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M94A3241.TXT
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1994-10-25
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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).