All animal experiments were conducted under protocol LC-070 approved by the Animal Care and Use Committee of the National Cancer Institute. For metastasis therapy studies, tumor cells were either orthotopically implanted into the mammary fat pad (4T1, EMT6, R3T, HRM1, 6DT1, D2A1, E0771, M6, MVT1), or delivered by tail-vein injection (TSAE1, F3II, MET1) into strain-matched mice. Unless otherwise indicated, primary tumors were resected when they reached 0.5–0.8 cm diameter. Mice were randomized to treatment groups of 10–15 mice/group, and treated with the neutralizing anti-TGF-β mouse monoclonal antibody 1D11 (Genzyme Corp, or BioXCell) or isotype control (13C4, Genzyme Corp. or MOPC21, BioXCell) at 5mg/Kg intraperitoneally 3x per week unless otherwise indicated. At resection or experimental endpoint, tumors were harvested for histology and molecular analyses. Metastatic burden in the lung was determined by counting histologically evident metastases in lung cross-sections. More details are available in Supplementary Methods .
Mopc 21
Manufactured by BioXCell
Sourced in United States
MOPC-21 is a mouse monoclonal antibody that recognizes an unspecified antigen. It is commonly used as an isotype control in immunological studies.
Automatically generated - may contain errors
Lab products found in correlation
Mar1 5a3, by BioXCell (11 mentions)
Miap410, by BioXCell (3 mentions)
Diphtheria toxin, by Merck Group (3 mentions)
C57bl 6 mice, by Charles River Laboratories (2 mentions)
Anti tgfβ antibody clone 1d11.16.8, by BioXCell (2 mentions)
Cd11b positive enrichment beads, by Miltenyi Biotec (2 mentions)
Bead based immunoassay for mouse cytokines and chemokines, by Eve Technologies (2 mentions)
Accuri c6, by BD (2 mentions)
Artificial csf, by Bio-Techne (2 mentions)
40kd mwco zeba spin desalting columns, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (2 mentions)
Polyclonal armenian hamster igg, by BioXCell (2 mentions)
Il 17a, by Thermo Fisher Scientific (2 mentions)
Anti tcr γδ uc7 13d5, by BioXCell (2 mentions)
Anti il 17a 17f3, by BioXCell (2 mentions)
Artificial csf, by Harvard Apparatus (2 mentions)
αcd4 gk1, by BioXCell (2 mentions)
Albumin dextrose catalase enrichment, by BD (1 mentions)
Middlebrook 7h9 broth, by Merck Group (1 mentions)
Tween 80, by Fujifilm (1 mentions)
71 protocols using mopc 21
1
Metastasis Inhibition by Anti-TGF-β Therapy
2
Antibody Therapy for Atopic Dermatitis
3
Mycobacterium and Klebsiella Infection Models
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Lyophilized M. bovis BCG (Tokyo strain) was purchased from Kyowa Pharmaceuticals and dissolved in 7H9 broth (Difco) supplemented with albumin–dextrose–catalase enrichment (BD Biosciences). Single colonies were grown with vigorous shaking at 37°C in Middlebrook 7H9 broth supplemented with 10% albumin–dextrose–catalase, 1% glycerol (Sigma-Aldrich), and 0.5% Tween 80 (Wako) until the optical density at 600 nm (OD600) reached 1. Bacteria were stored at −80°C in 50% glycerol as single-use aliquots. Mice were intratracheally infected with 1 × 106 CFUs of M. bovis BCG (Tokyo strain).
K. pneumoniae ATCC strain 43,816, serotype 2 (ATCC) was grown in Difco Nutrient Broth (Difco) for 18 h at 37°C with vigorous shaking. Bacteria were pelleted by centrifugation and stored at −80°C in 50% glycerol as single-use aliquots. 3-wk-old WT mice were intraperitoneally injected with 200 μg of 1C10-1F7 or mouse IgG1 isotype control mAbs (MOPC-21; Bioxcell) on −3 d, −1 h, and 3 d. These mice were intranasally inoculated with K. pneumoniae at 1 × 103 CFUs on day 0.
K. pneumoniae ATCC strain 43,816, serotype 2 (ATCC) was grown in Difco Nutrient Broth (Difco) for 18 h at 37°C with vigorous shaking. Bacteria were pelleted by centrifugation and stored at −80°C in 50% glycerol as single-use aliquots. 3-wk-old WT mice were intraperitoneally injected with 200 μg of 1C10-1F7 or mouse IgG1 isotype control mAbs (MOPC-21; Bioxcell) on −3 d, −1 h, and 3 d. These mice were intranasally inoculated with K. pneumoniae at 1 × 103 CFUs on day 0.
