Journal: Journal of Hematology & Oncology

Article Title: Improvement of the anticancer efficacy of PD-1/PD-L1 blockade via combination therapy and PD-L1 regulation

doi: 10.1186/s13045-022-01242-2

Figure Lengend Snippet: Completed clinical trials of approved PD-1/PD-L1 inhibitors in combination with other approved treatment strategy

Article Snippet: , , Ipilimumab , CTLA-4 antibody , Malignant pleural mesothelioma , Phase 2 , NCT03048474.

Techniques: Clinical Proteomics, Capsules, Virus, Irradiation

Journal: Frontiers in Immunology

Article Title: Combining Immunotherapy with Oncogene-Targeted Therapy: A New Road for Melanoma Treatment

doi: 10.3389/fimmu.2015.00046

Figure Lengend Snippet: Ongoing clinical studies combining immunotherapy and oncogene-targeted therapy .

Article Snippet: Ipilimumab (Yervoy, Bristol-Myers Squibb) is a monoclonal antibody (IgG1) directed against CTLA-4 that was developed for systemic anti-tumor immunotherapy.

Techniques: Biomarker Discovery, Inhibition, Mutagenesis, Injection, Activity Assay, Immunopeptidomics, Capsules, Vaccines, Derivative Assay, Expressing, Amplification, Ex Vivo

Journal: iScience

Article Title: Development of chimeric antigen receptor (CAR)-T cells targeting A56 viral protein implanted by oncolytic virus

doi: 10.1016/j.isci.2024.109256

Figure Lengend Snippet: Essential domain of A56 for its localization on cell plasma membrane (A) Confocal immunofluorescence (IF) microscopy of OVV-infected A549 tumor cells showing A56 expression; nuclei (blue), OVV (green), A56 (red); scale bar, 10 μm. (B) Full A56 construct design compared with those of various A56 mutant constructs that do not have one or more domains; Sg, signaling domain; IgV, IgV-like domain; TDR, tandem repeat domain; S, stalk region; TM, transmembrane domain; CT, cytoplasmic tail. (C) Merged confocal images of A56 localization on the plasma membranes of U2-OS tumor cells upon full A56-expressing plasmid transduction compared to proteins enclosed within the Golgi or ER from transduction with mutant A56 vectors; nuclei (blue), A56 (green), Golgi apparatus or endoplasmic reticulum (red); scale bar, 10 μm.

Article Snippet: For A56 detection on the plasma membrane, the cells were labeled with 1μg of anti-A56 antibody (Mouse IgG1 w/o kappa light chain) (LakePharma, Variant 3) diluted in PBS supplemented with 2% FBS for 30 min at 4°C.

Techniques: Clinical Proteomics, Membrane, Immunofluorescence, Microscopy, Infection, Expressing, Construct, Mutagenesis, Plasmid Preparation, Transduction

Journal: iScience

Article Title: Development of chimeric antigen receptor (CAR)-T cells targeting A56 viral protein implanted by oncolytic virus

doi: 10.1016/j.isci.2024.109256

Figure Lengend Snippet: Localization of A56 on solid tumor plasma membrane and distribution in tumor tissue (A) OTS-412 were treated in A549, HCT-116, HeLa cancer cells at 0.05 and 0.1 MOI for 24 h followed by detection of A56 expression via flow cytometry. Dead cells were excluded by Zombie Aqua (ZA) staining. For intracellular detection of A56, fixation/permeabilization was performed before anti-A56 staining. (B) Analysis of A56 expression compared to A27L (a vaccinia virus cytosolic protein) expression on tumor tissue collected 4 days post-OV injection of HT-29 tumor-bearing mice. Scale bar, 100 px (=438 μm). (C) IF analysis of tumor-specific A56 expression and non-expression on other tissues from VX2 tumor-bearing New Zealand White rabbits collected on day (D) 5, 14, and 28 after intravenous injection of OTS-412/HU; The data shown are representative of six and two individual rabbits from the treatment and control groups, respectively. Scale bar, 200 μm. (D) Quantitation of A56 expression detected from each tissue of Figure 2 C presented in mean intensity of fluorescence. Data are represented as mean ± SEM. (E) Distribution of OTS-412 in the OTS-412/HU combination group presented in mean viral copies per 25 mg of each tissue and 1 mL of PBMC analyzed until D28 post virus injection. Data are represented as mean ± SEM. BM; bone marrow, AG; adrenal gland.

Article Snippet: For A56 detection on the plasma membrane, the cells were labeled with 1μg of anti-A56 antibody (Mouse IgG1 w/o kappa light chain) (LakePharma, Variant 3) diluted in PBS supplemented with 2% FBS for 30 min at 4°C.

