title: Generation of representative primary virus isolates from blood plasma after isolation of HIV-1 with CD44 MicroBeads.
abstract: Infection of cell cultures with cell-free virus isolated from HIV-infected patients is notoriously difficult and results in a loss of viral variation. Here, we describe viral sequences from PBMC, U87.CD4.CCR5 and U87.CD4.CXCR4 cell cultures and compare them to those from blood plasma from 12 patients from whom virus particles were isolated using CD44 MicroBeads. In both PBMC and U87.CD4.CCR5 cultures, 66% of the plasma viral strains were retrieved after culturing. In addition, coreceptor use was predicted based on the env-V3 sequence and tested in U87.CD4 cells expressing either CCR5 or CXCR4. Recovery was lower for the CXCR4-using viruses. Only 50% of the virus clusters predicted to use CXCR4 could be retrieved from cell cultures, while 71% of CCR5-using strains were found in U87.CCR5 cultures. Therefore, isolation of primary viruses with CD44 MicroBeads results in a good representation in cell culture of the in vivo divergence.
authors.affiliation: Laboratory of Experimental Virology, Department of Medical Microbiology, Centre of Infection and Immunity Amsterdam (CINIMA), Academic Medical Centre of the University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Marion
authors.initials: M
authors.lastname: Cornelissen
authors.forename: Edwin J
authors.initials: EJ
authors.lastname: Heeregrave
authors.forename: Fokla
authors.initials: F
authors.lastname: Zorgdrager
authors.forename: Georgios
authors.initials: G
authors.lastname: Pollakis
authors.forename: William A
authors.initials: WA
authors.lastname: Paxton
authors.forename: Antoinette C
authors.initials: AC
authors.lastname: van der Kuyl
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D016000:Cluster Analysis; D005838:Genotype; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D008863:Microspheres; D008969:Molecular Sequence Data; D010802:Phylogeny; D010949:Plasma / Q000821:virology*; D011110:Polymorphism, Genetic; D015728:Receptors, HIV / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation; D054299:env Gene Products, Human Immunodeficiency Virus
chemical_list: C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D015728:Receptors, HIV; D054299:env Gene Products, Human Immunodeficiency Virus