Kinetics of human immunodeficiency virus type 1 (HIV) DNA integration in acutely infected cells as determined using a novel assay for detection of integrated HIV DNA

被引:87
作者
Vandegraaff, N
Kumar, R
Burrell, CJ
Li, P
机构
[1] Natl Ctr HIV Virol Res, Inst Med & Vet Sci, Infect Dis Labs, Adelaide, SA 5000, Australia
[2] Univ Adelaide, Dept Mol Biosci, Adelaide, SA, Australia
关键词
D O I
10.1128/JVI.75.22.11253-11260.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We have developed a novel linker-primer PCR assay for the detection and quantification of integrated human immunodeficiency virus type 1 (HIV) DNA. This assay reproducibly allowed the detection of 10 copies of integrated HIV DNA, in a background of 2 x 10(5) cell equivalents of human chromosomal DNA, without amplifying extrachromosomal HIV DNA. We have used this assay and a near-synchronous one-step T-cell infection model to investigate the kinetics of viral DNA accumulation following HIV infection. We report here that integrated HIV DNA started accumulating 1 h after the first appearance of extrachromosomal viral DNA and accounted for similar to 10% of the total HIV DNA synthesized in the first round of viral replication. These results highlight the efficient nature of integrase-mediated HIV integration in infected T cells.
引用
收藏
页码:11253 / 11260
页数:8
相关论文
共 34 条
[21]   TEMPORAL ASPECTS OF DNA AND RNA-SYNTHESIS DURING HUMAN IMMUNODEFICIENCY VIRUS-INFECTION - EVIDENCE FOR DIFFERENTIAL GENE-EXPRESSION [J].
KIM, SY ;
BYRN, R ;
GROOPMAN, J ;
BALTIMORE, D .
JOURNAL OF VIROLOGY, 1989, 63 (09) :3708-3713
[22]   REQUIREMENT OF ACTIVE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 INTEGRASE ENZYME FOR PRODUCTIVE INFECTION OF HUMAN T-LYMPHOID CELLS [J].
LAFEMINA, RL ;
SCHNEIDER, CL ;
ROBBINS, HL ;
CALLAHAN, PL ;
LEGROW, K ;
ROTH, E ;
SCHLEIF, WA ;
EMINI, EA .
JOURNAL OF VIROLOGY, 1992, 66 (12) :7414-7419
[23]   HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 DNA-SYNTHESIS, INTEGRATION, AND EFFICIENT VIRAL REPLICATION IN GROWTH-ARRESTED T-CELLS [J].
LI, GR ;
SIMM, M ;
POTASH, MJ ;
VOLSKY, DJ .
JOURNAL OF VIROLOGY, 1993, 67 (07) :3969-3977
[24]   Retroviral cDNA integration: Stimulation by HMG I family proteins [J].
Li, L ;
Yoder, K ;
Hansen, MST ;
Olvera, J ;
Miller, MD ;
Bushman, FD .
JOURNAL OF VIROLOGY, 2000, 74 (23) :10965-10974
[25]   SYNTHESIS OF HUMAN-IMMUNODEFICIENCY-VIRUS DNA IN A CELL-TO-CELL TRANSMISSION MODEL [J].
LI, P ;
BURRELL, CJ .
AIDS RESEARCH AND HUMAN RETROVIRUSES, 1992, 8 (02) :253-259
[26]   Human immunodeficiency virus type 1 preintegration complexes: Studies of organization and composition [J].
Miller, MD ;
Farnet, CM ;
Bushman, FD .
JOURNAL OF VIROLOGY, 1997, 71 (07) :5382-5390
[27]   PERSISTENT HUMAN IMMUNODEFICIENCY VIRUS TYPE-1 INFECTION OF MONOBLASTOID CELLS LEADS TO ACCUMULATION OF SELF-INTEGRATED VIRAL-DNA AND TO PRODUCTION OF DEFECTIVE VIRIONS [J].
PAUZA, CD ;
GALINDO, J .
JOURNAL OF VIROLOGY, 1989, 63 (09) :3700-3707
[28]   INTEGRATION IS ESSENTIAL FOR EFFICIENT GENE-EXPRESSION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 [J].
SAKAI, H ;
KAWAMURA, M ;
SAKURAGI, JI ;
SAKURAGI, S ;
SHIBATA, R ;
ISHIMOTO, A ;
ONO, N ;
UEDA, S ;
ADACHI, A .
JOURNAL OF VIROLOGY, 1993, 67 (03) :1169-1174
[29]   Human immunodeficiency virus type 1 replication is blocked prior to reverse transcription and integration in freshly isolated peripheral blood monocytes [J].
Sonza, S ;
Maerz, A ;
Deacon, N ;
Meanger, J ;
Mills, J ;
Crowe, S .
JOURNAL OF VIROLOGY, 1996, 70 (06) :3863-3869
[30]   HIV-1 REPLICATION IS CONTROLLED AT THE LEVEL OF T-CELL ACTIVATION AND PROVIRAL INTEGRATION [J].
STEVENSON, M ;
STANWICK, TL ;
DEMPSEY, MP ;
LAMONICA, CA .
EMBO JOURNAL, 1990, 9 (05) :1551-1560