Techniques: Clinical Proteomics, Membrane, Expressing, Flow Cytometry, Staining, Virus, Injection, Control, Quantitation Assay, Fluorescence

Journal: iScience

Article Title: Development of chimeric antigen receptor (CAR)-T cells targeting A56 viral protein implanted by oncolytic virus

doi: 10.1016/j.isci.2024.109256

Figure Lengend Snippet: In vitro and in vivo functionality of A56-specific A56 CAR-T cells (A) Real-time cytotoxicity of A56 CAR-T cells against HCT-116 cells infected with 0.05 MOI of OTS-412. (B) Quantification of A56 CAR-T cell cytotoxicity in various human cancer cell lines infected with OTS-412. (∗∗∗ p < 0.001, comparison of T cell groups; n = 2 for A549, HCT-116, HeLa, SK-MEL-5 and n = 3 for MCF7 and PC-3 by two-tailed unpaired t test). Data are represented as the mean value ± SEM. (C) Aggregation of A56 CAR-T cells targeting tumor cells observed every 12 h until day 4 in real-time. (D) Antigen-dependent anti-tumor efficacy of A56 CAR-T cells in HCT-116 tumor-bearing mice after intratumoral T cell injection following OTS-412 injection with or without HU (∗ p < 0.05, ∗∗ p < 0.01, comparison of T cell groups; saline, n = 5; treatment groups, n = 4, two-way ANOVA with multiple comparison). Data are represented as the mean value ± SD. (E) IHC analysis of A56 expression and infiltrated hCD3 + T cells in tumors isolated from HCT-116 tumor-bearing mice 21 days post-intravenous T cell treatment. Scale bar, 20 μm (left panel) and quantitation of tumor-infiltrating A56 CAR-T cells positive for hCD3 (right panel) (∗ p = 0.01, n = 3, two-tailed unpaired t test). Data are presented as the mean ± SD. MOI, multiplicity of infection; IHC, immunohistochemistry.

Article Snippet: For A56 detection on the plasma membrane, the cells were labeled with 1μg of anti-A56 antibody (Mouse IgG1 w/o kappa light chain) (LakePharma, Variant 3) diluted in PBS supplemented with 2% FBS for 30 min at 4°C.

Techniques: In Vitro, In Vivo, Infection, Comparison, Two Tailed Test, Injection, Saline, Expressing, Isolation, Quantitation Assay, Immunohistochemistry

Journal: iScience

Article Title: Development of chimeric antigen receptor (CAR)-T cells targeting A56 viral protein implanted by oncolytic virus

doi: 10.1016/j.isci.2024.109256

Figure Lengend Snippet:

Article Snippet: For A56 detection on the plasma membrane, the cells were labeled with 1μg of anti-A56 antibody (Mouse IgG1 w/o kappa light chain) (LakePharma, Variant 3) diluted in PBS supplemented with 2% FBS for 30 min at 4°C.

Techniques: Staining, Virus, Recombinant, Isolation, Saline, Plasmid Preparation, Cell Isolation, Expressing, Software

Journal: Nature Communications

Article Title: Virus-specific memory T cell responses unmasked by immune checkpoint blockade cause hepatitis

doi: 10.1038/s41467-021-21572-y

Figure Lengend Snippet: a Individualised treatment of melanoma is guided by tumour staging, presence of B-RAF mutations and fitness-for-toxicity. b Colitis, hepatitis and thyroiditis are common immune-related complications of dual therapy with Nivolumab plus Ipilimumab; 31.4% of patients experienced two or more of these immune-related adverse reactions ( n = 89). c Colitis, hepatitis and thyroiditis occurred independently and were not significantly associated with clinical response ( n = 89; F.E). d – h Patients who developed hepatitis of any grade following dual therapy lacked biochemical signs of liver inflammation before treatment. In particular, no clinically meaningful differences in plasma levels of d aspartate transaminase (AST; n = 87; M.W.; Bonferroni-corrected p -value, m = 5), e alanine transaminase (ALT; n = 89; M.W.; Bonferroni-corrected p -value, m = 5), f gamma glutamyl transaminase (γ-GT; n = 89; M.W.; Bonferroni-corrected p -value, m = 5), g total bilirubin ( n = 87; M.W.; Bonferroni-corrected p -value, m = 5), or h C-reactive protein (CRP; n = 85; M.W.; Bonferroni-corrected p -value, m = 5) were observed between patients who developed hepatitis and those who did not. Median values are indicated by a red line. i , j Biochemical markers of tumour burden were not different between patients who developed hepatitis and those who did not. i Pre-treatment levels of lactate dehydrogenase ( n = 89; M.W.; Bonferroni-corrected p -value, m = 4). Median values are indicated by a red line. j Pre-treatment levels of protein S100 ( n = 89; M.W.; Bonferroni-corrected p -value, m = 4). k – m No association was observed between seropositivity for k hepatitis B virus core antigen (HBcAg; n = 85; F.E.; Bonferroni-corrected p -value, m = 3), l hepatitis C virus (HCV; n = 85; F.E.; Bonferroni-corrected p -value, m = 3), or m hepatitis E virus (HEV; n = 67; F.E.; Bonferroni-corrected p -value, m = 3) and development of hepatitis following dual therapy. n No association was observed between rounds of αPD-1/αCTLA-4 administered and development of hepatitis ( n = 89; M.W.).

