Document 3241
 DOCN  M94A3241
 TI    Transformation of human diploid fibroblasts by a transmissible agent
       obtained from an HIV-infected patient.
 DT    9412
 AU    Dolei A; Serra C; Gentile M; Falcone V; Conaldi PG; Casalone R; Toniolo
       A; Institute of Microbiology, Univ. of Sassari, Italy.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(1):120 (abstract no. PA0099). Unique
       Identifier : AIDSLINE ICA10/94369334
 AB    OBJECTIVE: To study the effects of clinical HIV-1 isolates on human
       fibroblastoid cells. Exposure to one of the isolates was followed by
       immortalization of MRC-5 diploid fibroblasts. The cell-free supernatant
       of these cells was able to immortalize new batches of MRC-5 fibroblasts.
       Uninfected and HIV-1 infected MRC-5 control cultures did not undergo
       transformation upon repeated passage under the same culture conditions.
       METHODS: Transformed fibroblasts (lines DOR, D3A, D3B) were tested for
       the presence of viral agents common in AIDS patients and subjected to
       transformation assays in vitro. Genomic analysis was made by in situ
       hybridization with human centromere-specific probes, typing of six
       independent loci by PCR and hybridization, and chromosome analysis by
       standard techniques. RESULTS: DOR and D3B lines had lost the typical
       fibroblastoid morphology, showed a reduced dependence on serum and
       growth factors, and were capable of forming colonies in soft agar. All
       cells were aneuploid and showed specific alterations of chromosomes 1,
       5, and 10 that were absent in the parental line. Their human origin was
       assessed with human-specific probes; parental and transformed lines had
       a common origin as established by typing of HLA-DQA1, LDLR, GYPA, HBGG,
       D7S8, and GC loci (p < 0.001). Transformed lines were negative for the
       following agents: HSV-1/2, CMV, EBV, HHV-6, SV40, HIV-1, and
       adenoviruses. When tested by IIF, human sera of normal individuals were
       not reactive, whereas 10-15% of sera from HIV-infected individuals
       produced cytoplasmic staining at dilutions > or = 1:100. DISCUSSION AND
       CONCLUSION: The data suggest that a transmissible agent capable of
       transforming fibroblastoid cells was obtained from an HIV patient.
       Characterization of the putative agent may shed light on AIDS-associated
       neopalstic disease.
 DE    Acquired Immunodeficiency Syndrome/*MICROBIOLOGY  Aneuploidy  Biological
       Markers/ANALYSIS  Cell Line, Transformed  *Cell Transformation, Viral
       *Chromosome Aberrations  Culture Media, Conditioned  Diploidy
       Fibroblasts  Human  HIV-1/GENETICS/ISOLATION & PURIF/*PHYSIOLOGY  In
       Situ Hybridization  Polymerase Chain Reaction  Support, Non-U.S. Gov't
       MEETING ABSTRACT

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).