Limits...
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.

Show MeSH

Related in: MedlinePlus

VSV-gp160G selectively infects and replicates within CD4+ cells. (A) Schematic representation of VSV-XN2 construct and VSV-gp160G construct with gp160G fusion protein replacing VSV-G. (B) Graphical representation of VSV-XN2 and VSV-gp160G virions depicting the replacement of VSV-G with an HIV gp160–VSV-G fusion. The relevant glycoproteins are VSV-G, HIV-gp160, and the HIV-gp160–VSV-G fusion protein gp160G (left, right, bottom). (C) Bright-field microscopy of HeLa and HeLa CD4+ cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.01 at 24 hpi. (D) Immunoblot analysis results for viral glycoprotein expression of either construct in infected HeLa or HeLa CD4+ cells infected at an MOI of 0.01 at 24 hpi. (E) Growth kinetic assay results for HeLa or HeLa CD4+ 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 < 0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4645320&req=5

Figure 1: VSV-gp160G selectively infects and replicates within CD4+ cells. (A) Schematic representation of VSV-XN2 construct and VSV-gp160G construct with gp160G fusion protein replacing VSV-G. (B) Graphical representation of VSV-XN2 and VSV-gp160G virions depicting the replacement of VSV-G with an HIV gp160–VSV-G fusion. The relevant glycoproteins are VSV-G, HIV-gp160, and the HIV-gp160–VSV-G fusion protein gp160G (left, right, bottom). (C) Bright-field microscopy of HeLa and HeLa CD4+ cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.01 at 24 hpi. (D) Immunoblot analysis results for viral glycoprotein expression of either construct in infected HeLa or HeLa CD4+ cells infected at an MOI of 0.01 at 24 hpi. (E) Growth kinetic assay results for HeLa or HeLa CD4+ 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 < 0.001).

Mentions: Recombinant VSV with endogenous VSV-G has a wide tropism and is able to infect most mammalian cell types (49). In murine models, VSV can exhibit toxicity when administered at high doses due to the onset of neuropathy. To eliminate this potential problem and to create an oncolytic vector that targets only ATL cells, which are CD4+, we generated a VSV vector with the G protein substituted with a hybrid fusion protein containing extracellular and transmembrane domains from HIV1 gp160 and the cytoplasmic region of VSV-G. HIV-1 utilizes its glycoprotein, gp160, to gain entry into T cells through entry association with CD4 (Fig. 1A and B) (48). Due to the lack of human CD4 in the BHK-21-WI cells, VSV-gp160G was first recovered with the inclusion of a support VSV-G plasmid to VSV-G pseudotype the virions and enable infection of BHK-21-WI cells. Then, after successful recovery, VSV-gp160G was grown in HeLa CD4+ cells to amplify the progeny virions sans VSV-G.


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 selectively infects and replicates within CD4+ cells. (A) Schematic representation of VSV-XN2 construct and VSV-gp160G construct with gp160G fusion protein replacing VSV-G. (B) Graphical representation of VSV-XN2 and VSV-gp160G virions depicting the replacement of VSV-G with an HIV gp160–VSV-G fusion. The relevant glycoproteins are VSV-G, HIV-gp160, and the HIV-gp160–VSV-G fusion protein gp160G (left, right, bottom). (C) Bright-field microscopy of HeLa and HeLa CD4+ cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.01 at 24 hpi. (D) Immunoblot analysis results for viral glycoprotein expression of either construct in infected HeLa or HeLa CD4+ cells infected at an MOI of 0.01 at 24 hpi. (E) Growth kinetic assay results for HeLa or HeLa CD4+ 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 < 0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645320&req=5

Figure 1: VSV-gp160G selectively infects and replicates within CD4+ cells. (A) Schematic representation of VSV-XN2 construct and VSV-gp160G construct with gp160G fusion protein replacing VSV-G. (B) Graphical representation of VSV-XN2 and VSV-gp160G virions depicting the replacement of VSV-G with an HIV gp160–VSV-G fusion. The relevant glycoproteins are VSV-G, HIV-gp160, and the HIV-gp160–VSV-G fusion protein gp160G (left, right, bottom). (C) Bright-field microscopy of HeLa and HeLa CD4+ cells infected with either VSV-XN2 or VSV-gp160G at an MOI of 0.01 at 24 hpi. (D) Immunoblot analysis results for viral glycoprotein expression of either construct in infected HeLa or HeLa CD4+ cells infected at an MOI of 0.01 at 24 hpi. (E) Growth kinetic assay results for HeLa or HeLa CD4+ 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 < 0.001).
Mentions: Recombinant VSV with endogenous VSV-G has a wide tropism and is able to infect most mammalian cell types (49). In murine models, VSV can exhibit toxicity when administered at high doses due to the onset of neuropathy. To eliminate this potential problem and to create an oncolytic vector that targets only ATL cells, which are CD4+, we generated a VSV vector with the G protein substituted with a hybrid fusion protein containing extracellular and transmembrane domains from HIV1 gp160 and the cytoplasmic region of VSV-G. HIV-1 utilizes its glycoprotein, gp160, to gain entry into T cells through entry association with CD4 (Fig. 1A and B) (48). Due to the lack of human CD4 in the BHK-21-WI cells, VSV-gp160G was first recovered with the inclusion of a support VSV-G plasmid to VSV-G pseudotype the virions and enable infection of BHK-21-WI cells. Then, after successful recovery, VSV-gp160G was grown in HeLa CD4+ cells to amplify the progeny virions sans VSV-G.

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