Journal: PLOS Pathogens

Article Title: The CXCR6-CXCL16 axis mediates T cell control of polyomavirus infection in the kidney

doi: 10.1371/journal.ppat.1012969

Figure Lengend Snippet: (A) CXCL16 mRNA expression in the kidney during MuPyV infection. Expression is shown as fold change relative to sham infected samples. Data are from three independent experiments (n = 10-11). (B) Expression of NKCC2 [marks the ascending loop of Henle ], CD8, and CXCL16 in epithelia in sham-infected and 8 dpi kidneys; bottom right photomicrographs are merged images. Representative of two independent experiments. (C) CXCL16 expression in sham-infected, 4 dpi, and 8 dpi kidney lysates (left). Western blot image is representative of two independent experiments with each lane indicating protein lysate from kidneys of individual mice. Protein band intensity quantification for sCXCL16 was normalized to β-actin and analyzed by ImageLab and normalized to the loading control (right). Data are combined from two independent experiments (n = 3-5). (D) Experimental design of in vivo CXCL16 mAb administration. Mice were administered 250 µg of CXCL16 mAb or control rat IgG every two days from days 4-14 post infection and euthanized at 15 dpi. Numbers of CD45 mAb i.v.-negative, CD8 + CD44 + D b -LT359 tetramer + T cells and CD4 + CD44 + T cells in kidneys and spleens of infected mice given anti-CXCL16 or control rat IgG. Data were analyzed by one-way ANOVA (A and C) and by multiple Mann-Whitney tests (D).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit anti-CXCL16 (Bioss) or rabbit anti-β-actin (Cell Signaling Technology) in 1% milk in TBS-T. Blots were washed with TBS-T followed by a 1.5 h incubation with horseradish peroxidase conjugated secondary antibodies (BioLegend) in 1% milk in TBS-T. After another wash with TBS-T, blots were developed with SuperSignal West Pico PLUS Chemiluminescence Substrate (ThermoFisher Scientific).

Techniques: Expressing, Infection, Western Blot, Control, In Vivo, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: The CXCR6-CXCL16 axis mediates T cell control of polyomavirus infection in the kidney

doi: 10.1371/journal.ppat.1012969

Figure Lengend Snippet: (A) Dense lymphoplasmacytic infiltrate surrounding and infiltrating cortical tubules (H&E, x400). (B) Immunohistochemical staining for Large T antigen in nuclei of tubular epithelium (x250). (C) Expression of CXCR6 on infiltrating CD8 + cells. Photomicrographs (right) are enlarged images in the white squares (left); bottom right are merged images. No 1 o , no primary antibody. (D) Log-fold change of CXCL16 and CXCR6 across four independent studies, each comparing KTx biopsies from patients with PVAN and stable graft function. The error bars indicate the 95% confidence interval of log-fold change, with horizontal dashed grey line indicating log-fold change = 0, or no difference. GSE120495 is an RNA-seq study, while remaining studies are microarray based. Given the heterogeneity of the data, a Cauchy combination method with uniform weights was employed to integrate the p-values across these studies. The combined p-values for CXCL16 = 0.000337 and CXCR6 = 2.72x10 -05 . ns, non-significant at adjusted p-value > 0.05; *, statistical significance at adjusted p-value ≤ 0.05; **, statistical significance at adjusted p-value ≤ 0.01; ***, statistical significance at adjusted p-value ≤ 0.001.

Article Snippet: Membranes were incubated overnight at 4°C with rabbit anti-CXCL16 (Bioss) or rabbit anti-β-actin (Cell Signaling Technology) in 1% milk in TBS-T. Blots were washed with TBS-T followed by a 1.5 h incubation with horseradish peroxidase conjugated secondary antibodies (BioLegend) in 1% milk in TBS-T. After another wash with TBS-T, blots were developed with SuperSignal West Pico PLUS Chemiluminescence Substrate (ThermoFisher Scientific).

Techniques: Immunohistochemical staining, Staining, Expressing, RNA Sequencing, Microarray

Journal: PLOS Pathogens

Article Title: The CXCR6-CXCL16 axis mediates T cell control of polyomavirus infection in the kidney

doi: 10.1371/journal.ppat.1012969

Figure Lengend Snippet: Summary of differential gene expression of CXCL16 and CXCR6 in four studies of KTx biopsies with PVAN. Because of the heterogeneity of the data (i.e., one RNA-seq, three microarray), a Cauchy combination method with uniform weights was employed to integrate the p-values across these studies. P-values significant after false discover rate (FDR) correction are bolded and italicized in the Adj. P-value column. Cauchy combined p-values < 0.05 are bolded and italicized.

