Isolation and Initial Characterization of Multiple Species of T-Lymphocyte Subset cDNA Clones

A modified differential screening procedure was applied to analyze cDNA libraries of cloned helper T lymphocytes (Th) and cytolytic T lymphocytes (CTL). Negative and positive differential screening and RNA blot analysis were used to identify cDNA clones that were expressed preferentially in Th or CT...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1987-05, Vol.84 (9), p.2896-2900
Main Authors: Kwon, Byoung S., Kim, Gwan S., Prystowsky, Michael B., Lancki, David W., Sabath, Daniel E., Pan, Julian, Weissman, Sherman M.
Format: Article
Language:English
Subjects:
Analysis of the immune response. Humoral and cellular immunity
Animals
B lymphocytes
Biological and medical sciences
CDNA libraries
Cloning, Molecular
Complementary DNA
Complementary RNA
DNA - isolation & purification
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Genetic screening
Immunobiology
Lymphocyte Activation
Messenger RNA
Mice
Nucleic Acid Hybridization
Organs and cells involved in the immune response
RNA
Signal detection
T cell antigen receptors
T lymphocytes
T-Lymphocytes - classification
T-Lymphocytes - immunology
T-Lymphocytes, Cytotoxic - classification
T-Lymphocytes, Cytotoxic - immunology
T-Lymphocytes, Helper-Inducer - classification
T-Lymphocytes, Helper-Inducer - immunology
Transcription, Genetic
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Description
Summary:A modified differential screening procedure was applied to analyze cDNA libraries of cloned helper T lymphocytes (Th) and cytolytic T lymphocytes (CTL). Negative and positive differential screening and RNA blot analysis were used to identify cDNA clones that were expressed preferentially in Th or CTL. Seven clones corresponded to previously described T-cell genes, and 16 additional types of cDNA clones were isolated, 9 from Th and 7 from CTL. Of these, 3 were expressed in both Th and CTL, 7 were expressed in only Th, and 6 only in CTL. These clones were analyzed for induction after stimulation by interleukin 2 or Con A or after stimulation of the T-cell antigen receptor (TCR). Three different patterns of expression were seen: (i) induction only by Con A, (ii) induction by Con A and interleukin 2, and (iii) induction by Con A and TCR stimulation. The approach is potentially useful for analyzing paths of T-cell differentiation and detecting cDNA clones encoding unrecognized cytokines.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.84.9.2896