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Journal of Virology, March 2004, p. 2841-2852, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.2841-2852.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Phenotypic and Genotypic Comparisons of CCR5- and CXCR4-Tropic Human Immunodeficiency Virus Type 1 Biological Clones Isolated from Subtype C-Infected Individuals

Georgios Pollakis,1* Almaz Abebe,2 Aletta Kliphuis,1 Moustapha I. M. Chalaby,1,{dagger} Margreet Bakker,1 Yohannes Mengistu,3 Margreet Brouwer,4,{ddagger} Jaap Goudsmit,1,5,§ Hanneke Schuitemaker,4 and William A. Paxton1*

Department of Human Retrovirology, Academic Medical Center, University of Amsterdam,1 Center for Poverty-Related Communicable Diseases, 1105 AZ Amsterdam,5 Sanquin Research at CLB and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, The Netherlands,4 Ethiopian Health and Nutrition Research Institute (EHNRI),2 Faculty of Medicine, Department of Microbiology, Immunology and Parasitology, Addis Ababa University, Addis Ababa, Ethiopia3

Received 18 September 2003/ Accepted 4 November 2003

Individuals infected with human immunodeficiency virus type 1 (HIV-1) subtype C infrequently harbour X4 viruses. We studied R5 and X4 biological clones generated from HIV-1 subtype C-infected individuals. All subtype C R5 viruses demonstrated slower profiles of replication on CD4+ lymphocytes in comparison to subtype B viruses, whereas subtype C X4 viruses replicated with comparable efficiency to subtype B X4 viruses. No differences were identified in CC or CXC chemokine inhibitions (RANTES and SDF-1{alpha}, respectively) between subtype C and subtype B viruses. Immature dendritic cells were shown in coculture experiments to similarly enhance the infection of subtype C and subtype B R5 as well as X4 viruses. By amino acid sequence analysis, we showed that the R5 and X4 subtype C gp120 envelope gene alterations were similar to those for a switching subtype B virus, specifically with respect to the V3 charge and envelope N-linked glycosylation patterns. By phylogenetic analysis, we showed that one patient was infected with HIV-1 C' and the other was infected with HIV-1 C" and that one of the patients harbored a virus that was a recombinant in the gp120 env gene between an R5 and an X4 virus, with the resultant virus being R5. No differences were identified between the long terminal repeat regions of the subtype C R5 and X4 biological clones. These results indicate that even though R5 subtype C viruses are restrictive for virus replication, the R5-to-X4 phenotype switch can occur and does so in a manner similar to that of subtype B viruses.

* Corresponding author. Mailing address for Georgios Pollakis: Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: 31 20 566 8572. Fax: 31 20 961 6531. E-mail: g.pollakis{at}amc.uva.nl. Mailing address for William A. Paxton: Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: 31 20 566 4739. Fax: 31 20 961 6531. E-mail: w.a.paxton{at}amc.uva.nl.

{dagger} Present address: Genmab B.V., 3584 CK Utrecht, The Netherlands.

{ddagger} Present address: Center of Microbial and Plant Genetics (CMPG), Catholic University Leuven, B-3001 Heverlee-Leuven, Belgium.

§ Present address: Crucell Holland B.V., 2333 CN Leiden, The Netherlands.

Journal of Virology, March 2004, p. 2841-2852, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.2841-2852.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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