Estimation by limiting dilution analysis of human IL 2-secreting T cells: detection of IL 2 produced by single lymphokine-secreting T cells
Journal Article
·
· J. Immunol.; (United States)
OSTI ID:5805384
We present here a culture method for the estimation, in human blood, of the number of lymphocytes that can respond to mitogen by producing interleukin 2 (IL 2). T cells are cultured at limiting dilutions with PHA or Con A in the presence of Epstein Barr virus-transformed human lymphoblastoid cells (EB-LCL), and supernatants are tested 3 days later for IL 2 content by a cell proliferation assay. The distribution of negative wells follows the expected Poisson single-hit relationship, suggesting that the assay is sensitive to single cells of a single limiting cell type. On average, 16.3% of peripheral blood mononuclear cells can produce IL 2 in such clonal cultures (mean of 12 determinations; SD = 5.6%). Surprisingly, irradiation (up to 2000 rad) of the titrated responder cell population diminishes the estimated frequencies by less than 50%. The ability to detect IL 2 levels in cultures containing only a single, nonproliferating T lymphocyte allows us to estimate the amount of IL 2 generated by an individual effector cell during a 3-day culture interval after mitogen stimulation. The average responding, irradiated T cell generates 0.92 pg of IL 2 (median) within 3 days. The method presented provides a straightforward way to provide independent estimates of responding cell number and of lymphokine production per cell in a variety of clinical situations.
- Research Organization:
- Boston Univ. School of Medicine, MA
- OSTI ID:
- 5805384
- Journal Information:
- J. Immunol.; (United States), Journal Name: J. Immunol.; (United States) Vol. 9; ISSN JOIMA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
551001 -- Physiological Systems-- Tracer Techniques
560121* -- Radiation Effects on Cells-- External Source-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CULTURES
CELL PROLIFERATION
CLONING
CONNECTIVE TISSUE CELLS
DOSE-RESPONSE RELATIONSHIPS
GROWTH FACTORS
IRRADIATION
LEUKOCYTES
LYMPHOCYTES
LYMPHOKINES
MATERIALS
MITOGENS
ORGANIC COMPOUNDS
PROTEINS
SECRETION
SOMATIC CELLS
STEM CELLS
560121* -- Radiation Effects on Cells-- External Source-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CULTURES
CELL PROLIFERATION
CLONING
CONNECTIVE TISSUE CELLS
DOSE-RESPONSE RELATIONSHIPS
GROWTH FACTORS
IRRADIATION
LEUKOCYTES
LYMPHOCYTES
LYMPHOKINES
MATERIALS
MITOGENS
ORGANIC COMPOUNDS
PROTEINS
SECRETION
SOMATIC CELLS
STEM CELLS