Journal: Nature Communications

Article Title: Cancer immune control needs senescence induction by interferon-dependent cell cycle regulator pathways in tumours

doi: 10.1038/s41467-020-14987-6

Figure Lengend Snippet: a Individual follow-up of tumour volumes. CD8-depleted mice were subcutaneously (s.c.) engrafted with 1 × 10 6 RT2-, RT2. Stat1 −/− -, RT2. Cdkn2a −/− - or RT2.CRISPR- Cdkn2a -cancer cells. Treatment with isotype control mAbs (Ctr) or combined immune checkpoint inhibitors (ICB; anti-PD-L1 and anti-LAG-3) once per week was started when tumours were >3 mm in diameter. Cancer size was measured 2 times per week. Number of mice with regressing tumours and the total number of mice is given in parenthesis; RT2 Ctr N = 9, ICB N = 10, RT2. Stat1 −/− Ctr N = 6, ICB N = 8, RT2. Cdkn2a −/− Ctr N = 12, ICB N = 14, RT2.CRISPR- Cdkn2a Ctr N = 5, ICB N = 10. Each cell line was given a different lining. Black lines summarise the results for different treatment groups (as obtained from ANCOVA). p -values examine the question whether the treatment effect was different between two genotypes. Mice were killed either when tumours reached the critical diameter of 15–20 mm or ulcerated, or when mice developed signs of wasting. b Representative triple-staining for the senescence marker p16 Ink4a (red) and the proliferation marker Ki67 (blue) and for nuclei (green) of the s.c. tumour of individual mice treated as described in ( a ). Scale bar 2 µm. c Individual data points showing quantification of p16 Ink4a+ (left) or Ki67 + (right) cells. Each data point represents the total of three tumour slides measurements, tumours of three individual mice (described in a ) were analysed. In a significance was tested by using unequal variances t -test, p -values examines the treatment effect, comparing the ICB-treated RT2-cancers with each ICB-treated knock-out group.

Article Snippet: Cdkn2a −/− - or RT2.CRISPR- Cdkn2a -cancer cells were s.c. transplanted, three days after depletion of CD8 cells with 100 μg anti-CD8 mAb (Rm-CD8-2 AK, Core Facility mAb, Helmholtz-Zentrum München).

Techniques: CRISPR, Control, Staining, Marker, Knock-Out

Journal: International journal of radiation oncology, biology, physics

Article Title: AN ANTI-TUMOR IMMUNE RESPONSE IS EVOKED BY PARTIAL-VOLUME SINGLE DOSE RADIATION IN TWO MURINE MODELS

doi: 10.1016/j.ijrobp.2018.10.009

Figure Lengend Snippet: Quantification (A) and example images (B) of the immunofluorescence staining of CD8+ cells from the in-field and out-of-field hemi-irradiated tumors at 24 hours after irradiation with 10Gy RT (blue: DAPI, green: CD8). An average of 3 separate experiments is shown, with 21 mice per treatment group. (C) Quantification of CD8+ cells from: unirradiated controls, 100% irradiated and from the in-field and out-of-field halves of the hemi-irradiated tumors. A representative experiment is shown, with 5 mice per group. Experiment was performed 3 times, with a total of 15 mice per group. (D) The effects of CD8+ T cells depletion on 67NT tumor response to RT. CD8+ T cells were depleted by four IP injections of anti-CD8 antibody, at a dose of 200μg/injection, starting 3 days before RT and 0, +3, +6 days after RT. Experiment was done once, with 5 mice per treatment group.

Article Snippet: Antibody and drug treatment Anti-CD8-alpha antibody (Mab/hybridoma 53–6.72) was obtained from the Antibody and Bioresource Core Facility at MSKCC.

Techniques: Immunofluorescence, Staining, Irradiation, Injection

Journal: International journal of radiation oncology, biology, physics

Article Title: AN ANTI-TUMOR IMMUNE RESPONSE IS EVOKED BY PARTIAL-VOLUME SINGLE DOSE RADIATION IN TWO MURINE MODELS

doi: 10.1016/j.ijrobp.2018.10.009

Figure Lengend Snippet: (A) Representative images of ICAM expressed on blood vessels in the tumor (red: Meca-32, green: ICAM). An increase in ICAM expression was observed 24 hours post 10Gy RT in the out-of-field half of the tumor. (B) Quantitation of ICAM staining in tumors. An average of 3 separate experiments is shown, with a total of 15 mice per group. (C) The effects of blocking ICAM on 67NR tumor response to RT. IP injections of ICAM-blocking antibody were administered at a dose of 300μg/injection 2, 16 and 48 hours after RT. Experiment was done once, with 5 mice per group. (D) Effect of blocking ICAM on CD8+ T cell infiltration following 10Gy hemi- or total irradiation and treatment with anti-ICAM antibody. IP injections of ICAM-blocking antibody were administered at a dose of 300μg/injection, 2 and 16 hours after RT. After 24 hours tumors were collected and CD8+ cells were visualized using immunofluorescence. Experiment was done once, with 5 mice per group.

