Journal: Gene therapy

Article Title: Targeting virus entry and membrane fusion through specific peptide/MHC complexes using a high-affinity T-cell receptor.

doi: 10.1038/sj.gt.3302286

Figure Lengend Snippet: Figure 2 TCR-mediated virus-cell entry is highly specific and requires high-affinity interaction between the TCR and its cognate MHC/peptide complex. (a) Infection of EL4 cells by MV-eGFP, MV-T7, MV-m33, and MV-CD38 in the absence or presence of 5 106 M pOV8 (SIINFEKL) or SIYR peptide. Only the MV displaying the high-affinity m33 TCR infected these H-2b cells presenting the SIYR peptide. None of the viruses infected cells presenting the null peptide pOV8. Scale bar ¼ 300 mm. (b) Percentage of GFP positive after infection of SIYR or pOV8-loaded (3 106 M) EL4 (H-2b) and P815 (H-2d) cells with MV-m33. Infection of SIYR/EL4 cells by MV-m33 was efficiently inhibited by addition of anti-Kb antibody (50 mg/ml). Bars represent duplicate experiment.

Article Snippet: The EL4 T-cell lymphoma (H-2b), EL4 expressing chicken ovalbumin (E.G7.OVA, ATCC CRL 2113), EL4 expressing SIYRYYGL peptide (ETS-1), and P815 mastocytoma (H-2d, ATCC TIB-64) cells were grown in 10% FBS-DMEM.

Techniques: Virus, Infection

Journal: Gene therapy

Article Title: Targeting virus entry and membrane fusion through specific peptide/MHC complexes using a high-affinity T-cell receptor.

doi: 10.1038/sj.gt.3302286

Figure Lengend Snippet: Figure 3 MV-m33 efficiently infected and caused extensive cell-to-cell fusion in EL4 cells stably expressing SIYR peptide (ETS-1), but not cells expressing chicken ovalbumin (EG7 OVA). (a) Cells were incubated with MV-eGFP, MV-T7 or MV-m33 at an MOI of 0.5 or 2.0 and photographed 48 h later. Only MV displaying the high-affinity scTCR (m33) was able to efficiently infect and cause extensive cell-to-cell fusion in ETS-1 cells. None of the viruses infected EL4 cells expressing a null peptide (EG7 OVA). Scale bar ¼ 300 mm. (b) Infection was also carried out in the presence of a fusion inhibitory peptide (FIP) that prevents cell-to-cell fusion. The percentage of single infected cells was analyzed by flow cytometry and expressed as the number of GFP-positive cells/ml of virus. (c) Infection of ETS-1 cells was correlated with the amount of MV-m33 virus added, and was inhibited more than 90% by anti-Kb antibody (50 mg/ ml). In contrast, the virus did not infect EG7 OVA cells expressing an irrelevant peptide/MHC complex (OVA/Kb).

Article Snippet: The EL4 T-cell lymphoma (H-2b), EL4 expressing chicken ovalbumin (E.G7.OVA, ATCC CRL 2113), EL4 expressing SIYRYYGL peptide (ETS-1), and P815 mastocytoma (H-2d, ATCC TIB-64) cells were grown in 10% FBS-DMEM.

Techniques: Infection, Stable Transfection, Expressing, Incubation, Cytometry, Virus

Journal: Gene therapy

Article Title: Targeting virus entry and membrane fusion through specific peptide/MHC complexes using a high-affinity T-cell receptor.

doi: 10.1038/sj.gt.3302286

Figure Lengend Snippet: Figure 4 Infection of EL4 cells by MV-m33 is dependent on the dose of SIYR peptide. (a) EL4 cells were loaded with increasing concentrations of SIYR peptide and infected with MV-m33 at MOIs of 2.0, 5.0, and 10.0. The percentage of GFP-positive cells was determined 72 h post infection by flow cytometry. (b) Photograph of infected EL4 cells loaded with 3 108–3 106 M SIYR peptide. (c) Number of peptide/MHC complexes on EL4 cells after loading with increasing concentrations of SIYR peptide. The number of peptide/MHC complexes present on ETS-1 cells is also shown (arrow).

Article Snippet: The EL4 T-cell lymphoma (H-2b), EL4 expressing chicken ovalbumin (E.G7.OVA, ATCC CRL 2113), EL4 expressing SIYRYYGL peptide (ETS-1), and P815 mastocytoma (H-2d, ATCC TIB-64) cells were grown in 10% FBS-DMEM.

Techniques: Infection, Cytometry