Characterization of the requirements for human T cell mitogenesis by using suboptimal concentrations of phytohemagglutinin

Gordon Mills, J. W W Lee, R. K. Cheung, E. W. Gelfand

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

To characterize the requirements for T cell proliferation, we have studied the response of purified populations of human T cells to varying concentrations of the mitogen phytohemagglutinin (PHA). Concentrations of PHA which induce optimal proliferative responses induce increases in cytosolic free calcium ([Ca2+](i)), expression of interleukin 2 (IL 2) receptors, and production of IL 2. As the concentration of PHA is decreased, each of these processes decreases in parallel. At suboptimal concentrations of PHA, the addition of exogenous IL 2 reconstitutes both the proliferative response and the expression of the IL 2 receptor, as measured by immunofluorescence with antibodies directed against the TAC/IL 2 receptor molecule, but without reconstituting the increase in [Ca2+](i). Therefore, the concentration dependence of responses to PHA appears to be secondary to an absence of IL 2 production due to a failure to induce an increase in [Ca2+](i). The addition of the calcium ionophores A23187 and ionomycin or of accessory cells to low concentrations of PHA induces increases in [Ca2+](i) and subsequent proliferative responses, suggesting that the two events are linked. The proliferative response can be inhibited by antibodies directed towards IL 2 or the IL 2 receptor, indicating that the proliferative response was at least partially dependent on the production and action of IL 2. This suggests that, although increases in [Ca2+](i) are an integral event in the induction of proliferation by PHA, the increase in [Ca2+](i) is required for the production but not the action of IL 2. In addition, low concentrations of PHA deliver an additional signal to cells, independent of an increase in [Ca2+](i), which induces IL 2 receptor expression and allows a proliferative response in the presence of exogenous IL 2.

Original languageEnglish (US)
Pages (from-to)3087-3093
Number of pages7
JournalJournal of Immunology
Volume135
Issue number5
StatePublished - Dec 1 1985
Externally publishedYes

Fingerprint

Phytohemagglutinins
Interleukin-2
Interleukin-2 Receptors
T-Lymphocytes
Ionomycin
Calcium Ionophores
Antibodies
Calcimycin
Mitogens
Fluorescent Antibody Technique
Cell Proliferation
Calcium
Population

ASJC Scopus subject areas

  • Immunology

Cite this

Characterization of the requirements for human T cell mitogenesis by using suboptimal concentrations of phytohemagglutinin. / Mills, Gordon; Lee, J. W W; Cheung, R. K.; Gelfand, E. W.

In: Journal of Immunology, Vol. 135, No. 5, 01.12.1985, p. 3087-3093.

Research output: Contribution to journalArticle

@article{4680b3c7b22c49a2bed7e4a2e52f583f,
title = "Characterization of the requirements for human T cell mitogenesis by using suboptimal concentrations of phytohemagglutinin",
abstract = "To characterize the requirements for T cell proliferation, we have studied the response of purified populations of human T cells to varying concentrations of the mitogen phytohemagglutinin (PHA). Concentrations of PHA which induce optimal proliferative responses induce increases in cytosolic free calcium ([Ca2+](i)), expression of interleukin 2 (IL 2) receptors, and production of IL 2. As the concentration of PHA is decreased, each of these processes decreases in parallel. At suboptimal concentrations of PHA, the addition of exogenous IL 2 reconstitutes both the proliferative response and the expression of the IL 2 receptor, as measured by immunofluorescence with antibodies directed against the TAC/IL 2 receptor molecule, but without reconstituting the increase in [Ca2+](i). Therefore, the concentration dependence of responses to PHA appears to be secondary to an absence of IL 2 production due to a failure to induce an increase in [Ca2+](i). The addition of the calcium ionophores A23187 and ionomycin or of accessory cells to low concentrations of PHA induces increases in [Ca2+](i) and subsequent proliferative responses, suggesting that the two events are linked. The proliferative response can be inhibited by antibodies directed towards IL 2 or the IL 2 receptor, indicating that the proliferative response was at least partially dependent on the production and action of IL 2. This suggests that, although increases in [Ca2+](i) are an integral event in the induction of proliferation by PHA, the increase in [Ca2+](i) is required for the production but not the action of IL 2. In addition, low concentrations of PHA deliver an additional signal to cells, independent of an increase in [Ca2+](i), which induces IL 2 receptor expression and allows a proliferative response in the presence of exogenous IL 2.",
author = "Gordon Mills and Lee, {J. W W} and Cheung, {R. K.} and Gelfand, {E. W.}",
year = "1985",
month = "12",
day = "1",
language = "English (US)",
volume = "135",
pages = "3087--3093",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "5",

