cd8  (Cedarlane)

Journal: Science Advances

Article Title: ATR inhibition activates cancer cell cGAS/STING-interferon signaling and promotes antitumor immunity in small-cell lung cancer

doi: 10.1126/sciadv.ado4618

Figure Lengend Snippet: ( A ) Tumor growth curves (means ± SE) of vehicle, anti–PD-L1 antibody alone (300 μg per body, once weekly), ceralasertib alone (50 mg/kg, 5 of 7 days, once a day.), and ceralasertib plus anti–PD-L1 antibody groups in B6129F1 mice injected with RPP cells ( n = 4 in vehicle and anti–PD-L1 antibody group, n = 6 in the other groups). ( B to E ) RPP tumors were harvested on day 22 for immune profiling by flow cytometry ( n = 4 in vehicle and anti–PD-L1 antibody group, n = 6 in ceralasertib, n = 5 in ceralasertib plus anti–PD-L1 antibody group). Flow cytometry analysis of (B) CD45 + CD3 + T cells, (C) CD45 + CD3 + CD8 + T cells, (D) CD45 + CD3 + CD8 + ICOS + T cells, (E) effector memory CD8 + T cells: CD45 + CD3 + CD8 + CD44 hi CD62L lo . Percentages in (B), (C), and (E) were calculated relative to CD45 + cells, and percentages in (D) were calculated relative to CD45 + CD8 + cells. ( F ) CD8 immunohistochemistry was performed on tumor sections resected on day 22. Scale bar, 50 μm. ( G ) Tumor growth curves (means ± SE) of vehicle, anti–PD-L1 antibody alone, ceralasertib alone, and ceralasertib plus anti–PD-L1 antibody [same dose as (A)] groups in B6FVBF1/J mice injected with RPM cells ( n = 6 per group). ( H to K ) RPM tumors were harvested at day 18 for immune profiling by flow cytometry ( n = 5 in anti–PD-L1 group and n = 6 in the other groups). Flow cytometry analysis of (H) CD45 + CD3 + T cells, (I) CD45 + CD3 + CD8 + T cells, (J) CD45 + CD3 + CD8 + ICOS + T cells, and (K) effector memory CD8 + T cells: CD45 + CD3 + CD8 + CD44 hi CD62L lo . Percentages were the ratio to CD45 + cells. ( L ) CD8 immunohistochemistry of tumors resected on day 18. Scale bar, 50 μm. Cerala, ceralasertib. P values were calculated by one-way analysis of variance (ANOVA). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and by Student’s t test for comparing ceralasertib alone and ceralasertib plus anti–PD-L1 antibody (A). ab, antibody. ns, not significant.

Article Snippet: Paraffin blocks were sectioned at 5-μm thickness, and immunohistochemistry was performed with anti-CD8a antibody [Cedarlane, HS-3610003(Sy)].

Techniques: Injection, Flow Cytometry, Immunohistochemistry

Journal: BMC Immunology

Article Title: DX5 + NKT cells display phenotypical and functional differences between spleen and liver as well as NK1.1 - Balb/c and NK1.1 + C57Bl/6 mice

doi: 10.1186/1471-2172-12-26

Figure Lengend Snippet: FACS-Analysis of spleen DX5 + NKT cells revealed interstrain differences in several phenotypical markers between NK1.1 - Balb/c and NK1.1 + C57Bl/6 mice . In the NK1.1 + mice strain spleen DX5 + NKT cells expressed less CD4 but more CD8 (A). Upon activation the TCR α/β down-regulation was confirmed. In addition to NK.1.1 more spleen DX5 + NKT cells in C57Bl/6 mice were positive for CD94 (B). Maturation marker CD62L was slightly higher expressed in freshly-isolated spleen DX5 + NKT cells in C57Bl/6 mice compared to Balb/c, whereas CD103 was less (C). Functional marker CD154 and CD178 were only little expressed in freshly-isolated spleen DX5 + NKT cells in both mice strains (D). Upon stimulation spleen DX5 + NKT cells in NK1.1 + C57Bl/6 mice displayed less up-regulation of CD154 compared to Balb/c. Results are given as mean + SEM. Experiments were repeated at least three times (* p < 0.05).

