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Title: Estimation by limiting dilution analysis of human IL 2-secreting T cells: detection of IL 2 produced by single lymphokine-secreting T cells

Abstract

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 providemore » independent estimates of responding cell number and of lymphokine production per cell in a variety of clinical situations.« less

Authors:
;
Publication Date:
Research Org.:
Boston Univ. School of Medicine, MA
OSTI Identifier:
5805384
Resource Type:
Journal Article
Journal Name:
J. Immunol.; (United States)
Additional Journal Information:
Journal Volume: 9
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; 59 BASIC BIOLOGICAL SCIENCES; LYMPHOCYTES; CELL PROLIFERATION; LYMPHOKINES; SECRETION; MITOGENS; BIOLOGICAL EFFECTS; BLOOD; CELL CULTURES; CLONING; DOSE-RESPONSE RELATIONSHIPS; IRRADIATION; STEM CELLS; ANIMAL CELLS; BIOLOGICAL MATERIALS; BLOOD CELLS; BODY FLUIDS; CONNECTIVE TISSUE CELLS; GROWTH FACTORS; LEUKOCYTES; MATERIALS; ORGANIC COMPOUNDS; PROTEINS; SOMATIC CELLS; 560121* - Radiation Effects on Cells- External Source- (-1987); 551001 - Physiological Systems- Tracer Techniques

Citation Formats

Vie, H, and Miller, R A. Estimation by limiting dilution analysis of human IL 2-secreting T cells: detection of IL 2 produced by single lymphokine-secreting T cells. United States: N. p., 1986. Web.
Vie, H, & Miller, R A. Estimation by limiting dilution analysis of human IL 2-secreting T cells: detection of IL 2 produced by single lymphokine-secreting T cells. United States.
Vie, H, and Miller, R A. 1986. "Estimation by limiting dilution analysis of human IL 2-secreting T cells: detection of IL 2 produced by single lymphokine-secreting T cells". United States.
@article{osti_5805384,
title = {Estimation by limiting dilution analysis of human IL 2-secreting T cells: detection of IL 2 produced by single lymphokine-secreting T cells},
author = {Vie, H and Miller, R A},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/5805384}, journal = {J. Immunol.; (United States)},
number = ,
volume = 9,
place = {United States},
year = {Thu May 01 00:00:00 EDT 1986},
month = {Thu May 01 00:00:00 EDT 1986}
}