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Journal of Virology, April 2001, p. 3141-3151, Vol. 75, No. 7
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.7.3141-3151.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Mouse-Human Heterokaryons Support Efficient Human Immunodeficiency Virus Type 1 Assembly

Roberto Mariani,1 Beth A. Rasala,1 Gabriel Rutter,2 Klaus Wiegers,2 Stephanie M. Brandt,1 Hans-Georg Kräusslich,2,dagger and Nathaniel R. Landau1,*

Infectious Disease Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037,1 and Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität Hamburg, D-20251 Hamburg, Germany2

Received 25 October 2000/Accepted 11 January 2001

Murine cells do not support human immunodeficiency virus type 1 (HIV-1) replication because of blocks to virus entry, proviral expression, and virion assembly. In murine 3T3 fibroblasts, the block to HIV-1 entry is relieved by the introduction of human CD4 and CCR5 or CXCR4, and proviral expression is increased by the introduction of the Tat cofactor, human cyclin T1; however, because of the assembly block, virus fails to spread. A panel of rodent cell lines expressing human CD4, CCR5, and cyclin T1 was established and studied for the ability to support virus replication. Mus musculus lymphoid cell lines EL4 and L1-2 and Mus dunni fibroblasts supported only low levels of virus assembly and released small amounts of infectious virus. CHO and Rat2 cell lines produced more infectious virus, but this production was still 40-fold lower than production in human cells. Only CHO cells expressing the three human cofactors were partially permissive for HIV-1 replication. To investigate the basis of the block to HIV-1 assembly, mouse-human heterokaryons were tested for ability to assemble and release virus. Fusion of human cells to HIV-1-infected mouse cells expressing CD4, CCR5, and cyclin T1 caused a 12-fold increase in virion release and a 700-fold increase in infectious virus production. Fusion of HIV-1-infected M. dunni tail fibroblasts to uninfected human cells caused a similar increase in virus release. More efficient virus release was not caused by increased proviral transcription or increased synthesis of virion components. Analysis of reciprocal heterokaryons suggested the absence of an inhibitor of virus assembly. Taken together, the results suggested that murine fibroblasts lack a cofactor that is required for efficient virus assembly and release.

* Corresponding author. Mailing address: The Salk Institute for Biological Studies, Infectious Disease Laboratory, 10010 N. Torrey Pines Rd., La Jolla, CA 92037. Phone: (858) 453-4100. Fax: (858) 554-0341. E-mail: Landau{at}salk.edu.

dagger Present address: Abteilung Virologie, Universität Heidelberg, D-69120 Heidelberg, Germany.

Journal of Virology, April 2001, p. 3141-3151, Vol. 75, No. 7
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.7.3141-3151.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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