Article Snippet: The following reagents were used for cell surface labeling in multiparameter flow cytometric analysis (FACS Calibur, BD Bioscience): FITC or Alexa Fluoar 647-conjugated anti-mouse CD3 (clone: 17A2, rat IgG2b), APC-conjugated anti-mouse-CD4 (clone: GK1.5, rat IgG2b), APC-conjugated anti-mouse-CD8a (clone: 53-6.7, rat IgG2a), PE-conjugated anti-mouse-CD38 (clone: 90, rat IgG2a), PE-conjugated anti-mouse-CD49b (clone: DX5, rat IgM), APC-conjugated anti-mouse-CD62L (clone: MEL-14, rat IgG2a), PE-conjugated anti-mouse-CD103 (clone: M290, rat IgG2a), PE-conjugated anti-mouse-CD178 (Fas-Ligand) (clone: MFL3, hamster IgG1), PE-conjugated anti-mouse-Vβ 8.1, 8.2 TCR (clone: MR5-2, rat IgG2a) all BD Biosciences.

Techniques: Activation Assay, Marker, Isolation, Functional Assay

Journal: PLoS ONE

Article Title: Angiotensin II Is a New Component Involved in Splenic T Lymphocyte Responses during Plasmodium berghei ANKA Infection

doi: 10.1371/journal.pone.0062999

Figure Lengend Snippet: C57BL/6 mice were infected with P. berghei ANKA and treated with vehicle, losartan or captopril by gavage. T cells were isolated at day 6 post infection, stained with fluorescent antibodies, and analyzed by flow cytometry. (A) Representative dot plots of CD4 + CD69 + and CD8 + CD69 + T cells obtained from gated CD3 + cells. Percentage of CD4 + CD69 + T cells (B) and CD8 + CD69 + T cells (C). Parasitemia was determined by Giemsa-stained blood smears in vehicle- (solid line), losartan- (dotted line) or captopril- (dash line) treated mice (D). Mortality was checked daily in vehicle- (▪), losartan- (▴) or captopril- (▾) treated mice (E). Behavioral analysis was assessed as described in Materials and Methods (F). Evans blue dye extraction from the brain tissue to evaluate blood-brain barrier disruption (G). The results are expressed as means±SD. Statistically significant compared with values for *naive mice ( p <0.05) and #vehicle-treated mice infected with P. berghei ANKA ( p <0.05). Statistically significant compared with values for vehicle-treated infected mice at a day 5, b day 7 or day c 10 post-infection.

Article Snippet: Fluorescein isothiocyanate (FITC)-, phycoerythrin (PE)- and PE-Cy5.5-conjugated hamster IgG1 anti-murine CD3 (145-2C11), PE- and PerCP-conjugated rat IgG2a anti-murine CD4 (RM4-5), PE-conjugated rat IgG2a anti-murine CD8a (53–6.7), FITC-conjugated rat IgG2a anti-murine CD45R/B220 (RA3-6B2), PE-Cy5-conjugated rat IgG2b anti-murine CD44 (IM7), FITC-conjugated rat IgG2a anti-murine CD62L (MEL-14), PE-conjugated rat IgG1 anti-murine IFN-γ (XMG1.2), PE-conjugated rat IgG1 anti-murine IL-17 (TC11-18H10), PE-conjugated rat IgG2b anti-murine IL-10 (JES5-16E3), PE-conjugated rat IgG2b anti-murine IL-4 (BVD4-1D11), biotin-conjugated rat IgG2a anti-murine CD11a (MD17/4), PE-conjugated streptavidin, PerCP/PE/FITC-conjugated hamster IgG1 and IgG2, and goat IgG2a isotype controls were all purchased from BD Pharmingen (San Diego, CA, USA).

