Rhesus M1.3S Cells Suitable for Biological Evaluation of Macaque-Tropic HIV/SIV Clones

Doi, Naoya; Fujiwara, Sachi; Adachi, Akio; Nomaguchi, Masako

doi:10.3389/fmicb.2011.00115

Cited by 13 publications

(16 citation statements)

References 16 publications

(21 reference statements)

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“…An RhM lymphocyte cell line, M1.3S (25), was maintained in RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum (FBS) and 50 units/ml of recombinant human interleukin-2 (IL-2) (Bio-Rad Laboratories Inc., Hercules, CA). The human monolayer cell lines 293T (27) and HEp2 (ATCC CCL-23) and the RhM kidney cell line LLC-MK2 (ATCC CCL-7) were maintained in Eagle's minimal essential medium (MEM) containing 10% FBS.…”

Section: Methodsmentioning

confidence: 99%

“…The construction of MN4Rh-3, SIVmac239 clone MA239N, and its nef-deficient variant MA239N-⌬N was described previously (19,25). CA alterations of HIV-1mt proviral clones were performed by replacing target sites of HIV-1mt CA with the corresponding sites of SIVmac239 (both codons and amino acids were changed to those of SIVmac239) by using the QuikChange site-directed mutagenesis kit (Agilent Technologies Inc., Santa Clara, CA).…”

Section: Methodsmentioning

confidence: 99%

“…In this study, we generated new HIV-1mt clones from MN4Rh-3 that potentially possess an improved capacity to antagonize RhM TRIM5␣ and tetherin proteins. First, we constructed a number of MN4Rh-3 CA mutant clones; examined their growth potentials in an RhM lymphocyte cell line, M1.3S (25); and selected a new HIV-1mt clone, MN4/LSDQ, carrying only three amino acid substitutions in MN4Rh-3 CA (Fig. 1).…”

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confidence: 99%

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Generation of Rhesus Macaque-Tropic HIV-1 Clones That Are Resistant to Major Anti-HIV-1 Restriction Factors

Nomaguchi

Yokoyama

Kono

et al. 2013

J Virol

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h Human immunodeficiency virus type 1 (HIV-1) replication in macaque cells is restricted mainly by antiviral cellular APOBEC3, TRIM5␣/TRIM5CypA, and tetherin proteins. For basic and clinical HIV-1/AIDS studies, efforts to construct macaque-tropic HIV-1 (HIV-1mt) have been made by us and others. Although rhesus macaques are commonly and successfully used as infection models, no HIV-1 derivatives suitable for in vivo rhesus research are available to date. In this study, to obtain novel HIV-1mt clones that are resistant to major restriction factors, we altered Gag and Vpu of our best HIV-1mt clone described previously. First, by sequence-and structure-guided mutagenesis, three amino acid residues in Gag-capsid (CA) (M94L/R98S/G114Q) were found to be responsible for viral growth enhancement in a macaque cell line. Results of in vitro TRIM5␣ susceptibility testing of HIV-1mt carrying these substitutions correlated well with the increased viral replication potential in macaque peripheral blood mononuclear cells (PBMCs) with different TRIM5 alleles, suggesting that the three amino acids in HIV-1mt CA are involved in the interaction with TRIM5␣. Second, we replaced the transmembrane domain of Vpu of this clone with the corresponding region of simian immunodeficiency virus SIVgsn166 Vpu. The resultant clone, MN4/LSDQgtu, was able to antagonize macaque but not human tetherin, and its Vpu effectively functioned during viral replication in a macaque cell line. Notably, MN4/ LSDQgtu grew comparably to SIVmac239 and much better than any of our other HIV-1mt clones in rhesus macaque PBMCs. In sum, MN4/LSDQgtu is the first HIV-1 derivative that exhibits resistance to the major restriction factors in rhesus macaque cells.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Generation of Rhesus Macaque-Tropic HIV-1 Clones That Are Resistant to Major Anti-HIV-1 Restriction Factors

Nomaguchi

Yokoyama

Kono

et al. 2013

J Virol

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“…Our results here show that the Gag-MA 66SR mutation positively acts on singlecycle viral infectivity. (14) was maintained in RPMI1640 medium containing 10% heat-inactivated fetal bovine serum and 50 units/ml of recombinant human interleukin-2 (Bio-Rad Laboratories Inc.). Virus stocks for infection experiments were prepared from 293T cells transfected with various proviral clones by the calcium-phosphate co-precipitation method (15), and their virionassociated RT activities were determined as previously described (11,16).…”

Section: Introductionmentioning

confidence: 99%

Single-amino acid mutation 66SR in Gag-matrix enhances viral single-cycle infectivity of R5-tropic HIV-1rmt

et al. 2015

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“…Since the lymphocyte cell lines immortalized by HVS do not lose their original characteristics as primary lymphocytes in most cases and are readily maintained for experiments, to biologically characterize viruses like HIV-1, it is quite important for laboratory researchers to have HVS-immortalized cell lines. In our laboratory, cynomolgus HSC-F (Akari et al, 1996;Fujita et al, 2003) and rhesus M1.3S (Doi et al, 2011) Table 4. Cell lines for virological evaluation of HIV-1mt.…”

Section: Generation and Characterization Of Various Hiv-1mt Clonesmentioning

confidence: 99%

Macaque-Tropic HIV-1 Derivatives: A Novel Experimental Approach to Understand Viral Replication and Evolution in Vivo

Nomaguchi¹,

Doi²,

Fujiwara³

et al. 2011

HIV-Host Interactions

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Rhesus M1.3S Cells Suitable for Biological Evaluation of Macaque-Tropic HIV/SIV Clones

Cited by 13 publications

References 16 publications

Generation of Rhesus Macaque-Tropic HIV-1 Clones That Are Resistant to Major Anti-HIV-1 Restriction Factors

Generation of Rhesus Macaque-Tropic HIV-1 Clones That Are Resistant to Major Anti-HIV-1 Restriction Factors

Single-amino acid mutation 66SR in Gag-matrix enhances viral single-cycle infectivity of R5-tropic HIV-1rmt

Macaque-Tropic HIV-1 Derivatives: A Novel Experimental Approach to Understand Viral Replication and Evolution in Vivo

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