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Retargeting Oncolytic Vesicular Stomatitis Virus to Human T-Cell Lymphotropic Virus Type 1-Associated Adult T-Cell Leukemia.

Betancourt D, Ramos JC, Barber GN - J. Virol. (2015)

Bottom Line: VSV-gp160G was further noted to be highly attenuated and did not replicate efficiently in or induce significant cell death of primary CD4(+) T cells.Importantly, VSV-gp160G effectively exerted potent oncolytic activity in patient-derived ATL transplanted into NSG mice and facilitated a significant survival benefit.This effect greatly reduced neurotoxic risk associated with VSV infection while still allowing VSV to effectively target ATL cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Miami School of Medicine, Miami, Florida, USA.

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Related in: MedlinePlus

VSV-gp160G is specific for transformed CD4+ ATL cells. (A) Bright–field microscopy of VSV-XN2- and VSV-gp160G-infected ATL (MT-2, MT-4, and TLO-m1) and BJAB cells at an MOI of 0.01 at 24 hpi. (B) Immunoblot analysis for viral glycoprotein expression in ATL and BJAB cells infected with VSV-XN2 or VSV-gp160G at MOIs of 0.01 or 0.1 at 24 hpi. (C) Growth kinetic assay results for ATL or BJAB cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.001. Supernatant was analyzed by a standard plaque assay using HeLa CD4+ cells. (Student t test, two tailed, equal variance; *, P < 0.01; **, P < 0.001).
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Figure 3: VSV-gp160G is specific for transformed CD4+ ATL cells. (A) Bright–field microscopy of VSV-XN2- and VSV-gp160G-infected ATL (MT-2, MT-4, and TLO-m1) and BJAB cells at an MOI of 0.01 at 24 hpi. (B) Immunoblot analysis for viral glycoprotein expression in ATL and BJAB cells infected with VSV-XN2 or VSV-gp160G at MOIs of 0.01 or 0.1 at 24 hpi. (C) Growth kinetic assay results for ATL or BJAB cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.001. Supernatant was analyzed by a standard plaque assay using HeLa CD4+ cells. (Student t test, two tailed, equal variance; *, P < 0.01; **, P < 0.001).

Mentions: After confirming VSV-gp160G specificity for HeLa CD4+ cells, we determined the virus's ability to selectively infect and replicate within ATL lines. The CD4+ ATL lines MT-2, MT-4, and TLO-m1 and the CD4− Burkitt's B cell lymphoma cell line BJAB were all infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.01 and analyzed for their respective CPEs at 24 hpi. Again, as expected, VSV-XN2 induced cell rounding against all cells lines, whereas VSV-gp160G induced syncytium formation only against the CD4+ ATL lines (Fig. 3A). MT-2, MT-4, TLO-m1, and BJAB cells were infected with either VSV-XN2 or VSV-gp160G at the indicated MOIs and then harvested at 24 hpi for immunoblotting to check for viral glycoprotein expression. HIV-1 glycoproteins gp120 and gp41 were only detected in ATL cell lines infected with VSV-gp160G, and BJAB cells did not express HIV-1 glycoproteins. Using α-HIVgp41 antibody, full-length gp160G and the gp41G fragments were detected in all CD4+ cells infected with VSV-gp160G (Fig. 3B). However, the gp41G subunit was not readily detected in MT-4 cells for reasons that remain unclear but may involve aberrant glycosylation events that could occur in these cells and that may affect antibody recognition. The detection of the heavy and light bands using α-VSV-G antibody in VSV-gp160G-infected cells correspond to recognition of the cytoplasmic tail of the VSV-G protein that was fused to gp160.


Retargeting Oncolytic Vesicular Stomatitis Virus to Human T-Cell Lymphotropic Virus Type 1-Associated Adult T-Cell Leukemia.

Betancourt D, Ramos JC, Barber GN - J. Virol. (2015)

VSV-gp160G is specific for transformed CD4+ ATL cells. (A) Bright–field microscopy of VSV-XN2- and VSV-gp160G-infected ATL (MT-2, MT-4, and TLO-m1) and BJAB cells at an MOI of 0.01 at 24 hpi. (B) Immunoblot analysis for viral glycoprotein expression in ATL and BJAB cells infected with VSV-XN2 or VSV-gp160G at MOIs of 0.01 or 0.1 at 24 hpi. (C) Growth kinetic assay results for ATL or BJAB cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.001. Supernatant was analyzed by a standard plaque assay using HeLa CD4+ cells. (Student t test, two tailed, equal variance; *, P < 0.01; **, P < 0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 3: VSV-gp160G is specific for transformed CD4+ ATL cells. (A) Bright–field microscopy of VSV-XN2- and VSV-gp160G-infected ATL (MT-2, MT-4, and TLO-m1) and BJAB cells at an MOI of 0.01 at 24 hpi. (B) Immunoblot analysis for viral glycoprotein expression in ATL and BJAB cells infected with VSV-XN2 or VSV-gp160G at MOIs of 0.01 or 0.1 at 24 hpi. (C) Growth kinetic assay results for ATL or BJAB cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.001. Supernatant was analyzed by a standard plaque assay using HeLa CD4+ cells. (Student t test, two tailed, equal variance; *, P < 0.01; **, P < 0.001).
Mentions: After confirming VSV-gp160G specificity for HeLa CD4+ cells, we determined the virus's ability to selectively infect and replicate within ATL lines. The CD4+ ATL lines MT-2, MT-4, and TLO-m1 and the CD4− Burkitt's B cell lymphoma cell line BJAB were all infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.01 and analyzed for their respective CPEs at 24 hpi. Again, as expected, VSV-XN2 induced cell rounding against all cells lines, whereas VSV-gp160G induced syncytium formation only against the CD4+ ATL lines (Fig. 3A). MT-2, MT-4, TLO-m1, and BJAB cells were infected with either VSV-XN2 or VSV-gp160G at the indicated MOIs and then harvested at 24 hpi for immunoblotting to check for viral glycoprotein expression. HIV-1 glycoproteins gp120 and gp41 were only detected in ATL cell lines infected with VSV-gp160G, and BJAB cells did not express HIV-1 glycoproteins. Using α-HIVgp41 antibody, full-length gp160G and the gp41G fragments were detected in all CD4+ cells infected with VSV-gp160G (Fig. 3B). However, the gp41G subunit was not readily detected in MT-4 cells for reasons that remain unclear but may involve aberrant glycosylation events that could occur in these cells and that may affect antibody recognition. The detection of the heavy and light bands using α-VSV-G antibody in VSV-gp160G-infected cells correspond to recognition of the cytoplasmic tail of the VSV-G protein that was fused to gp160.

Bottom Line: VSV-gp160G was further noted to be highly attenuated and did not replicate efficiently in or induce significant cell death of primary CD4(+) T cells.Importantly, VSV-gp160G effectively exerted potent oncolytic activity in patient-derived ATL transplanted into NSG mice and facilitated a significant survival benefit.This effect greatly reduced neurotoxic risk associated with VSV infection while still allowing VSV to effectively target ATL cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Miami School of Medicine, Miami, Florida, USA.

Show MeSH
Related in: MedlinePlus