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. 2011 May 23:2:115.
doi: 10.3389/fmicb.2011.00115. eCollection 2011.

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

Naoya Doi  1 , Sachi Fujiwara, Akio Adachi, Masako Nomaguchi
Affiliations

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

Naoya Doi et al. Front Microbiol. .
No abstract available

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Figures

Figure 1
Figure 1
Virological characteristics of HIV-1/SIVmac clones. (A) Proviral genome structure of various viruses used for infection of M1.3S. Genomes of various HIV-1mt clones generated in our laboratory are schematically illustrated. Parental clones HIV-1 NL4-3 (Adachi et al., 1986) and SIVmac MA239N (Doi et al., 2010) are shown at the top. Accessory gene-inactivated mutants of SIVmac MA239N (circled) were constructed by substituting an internal stop codon in the nef gene of MA239 mutants (Shibata et al., 1991) with a Glu codon as reported for construction of MA239N (Doi et al., 2010). White and gray areas indicate the genomic regions of NL4-3 and MA239N, respectively. An arrow and arrowheads indicate the site of the four mutations in gag, pol, and env genes, respectively. The dotted region contains additional single-nucleotide mutations relative to the other clones. Enlarged Gag-CA region is shown on the right. For details, see the text. (B) Replication kinetics of the viruses in M1.3S cells. Cell-free viruses were prepared from 293T cells transfected with proviral clones indicated, and inoculated into 1 × 106 cells of M1.3S. Viral replication was monitored at intervals by reverse transcriptase (RT) activity in the culture supernatants. The experiments were done as described previously (Kamada et al., ; Doi et al., 2010). For infection, 6.3 × 104 and 2.4 × 106 RT units of MA239N and HIV-1mt clones (left), respectively, and 5.0 × 104 RT units of MA239N clones (right) were used. The representative data in two independent experiments are shown. Replication of NL-DT5RS, MN4–8, and MN4–8S was not detected as observed for NL-DT5R.
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