Article Snippet: Stage IV patients with unresectable metastatic disease who received first- or second-line checkpoint inhibitor therapy were initially treated with Nivolumab (αPD-1; Bristol-Myers Squibb) and Ipilimumab (αCTLA-4; Bristol-Myers Squibb) for four cycles, and thereafter with Nivolumab maintenance therapy (3 mg/kg at 3-week intervals).

Techniques:

Journal: Nature Communications

Article Title: Virus-specific memory T cell responses unmasked by immune checkpoint blockade cause hepatitis

doi: 10.1038/s41467-021-21572-y

Figure Lengend Snippet: a Peripheral blood samples were collected from melanoma patients with metastatic disease receiving αPD-1/αCTLA-4 therapy immediately before administration of the first dose ( n = 89). Leucocyte subsets differentially represented in patients with or without hepatitis were identified in a randomly assigned training set (B.H.-corrected t -tests; n = 44; m = 50; FDR = 0.25). Red dots indicate significantly differently represented subsets. Example gating strategies for analysis of flow cytometry data are provided as Supplementary Figs. – . b CD4 + T EM % in training set patients with or without treatment-related hepatitis ( n = 44; M.W.). Median values are indicated by a red line. c CD4 + T EM % in validation set patients with or without treatment-related hepatitis ( n = 45; M.W.). d ROC analysis of CD4 + T EM % as a discriminatory marker for treatment-related hepatitis in the validation set ( n = 45). e Comparison of the bimodal distribution of CD4 + T EM % in patients with unresectable metastatic disease ( n = 107) and the normal distribution ( n = 49; K2 = 2.79; p = 0.248) of CD4 + T EM % in patients with completely resected tumours. A cut-off of CD4 + T EM ≥ 21% was set (indicated by a dashed red line) below which 99% of completely resected tumour cases should fall. Four pink points represent CD4 + T EM ≥21% patients with metastatic disease who were electively treated with αPD-1 monotherapy. f In the validation set, 68.9% patients were correctly classified using a cut-off of CD4 + T EM ≥ 21 %, whereas 55.6% were correctly classified under the no-information model ( n = 45; F.E.). g CD4 + T EM ≥ 21 % is not a marker of predisposition to αPD-1/αCTLA-4-related colitis ( n = 89; F.E.). h CD4 + T EM ≥21% patients did not experience more severe hepatitis than CD4 + T EM <21% patients ( n = 38; F.E.). Dashed red line indicates a cut-off of CD4 + T EM ≥ 21%. i Time-to-first presentation of hepatitis was not different between CD4 + T EM ≥21% ( n = 12) and T EM <21% ( n = 26) patients (log-rank). j Twelve of 12 patients with unresectable metastatic melanoma and CD4 + T EM ≥ 21% developed hepatitis after αPD-1/αCTLA-4 dual therapy. By contrast, 3 of 4 CD4 + T EM ≥21% patients treated with αPD-1 monotherapy did not develop hepatitis (F.E.; p = 0.007).

Article Snippet: Stage IV patients with unresectable metastatic disease who received first- or second-line checkpoint inhibitor therapy were initially treated with Nivolumab (αPD-1; Bristol-Myers Squibb) and Ipilimumab (αCTLA-4; Bristol-Myers Squibb) for four cycles, and thereafter with Nivolumab maintenance therapy (3 mg/kg at 3-week intervals).

