Inhibition of the Replication of Feline Immunodeficiency Virus by Lentiviral Vector-Mediated RNA Interference in Feline Cell Lines

ABSTRACT The Ref1 and Lv1 postentry restrictions in human and monkey cells have been analyzed for lentiviruses in the primate and ungulate groups, but no data exist for the third (feline) group. We compared feline immunodeficiency virus (FIV) to other restricted (human immunodeficiency virus type 1 [HIV-1], equine infectious anemia virus [EIAV]) and unrestricted (NB-tropic murine leukemia virus [NB-MLV]) retroviruses across wide ranges of viral inputs in cells from multiple primate and nonprimate species. We also characterized restrictions conferred to permissive feline and canine cells engineered to express rhesus and human TRIM5α proteins and performed RNA interference (RNAi) against endogenous TRIM5α. We find that expression of rhesus or human TRIM5α proteins in feline cells restricts FIV, impairing pseudotyped vector transduction and viral replication, but rhesus TRIM5α is more restricting than human TRIM5α. Notably, however, canine cells did not support restriction by human TRIM5α and supported minimal restriction by rhesus TRIM5α, suggesting that these proteins may not function autonomously or that a canine factor interferes. Stable RNAi knockdown of endogenous rhesus TRIM5α resulted in marked increases in FIV and HIV-1 infectivities while having no effect on NB-MLV. A panel of nonprimate cell lines varied widely in susceptibility to lentiviral vector transduction, but normalized FIV and HIV-1 vectors varied concordantly. In contrast, in human and monkey cells, relative restriction of FIV compared to HIV-1 varied from none to substantial, with the greatest relative infectivity deficit for FIV vectors observed in human T-cell lines. Endogenous and introduced TRIM5α restrictions of FIV could be titrated by coinfections with FIV, HIV-1, or EIAV virus-like particles. Arsenic trioxide had complex and TRIM5α-independent enhancing effects on lentiviral but not NB-MLV infection. Implications for human gene therapy are discussed.

Download Full-text

Application of RNA interference for inhibiting the replication of feline immunodeficiency virus in chronically infected cell lines

Veterinary Microbiology ◽

10.1016/j.vetmic.2006.10.033 ◽

2007 ◽

Vol 120 (3-4) ◽

pp. 207-216 ◽

Cited By ~ 2

Author(s):

Kenji Baba ◽

Fuminori Mizukoshi ◽

Yuko Goto-Koshino ◽

Asuka Setoguchi-Mukai ◽

Yasuhito Fujino ◽

...

Keyword(s):

Rna Interference ◽

Infected Cell ◽

Cell Lines ◽

Feline Immunodeficiency Virus ◽

Immunodeficiency Virus

Download Full-text

Titration of Feline Immunodeficiency Virus-Based Lentiviral Vector Preparations

Cold Spring Harbor Protocols ◽

10.1101/pdb.prot067561 ◽

2011 ◽

Vol 2012 (1) ◽

pp. pdb.prot067561-pdb.prot067561

Author(s):

D. T. Saenz ◽

R. Barraza ◽

N. Loewen ◽

W. Teo ◽

E. M. Poeschla

Keyword(s):

Feline Immunodeficiency Virus ◽

Lentiviral Vector ◽

Immunodeficiency Virus

Download Full-text

Differences in Env and Gag protein expression patterns and epitope availability in feline immunodeficiency virus infected PBMC compared to infected and transfected feline model cell lines

Virus Research ◽

10.1016/j.virusres.2016.11.008 ◽

2017 ◽

Vol 227 ◽

pp. 249-260

Author(s):

Inge D.M. Roukaerts ◽

Chris K. Grant ◽

Sebastiaan Theuns ◽

Isaura Christiaens ◽

Delphine D. Acar ◽

...

Keyword(s):

Protein Expression ◽

Cell Lines ◽

Feline Immunodeficiency Virus ◽

Expression Patterns ◽

Gag Protein ◽

Immunodeficiency Virus ◽

Model Cell ◽

Feline Model

Download Full-text

Comparison of the Rev Transactivation of Feline Immunodeficiency Virus in Feline and Non-Feline Cell Lines.

Journal of Veterinary Medical Science ◽

10.1292/jvms.56.199 ◽

1994 ◽

Vol 56 (1) ◽

pp. 199-201 ◽

Cited By ~ 13

Author(s):

Keizo TOMONAGA ◽

Takayuki MIYAZAWA ◽

Yasushi KAWAGUCHI ◽

Mariko KOHMOTO ◽

Yasuo INOSHIMA ◽

...

