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M94A3242.TXT
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Document 3242
DOCN M94A3242
TI Establishment of HIV-1-producing T cells from peripheral lymphocytes:
participation of human complement factor B.
DT 9412
AU Nozaki-Renard J; Hirabuki N; Mizuno F; Iino T; Tada T; Dept. Microbiol.
Tokyo Medical College.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):120 (abstract no. PA0098). Unique
Identifier : AIDSLINE ICA10/94369333
AB OBJECTIVE: Research on the death and on the survival of HIV-1-infected
CD4+ T cells in human serum as a milieu supporting viral target cells.
STUDY SUBJECTS AND METHODS: 14 HIV-1-seroconverted individuals aged 8 to
65 (2 AC, 9 ARC and 3 AIDS cases). 4 male laboratory workers, as healthy
peripheral mononuclear cells (PMC) or normal human serum (NHS) donors.
PMC were prepared with Lymphoprep and adjusted to 2 x 10(6) cells/ml of
RPMI1640 supplemented with 10% heat-inactivated fetal calf serum.
Cultivation of 2 x 10(6) total cells/ml of mixed PMC from seroconverted
and healthy subjects was started in 24-well plates with 10% NHS: fresh,
heat-treated at 56 degrees C-30 min or at 50 degrees C-30 min (the
inactivating condition of complement factor B), and maintained for 6-8
weeks. Before multiple cloning procedure, p24 antigen in the culture
supernatant was examined by ELISA to determine the appearance of
HIV-producing cellular colonies. RESULTS: HIV-1-producing colonies were
obtained (12/14 subjects: 85.7%) from the wells cultured with fresh NHS;
no HIV+ colony could be maintained in other wells, with either
heat-treated NHS or fetal calf serum. But, 50 degrees C-30 min-treated
NHS, reconstructed with human factor B, recovered the ability to rescue
HIV+ cells from viral cytopathogenicity. Established clones originated
from PMC of seroconverted subjects since DNA sequences of
displacement-loop region of mitochondria (transmitted by maternal
lineage) were identical. Northern hybridization showed the clones had
TCR beta chain. DISCUSSION: HIV-1-producing T cells, in longum
divisible, were easily obtained with human complement factor B from
cocultured PMC and, as previously reported, from leukemic T cell lines
infected with HIV in vitro. This suggests that factor B is instrumental
in rescuing HIV-1-positive T cells, which might explain the latent phase
in natural infection.
DE Acquired Immunodeficiency Syndrome/*IMMUNOLOGY Adolescence Adult Aged
AIDS-Related Complex/*IMMUNOLOGY Blood Donors Cells, Cultured Child
Comparative Study Enzyme-Linked Immunosorbent Assay Human HIV Core
Protein p24/ANALYSIS/BIOSYNTHESIS HIV Seropositivity/*IMMUNOLOGY
HIV-1/DRUG EFFECTS/*PHYSIOLOGY Male Middle Age Properdin Factor
B/PHARMACOLOGY/*PHYSIOLOGY Reference Values T4
Lymphocytes/CYTOLOGY/*MICROBIOLOGY/PATHOLOGY *Virus Replication/DRUG
EFFECTS MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).