title: Isolation and characterization of a replication-competent molecular clone of an HIV-1 circulating recombinant form (CRF33_01B).
abstract: A growing number of emerging HIV-1 recombinants classified as circulating recombinant forms (CRFs) have been identified in Southeast Asia in recent years, establishing a molecular diversity of increasing complexity in the region. Here, we constructed a replication-competent HIV-1 clone for CRF33_01B (designated p05MYKL045.1), a newly identified recombinant comprised of CRF01_AE and subtype B. p05MYKL045.1 was reconstituted by cloning of the near full-length HIV-1 sequence from a newly-diagnosed individual presumably infected heterosexually in Kuala Lumpur, Malaysia. The chimeric clone, which contains the 5' LTR (long terminal repeat) region of p93JP-NH1 (a previously isolated CRF01_AE infectious clone), showed robust viral replication in the human peripheral blood mononuclear cells. This clone demonstrated robust viral propagation and profound syncytium formation in CD4+, CXCR4-expressing human glioma NP-2 cells, indicating that p05MYKL045.1 is a CXCR4-using virus. Viral propagation, however, was not detected in various human T cell lines including MT-2, M8166, Sup-T1, H9, Jurkat, Molt-4 and PM1. p05MYKL045.1 appears to proliferate only in restricted host range, suggesting that unknown viral and/or cellular host factors may play a role in viral infectivity and replication in human T cell lines. Availability of a CRF33_01B molecular clone will be useful in facilitating the development of vaccine candidates that match the HIV-1 strains circulating in Southeast Asia.
authors.affiliation: Laboratory of Molecular Virology and Epidemiology, AIDS Research Center, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan.
authors.forename: Kok Keng
authors.initials: KK
authors.lastname: Tee
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kusagawa
authors.forename: Xiao-Jie
authors.initials: XJ
authors.lastname: Li
authors.forename: Narumi
authors.initials: N
authors.lastname: Onogi
authors.forename: Maya
authors.initials: M
authors.lastname: Isogai
authors.forename: Saiki
authors.initials: S
authors.lastname: Hase
authors.forename: Rie
authors.initials: R
authors.lastname: Uenishi
authors.forename: Huanan
authors.initials: H
authors.lastname: Liao
authors.forename: Adeeba
authors.initials: A
authors.lastname: Kamarulzaman
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Takebe
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D010802:Phylogeny; D011995:Recombination, Genetic; D014779:Virus Replication
chemical_list: D017931:DNA Primers
references.citation: AIDS. 2006 Oct 24;20(16):W13-23
references.pmid: 17053344
references.citation: AIDS Rev. 2007 Apr-Jun;9(2):75-87
references.pmid: 17694675
references.citation: Biochem Biophys Res Commun. 1999 May 10;258(2):313-21
references.pmid: 10329384
references.citation: J Acquir Immune Defic Syndr. 2006 Dec 15;43(5):523-9
references.pmid: 17031320
references.citation: J Virol. 1995 Jun;69(6):3949-54
references.pmid: 7745752
references.citation: J Virol. 1986 Aug;59(2):284-91
references.pmid: 3016298
references.citation: Adv Pharmacol. 2008;56:1-25
references.pmid: 18086407
references.citation: Virology. 2000 Dec 5;278(1):103-10
references.pmid: 11112486
references.citation: Nature. 1995 Mar 9;374(6518):124-6
references.pmid: 7877682
references.citation: AIDS Res Hum Retroviruses. 2002 May 20;18(8):585-9
references.pmid: 12036488
references.citation: Mol Biol Evol. 2007 Aug;24(8):1596-9
references.pmid: 17488738
references.citation: AIDS Res Hum Retroviruses. 2007 Jun;23(6):829-33
references.pmid: 17604547
references.citation: AIDS Res Hum Retroviruses. 2002 Jan 20;18(2):115-22
references.pmid: 11839144
references.citation: AIDS Res Hum Retroviruses. 2003 Jul;19(7):561-7
references.pmid: 12908933
references.citation: J Virol. 1999 Jan;73(1):152-60
references.pmid: 9847317
references.citation: AIDS Res Hum Retroviruses. 2008 Feb;24(2):251-7
references.pmid: 18284324
references.citation: J Virol. 1998 Jul;72(7):5680-98
references.pmid: 9621027
references.citation: J Virol. 1999 Jul;73(7):5520-6
references.pmid: 10364300
references.citation: Science. 1985 Aug 9;229(4713):563-6
references.pmid: 2992081
references.citation: J Virol. 2003 Oct;77(20):11193-200
references.pmid: 14512567