Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Jan;3(1):55-61.
doi: 10.4103/2141-9248.109490.

Increased Virus Replication and Cytotoxicity of Non-pathogenic Simian Human Immuno Deficiency Viruses-NM-3rN After Serial Passage in a Monkey-Derived Cell Line

Tb Kwofie  1 , T Miura
Affiliations

Increased Virus Replication and Cytotoxicity of Non-pathogenic Simian Human Immuno Deficiency Viruses-NM-3rN After Serial Passage in a Monkey-Derived Cell Line

Tb Kwofie et al. Ann Med Health Sci Res. 2013 Jan.

Abstract

Background: Infection and disease induction of variants of HIV type 1 (HIV-1) in vivo, especially their persistence, replication and rate of disease progression, have been found to depend on phenotypic characteristics. However, the mechanism (s) underlying these diverse phenotypic characteristics remain poorly understood.

Aim: It was aimed at determining whether a SHIV that had been adapted to a monkey-derived cell line could be used to explain the mechanism that underlies adaptive evolution of a virus to its host cell environment.

Materials and methods: Standard procedures in virology such as cell culturing, FACS analysis and ELISA were employed to measure virus replication and growth kinetics, cell viability, reverse transcriptase (RT) activity assay and CD4 cells down-regulation.

Results: After about 20 passages, LT efficiently adapted to the monkey-derived cell line and replicated much better than the parent virus. LT accumulated a number of mutations in its entire genome with a majority of them being monkey cell-specific.

Conclusion: Thus we think we have obtained a virus that may enable studies to determine which of these mutations are specifically related to in vitro viral replication and which are specifically related to cytotoxicity so as to explain the mechanism associated with viral cytotoxicity and host cell specificity.

Keywords: Cell culture; Evolution; HIV/SIV; Monkey; Replication.

PubMed Disclaimer

Conflict of interest statement

Conflict of Interest: None declared.

Figures

Figure 1
Figure 1
Replication kinetics of NM-3rN(M) (O) and NM-3rN(LT) (O) viruses, expressed as reverse transcriptase activity, in M8166 cells (a), HSC-F cells (b), Human PBMC (c) and Monkey PBMC (d). Duplicate cultures of 5 × 105 of these cells were infected with 100 TCID50 of each virus in a 96-well micro-plate as described in materials and methods. The kinetics shown for the viruses is the average of the duplicate cultures
Figure 2
Figure 2
This is a figure showing the down-regulation of the CD4+ cell population by the viruses {NM-3rN(M) and NM-3rN(LT)} in HSC-F cells. The CD4+ cell populations at the indicated time points 3, 6, 9 and 12 days post infection were monitored by FACS analysis as explained in the materials and methods
Figure 3
Figure 3
This show the reduction of viable cells by the viruses NM-3rN (M) and (LT) in monkey PBMC and HSC-F cells 3 days after infecting these cells with the viruses as described in materials and methods
Figure 4
Figure 4
These figures show the relationship between the infectivity of the viruses NM-3rN (M) and (LT) and their corresponding down-regulation of the CD4+ cells monitored in HSC-F cells as described in the materials and methods. (a) the percentage of the cells that were infected at the indicated time points while (b) the percentage of the CD4+ cell remaining in the culture at the same time points
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Rudensey LM, Papenhausen MD, Overbaugh J. Replication and persistence of simian immunodeficiency virus variants after passage in macaque lymphocytes and established human cell lines. J Virol. 1993;67:1727–33. - PMC - PubMed
    1. Fenyo EM, Albert J, Asjo B. Replicative capacity, cytopathic effect and cell tropism of HIV. AIDS. 1989;3:5–12. - PubMed
    1. Greenier JL, Miller CJ, Lu D, Dailey PJ, Lu FX, Kunstman KJ, et al. Route of simian immunodeficiency virus inoculation determines the complexity but not the identity of viral variant populations that infect rhesus macaques. J Virol. 2001;75:3753–65. - PMC - PubMed
    1. Kestler HWD, Ringler DJ, Mori K, Panicali DL, Sehgal PK, Daniel MD, et al. Importance of the nef gene for maintenance of high virus loads and for development of AIDS. Cell. 1991;65:651–62. - PubMed
    1. Baba TW, Jeong YS, Pennick D, Bronson R, Greene MF, Ruprecht RM. Pathogenicity of live attenuated SIV after mucosal infection of neonatal macaques. Science. 1995;267:1820–5. - PubMed