4
Regulatory T Cell Suppression Assay
5
Identification and Characterization of Neutralizing Antibodies
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The unmutated ancestor sequences (WEN-1UA, WEN-3UA) of the rCl13-neutralizing monoclonal antibodies WEN-1 and WEN-3 (Figure S1 ; [Eschli et al., 2007 (link), Seiler et al., 1998 (link)]) were identified by IgBlast (IgBlast) sequence analysis. All mutations deviating from the closest germline V(D)J heavy (HC) and light chain (LC) sequences were reverted. The corresponding cDNAs were synthesized (Genscript) and introduced into HC and LC expression cassettes for recombinant expression in a mouse IgG1 format (provided by Dr. Shozo Izui, University of Geneva). The antibodies were produced by transient co-transfection of the HC and LC expression plasmids in CHO cells (Protein Expression Core Facility, PECF, of the Swiss Federal Technical Highschool, EPFL, Lausanne, Switzerland). The antibodies were purified on an ÄKTAprime plus purification system using Protein G columns (GE healthcare). After 24 hours of PBS dialysis, the purified antibodies were quantified by IgG ELISA. MOPC-21 (BioXcell) served as isotype control antibody. The KL25-W104L, KL25-F106L and KL25-Y108S antibodies as well as a matching KL25 wild-type control antibody (Bruns et al., 1983 (link)) were obtained by analogous procedures, except that a mouse IgG2a expression format was used for these experiments.
6
Intranasal Asp/Ova Challenge in Mice
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Mice were anesthetized with isoflurane (Piramal, Bethlehem, PA, USA) and were intranasally administered with a mixture of 7 μg of proteinase extracted from Aspergillus melleus (Sigma, St Louis, MO, USA) and 20 μg of Ovalbumin (Ova; Grade V, Sigma, St Louis, MO, USA) (Asp/Ova) in 50 μl of PBS (Katy, TX, USA) every two days for a total of five times (day 0, 2, 4, 6, 8). Sixteen hours after the last challenge, all mice were euthanized and the bronchial lymph nodes, superficial cervical lymph nodes and sera were obtained for further analysis. For TGF-β neutralization experiments, mice were injected intraperitoneally with 200 μg of an anti-TGF-β neutralizing antibody (1D11, BioXCell, West Lebanon, NH, USA) or their corresponding IgG1 control (MOPC-21, BioXCell, West Lebanon, NH, USA) three times every two days (day 0, 2, 4). For STAT3 inhibition experiments, mice were treated with intraperitoneal injections of 0.5 mg/kg STA-21 (Santa Cruz Bio-technology, Santa Cruz, CA, USA) or vehicle every two days for 9 days (day 0, 2, 4, 6, 8) and were treated with intranasal injections of 0.25 mg/kg STA-21 or vehicle every other day for 9 days (day 1, 3, 5, 7).
7
Evaluating IL-2 Variants in PD-1 Therapy
8
Anti-CD22 Treatment in Aged Mice
9
Investigating Anti-PD-1 and Anti-gp49 Effects on Tumor Metastasis
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Regarding tumor metastases, LLC (1.5 × 106 cells/mouse) and B16F10 (5 × 105 cells/mouse) cells were inoculated into B6 and gp49B−/− mice by intravenous injection. After inoculation, LLC- and B16F10-bearing mice were sacrificed on day 30 and day 21, respectively, and lung and liver tissues from tumor-bearing mice were harvested for analysis of tumor metastasis by hematoxylin–eosin (H&E) staining or counting of surface nodules. For antibody treatment, the B6 mice were inoculated intravenously at a low dose of 5 × 105 LLC-Luc2 cells or 5 × 104 B16F10 cells. At 3 days post-tumor inoculation, mice were treated with anti-PD-1 (RPM1-14, Bio X Cell; 200 μg/mouse), anti-gp49 (H1.1-Amenian hamster IgG from hybridoma cells26 (link) or H1.1-mIgG1, 200 μg/mouse), or isotype matched control antibody (2A3 (Bio X cell), HTK888 (Biolegend) or MOPC-21 (Bio X Cell); 200 μg/mouse) by intraperitoneal injection six times at 3-day intervals. LLC-Luc2 tumor-bearing mice with antibody treatment were imaged using the IVIS Spectrum In Vivo Imaging System (Perkin Elmer) at three weeks after tumor inoculation, and sacrificed at five weeks after tumor injection for detection of tumor metastases in lung and liver by H&E staining.
10
Modulating Immune Pathways in Infection
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To block the PD-1/PD-L1 pathway, infected WT and IL-17RA KO mice were injected i.p. with 200 μg rat anti-mouse PD-L1 antibody (10F.9G2, BioXcell) or total rat IgG isotype control (Jackson Research) every 3 days from 15 dpi until sacrifice at day 21 dpi. Infection matched WT mice were assayed in parallel as controls.
For the in vivo neutralization of IL-17A, infected WT mice were injected i.p. with 200 μg rat anti-mouse IL-17 antibody (17F3, BioXcell) or rat total IgG1 isotype control (MOPC-21, BioXcell) every 3 days from day 13 dpi until sacrifice at 21 dpi. Infection-matched IL-17RA KO mice were assayed in parallel as controls.
For the in vivo neutralization of IL-17A, infected WT mice were injected i.p. with 200 μg rat anti-mouse IL-17 antibody (17F3, BioXcell) or rat total IgG1 isotype control (MOPC-21, BioXcell) every 3 days from day 13 dpi until sacrifice at 21 dpi. Infection-matched IL-17RA KO mice were assayed in parallel as controls.
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