cxcl10  (R&D Systems)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Age-related dysregulation of CXCL9/10 in monocytes is linked to impaired innate immune responses in a mouse model of Staphylococcus aureus osteomyelitis

doi: 10.1007/s00018-024-05311-2

Figure Lengend Snippet: The expression of CXCL9 and CXCL10 are highly activated by S. aureus challenge in bone marrow monocytes of 8-week-old mice. ( a ) Gene ontology (GO) enrichment analysis of biological processes for upregulated differentially expressed genes (DEGs). The transcriptome data of S. aureus -infected bone and control ones in young mice on day 3 after surgery (GSE166522) were analyzed by bioinformatics. ( b ) Venn diagram of overlapping DEGs between cytokine-mediated signaling pathway and response to molecule of bacterial origin. ( c ) Heatmap of 12 overlapping DEGs. ( d ) mRNA expression of the top 3 highly up-regulated genes (CXCL9, IL-1β, CXCL10) in bone tissue on day 3 after surgery. n = 4/ group. One-way ANOVA with Dunnett’s T3 post hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001. ( e ) Representative images of immunohistochemical staining for CXCL9 in S. aureus -infected femurs and controls on day 3 after surgery, scale bar = 50 μm. ( f ) The percentages of CXCL9-positive stained area in the area of field of view (FOV) in bone marrow. n = 5/ group. One-way ANOVA with Dunnett’s T3 post hoc test, ** p < 0.01. ( g ) Representative images of CXCL10 immunohistochemical staining in bone tissue on day 3 after surgery, scale bar = 50 μm. ( h ) The percentages of CXCL10-positive stained area in the area of FOV in bone marrow. n = 5/ group. One-way ANOVA with Bonferroni post hoc test, *** p < 0.001. mRNA expression of CXCL9 ( i ) and CXCL10 ( j ) in monocytes, macrophages and neutrophils isolated from bone marrow of 8-week-old mice and 10-month-old mice. Cells were treated with various MOI (0, 2, 10 and 50) of S. aureus , and total RNA were collected after 12 h. n = 3/group, One-way ANOVA with Bonferroni post hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: To activate CXCR3 signaling in middle-aged mice with S. aureus osteomyelitis, 10-month-old mice were injected with 100 ng recombinant mouse CXCL9 protein (#C600269, BBI Life Science, Shanghai, China) or recombinant mouse CXCL10 protein (#HY-P722, MedChemExpress, USA) into the bone marrow cavity using a microsyringe, while the control group was injected with the same volume of PBS.

Techniques: Expressing, Infection, Control, Immunohistochemical staining, Staining, Isolation

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Age-related dysregulation of CXCL9/10 in monocytes is linked to impaired innate immune responses in a mouse model of Staphylococcus aureus osteomyelitis

doi: 10.1007/s00018-024-05311-2

Figure Lengend Snippet: Monocyte-derived CXCL9/10 evoked by S. aureus enhance the bactericidal function of neutrophils and macrophages. ( a ) Representative images of the remaining extracellular and intracellular S. aureus colonies inoculated in TSA plates, and quantitative analysis of extracellular ( b ) and intracellular ( c ) bacterial colonies of neutrophils culture. After neutrophils were pre-stimulated with 100 ng/ml of recombinant CXCL9 (r-CXCL9), recombinant CXCL10 (r-CXCL10), or a combination of them for 4 h, cells were challenged with S. aureus at MOI of 10 for 30 min. n = 3/ group. One-way ANOVA with Bonferroni post hoc test, ** p < 0.01, *** p < 0.001. ( d ) Representative images of intracellular S. aureus of BMDMs inoculated in TSA and ( e ) quantification of bacteria colonies of BMDMs pre-treated with r-CXCL9, r-CXCL10, and a combination of them. n = 3/ group. One-way ANOVA with Bonferroni post hoc test, *** p < 0.001. ( f ) Representative images of the remaining extracellular and intracellular S. aureus colonies inoculated in TSA plates from neutrophils culture, and quantitative analysis of extracellular ( g ) and intracellular ( h ) bacterial colonies. After neutrophils were pre-stimulated for 4 h with CM-8 W Mono S. aureus that had been treated with recombinant siRNA for CXCL9 (si-CXCL9), CXCL10 (si-CXCL10), a combination of si-CXCL9 and si-CXCL10, or negative control (si-NC), cells were then challenged with S. aureus at MOI of 10 for 30 min. CM-8 W Mono- S. aureus represents the CM of monocytes isolated from the bone marrow of 8-week-old and challenged by S. aureus . n = 3/ group. One-way ANOVA with Bonferroni post hoc test, * p < 0.05, ** p < 0.01. ( i ) Representative images of intracellular S. aureus colonies of BMDMs and ( j ) quantification of bacterial colonies of BMDMs pretreated with CM-8 W Mono- S. aureus with the knockdown of CXCL9 and CXCL10. n = 3/ group. One-way ANOVA with Bonferroni post hoc test, * p < 0.05

