Journal: Journal for immunotherapy of cancer

Article Title: Systemic and intravesical adoptive cell therapy of tumor-reactive T cells can decrease bladder tumor growth in vivo.

doi: 10.1136/jitc-2020-001673

Figure Lengend Snippet: Figure 5 CXCR3 mediates T cell infiltration into orthotopic ovalbumin expressing MB49 tumor cell line (MB49OVA) bladder tumor from intravesical space. (A) RNA was isolated from MB49 and MB49OVA tumor cells and analyzed for expression of chemokines CXCL9, CXCL10 and CXCL11 as determined by PCR. (B) OT-I T cells were isolated from spleen of transgenic mice and tumor-infiltrating lymphocytes (TILs) were isolated from MB49 orthotopic tumors. Expression of CXCR3 was determined on these cell populations by flow cytometry. (C) TIL samples expanded from patient bladder tumors were analyzed for CXCR3 expression by flow cytometry. In vivo, mice were treated with OT-I T cells coated with a CXCR3 blocking antibody or rat IgG (rIgG). (D) Tumors were collected 3 and 24 hours after infusion and analyzed for CD45.1+ T cell infiltration by flow cytometry. (E) Tumor volume was measured by ultrasound. Precoating OT-I T cells with αCXCR3 decreased T cell infiltration into the tumor and abrogated the decrease in tumor growth seen with OT-I T cells coated with a control rIgG antibody. N=5 per group. Repeated 2 times. ns, not significant.

Article Snippet: OT- I T cells were isolated as described and incubated for 30 min on ice with either normal rat IgG isotype control or CXCR3 blocking antibody (500 μg, catalog# BE0249, BioXcell).

Techniques: Expressing, Isolation, Transgenic Assay, Flow Cytometry, In Vivo, Blocking Assay, Control

Journal: Journal of acquired immune deficiency syndromes (1999)

Article Title: Simian immunodeficiency Virus Infection Potently Modulates Chemokine Networks and Immune Environments in Hilar Lymph Nodes of Cynomolgus Macaques

doi: 10.1097/QAI.0b013e31828ac85f

Figure Lengend Snippet: Induction of chemokine and TLR3 mRNAs following stimulation of HiLN cells with IFN-γ, LPS or poly I:C. Using real-time RT-PCR, the relative mRNA levels of the indicated genes were detected in HiLN cells following exposure to IFN-γ, LPS or Poly I:C. In (A-C) are shown the induction of CXCR3 ligand mRNAs with IFN-γ at 6 and 24 hr post-stimulation. In (D-F) are shown the induction of CXCR3 and CCR5 ligand mRNAs with LPS at 6 and 24 hr post-stimulation. In (G-I) are shown the induction of CXCR3 and CCR5 ligand and TLR3 mRNAs with poly I:C at 6 and 24 hr post-stimulation. These data are combined analyses of HiLN cells from three different uninfected animals (only two for TLR3). Open symbols represent the untreated control cells and grey-filled symbols represented the paired treated cells. The mean values are shown superimposed upon the individual data points. Paired t-tests were used to compare the control from treated cultures and all p-values < 0.05 are provided.

Article Snippet: Antibodies used for IHC were specific for CD209/DC-SIGN (BD Pharmingen, DCN46) or CD183/CXCR3 (BD, 1C6).

Techniques: Quantitative RT-PCR

Journal: Journal of acquired immune deficiency syndromes (1999)

Article Title: Simian immunodeficiency Virus Infection Potently Modulates Chemokine Networks and Immune Environments in Hilar Lymph Nodes of Cynomolgus Macaques

doi: 10.1097/QAI.0b013e31828ac85f

Figure Lengend Snippet: Measurement of Host Response mRNA Expression Levels in HiLNs by Real-time RT-PCR

Article Snippet: Antibodies used for IHC were specific for CD209/DC-SIGN (BD Pharmingen, DCN46) or CD183/CXCR3 (BD, 1C6).

Techniques: Expressing

Journal: Journal of acquired immune deficiency syndromes (1999)

Article Title: Simian immunodeficiency Virus Infection Potently Modulates Chemokine Networks and Immune Environments in Hilar Lymph Nodes of Cynomolgus Macaques

doi: 10.1097/QAI.0b013e31828ac85f

Figure Lengend Snippet: Flow cytometric and IHC measurement of cell types in HiLNs from SIV-infected and uninfected macaques. A: Flow cytometry was performed on available samples of viably cryopreserved HiLN cells using fluorophore-conjugated antibodies to CD3, CD20, CD4, CD8, CCR5 and/or CXCR3. Shown are the percentages of CD3+/CD20− CD4+ or CD8+ lymphocytes that were also either CCR5+ or CXCR3+ from individual animals, with the group medians and interquartile ranges. Groups were compared using the Mann-Whitney nonparametric test. B: Immunohistochemistry (IHC) was performed to detect CXCR3+ cells in HiLN tissue sections, with antigen-expressing cells staining brown. Original magnification, X100 and X200 (C). C: IHC was performed to detect DC-SIGN+ cells in HiLN tissue sections (original magnifications x200). In addition, ISH for DC-SIGN mRNA was performed simultaneously on HiLN sections stained for either CD68 or MHC-II (original magnificaitons×400). The average density of DC-SIGN+ cells per 0.3mm2 was determined for individual HiLNs from the indicated disease states, with the median and interquartile ranges denoted (to the right). Group comparisons were made using the Mann-Whitney nonparametric test.

