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M9480500.TXT
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1994-08-20
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Document 0500
DOCN M9480500
TI Evaluation of human immunodeficiency virus type 1 (HIV-1)-specific
cytotoxic T-lymphocyte responses utilizing B-lymphoblastoid cell lines
transduced with the CD4 gene and infected with HIV-1.
DT 9410
AU McElrath MJ; Rabin M; Hoffman M; Klucking S; Garcia JV; Greenberg PD;
Department of Medicine, University of Washington School of; Medicine,
Seattle.
SO J Virol. 1994 Aug;68(8):5074-83. Unique Identifier : AIDSLINE
MED/94309173
AB Analysis of major histocompatibility complex-restricted cytotoxic T
lymphocytes (CTL) capable of killing human immunodeficiency virus type 1
(HIV-1)-infected targets is essential for elucidating the basis for
HIV-1 disease progression and the potential efficacy of candidate
vaccines. The use of primary CD4+ T cells with variable infectivity as
targets for such studies has significant limitations, and immortal
autologous cells with high levels of CD4 expression that can be
consistently infected with HIV-1 would be of much greater utility.
Therefore, we transduced Epstein-Barr-virus-transformed B-lymphoblastoid
cell lines (LCL) with a retroviral vector, LT4SN, containing the human
CD4 gene. Stable LCL in which more than 95% of cells expressed membrane
CD4 were obtained. Aliquots were infected with HIV-1, and, after 4 to 7
days, nearly all of the cells contained cytoplasmic gag and produced
high levels of p24 antigen. The ability of major histocompatibility
complex-restricted CD8+ CTL to lyse such HIV-1-infected CD4-transduced
LCL (LCL-CD4HIV-1) was evaluated. These autologous targets were lysed by
CTL generated from an HIV-1-uninfected vaccinee over a broad range of
effector-to-target ratios. Similarly, the LCL-CD4HIV-1 were efficiently
lysed by fresh circulating CTL from HIV-1-infected individuals, as well
as by CTL activated by in vitro stimulation. Both HIV-1 env- and
gag-specific CTL effectors lysed LCL-CD4HIV-1, consistent with the
cellular expression of both HIV-1 genes. The LCL-CD4HIV also functioned
as stimulator cells, and thus are capable of amplifying CTL against
multiple HIV-1 gene products in HIV-1-infected individuals. The ability
to produce HIV-1-susceptible autologous immortalized cell lines that can
be employed as target cells should enable a more detailed evaluation of
vaccine-induced CTL against both homologous and disparate HIV-1 strains.
Furthermore, the use of LCL-CD4HIV-1 should facilitate the analysis of
the range of HIV-1 gene products recognized by CTL in seropositive
persons.
DE Antigens, CD4/GENETICS/*IMMUNOLOGY AIDS Vaccines/IMMUNOLOGY
B-Lymphocytes/CYTOLOGY/*IMMUNOLOGY/MICROBIOLOGY Cell Line, Transformed
Human HIV Seropositivity/IMMUNOLOGY HIV-1/*IMMUNOLOGY/PHYSIOLOGY
Support, U.S. Gov't, P.H.S. T-Lymphocytes, Cytotoxic/*IMMUNOLOGY
Transduction, Genetic Virus Replication JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).