Article Snippet: To activate CXCR3 signaling in middle-aged mice with S. aureus osteomyelitis, 10-month-old mice were injected with 100 ng recombinant mouse CXCL9 protein (#C600269, BBI Life Science, Shanghai, China) or recombinant mouse CXCL10 protein (#HY-P722, MedChemExpress, USA) into the bone marrow cavity using a microsyringe, while the control group was injected with the same volume of PBS.

Techniques: Derivative Assay, Recombinant, Bacteria, Negative Control, Isolation, Knockdown

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Age-related dysregulation of CXCL9/10 in monocytes is linked to impaired innate immune responses in a mouse model of Staphylococcus aureus osteomyelitis

doi: 10.1007/s00018-024-05311-2

Figure Lengend Snippet: CXCR3 signaling in neutrophils and macrophages promotes their bactericidal function. ( a ) Representative images of the remaining extracellular and intracellular S. aureus colonies of neutrophils inoculated in TSA, and quantitative analysis of extracellular ( b ) and intracellular ( c ) bacterial colonies. Neutrophils were pre-stimulated with AMG487 (500 µM), an inhibitor of CXCR3, for 1 h, and then cultured in CM-8 W Mono- S. aureus for 4 h. Next, cells were challenged with S. aureus (MOI = 10) for 30 min, and bactericidal function were evaluated. n = 3/ group. Student’s t test, * p < 0.05, ** p < 0.01. ( d ) Representative images of the remaining intracellular S. aureus colonies of BMDMs inoculated in TSA, and ( e ) quantification of bacterial colonies. BMDMs were pre-stimulated with 500 µM AMG487 for 1 h, and then further stimulated with CM-8 W Mono- S. aureus for 4 h. Next, cells were challenged with S. aureus (MOI = 10) for 1 h. After removal of the extracellular S. aureus , culture of the BMDMS was continued with CM-8 W Mono- S. aureus and AMG487 for 12 h and bacterial loading were evaluated. n = 3/ group. Student’s t test, * p < 0.05. ( f and i ) Representative images of the remaining extracellular and intracellular S. aureus colonies of neutrophils inoculated in TSA, and quantitative analysis of extracellular ( g and j ) and intracellular ( h and k ) bacterial colonies. Neutrophils were pre-stimulated with 500 µM AMG487 for 1 h, and then further treated with 100 ng/ml of r-CXCL9 or r-CXCL10 for 4 h. Next, S. aureus (MOI = 10) was co-cultured with pre-stimulated neutrophils for 30 min. Finally, the extracellular and intracellular bacterial loading were evaluated. n = 3/ group. Student’s t test, * p < 0.05, *** p < 0.001. ( l ) Representative images of remaining intracellular bacteria of BMDMs inoculated on TSA and (m and n) quantification of bacterial colonies in BMDMs. BMDMs were pre-stimulated with 500 µM AMG487 for 1 h, and then further stimulated with 100 ng/ml of r-CXCL9 or r-CXCL10 for 4 h. Next, cells were challenged with S. aureus (MOI = 10) for 1 h. After removal of the extracellular S. aureus , culture of the BMDMS was continued with AMG487 and with the presence or absence of r-CXCL9 or r-CXCL10 for 12 h. n = 3/ group. Student’s t test, * p < 0.05, ** p < 0.01

Article Snippet: To activate CXCR3 signaling in middle-aged mice with S. aureus osteomyelitis, 10-month-old mice were injected with 100 ng recombinant mouse CXCL9 protein (#C600269, BBI Life Science, Shanghai, China) or recombinant mouse CXCL10 protein (#HY-P722, MedChemExpress, USA) into the bone marrow cavity using a microsyringe, while the control group was injected with the same volume of PBS.

