invivo mab rat igg2a isotype control antibody clone 2a3  (Bio X Cell)

Journal: Cell Reports Medicine

Article Title: Fc-optimized anti-CTLA-4 antibodies increase tumor-associated high endothelial venules and sensitize refractory tumors to PD-1 blockade

doi: 10.1016/j.xcrm.2025.102141

Figure Lengend Snippet: Fc-enhanced mouse anti-CTLA-4 antibodies prune TA-HEVs in the absence of concurrent PD-1 blockade (A) Treatment schedule. i.p., intraperitoneal; control, isotype control antibodies. (B and C) Mean and individual tumor growth. Data were obtained from two independent experiments ( n = 10 mice per group). (D–G) Frequency of FOXP3 + Tregs in tumor, numbers of tumor-infiltrating FOXP3 + Tregs and FOXP3 − Tconv, and ratio of Tconv to Treg cell numbers in tumors following indicated treatments. Each symbol represents an individual mouse ( n = 10, two independent experiments). (H–M) Frequencies of CD45 − CD31 high endothelial cells, TA-HECs, MECA-79 + CD62P + TA-HECs, numbers of TA-ECs and TA-HECs, and mean fluorescence intensity (MFI) of MECA-79 and CD62P in TA-HECs, following indicated treatments, quantified by flow cytometry. Each symbol represents an individual mouse ( n = 10, two independent experiments). (N) Histograms showing expression of PD-L1 in CD45 − CD31 high tumor endothelial cells following indicated treatments, quantified by flow cytometry. MFI of PD-L1 is quantified. Each symbol represents an individual mouse ( n = 4–5). (O) Treatment schedule. i.p., intraperitoneal; control, isotype control antibodies. (P and Q) Mean and individual tumor growth. Data were obtained from two independent experiments (control, n = 8 mice; 9D9-IgG2a + anti-PD-1, n = 10 mice). (R–T) Numbers of tumor-infiltrating FOXP3 + Tregs and FOXP3 − Tconv, and ratio of Tconv to Tregs cell numbers in tumors following indicated treatments. Each symbol represents an individual mouse ( n = 8–10, two independent experiments). (U) Frequency of TA-HECs. Representative dot plots of TA-HECs are shown. Each symbol represents an individual mouse ( n = 8–10, two independent experiments). Data are shown as mean ± SEM. All p values were determined by unpaired two-tailed Student’s t test except for graph in (N) for which p values were determined by one-way ANOVA with Tukey’s multiple comparison test, and for (B) and (P) for which p values were determined by two-way ANOVA. See also .

Article Snippet: InVivo MAb Rat IgG2a isotype control (clone 2A3) , Bio X Cell , Cat# BE0089; RRID: AB_1107769.

Techniques: Control, Fluorescence, Flow Cytometry, Expressing, Two Tailed Test, Comparison

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NECTIN4 regulates the cell surface expression of CD155 in non-small cell lung cancer cells and induces tumor resistance to PD-1 inhibitors

doi: 10.1007/s00262-025-04079-z

Figure Lengend Snippet: Evaluation of NECTIN4 and CD155 interactions and colocalization. A , Co-immunoprecipitation (Co-IP) experiments were performed using HEK293T cells transfected with CD155 and Flag-NECTIN4 expression vectors. Cell lysates were immunoprecipitated using Flag antibody or CD155 antibody, and immunoblotting was performed. Mouse IgG was used as a control for immunoprecipitation. B , A proximity ligation assay (PLA) targeting CD155 and NECTIN4 was performed in empty vector control and NECTIN4 overexpressing H322 and EBC1 cell lines. The close colocalization of CD155 and NECTIN4 was detected by red fluorescent signals. Nuclei were stained with DAPI. Scale bars, 20 μm. The number of PLA signals indicating colocalization of CD155 and NECTIN4 was determined from z-projection images generated by z-stacking of optical sections for individual cells. C , Schematic diagram of domain-deleted Flag-NECTIN4. D , HEK293T cells were co-transfected with the CD155 expression vector and full-length Flag-NECTIN4 or domain-deleted Flag-NECTIN4 expression vectors, followed by co-immunoprecipitation. As references, input samples were analyzed by immunoblotting, and samples immunoprecipitated with CD155 antibody were detected with Flag antibody by immunoblotting. Statistical analysis was conducted using Student’s t test. * p < 0.05, **p < 0.01. OE, overexpression

