were incubated overnight at 4 ◦C with primary antibodies specific for CD8 (1:500, abcam, AB199016, UK), CCL5 (1:300, Origene, AA01152, USA), and CXCL10 (1:300, Proteintech, 10937-1-AP, China).

Techniques: Knock-Out, Chemotaxis Assay, Expressing, Activation Assay, Control, Quantitative RT-PCR, Flow Cytometry, Multiplex Assay, Cell Culture, In Situ

Journal: Cancer letters

Article Title: PARP inhibitor plus radiotherapy reshape the immune suppressive microenvironment and potentiate the efficacy of immune checkpoint inhibitors in tumors with IDH1 mutation.

doi: 10.1016/j.canlet.2024.216676

Figure Lengend Snippet: Fig. 9. IDH mutant tumors have fewer tumor-infiltrating CD8+ T cells and reduced PD-L1 expression compared with their wild type (WT) counterparts. The key chemokines CXCL9, CXCL10, and CCL5, in tandem with a decrease in CD8+ T cell infiltration into and an attenuation of the IFN pathway within these tumors. Treating IDH1mut tumors with a combination of the PARP inhibitor veliparib and RT reversed these phenotypes, resulting in increased CD8+ T cell tumor infiltration, apoptosis and ICD, and checkpoint inhibitor sensitivity. The effects observed from RT + PARPi were superior to either modality alone. We also observed heightened activation of cGAS-STING signaling, a key DDR pathway. Critically, cGAS KO abolished the enhanced therapeutic effects of RT + PPARi.

Article Snippet: Subsequently, slides were incubated overnight at 4 ◦C with primary antibodies specific for CD8 (1:500, abcam, AB199016, UK), CCL5 (1:300, Origene, AA01152, USA), and CXCL10 (1:300, Proteintech, 10937-1-AP, China).

Techniques: Mutagenesis, Expressing, Activation Assay

Journal: Cells

Article Title: Increased Adipose Tissue Expression of IL-23 Associates with Inflammatory Markers in People with High LDL Cholesterol.

doi: 10.3390/cells11193072

Figure Lengend Snippet: Figure 4. Increased adipose CCL5 protein expression in people with high LDL-c and positively correlated with IL-23 protein. CCL5 protein expression was determined in people with low LDL-c (L-LDL-c) and high LDL-c (H-LDL-c), five each, using IHC. (A) The representative IHC images obtained from three independent determinations with similar results show the increased adipose tissue TNF-α expression (40× or inset ×100 magnification) in people with H-LDL-c (≥2.9 mmol/L) compared to those with L-LDL-c (<2.9 mmol/L). (B) Increased CCL5 protein expression is shown in the H-LDL-c group compared to the L-LDL-c group (p = 0.0053). Staining intensity shown as arbitrary units (AU). (C) IL-23 protein correlated with CCL5 protein in adipose tissue. p-values < 0.05 were considered as statistically significant. ** Highly significant.

Article Snippet: Briefly, adipose tissue sections were incubated with primary antibodies, i.e., 1:200 dilution of rabbit polyclonal anti-IL-23 antibody (Abcam® ab115759; Waltham, MA, USA), 1:200 dilution of rabbit polyclonal anti-TNF antibody (Novus Biologicals Centennial, CO, USA; NBP1–19532) and 1:500 dilution of rabbit polyclonal anti-CCL5 antibody (R&D Systems AF478) overnight at room temperature.

Techniques: Expressing, Staining

Journal: Cell reports

Article Title: Arthritis flares mediated by tissue-resident memory T cells in the joint

doi: 10.1016/j.celrep.2021.109902

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-CCL5 antibody (Origene Cat# PP1081P1, RRID:AB_1006884) was used for the neutralization of CCL5 chemokine.

Techniques: Virus, Expressing, Recombinant, Methylation, Plasmid Preparation, Cell Isolation, Multiplex Assay, Gene Expression, Microarray, Software

Journal: Cell reports

Article Title: Arthritis flares mediated by tissue-resident memory T cells in the joint

doi: 10.1016/j.celrep.2021.109902

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-CCL5 antibody (Origene Cat# PP1081P1, RRID:AB_1006884) was used for the neutralization of CCL5 chemokine.

Techniques: Virus, Expressing, Recombinant, Methylation, Plasmid Preparation, Cell Isolation, Multiplex Assay, Gene Expression, Microarray, Software

Journal: Cell reports

Article Title: Arthritis flares mediated by tissue-resident memory T cells in the joint

doi: 10.1016/j.celrep.2021.109902

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-CCL5 antibody\ , Origene , Cat# PP1081P1; RRID: AB_1006884.

Techniques: Expressing, Recombinant, Methylation, Plasmid Preparation, Cell Isolation, Multiplex Assay, Microarray, Software

Journal: Cell reports

Article Title: Arthritis flares mediated by tissue-resident memory T cells in the joint

doi: 10.1016/j.celrep.2021.109902

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-CCL5 antibody\ , Origene , Cat# PP1081P1; RRID: AB_1006884.

Techniques: Expressing, Recombinant, Methylation, Plasmid Preparation, Cell Isolation, Multiplex Assay, Microarray, Software