Techniques: Cell Culture, Bacteria

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Age-related dysregulation of CXCL9/10 in monocytes is linked to impaired innate immune responses in a mouse model of Staphylococcus aureus osteomyelitis

doi: 10.1007/s00018-024-05311-2

Figure Lengend Snippet: CXCL9/CXCL10-CXCR3 signaling mediates age-related lesions in the acute phase of S. aureus osteomyelitis in mice. ( a ) Representative images of immunofluorescence staining for S. aureus and ( b ) quantification of S. aureus -positive stained area per area of field of view (FOV). ( c ) Representative images of H&E staining and ( d ) quantification of necrotic area per area of FOV. 10-month-old mice were injected with 1 µl r-CXCL9 (100 ng/µl) into the bone marrow where the implant was placed and S. aureus was injected, and right femurs were collected by day 3 after surgery for further analysis. n = 3/group. Scale bar = 50 μm. ( e ) Representative images of immunofluorescence staining for S. aureus and ( f ) quantification of S. aureus -positive stained area per FOV area. ( g ) Representative images of H&E staining and ( h ) quantification of necrotic area per area of FOV. 10-month-old mice were injected with 1 µl r-CXCL10 (100 ng/µl) into the bone marrow where the implant was placed and S. aureus was injected, and right femurs were collected by day 3 after surgery for further analysis. n = 3/group. Scale bar = 50 μm. ( i ) Representative images of immunofluorescence staining for S. aureus and ( j ) quantification of S. aureus -positive stained area per FOV area. ( k ) Representative images of H&E staining and ( l ) quantification of necrotic area per area of FOV. After being implanted and infected with S. aureus or treated with vehicle in right femurs, 8-week-old mice were injected subcutaneously with AMG487 (5 mg/kg, twice a day), right femurs were collected for further analysis by day 3 after surgery. n = 3/group, Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001. Red fluorescence indicates S. aureus positive staining, and blue DAPI stained nucleus. The red star indicates the cellular lytic changes, the green arrow nuclear fragmentation

Article Snippet: To activate CXCR3 signaling in middle-aged mice with S. aureus osteomyelitis, 10-month-old mice were injected with 100 ng recombinant mouse CXCL9 protein (#C600269, BBI Life Science, Shanghai, China) or recombinant mouse CXCL10 protein (#HY-P722, MedChemExpress, USA) into the bone marrow cavity using a microsyringe, while the control group was injected with the same volume of PBS.

Techniques: Immunofluorescence, Staining, Injection, Infection, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: Unveiling the improved targeting migration of mesenchymal stem cells with CXC chemokine receptor 3-modification using intravital NIR-II photoacoustic imaging

doi: 10.1186/s12951-022-01513-7

Figure Lengend Snippet: Upregulation of Cxcl10 in inflamed ears of CHS mice. a Representative photographs of mouse ears challenged with vehicle (Control) or DNFB (Inflamed). Photos were taken 24 h after challenging. b Thickness of mice ears challenged with vehicle (Control) or DNFB (Inflamed) before (Pre), 24 h and 72 h after challenging (n = 3–4 per group). c mRNA expression of Cxcl10 transcript in mouse ears challenged with vehicle (Control) or DNFB (Inflamed) (n = 3 per group). *** P < 0.001; ns, no significance; error bars, SEM

Article Snippet: Rabbit polyclonal antibody against Cxcr3 (NB100-56404), recombinant Cxcl10 protein (466-CR) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Control, Expressing

Journal: Journal of Nanobiotechnology

Article Title: Unveiling the improved targeting migration of mesenchymal stem cells with CXC chemokine receptor 3-modification using intravital NIR-II photoacoustic imaging

doi: 10.1186/s12951-022-01513-7

Figure Lengend Snippet: Overexpression of Cxcr3 in MSCs. a Representative bright field (BF) and green fluorescence images of MSC eGFP and MSC Cxcr3 . MSCs were transduced with lentivirus to express eGFP alone (MSC eGFP ) or in combination with Cxcr3 (MSC Cxcr3 ). b Analysis of Cxcr3 mRNA expression in MSC eGFP and MSC Cxcr3 . c Western blot of Cxcr3 in total cell lysates of MSC eGFP and MSC Cxcr3 . CypB, cyclophilin B, used as housekeeping gene. d Cxcr3 overexpression promoted the in vitro migration of MSCs toward Cxcl10. Representative images and quantification analysis of transwell migration assay were shown. e Quantification analysis of transwell migration assay of MSC Cxcr3 after incubating with vehicle (PBS) or TAT-CPNPs (NPs). Scale bars, 100 µm. * P < 0.05, *** P < 0.001; error bars, SEM

Article Snippet: Rabbit polyclonal antibody against Cxcr3 (NB100-56404), recombinant Cxcl10 protein (466-CR) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Over Expression, Fluorescence, Transduction, Expressing, Western Blot, In Vitro, Migration, Transwell Migration Assay