Article Snippet: One week after transplantation, tumor-bearing mice were treated intraperitoneally with monoclonal antibodies (mAbs) as follows: 200 μg of anti-PD-1 mAb (RMP1-14, BioXcell, #BE0146), 150 μg of anti-TIGIT mAb (1G9, BioXcell, #BE0274), 200 μg of control IgG2a mAb (2A3, BioXcell, #BE0089), and 150 μg of control IgG1 mAb (MOPC-21, BioXcell, #BE0083).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, Expressing, Western Blot, Control, Proximity Ligation Assay, Plasmid Preparation, Staining, Generated, Over Expression

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NECTIN4 regulates the cell surface expression of CD155 in non-small cell lung cancer cells and induces tumor resistance to PD-1 inhibitors

doi: 10.1007/s00262-025-04079-z

Figure Lengend Snippet: Overexpression of NECTIN4 confers resistance to PD-1 inhibitors. A , Schematic representation of the experiment: the subcutaneous transplantation of 4T1 EV control or NECTIN4-overexpressing cells, followed by treatment with anti-PD-1 antibody. The treatment was initiated on day 7 after tumor transplantation and administered every three days for a total of three times. Anti-PD-1 mAb, or IgG2a isotype were administered at a dose of 200 µg. B , Graph showing tumor volumes after transplantation. Tumor volumes were calculated using the formula: length × width × width/2. C , Individual tumor volume progression in each group. D , Analysis of tumor-infiltrating lymphocytes (TILs) by flow cytometry. Experiments were conducted with five mice per group and the data are presented as the mean ± SD. All in vivo experiments were performed in duplicates, with similar results. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test (B) or Student’s t-test (D). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. EV, empty vector; OE, overexpression

Article Snippet: One week after transplantation, tumor-bearing mice were treated intraperitoneally with monoclonal antibodies (mAbs) as follows: 200 μg of anti-PD-1 mAb (RMP1-14, BioXcell, #BE0146), 150 μg of anti-TIGIT mAb (1G9, BioXcell, #BE0274), 200 μg of control IgG2a mAb (2A3, BioXcell, #BE0089), and 150 μg of control IgG1 mAb (MOPC-21, BioXcell, #BE0083).

Techniques: Over Expression, Transplantation Assay, Control, Flow Cytometry, In Vivo, Plasmid Preparation

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NECTIN4 regulates the cell surface expression of CD155 in non-small cell lung cancer cells and induces tumor resistance to PD-1 inhibitors

doi: 10.1007/s00262-025-04079-z

Figure Lengend Snippet: Overcoming anti-PD-1 antibody resistance in NECTIN4-overexpressing 4T1 cells by combination therapy with anti-TIGIT antibody. A , Schematic representation of the experiment: the subcutaneous transplantation of 4T1 EV control or NECTIN4-overexpressing cells, followed by treatment with anti-PD-1 antibody alone or in combination with anti-TIGIT antibody. The treatment was initiated on day 7 after tumor transplantation and administered every three days for a total of three times. Anti-PD-1 mAb, or IgG2a isotype were administered at a dose of 200 µg, and anti-TIGIT mAb or IgG1 isotype were administered at a dose of 150 µg. B , Graph showing tumor volume progression after transplantation. C , Comparison of tumor weights in each group, measured post-excision. D , Photographs of tumors excised two days after the completion of treatment. E , Analysis of tumor-infiltrating lymphocytes (TILs) by flow cytometry. Experiments were conducted with five mice per group and the data are presented as the mean ± SD. All in vivo experiments were performed in duplicates, with similar results. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test (B, C) or Student’s t-test (E). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. EV, empty vector; OE, overexpression

Article Snippet: One week after transplantation, tumor-bearing mice were treated intraperitoneally with monoclonal antibodies (mAbs) as follows: 200 μg of anti-PD-1 mAb (RMP1-14, BioXcell, #BE0146), 150 μg of anti-TIGIT mAb (1G9, BioXcell, #BE0274), 200 μg of control IgG2a mAb (2A3, BioXcell, #BE0089), and 150 μg of control IgG1 mAb (MOPC-21, BioXcell, #BE0083).