Techniques: Infection, Isolation, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Angiotensin II Is a New Component Involved in Splenic T Lymphocyte Responses during Plasmodium berghei ANKA Infection

doi: 10.1371/journal.pone.0062999

Figure Lengend Snippet: T cells from naive mice and mice infected with P. berghei ANKA treated with vehicle, losartan or captopril by gavage were isolated at day 6 post infection, stained with fluorescent antibodies and analyzed by flow cytometry. (A) Representative dot plots of perforin + CD8 + T cells obtained from gated CD3 + cells. The percentage (B) and absolute number ( C ) of perforin + CD8 + T cells were determined. The results are expressed as means±SD. Statistically significant compared with values for *naive mice ( p <0.05) and #vehicle-treated mice infected with P. berghei ANKA ( p <0.05).

Article Snippet: Fluorescein isothiocyanate (FITC)-, phycoerythrin (PE)- and PE-Cy5.5-conjugated hamster IgG1 anti-murine CD3 (145-2C11), PE- and PerCP-conjugated rat IgG2a anti-murine CD4 (RM4-5), PE-conjugated rat IgG2a anti-murine CD8a (53–6.7), FITC-conjugated rat IgG2a anti-murine CD45R/B220 (RA3-6B2), PE-Cy5-conjugated rat IgG2b anti-murine CD44 (IM7), FITC-conjugated rat IgG2a anti-murine CD62L (MEL-14), PE-conjugated rat IgG1 anti-murine IFN-γ (XMG1.2), PE-conjugated rat IgG1 anti-murine IL-17 (TC11-18H10), PE-conjugated rat IgG2b anti-murine IL-10 (JES5-16E3), PE-conjugated rat IgG2b anti-murine IL-4 (BVD4-1D11), biotin-conjugated rat IgG2a anti-murine CD11a (MD17/4), PE-conjugated streptavidin, PerCP/PE/FITC-conjugated hamster IgG1 and IgG2, and goat IgG2a isotype controls were all purchased from BD Pharmingen (San Diego, CA, USA).

Techniques: Infection, Isolation, Staining, Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Murine DX5 + NKT Cells Display Their Cytotoxic and Proapoptotic Potentials against Colitis-Inducing CD4 + CD62L high T Cells through Fas Ligand

doi: 10.1155/2018/8175810

Figure Lengend Snippet: Flow cytometry analysis of the spleen CD3 + DX5 + NKT, CD8 + T, CD4 + CD62L high , and CD4 + CD62L low cells of Balb/c mice after separation by MACS and FACS sorting (a). Proliferation (b) and proliferation index (c) of CFSE-labeled CD4 + CD62L high cells after 48 h and 96 h of monoculture or coculture with CD8 + T cells or CD3 + DX5 + NKT. Results are given as mean + SEM. Experiments were repeated at least three times ( ∗ P < 0.05).

Article Snippet: The following reagents were used for cell labeling in multiparameter flow cytometric analysis (FACS Calibur, BD Biosciences): PE or FITC-conjugated anti-mouse CD4 (clone: RM4-5, Rat IgG2b) Alexa 648 or FITC-conjugated anti-mouse CD3 (clone: 17A2, rat IgG2b), Alexa 648 or FITC-conjugated anti-mouse-CD8a (clone: 53–6.7, rat IgG2a), PE-conjugated anti-mouse-CD49b (clone: DX5, rat IgM), FITC- or PE-conjugated anti-mouse-CD62L (clone: MEL-14, rat IgG2a), and FITC- or PE-conjugated anti-mouse-CD178 (Fas ligand) (clone: MFL3, hamster IgG1); all are from BD Biosciences.

