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).