Techniques: Transplantation Assay, Control, Comparison, Flow Cytometry, In Vivo, Plasmid Preparation, Over Expression

Journal: The Journal of Clinical Investigation

Article Title: Cxcr3 promotes protection from colorectal cancer liver metastasis by driving NK cell infiltration and plasticity

doi: 10.1172/JCI184036

Figure Lengend Snippet: ( A ) Experimental scheme of macrophage-NK cell coculture in vitro. ( B ) Lef,: representative contour plot of CD49a and CD49b expression by NK cells upon 72 hours coculture with macrophages purified from HL and LM with or without TGF-β-RI inhibitor (SB431542, 2.5 nM). Right, fold change (F.C.) of frequency ± SEM of CD49a + cells among NK cells cultured with macrophages from HL, MFL, or LM relative to NK cells cultured alone (IL-15 * P = 0.03, IL-15+TGF-β-RI inhibitor * P = 0.044, 2-tailed Student’s t test in HL Mac versus LM Mac in IL-15 and LM Mac in IL-15 versus IL-15+ TGF-β-RI inhibitor). 3 experiments were performed in duplicate. ( C ) Left, percent of migration of splenic NK cells in response to supernatants collected from MC38- or SL4-derived LM macrophages and no chemokine control (NC) (n > 3, * P = 0.01, **** P < 0.0001, 1-way ANOVA). Right, percent of migration in response to MC38-derived MFL, LM macrophage supernatants and NC ( n > 3, **** P < 0.0001, 1-way ANOVA). ( D ) Fold change of CD49a (left) and frequency of CD49a + CD69 + cells (right) in NK cells cultured alone or with LM-derived macrophages from MC38 and SL4 LM ( n = 5, * P < 0.02, **** P < 0.0001, 1-way ANOVA). ( E and F ) Top, experimental workflow of in vivo treatment with anti-CSF1R and with MC-21. ( E ) Bottom, representative contour plots show ILC1, CD49a + and CD49a – NK cells in LM of control mAb (Rat IgG2a)-treated and anti-CSF1R-treated tumor-bearing mice. Numbers in plots indicate frequency among NK1.1 + NKp46 + cells. Histogram plots show mean frequency ± SEM ( n = 9 total mice in 2 independent experiments, CD49a + NK ** P = 0.003, CD49a – NK * P = 0.02, 2-tailed Student’s t test). ( F ) Bottom, representative contour plots show ILC1, CD49a + , and CD49a – NK cells in LM of control mAb (Rat IgG2a)-treated and MC-21–treated tumor-bearing mice. Numbers in plots indicate frequency among NK1.1 + NKp46 + cells. Histogram plots show mean frequency ± SEM ( n = 4, 2 independent experiments were performed).

Article Snippet: To deplete F4/80 hi MAM tumor-bearing mice were treated i.p. with anti-CSF1R (CD115, clone AFS98, BioXcell) or control Rat IgG2a (clone 2A3, BioXcell) mAbs as following: 60 mg/kg at day +7 intrasplenic injection, 30 mg/kg at days +10 and +13, and 20 mg/kg at day +16.

Techniques: In Vitro, Expressing, Purification, Cell Culture, Migration, Derivative Assay, Control, In Vivo

Journal: Cell reports

Article Title: Clostridioides difficile toxin B subverts germinal center and antibody recall responses by stimulating a drug-treatable CXCR4-dependent mechanism

doi: 10.1016/j.celrep.2024.114245

Figure Lengend Snippet:

Article Snippet: InVivoPlus TM isotype control mAb, clone 2A3 , BioXCell , Cat# BP0089 RRID: AB_1107769.

Techniques: Control, Virus, Recombinant, Expressing, Suspension, Protease Inhibitor, SYBR Green Assay, Cell Isolation, Binding Assay, Bradford Protein Assay, Cell Counting, Enzyme-linked Immunosorbent Assay, Gene Expression, Software, Simple Western, Protein Extraction