Construction and characterization of a full-length infectious clone from a fast-replicating, X4-tropic HIV-1 subtype B' isolate.
In HIV-1 epidemics in China, HIV-1 subtype B' is the most predominant subtype circulating in intravenous drug users. In this study, we constructed an HIV-1 full-length infectious molecular clone based on the primary virus LWJ, which was isolated from an HIV-infected patient in Fujian Province, China. Phylogenetic and bootscanning analysis of the viral sequence revealed that the isolate LWJ belonged to HIV-1 subtype B'. The infectious clone was designated as "pLWJ". The virus (LWJ-c) produced from this infectious clone by in vitro transfection of 293T cells could infect both human peripheral blood mononuclear cells (PBMCs) and human the T cell line MT4. Interestingly, the cloned LWJ-c virus utilized CXCR4 as its co-receptor and could replicate in vitro with similar efficiency and kinetics compared to its parental primary isolate LWJ as well as the clade B reference virus NL4-3. The LWJ-c virus could also cause cytopathic effects in both PBMCs and MT cells. Sequence analysis of the envelope glycoprotein of pLWJ showed that a conserved GPGR motif and an arginine at position 11 were present in the V3 loop, which was consistent with previous reports regarding CXCR4 co-receptor usage and syncytium-inducing (SI) phenotype. Thus, the infectious clone represents a fast-replicating, high-producing, CXCR4-tropic and syncytium-inducing isolate. Given the prevalence of HIV-1 subtype B' in China, this infectious clone can be a very useful tool to provide a versatile molecular model for research focusing on the biological properties of this subtype.
Wu
Shou-li
SL
Fujian Medical University, Fuzhou, Fujian, China.
Yan
Yan-sheng
YS
Yan
Ping-ping
PP
Huang
Hai-long
HL
Wang
Hui-rong
HR
D020816:Amino Acid Motifs; D001665:Binding Sites; D002478:Cells, Cultured; D002681:China; D005818:Genetic Engineering; D005838:Genotype; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D008297:Male; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics; D015728:Receptors, HIV / Q000502:physiology; D017422:Sequence Analysis, DNA; D013601:T-Lymphocytes / Q000821:virology; D014773:Virology / Q000379:methods*; D053586:Virus Internalization; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics
D012367:RNA, Viral; D015728:Receptors, HIV; D054299:env Gene Products, Human Immunodeficiency Virus