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: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet: CyTOF antibody panel

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques:

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet: Selected genes involved in Tfh cell biology

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques: Activation Assay

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet: Distinct CD4 + T cell subsets contribute to the generation of Tfh with heterogeneous functional profiles (A) Mean fluorescence intensity of the CXCR5 marker expressed by CXCR5 + PD-1 + cells derived from (1) naive, (2) MemPD-1 neg , (3) MemPD-1 neg and Tfh. (B) Frequency of IL-21- and/or IFNγ-positive cells among CXCR5 + PD-1 + cells at day 3. (C – E) Representative flow plots showing CXCR3, ICOS, and CD40L expression by CXCR5 + PD-1 + cells (left panel) and frequency of CXCR3-, ICOS-, and CD40L-positive cells among CXCR5 + PD-1 + cells at day 3 (right panel). (F) Ex vivo cells or their respective Tfh D3 counterparts obtained after 3 days of splenocyte culture were co-cultured with autologous B cells for 7 days. (G) Box plots represent the frequency of CD27 + CD38 + cells among CD19 + cells, the concentration of total immunoglobulins and the absolute number of live B cells after co-culture. (H) Quantification of IgG1, IgG4, and IgA in the co-culture supernatants. Each symbol (A–H) represents an individual donor. (A–H) A Wilcoxon matched pairs test was performed, ∗, p < 0.05; ∗∗, p < 0.005; ∗∗∗, p < 0.001.

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques: Functional Assay, Fluorescence, Marker, Derivative Assay, Expressing, Ex Vivo, Cell Culture, Concentration Assay, Co-Culture Assay

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet:

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques: Cell Analysis, Purification, Virus, Recombinant, Blocking Assay, Antibody Labeling, Transfection, Software

Journal:

Article Title: Normal human pregnancy is associated with an elevation in the immune suppressive CD25 + CD4 + regulatory T-cell subset

doi: 10.1111/j.1365-2567.2004.01869.x

Figure Lengend Snippet: Three-colour flow cytometry analysis of CD4+ CD25+ cells during normal pregnancy. (a) Dot plot for fluorescence intensity of CD4 versus side light scatter showing gating for identification of CD4+ lymphocytes. (b) Dot plot for forward versus side light scatter of gated CD4+ lymphocytes demonstrating small size of these cells during pregnancy (black) and large size following mitogen stimulation (grey). (c) Dot plot for CD3 PE versus CD25 FITC of gated CD4+ lymphocytes demonstrating that all of these cells are T cells (CD3+) and that 22% during pregnancy (black) have weak CD25 expression (upper right quadrant) whilst all (grey) have strong CD25 expression after activation by mitogen. The quadrants are set according to isotypematched, negative control antibodies. (d–g, i) Dot plots for CD25 PE versus CD69 FITC, DR FITC, CD45RA FITC, CD45RO FITC, CD11a FITC, respectively, for gated CD4+ lymphocytes. (h) Dot plot for CD25 FITC versus CXCR3 PE for gated CD4+ lymphocytes.

Article Snippet: The whole blood fluorescence-activated cell sorter (FACS) lysis method was used with phycoerythrin- (PE), fluorescein isothiocyanate- (FITC) and peridinin chlorophyll protein (PerCP) fluorochrome-conjugated mouse monoclonal antibodies to CD3, CD4, CD8, CD11a, CD25, CD45RO, CD45RA, CD69, HLA-DR (Becton Dickinson, Oxford UK) and CXCR3 (R & D Systems, Oxford, UK).

Techniques: Flow Cytometry, Fluorescence, Expressing, Activation Assay, Negative Control

Journal: Advanced Science

Article Title: Interior Decorative VOCs Elevate T Cell‐Mediated Obstructive Lung Disease Risks via Osteogenesis‐Driven Lymphoid‐Biased Hematopoiesis

doi: 10.1002/advs.202512663

Figure Lengend Snippet: VOCs exposure enhances T cell maturation and their recruitment to lung tissues. Mice were exposed to CON or VOCs for 8‐week using the whole‐body inhalation exposure model. A) Representative images of H&E‐stained thymic sections. Black dashed lines indicate the cortical thickness of thymus. Scale bar: 100 µm. Immunohistochemical analysis for B) CD3 and C) CD8 in thymic sections, and the percentage of positive area. Yellow arrows indicate the positive expression. Scale bar: 40 µm. D) Representative flow cytometric dot plots and quantitative analysis of E) CD4 + T and F) CD8 + T cells in the PB. G) Immunostaining for the colocalization of CXCR3 (red) and CD8 (purple) in lung sections, and the corresponding quantitative analysis. Yellow arrows indicate CXCR3 colocalization with CD8. Scale bar: 20 µm. H) Immunostaining for the colocalization of CCR5 (green) and CD8 (purple) in lung sections, and the corresponding quantitative analysis. Yellow arrows indicate CCR5 colocalization with CD8. Scale bar: 20 µm. I) Quantitative analysis of CXCL9, CXCL10, and CXCL11 in the PB by ELISA assay. Protein expressions of CXCL10 in J) BALF and K) LI. L) A schematic representation summarizing the T cell maturation and recruitment to lung tissue. Statistical analysis was performed using two‐tailed unpaired t ‐test (A–C and E–K). * p < 0.05 and ** p < 0.01. Data are expressed as mean ± SEM ( n ≥ 3).

Article Snippet: Similarly, mouse lung sections were incubated with primary antibodies against CXCR3 (Bosterbio, China), CCR5 (Servicebio, China), and CD8 (Servicebio, China) at 4 °C overnight, followed by the application of secondary antibodies.

Techniques: Staining, Immunohistochemical staining, Expressing, Immunostaining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: iScience

Article Title: Defects in NK cell immunity of pediatric cancer patients revealed by deep immune profiling

doi: 10.1016/j.isci.2024.110837

Figure Lengend Snippet:

Article Snippet: Anti-CXCR3 (clone G025H7, conjugated to 163Dy) , Standard BioTools , Cat#3163004B.

Techniques: Purification, Clinical Proteomics, Recombinant, Blocking Assay, Staining, Saline, Mass Cytometry, Software, Cytometry