Keyword(s):

Cell Lines ◽

Feline Immunodeficiency Virus ◽

Immunodeficiency Virus

Download Full-text

Productive Infection of Human Peripheral Blood Mononuclear Cells by Feline Immunodeficiency Virus: Implications for Vector Development

Journal of Virology ◽

10.1128/jvi.73.3.2491-2498.1999 ◽

1999 ◽

Vol 73 (3) ◽

pp. 2491-2498 ◽

Cited By ~ 41

Author(s):

James Johnston ◽

Christopher Power

Keyword(s):

Cell Lines ◽

Human Cell ◽

Feline Immunodeficiency Virus ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Human Cell Lines ◽

Peripheral Blood Mononuclear ◽

Human Pbmc ◽

Immunodeficiency Virus ◽

Blood Mononuclear Cells

ABSTRACT Feline immunodeficiency virus (FIV) is a lentivirus causing immune suppression and neurological disease in cats. Like primate lentiviruses, FIV utilizes the chemokine receptor CXCR4 for infection. In addition, FIV gene expression has been demonstrated in immortalized human cell lines. To investigate the extent and mechanism by which FIV infected primary and immortalized human cell lines, we compared the infectivity of two FIV strains, V1CSF and Petaluma, after cell-free infection. FIV genome was detected in infected human peripheral blood mononuclear cells (PBMC) and macrophages at 21 and 14 days postinfection, respectively. Flow cytometry analysis of FIV-infected human PBMC indicated that antibodies to FIV p24 recognized 12% of the cells. Antibodies binding the CCR3 chemokine receptor maximally inhibited infection of human PBMC by both FIV strains compared to antibodies to CXCR4 or CCR5. Reverse transcriptase levels increased in FIV-infected human PBMC, with detection of viral titers of 101.3 to 102.1 50% tissue culture infective doses/106 cells depending on the FIV strain examined. Cell death in human PBMC infected with either FIV strain was significantly elevated relative to uninfected control cultures. These findings indicate that FIV can productively infect primary human cell lines and that viral strain specificity should be considered in the development of an FIV vector for gene therapy.

Download Full-text

737. Development of a Feline Immunodeficiency Virus-Based Lentiviral Vector for Genetic Modification of CNS Neurons

Molecular Therapy ◽

10.1016/j.ymthe.2004.06.679 ◽

2004 ◽

Vol 9 ◽

pp. S280

Keyword(s):

Genetic Modification ◽

Feline Immunodeficiency Virus ◽

Lentiviral Vector ◽

Immunodeficiency Virus ◽

Cns Neurons

Download Full-text

Optimization of Feline Immunodeficiency Virus Vectors for RNA Interference

Journal of Virology ◽

10.1128/jvi.00958-06 ◽

2006 ◽

Vol 80 (19) ◽

pp. 9371-9380 ◽

Cited By ~ 19

Author(s):

Scott Q. Harper ◽

Patrick D. Staber ◽

Christine R. Beck ◽

Sarah K. Fineberg ◽

Colleen Stein ◽

...

Keyword(s):

Gene Expression ◽

Rna Interference ◽

Reporter Gene ◽

Feline Immunodeficiency Virus ◽

Expression Cassette ◽

Expression Vectors ◽

Pol Ii ◽

Rev Response Element ◽

Immunodeficiency Virus ◽

In Cells

ABSTRACT RNA interference (RNAi) occurs naturally in plant and animal cells as a means for modulating gene expression. This process has been experimentally manipulated to achieve targeted gene silencing in cells, tissues, and animals, using a variety of vector systems. Here, we tested the hypothesis that vectors based on feline immunodeficiency virus (FIV) could be used for coexpression of reporter constructs and RNAi expression cassettes. We found, unexpectedly, in our initial constructs that placement of RNAi expression cassettes downstream from a polymerase II (pol II)-expressed reporter gene inhibited reporter expression but not vector titer. Through a series of intermediate vector constructs, we found that placement of the RNAi expression cassette relative to the Rev response element and the pol II expression cassette was critical for efficient RNAi and reporter gene expression. These results suggested that steric factors, including RNA structure and recruitment of competing transcriptional machinery, may affect gene expression from FIV vectors. In a second series of studies, we show that target sequence silencing can be achieved in cells transduced by FIV vectors coexpressing reporter genes and 3′ untranslated region resident microRNAs. The optimized FIV-based RNAi expression vectors will find broad use given the extensive tropism of pseudotyped FIV vectors for many cell types in vitro and in vivo.

Download Full-text