Article Snippet: Antibodies used for IHC were specific for CD209/DC-SIGN (BD Pharmingen, DCN46) or CD183/CXCR3 (BD, 1C6).

Techniques: Infection, Flow Cytometry, MANN-WHITNEY, Immunohistochemistry, Expressing, Staining

Journal: Journal of leukocyte biology

Article Title: Hyperforin down-regulates effector function of activated T lymphocytes and shows efficacy against Th1-triggered CNS inflammatory-demyelinating disease.

doi: 10.1189/jlb.0707469

Figure Lengend Snippet: Fig. 2. Hyp-DCHA down-regulates CXCR3 expression on activated T cells. Surface expression of CXCR3 on the membrane of 72 h PHA and IL-2- activated T cells as detected by FACS is shown. CXCR3 expression was evaluated on cells treated 24 h with 2.5 M Hyp-DCHA (upper panel); the gray peak shows the fluorescence of cells incubated with FITC-conjugated isotype control mAb (anti-IgG1). Unimodal distribution with a statistically significant shift (down-regulation) between a Hyp-DCHA-treated and untreated sample is observed. The down-regulation of CCR3 after 24 h of treatment is not significant (lower panel). Representative results of four independent experi- ments are shown.

Article Snippet: The cells were treated with commercially available FITC- or PE-conjugated mAb: anti-CD3, -CD4, and -CD8, isotype-matched controls (BD PharMingen, San Diego, CA, USA), anti-IL-4 and -IFN- (BD PharMingen), FITC-conjugated anti-human CXCR3 (clone 49801.111), and anti-human CCR3 (R&D Systems Inc., Minneapolis, MN, USA).

Techniques: Expressing, Membrane, Incubation, Control

Journal: Clinical Cancer Research

Article Title: Role of CXCR3 Ligands in IL-7/IL-7Rα-Fc–Mediated Antitumor Activity in Lung Cancer

doi: 10.1158/1078-0432.ccr-10-3346

Figure Lengend Snippet: Figure 4. CXCR3 pathway is required for IL-7/IL-7Ra-Fc–mediated antitumor activity. For in vivo neutralizations, mice bearing 5-day established subcutaneous tumors were treated with IL-7/IL7Ra-Fc on days 5 and 10. Twenty four hours prior to IL-7/IL7Ra-Fc treatment, and then 3 times per week, mice were injected i.p. individually with the respective specific or appropriate control antibodies for the duration of the experiment. A and B, neutralization of CXCL9 (A) or CXCL10 (B) or IFNg (B) reversed the antitumor benefit of IL-7/IL-7Ra-Fc [bottom, photograph of tumors from diluents, IL-7/IL-7Ra-Fc, and IL-7/IL-7Ra-Fc þ anti-CXCL9, or anti-CXCL10 or anti-IFNg (IL-7/IL-7Ra-Fc þ control Ab was the same as IL-7/IL-7Ra-Fc treated group, data not shown)]. C, CXCL9, CXCL10, and IFNg were reduced in tumors (day 15) following treatment of the tumor bearing mice with IL-7/IL-7Ra-Fc and the respective neutralizing antibodies: anti-CXCL9, or anti-CXCL10 or anti-IFNg antibody in comparison to control Abs. D, neutralization of CXCL9, CXCL10, or IFNg reduced the frequency of CXCR3 activated CD8 T cells in the tumor (day 15). Data; mean SEM; *, P < 0.01 for IL-7/IL-7Ra-Fc þ control Ab compared with IL-7/IL- 7Ra-Fc þ anti-cytokine Ab or Diluent groups (n ¼ 6/group).

Article Snippet: Anti-mouse mAbs to CXCR3 (220803), IL-7Ra-Fc chimeric molecule (747-MR; endotoxin level: less than 1.0 EU/mg of protein by LAL method), ELISA antibody pairs for murine IFNg , CXCL9, CXCL10, and IL12 were purchased from R&D Systems, sensitivity of 3 to 5 pg/mL.

Techniques: Activity Assay, In Vivo, Injection, Control, Neutralization, Comparison