Article Snippet: Membranes were incubated overnight at 4°C with rabbit anti-CXCL16 (Bioss) or rabbit anti-β-actin (Cell Signaling Technology) in 1% milk in TBS-T. Blots were washed with TBS-T followed by a 1.5 h incubation with horseradish peroxidase conjugated secondary antibodies (BioLegend) in 1% milk in TBS-T. After another wash with TBS-T, blots were developed with SuperSignal West Pico PLUS Chemiluminescence Substrate (ThermoFisher Scientific).

Techniques: Gene Expression, Microarray

Journal: Oncotarget

Article Title: Soluble chemokine (C-X-C motif) ligand 16 (CXCL16) in urine as a novel biomarker candidate to identify high grade and muscle invasive urothelial carcinomas

doi: 10.18632/oncotarget.20737

Figure Lengend Snippet: Comparison of normalized CXCL16 values (pg/mg creatinine) according to patient and sample characteristics

Article Snippet: The following primary antibodies were used: CXCL16 (1:250; Acris Antibodies) or CXCR6 (1:750; Abcam).

Techniques: Comparison

Journal: Oncotarget

Article Title: Soluble chemokine (C-X-C motif) ligand 16 (CXCL16) in urine as a novel biomarker candidate to identify high grade and muscle invasive urothelial carcinomas

doi: 10.18632/oncotarget.20737

Figure Lengend Snippet: Box plots of creatinine-normalized CXCL16 concentration in urine of all participants. CXCL16 concentration in urine of UC patients compared to population and hospital controls (A) . CXCL16 in urine of primary UC patients compared to recurrent UC patients (B) . Urinary CXCL16 levels in hospital controls versus low-grade and high-grade UC patients (C) . Comparison of muscle invasive (>pT1) with non-muscle invasive (≤pT1) UC patients (D) . Non-parametric Wilcoxon rank sum test were performed to examine differences between the groups.

Article Snippet: The following primary antibodies were used: CXCL16 (1:250; Acris Antibodies) or CXCR6 (1:750; Abcam).

Techniques: Concentration Assay, Comparison

Journal: Oncotarget

Article Title: Soluble chemokine (C-X-C motif) ligand 16 (CXCL16) in urine as a novel biomarker candidate to identify high grade and muscle invasive urothelial carcinomas

doi: 10.18632/oncotarget.20737

Figure Lengend Snippet: Influence of study group (population controls, hospital controls, low-grade UC patients, high-grade UC patients) and group characteristics on log(CXCL16/Creatinine) determined via multiple linear regression analysis

Article Snippet: The following primary antibodies were used: CXCL16 (1:250; Acris Antibodies) or CXCR6 (1:750; Abcam).

Techniques:

Journal: Oncotarget

Article Title: Soluble chemokine (C-X-C motif) ligand 16 (CXCL16) in urine as a novel biomarker candidate to identify high grade and muscle invasive urothelial carcinomas

doi: 10.18632/oncotarget.20737

Figure Lengend Snippet: CXCR 6 and CXCL16 immunoreaction in acute urocystitis (A) , low grade (B) and high urothelial carcinoma (C) . CXCR 6 and CXCL16 are expressed on the surface as well as in the cytosol of epithelial cells. Moreover, in certain areas there is a strong co-localisation of the receptor-ligand pair observable. For detection of negative immunoreactivity (NC) within the same tissue section a consecutive section was stained. The presented slides reveal representative examples of the CXCL16 and CXCR6 immunoreactivity. Magnification, x40; bar 100 μm.

Article Snippet: The following primary antibodies were used: CXCL16 (1:250; Acris Antibodies) or CXCR6 (1:750; Abcam).

Techniques: Staining

Journal: Oncotarget

Article Title: Soluble chemokine (C-X-C motif) ligand 16 (CXCL16) in urine as a novel biomarker candidate to identify high grade and muscle invasive urothelial carcinomas

doi: 10.18632/oncotarget.20737

Figure Lengend Snippet: Semiquantitative analysis of CXCR6 and CXCL16 immunoreactivity in tissue specimen of patients with primary urothelial cancer (UC; n=13 low grade, n=4 high grade) or an inflammation of the bladder (Uro)

Article Snippet: The following primary antibodies were used: CXCL16 (1:250; Acris Antibodies) or CXCR6 (1:750; Abcam).

Techniques: Staining

Journal: Oncotarget

Article Title: Soluble chemokine (C-X-C motif) ligand 16 (CXCL16) in urine as a novel biomarker candidate to identify high grade and muscle invasive urothelial carcinomas

doi: 10.18632/oncotarget.20737

Figure Lengend Snippet: Staining of CXCL16 in patient tissue was categorized as absent, low, intermediate or strong according to method 1 (Table ). Normalized CXCL16 concentrations in urine supernatant were compared to the immunoreactivity signal found in the matching tissues.

Article Snippet: The following primary antibodies were used: CXCL16 (1:250; Acris Antibodies) or CXCR6 (1:750; Abcam).

Techniques: Staining