Document 1173
 DOCN  M9591173
 TI    A simple method to selectively expand HIV-1 specific cytotoxic T
       lymphocytes in vitro.
 DT    9509
 AU    Shankar P; Fabry J; Lieberman J; Dept. of Medicine, New England Medical
       Center, Boston, MA 02111,; USA.
 SO    Immunol Invest. 1995 Mar;24(3):489-97. Unique Identifier : AIDSLINE
       MED/95309983
 AB    Cytotoxic T lymphocytes (CTL) may play a critical role in controlling
       the progression of HIV-1 disease. Conventional assays for demonstration
       of CTL against HIV-1 have used either fresh PBMC or T cell lines and
       clones generated by non-specific stimulation. These methods are limited
       in their sensitivity since without specific secondary stimulation in
       vitro, epitopes recognized at low frequency may not be detected.
       Moreover, derivation of CTL clones is labor intensive and not practical
       for studying a large number of patients. We have developed a simple
       method to enrich HIV-1 specific CTL in vitro. Autologous antigen
       presenting cells (APC), either adherent macrophages or EBV transformed
       B-lymphoblastoid cells, are infected with recombinant vaccinia virus
       encoding individual HIV-1 proteins and after overnight culture the
       vaccinia virus is inactivated by uv irradiation in the presence of
       psoralin. The infected APC are then cultured with patient's T cells and
       CTL activity determined 10-14 days later. We have used this method to
       stimulate patients' T cells obtained directly from PBMC and also after
       mitogenic stimulation. In both systems, the HIV-1 specific response
       could be enhanced up to five to ten fold. This enhancement is comparable
       to CTL selection by exposure to HIV-1 immunodominant peptide incubated
       APC. In some patients, viral-specific CTL could be detected after
       HIV-vaccinia selection even though the mitogen stimulated cultures had
       no demonstrable antiviral CTL activity. Selective expansion of CTL
       directed against multiple HIV-1 proteins (env, gag and RT) could be
       obtained from PBMC as well as from mitogen-stimulated lines from
       individual patients. As these lines are predominantly CD8+ T cells by
       flow cytometric analysis and are free of vaccinia virus as ascertained
       by the lack of cytopathic effect in culture, in vitro vaccinia selection
       might also be useful to generate CTL lines for adoptive immunotherapy.
 DE    Antigen-Presenting Cells/IMMUNOLOGY/VIROLOGY  Cell Line  Clone
       Cells/IMMUNOLOGY  Cytotoxicity Tests, Immunologic  Genetic
       Vectors/GENETICS/IMMUNOLOGY  Human  HIV Infections/*IMMUNOLOGY
       HIV-1/*IMMUNOLOGY  Lymphocyte Transformation/*IMMUNOLOGY  Support,
       Non-U.S. Gov't  Support, U.S. Gov't, P.H.S.  T-Lymphocytes,
       Cytotoxic/*IMMUNOLOGY  Vaccinia Virus/GENETICS/IMMUNOLOGY  JOURNAL
       ARTICLE

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       protected by Copyright Law (Title 17, U.S.Code).