Article Snippet: Antibody and drug treatment Anti-CD8-alpha antibody (Mab/hybridoma 53–6.72) was obtained from the Antibody and Bioresource Core Facility at MSKCC.

Techniques: Expressing, Quantitation Assay, Staining, Blocking Assay, Injection, Irradiation, Immunofluorescence

Journal: International journal of radiation oncology, biology, physics

Article Title: AN ANTI-TUMOR IMMUNE RESPONSE IS EVOKED BY PARTIAL-VOLUME SINGLE DOSE RADIATION IN TWO MURINE MODELS

doi: 10.1016/j.ijrobp.2018.10.009

Figure Lengend Snippet: (A) Effect of FTY720 treatment on tumor response. A representative experiment is shown, with 8 mice per group. Experiment was performed 3 times, with a total of 18 mice per group. (B) Effect of FTY720 treatment on CD8+ T cells levels in the blood following 10Gy hemi- or total irradiation. A representative experiment is shown, with 3 mice per group. Experiment was performed 2 times, with a total of 6 mice per group. Measurement of the level of CD8+ T cells in the blood confirmed FTY720 activity.

Article Snippet: Antibody and drug treatment Anti-CD8-alpha antibody (Mab/hybridoma 53–6.72) was obtained from the Antibody and Bioresource Core Facility at MSKCC.

Techniques: Irradiation, Activity Assay

Journal: International journal of radiation oncology, biology, physics

Article Title: AN ANTI-TUMOR IMMUNE RESPONSE IS EVOKED BY PARTIAL-VOLUME SINGLE DOSE RADIATION IN TWO MURINE MODELS

doi: 10.1016/j.ijrobp.2018.10.009

Figure Lengend Snippet: (A) Tumor growth delay curves for C57BL/6 mice bearing flank subcutaneous LLC lung tumors after receiving hemi- or total irradiation with 15 or 20Gy. A representative experiment is shown, with 5 mice per treatment group. Experiment was performed 3 times, with 15 mice per treatment group. (B) Same as (A) but showing the effects of CD8+ T cells depletion on LLC tumor response to RT. CD8+ T cells were depleted by four IP injections of anti-CD8 antibody, at a dose of 200 μg/injection, starting 3 days before RT and 0, +3, +6 days after RT. Experiment was done once, with 5 mice per group. (C) Same as (A) but showing the effects of blocking ICAM on LLC tumor response to RT. IP injections of ICAM-blocking antibody were administered at a dose of 300μg/injection 2, 16 and 48 hours after RT. Experiment was done once, with 5 mice per group.

Article Snippet: Antibody and drug treatment Anti-CD8-alpha antibody (Mab/hybridoma 53–6.72) was obtained from the Antibody and Bioresource Core Facility at MSKCC.

Techniques: Irradiation, Injection, Blocking Assay

Journal: PLoS ONE

Article Title: Anti-CD154 mAb and Rapamycin Induce T Regulatory Cell Mediated Tolerance in Rat-to-Mouse Islet Transplantation

doi: 10.1371/journal.pone.0010352

Figure Lengend Snippet: Cryostat sections were stained by anti-mouse CD4 ( A – D ), CD8 ( B – D ) and macrophage antibodies ( C – E ) in Group 1 ( A – B – C ) and Group 4 at 200 days post transplantation. In Group 1 of rejecting mice, immunohistology for cellular immune responses to concordant islet xenografts at time of rejection has detected mixed cellular infiltrates with presence of CD4+ ( A ), CD8+ ( B ), and macrophages ( C ). In Group 4 of tolerant mice, immunohistology for cellular immune responses to concordant islet xenografts of tolerant mice at 200 days post-transplantation detected only minimal cellular infiltrate with absence of CD4+ ( D ), CD8+ ( E ), and macrophages ( F ). (Magnification in A – C (200x), Magnification in D – F (100x)).

Article Snippet: Briefly serial frozen sections were stained for characterization of infiltrating cells with anti CD4 mAb and anti-CD8 mAb and anti mac-1 (Morphosys AbD Düsseldorf, Germany).

Techniques: Staining, Transplantation Assay