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.
Highlights
The ability of the human immunodeficiency virus type 1 (HIV-1) to recombine at high frequency is a critical evolutionary feature that dictates the extensive diversification of HIV-1 [1,2]
Intersubtype recombination among distinct HIV-1 strains generating various types of circulating recombinant form (CRF) have been documented worldwide [3,4] in which CRF01_AE and CRF02_AG are two major representative recombinants that are found in Southeast Asia and West Africa, respectively
We reconstituted a chimeric infectious molecular clone of CRF33_01B, which contains the 59 LTR derived from p93JP-NH1
Summary
The ability of the human immunodeficiency virus type 1 (HIV-1) to recombine at high frequency is a critical evolutionary feature that dictates the extensive diversification of HIV-1 [1,2]. Using the RT-positive culture supernatant, growth kinetics of each clone was assessed in CXCR4- or CCR5-expressing CD4+ human glioma NP-2 cells [14] to determine: (1) virus infectivity, and (2) coreceptor usage. Replication kinetics in human PBMCs and T cell lines To test the replication kinetics and coreceptor tropism, viral stock was prepared by transfecting 5 mg HIV-1 clone into 66105
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