Journal: Cancer gene therapy

Article Title: Redirected cellular cytotoxicity by infection of effector cells with a recombinant vaccinia virus encoding a tumor-specific monoclonal antibody.

doi: 10.1038/sj.cgt.7700161

Figure Lengend Snippet: Figure 4. mAb CO17-1A-mediated ADCC of P815-GA733-2 cells by GM- CSF-activated macrophages infected with VVs encoding for the secreted (Œ, VVTG1142) or membrane-an- chored (E, VVTG2179) CO17-1A mAb. The results, which are ex- pressed as a percentage of specific 51Cr release, are the means of three separate experiments, with data points measured in triplicate. The SD was never .5% of the reported val- ues (data not shown). As a positive control, CO17-1A mAb (, 10 mg/mL) preincubated macrophages were used. VVTG186 (f) infected macro- phages and irrelevant isotype- matched mAb preincubated macro- phages (Cont Ab, 3) were used as negative controls.

Article Snippet: Murine P815 cells (DBA2 mouse mastocytoma (H2Dd)) transfected with the human GA733-2 cDNA were established as described previously.22,23 Murine RMA cells (C57BL/6 mouse T-cell lymphoma; American Type Culture Collection, Manassas, Va; (H2Db)) were used to stimulate murine CTLs.

Techniques: Infection, Membrane, Positive Control

Journal: Cancer gene therapy

Article Title: Redirected cellular cytotoxicity by infection of effector cells with a recombinant vaccinia virus encoding a tumor-specific monoclonal antibody.

doi: 10.1038/sj.cgt.7700161

Figure Lengend Snippet: Figure 5. mAb CO17-1A-mediated ADCC by Ag-activated murine CTLs infected with VVs coding for secreted (Œ, VVTG1142) or membrane-anchored (E, VVTG2179) CO17-1A mAb. A: P815-GA733-2 target cells; B: RMA target cells; C: P815 target cells. The results, which are expressed as a percentage of specific 51Cr release, are the means of two separate experiments done in triplicate. The SD was never .5% of the reported values. Alloreactive murine CTLs infected with VVTG18663 were used as negative controls.

Article Snippet: Murine P815 cells (DBA2 mouse mastocytoma (H2Dd)) transfected with the human GA733-2 cDNA were established as described previously.22,23 Murine RMA cells (C57BL/6 mouse T-cell lymphoma; American Type Culture Collection, Manassas, Va; (H2Db)) were used to stimulate murine CTLs.

Techniques: Infection, Membrane

Journal:

Article Title: Vaccination with Trypomastigote Surface Antigen 1-Encoding Plasmid DNA Confers Protection against Lethal Trypanosoma cruzi Infection

doi:

Figure Lengend Snippet: Induction of long-lasting TSA-1515–522-specific, CD8+-T-cell-dependent, MHC class I-restricted CTL in TSA-1 DNA-immunized B6 mice. Four weeks following the first 100-μg intramuscular dose of plasmid DNA, B6 (H-2b) mice were injected via the same route with an identical dose of the priming vaccine, consisting of either control VR1012 plasmid or the TSA-1-expressing constructs VR1012 TSA1.7 and VR1012 TSA2.1. (A) Immune SCs were obtained 2 weeks after the second immunization and stimulated in vitro with peptide TSA-1515–522 (1 μM). After 6 days, recall CTL activity of undepleted (Undepl.) responder cultures was assessed in a 5-h 51Cr release assay against RMA-S (H-2b) and P815 (H-2d) target cells sensitized with TSA-1515–522 peptide (0.05 μM) at the indicated E/T ratios. RMA-S cells pulsed with OVA257–264 peptide (0.05 μM) were used as negative-control target cells. CTL activity of effector cells depleted (Depl.) of CD4+ or CD8+ T cells was measured at a 50:1 E/T ratio against TSA-1515–522-sensitized (0.05 μM) target cells. (B) Immune SCs from DNA-vaccinated or T. cruzi-infected mice were obtained 7 and 6 months after the second immunization or parasite challenge, respectively, and processed as described for panel A.

Article Snippet: P815 cells ( H-2 d ; mastocytoma cells; ATCC TIB 64), J774 cells ( H-2 d ; macrophages; ATCC TIB 67), 3T3 cells ( H-2 d ; fibroblasts; ATCC CCL 163), and Vero cells (African Green monkey kidney cells; ATCC CCL 81) (all from the American Type Culture Collection, Rockville, Md.); RMA-S cells (peptide TAP.2 transporter-deficient, low H-2 b expressor mutant of the RBL-5 Rauscher virus-induced T-cell lymphoma; provided by H.-G. Ljundggren, Karolinska Institute, Stockholm, Sweden); and 5A.Kb.α3 cells ( H-2 k fibroblasts stably transfected with the K b gene; provided by S. Jameson, University of Minnesotta, Minneapolis) were maintained in complete RPMI 1640 (Mediatech, Herndon, Va.) medium (CR) containing 10% heat-inactivated fetal bovine serum (HyClone, Logan, Utah), 20 mM HEPES, 2 mM l -glutamine, 1 mM sodium pyruvate, 0.1 mM nonessential amino acids and 50 μg of gentamicin per ml (all from Gibco BRL, Gaithersburg, Md.).

Techniques: Plasmid Preparation, Injection, Expressing, Construct, In Vitro, Activity Assay, Release Assay, Negative Control, Infection