Document 0688
 DOCN  M9460688
 TI    The influence of cell culture and storage conditions on HIV-1
       infectivity and fusogenic activity.
 DT    9404
 AU    Ongradi J; Ceccherini-Nelli L; Szilagyi JF; Specter S; Pistello M; Laird
       HM; Bendinelli M; Department of Biomedicine, University of Pisa, Italy.
 SO    Acta Microbiol Hung. 1992;39(3-4):207-21. Unique Identifier : AIDSLINE
       MED/94160675
 AB    We have previously demonstrated that acidic medium inhibits the
       replication of HIV-1. The present study was designed to examine the
       effects of other growth conditions and infection of fibroblasts by
       coculture with HIV infected lymphoid cells. Several lymphoblastoid cell
       lines normally grown in RPMI-1640 were grown in Eagle's MEM. These cells
       supported virus replication to higher titres than did RPMI-1640. Peak
       viral titres were achieved within 24-48 h after newly infected or
       chronically infected cells were placed in fresh medium. When virus was
       stored in liquid medium either frozen or at higher temperatures, virus
       titres were retained for several months while frozen but decreased upon
       storage at 4 degrees C or higher. If cells were passaged after
       trypsinization in Ca(++)-depleted medium, then a decreased
       susceptibility of cells for HIV-1 by 2 log10 at 24 h post infection was
       observed. Infectivity of cell-free and cell-associated HIV-1 was
       measured using syncytium formation, reverse transcriptase activity and
       p24 antigen. No fusion between HIV-1 infected CD4+ lymphoblasts and CD4-
       fibroblasts was observed but HIV-1 infected lymphoid cells, even in the
       absence of syncytium formation, exerted a strong toxic effect on
       fibroblasts. This study extends previous findings that medium acidity
       was inhibitory to virus replication and survival. Thus, conditions for
       study of HIV must be well controlled in buffered medium so that
       misleading results are not obtained regarding virus multiplication and
       possibly regarding transmission to and pathogenesis in CD4- cells.
 DE    Animal  Cell Fusion  Cell Line  Culture Media  Freezing  Human  HIV Core
       Protein p24/METABOLISM  HIV-1/GROWTH &
       DEVELOPMENT/IMMUNOLOGY/*PHYSIOLOGY  Lymphocytes  Reverse
       Transcriptase/METABOLISM  Support, Non-U.S. Gov't  Temperature  Virus
       Cultivation/METHODS  *Virus Replication  JOURNAL ARTICLE

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