Techniques: Flow Cytometry, Labeling

Journal: Journal of Immunology Research

Article Title: Murine DX5 + NKT Cells Display Their Cytotoxic and Proapoptotic Potentials against Colitis-Inducing CD4 + CD62L high T Cells through Fas Ligand

doi: 10.1155/2018/8175810

Figure Lengend Snippet: Intracellular flow cytometry detection of IFN- γ in CD4 + CD62L high (a) and CD4 + CD62L low cells (b) after 4 h and 10 h of monoculture or coculture with CD8 + T cells or CD3 + DX5 + NKT. Results are given as mean + SEM. Experiments were repeated at least three times ( ∗ P < 0.05).

Article Snippet: The following reagents were used for cell labeling in multiparameter flow cytometric analysis (FACS Calibur, BD Biosciences): PE or FITC-conjugated anti-mouse CD4 (clone: RM4-5, Rat IgG2b) Alexa 648 or FITC-conjugated anti-mouse CD3 (clone: 17A2, rat IgG2b), Alexa 648 or FITC-conjugated anti-mouse-CD8a (clone: 53–6.7, rat IgG2a), PE-conjugated anti-mouse-CD49b (clone: DX5, rat IgM), FITC- or PE-conjugated anti-mouse-CD62L (clone: MEL-14, rat IgG2a), and FITC- or PE-conjugated anti-mouse-CD178 (Fas ligand) (clone: MFL3, hamster IgG1); all are from BD Biosciences.

Techniques: Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Murine DX5 + NKT Cells Display Their Cytotoxic and Proapoptotic Potentials against Colitis-Inducing CD4 + CD62L high T Cells through Fas Ligand

doi: 10.1155/2018/8175810

Figure Lengend Snippet: Intracellular flow cytometry analysis of caspase-3 in CD4 + CD62L high cells after 10 h and 48 h of monoculture or coculture with CD8 + T or CD3 + DX5 + NKT cells. Results are given as mean + SEM. Experiments were repeated at least three times ( ∗ P < 0.05).

Article Snippet: The following reagents were used for cell labeling in multiparameter flow cytometric analysis (FACS Calibur, BD Biosciences): PE or FITC-conjugated anti-mouse CD4 (clone: RM4-5, Rat IgG2b) Alexa 648 or FITC-conjugated anti-mouse CD3 (clone: 17A2, rat IgG2b), Alexa 648 or FITC-conjugated anti-mouse-CD8a (clone: 53–6.7, rat IgG2a), PE-conjugated anti-mouse-CD49b (clone: DX5, rat IgM), FITC- or PE-conjugated anti-mouse-CD62L (clone: MEL-14, rat IgG2a), and FITC- or PE-conjugated anti-mouse-CD178 (Fas ligand) (clone: MFL3, hamster IgG1); all are from BD Biosciences.

Techniques: Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Murine DX5 + NKT Cells Display Their Cytotoxic and Proapoptotic Potentials against Colitis-Inducing CD4 + CD62L high T Cells through Fas Ligand

doi: 10.1155/2018/8175810

Figure Lengend Snippet: Flow cytometry analysis FasL expression of CD8 T and CD3 + DX5 + NKT cells after 4 h and 10 h monoculture or coculture with CD4 + CD62L high cells. Results are given as mean + SEM. Experiments were repeated at least three times ( ∗ P < 0.05).

Article Snippet: The following reagents were used for cell labeling in multiparameter flow cytometric analysis (FACS Calibur, BD Biosciences): PE or FITC-conjugated anti-mouse CD4 (clone: RM4-5, Rat IgG2b) Alexa 648 or FITC-conjugated anti-mouse CD3 (clone: 17A2, rat IgG2b), Alexa 648 or FITC-conjugated anti-mouse-CD8a (clone: 53–6.7, rat IgG2a), PE-conjugated anti-mouse-CD49b (clone: DX5, rat IgM), FITC- or PE-conjugated anti-mouse-CD62L (clone: MEL-14, rat IgG2a), and FITC- or PE-conjugated anti-mouse-CD178 (Fas ligand) (clone: MFL3, hamster IgG1); all are from BD Biosciences.

Techniques: Flow Cytometry, Expressing