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Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees.

Publication ,  Journal Article
Takehisa, J; Kraus, MH; Decker, JM; Li, Y; Keele, BF; Bibollet-Ruche, F; Zammit, KP; Weng, Z; Santiago, ML; Kamenya, S; Wilson, ML; Pusey, AE ...
Published in: Journal of virology
July 2007

Studies of simian immunodeficiency viruses (SIVs) in their endangered primate hosts are of obvious medical and public health importance, but technically challenging. Although SIV-specific antibodies and nucleic acids have been detected in primate fecal samples, recovery of replication-competent virus from such samples has not been achieved. Here, we report the construction of infectious molecular clones of SIVcpz from fecal viral consensus sequences. Subgenomic fragments comprising a complete provirus were amplified from fecal RNA of three wild-living chimpanzees and sequenced directly. One set of amplicons was concatenated using overlap extension PCR. The resulting clone (TAN1.24) contained intact genes and regulatory regions but was replication defective. It also differed from the fecal consensus sequence by 76 nucleotides. Stepwise elimination of all missense mutations generated several constructs with restored replication potential. The clone that yielded the most infectious virus (TAN1.910) was identical to the consensus sequence in both protein and long terminal repeat sequences. Two additional SIVcpz clones were constructed by direct synthesis of fecal consensus sequences. One of these (TAN3.1) yielded fully infectious virus, while the second one (TAN2.69) required modification at one ambiguous site in the viral pol gene for biological activity. All three reconstructed proviruses produced infectious virions that replicated in human and chimpanzee CD4(+) T cells, were CCR5 tropic, and resembled primary human immunodeficiency virus type 1 isolates in their neutralization phenotype. These results provide the first direct evidence that naturally occurring SIVcpz strains already have many of the biological properties required for persistent infection of humans, including CD4 and CCR5 dependence and neutralization resistance. Moreover, they outline a new strategy for obtaining medically important "SIV isolates" that have thus far eluded investigation. Such isolates are needed to identify viral determinants that contribute to cross-species transmission and host adaptation.

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Published In

Journal of virology

DOI

EISSN

1098-5514

ISSN

0022-538X

Publication Date

July 2007

Volume

81

Issue

14

Start / End Page

7463 / 7475

Related Subject Headings

  • Virus Replication
  • Virology
  • Simian immunodeficiency virus
  • Simian Immunodeficiency Virus
  • Sequence Homology, Amino Acid
  • Phylogeny
  • Pan troglodytes
  • Neutralization Tests
  • Molecular Sequence Data
  • Gene Products, pol
 

Citation

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MLA
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Takehisa, J., Kraus, M. H., Decker, J. M., Li, Y., Keele, B. F., Bibollet-Ruche, F., … Hahn, B. H. (2007). Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees. Journal of Virology, 81(14), 7463–7475. https://doi.org/10.1128/jvi.00551-07
Takehisa, Jun, Matthias H. Kraus, Julie M. Decker, Yingying Li, Brandon F. Keele, Fréderic Bibollet-Ruche, Kenneth P. Zammit, et al. “Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees.Journal of Virology 81, no. 14 (July 2007): 7463–75. https://doi.org/10.1128/jvi.00551-07.
Takehisa J, Kraus MH, Decker JM, Li Y, Keele BF, Bibollet-Ruche F, et al. Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees. Journal of virology. 2007 Jul;81(14):7463–75.
Takehisa, Jun, et al. “Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees.Journal of Virology, vol. 81, no. 14, July 2007, pp. 7463–75. Epmc, doi:10.1128/jvi.00551-07.
Takehisa J, Kraus MH, Decker JM, Li Y, Keele BF, Bibollet-Ruche F, Zammit KP, Weng Z, Santiago ML, Kamenya S, Wilson ML, Pusey AE, Bailes E, Sharp PM, Shaw GM, Hahn BH. Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees. Journal of virology. 2007 Jul;81(14):7463–7475.

Published In

Journal of virology

DOI

EISSN

1098-5514

ISSN

0022-538X

Publication Date

July 2007

Volume

81

Issue

14

Start / End Page

7463 / 7475

Related Subject Headings

  • Virus Replication
  • Virology
  • Simian immunodeficiency virus
  • Simian Immunodeficiency Virus
  • Sequence Homology, Amino Acid
  • Phylogeny
  • Pan troglodytes
  • Neutralization Tests
  • Molecular Sequence Data
  • Gene Products, pol