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

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

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

Jun Takehisa,Matthias H Kraus,Julie M Decker,Yingying Li,Brandon F Keele, Bibollet-Ruche,Kenneth P Zammit,Zhiping Weng,Mario L Santiago,Shadrack Kamenya,Michael L Wilson,Anne E Pusey,Elizabeth Bailes,Paul M Sharp,George M Shaw,Beatrice H Hahn,

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.

Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees. Publishing Authors By Initials

J Takehisa,MH Kraus,JM Decker,Y Li,BF Keele,F Bibollet-Ruche,KP Zammit,Z Weng,ML Santiago,S Kamenya,ML Wilson,AE Pusey,E Bailes,PM Sharp,GM Shaw,BH Hahn,

For similar biological phenomena, cell phenomena, and immunity: biological phenomena: microbiologic phenomena: viral physiology: virus replication research abstracts see: biological phenomena, cell phenomena, and immunity: biological phenomena: microbiologic phenomena: viral physiology: virus replication research

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MEDLINE DATE:

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

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of virology

VOLUME: 81

Page Numbers: 7463-75

Journal Abbreviation: J. Virol.

ISSN: 0022-538X

DAY: 9

MONTH: 05

YEAR: 2007

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

Number of References:

LANGUAGE: eng

NlmUniqueID: 113724

Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees. Keywords Mesh Terms:

KEYWORDS: Virus Replication

MESH TERMS: genetics

Chemical & Substance for Abstract: Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees. Information

Substance Name: Gene Products, pol

Registry Number: 0

Grant and Affiliation Information for Generation of infectious molecular clones of simian immunodeficiency virus from fecal consensus sequences of wild chimpanzees.

AFFILIATION: Department of Medicine, University of Alabama at Birmingham, 720 20th Street South, Kaul 816, Birmingham, AL 35294, USA.

Country: United States

AGENCY: United States NIAID

GRANT: R01 AI58715

ACRONYM: AI

MEDLINETA: J Virol

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