Techniques: Flow Cytometry, Marker

Journal: Nature Communications

Article Title: Virus-specific memory T cell responses unmasked by immune checkpoint blockade cause hepatitis

doi: 10.1038/s41467-021-21572-y

Figure Lengend Snippet: a Seasonal presentation of CD4 + T EM ≥21% patients between 2017 and 2020 ( n = 103; F.E.). Dashed red line indicates a cut-off of CD4 + T EM ≥ 21%. b CD4 + T EM ≥21% status was associated with high serum levels of anti-CMV IgG antibodies ( n = 100; F.E.; p = 1.2 ⨯ 10 −5 ). Dashed red line indicates a cut-off of CD4 + T EM ≥ 21%. c CD4 + T EM ≥21% status was associated with CMV-reactivity in pp65 ELISPOT ( n = 53; F.E.; p = 9.0 ⨯ 10 −5 ). Dashed red line indicates a cut-off of CD4 + T EM ≥ 21%. d Development of hepatitis was associated with CMV-seropositivity and CD4 + T EM ≥21% status ( n = 89). Median values are indicated by a red line. e ROC analysis showing CD4 + T EM % is a superior discriminator of patients at risk of hepatitis when considering only CMV IgG + cases as opposed to all cases. f Classification of patients with unresectable metastatic melanoma who did or did not develop αPD-1/αCTLA-4-related hepatitis according to CMV IgG status and baseline CD4 + T EM cell frequency using a revised cut-off of CD4 + T EM ≥ 16%. Red boxes indicate 34 of 40 (85%) cases correctly classified by our model ( n = 40; F.E.; p = 1.4 ⨯ 10 −5 ). Green box indicates 2 of 17 (11.7%) cases predicted to develop hepatitis who did not. Pink box indicates 4 of 23 (17.4%) cases predicted not to develop hepatitis who did. Blue box indicates 19 of 49 CMV IgG − patients not considered by our model who developed hepatitis.

Article Snippet: Stage IV patients with unresectable metastatic disease who received first- or second-line checkpoint inhibitor therapy were initially treated with Nivolumab (αPD-1; Bristol-Myers Squibb) and Ipilimumab (αCTLA-4; Bristol-Myers Squibb) for four cycles, and thereafter with Nivolumab maintenance therapy (3 mg/kg at 3-week intervals).

Techniques: Enzyme-linked Immunospot

Journal: Nature Communications

Article Title: Virus-specific memory T cell responses unmasked by immune checkpoint blockade cause hepatitis

doi: 10.1038/s41467-021-21572-y

Figure Lengend Snippet: A 54-year-old male patient who received αPD-1 (Nivolumab) plus αCTLA-4 (Ipilimumab) dual therapy for metastatic melanoma presented with late-onset hepatitis. a Course of treatment. b Change in hepatitis-related parameters over time and their association with introduction, withdrawal and re-introduction of valganciclovir treatment. c The patient presented at the end of week 40 with recrudescent hepatitis and was treated with 900 mg/day valganciclovir for 2 days prior to liver biopsy. Histopathological image of the liver biopsy (H&E staining; scale bar 500 µm). d Generally, the liver parenchyma appeared normal (H&E staining; scale bar 100 µm). e Only a few lymphocytes and sparse necrotic hepatocytes were observed in portal areas with small bile plugs (arrows) with no signs of hepatitis (H&E staining; scale bar 100 µm).

Article Snippet: Stage IV patients with unresectable metastatic disease who received first- or second-line checkpoint inhibitor therapy were initially treated with Nivolumab (αPD-1; Bristol-Myers Squibb) and Ipilimumab (αCTLA-4; Bristol-Myers Squibb) for four cycles, and thereafter with Nivolumab maintenance therapy (3 mg/kg at 3-week intervals).

Techniques: Staining

Journal: Nature Communications

Article Title: Virus-specific memory T cell responses unmasked by immune checkpoint blockade cause hepatitis

doi: 10.1038/s41467-021-21572-y

Figure Lengend Snippet: A 49-year-old male patient who received αPD-1 (Nivolumab) plus αCTLA-4 (Ipilimumab) dual therapy for metastatic melanoma presented with late-onset hepatitis. a Course of treatment. b Change in hepatitis-related parameters over time and their association with introduction of valganciclovir treatment. c Histopathological image of a liver biopsy taken at the start of week 72 showing signs consistent with drug toxicity, autoimmunity or viral infection (H&E staining; scale bar 500 µm). d Extensive centrilobular necrosis (arrows) involving 30–40% of hepatocytes was observed (H&E staining; scale bar 100 µm). e Dense inflammatory infiltration of lymphocytes, eosinophils and neutrophils was seen in portal areas (H&E staining; scale bar 100 µm).

Article Snippet: Stage IV patients with unresectable metastatic disease who received first- or second-line checkpoint inhibitor therapy were initially treated with Nivolumab (αPD-1; Bristol-Myers Squibb) and Ipilimumab (αCTLA-4; Bristol-Myers Squibb) for four cycles, and thereafter with Nivolumab maintenance therapy (3 mg/kg at 3-week intervals).

Techniques: Infection, Staining