}

TY - JOUR

T1 - Characterization of the requirements for human T cell mitogenesis by using suboptimal concentrations of phytohemagglutinin

AU - Mills, Gordon

AU - Lee, J. W W

AU - Cheung, R. K.

AU - Gelfand, E. W.

PY - 1985/12/1

Y1 - 1985/12/1

N2 - To characterize the requirements for T cell proliferation, we have studied the response of purified populations of human T cells to varying concentrations of the mitogen phytohemagglutinin (PHA). Concentrations of PHA which induce optimal proliferative responses induce increases in cytosolic free calcium ([Ca2+](i)), expression of interleukin 2 (IL 2) receptors, and production of IL 2. As the concentration of PHA is decreased, each of these processes decreases in parallel. At suboptimal concentrations of PHA, the addition of exogenous IL 2 reconstitutes both the proliferative response and the expression of the IL 2 receptor, as measured by immunofluorescence with antibodies directed against the TAC/IL 2 receptor molecule, but without reconstituting the increase in [Ca2+](i). Therefore, the concentration dependence of responses to PHA appears to be secondary to an absence of IL 2 production due to a failure to induce an increase in [Ca2+](i). The addition of the calcium ionophores A23187 and ionomycin or of accessory cells to low concentrations of PHA induces increases in [Ca2+](i) and subsequent proliferative responses, suggesting that the two events are linked. The proliferative response can be inhibited by antibodies directed towards IL 2 or the IL 2 receptor, indicating that the proliferative response was at least partially dependent on the production and action of IL 2. This suggests that, although increases in [Ca2+](i) are an integral event in the induction of proliferation by PHA, the increase in [Ca2+](i) is required for the production but not the action of IL 2. In addition, low concentrations of PHA deliver an additional signal to cells, independent of an increase in [Ca2+](i), which induces IL 2 receptor expression and allows a proliferative response in the presence of exogenous IL 2.

AB - To characterize the requirements for T cell proliferation, we have studied the response of purified populations of human T cells to varying concentrations of the mitogen phytohemagglutinin (PHA). Concentrations of PHA which induce optimal proliferative responses induce increases in cytosolic free calcium ([Ca2+](i)), expression of interleukin 2 (IL 2) receptors, and production of IL 2. As the concentration of PHA is decreased, each of these processes decreases in parallel. At suboptimal concentrations of PHA, the addition of exogenous IL 2 reconstitutes both the proliferative response and the expression of the IL 2 receptor, as measured by immunofluorescence with antibodies directed against the TAC/IL 2 receptor molecule, but without reconstituting the increase in [Ca2+](i). Therefore, the concentration dependence of responses to PHA appears to be secondary to an absence of IL 2 production due to a failure to induce an increase in [Ca2+](i). The addition of the calcium ionophores A23187 and ionomycin or of accessory cells to low concentrations of PHA induces increases in [Ca2+](i) and subsequent proliferative responses, suggesting that the two events are linked. The proliferative response can be inhibited by antibodies directed towards IL 2 or the IL 2 receptor, indicating that the proliferative response was at least partially dependent on the production and action of IL 2. This suggests that, although increases in [Ca2+](i) are an integral event in the induction of proliferation by PHA, the increase in [Ca2+](i) is required for the production but not the action of IL 2. In addition, low concentrations of PHA deliver an additional signal to cells, independent of an increase in [Ca2+](i), which induces IL 2 receptor expression and allows a proliferative response in the presence of exogenous IL 2.

UR - http://www.scopus.com/inward/record.url?scp=0022202935&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022202935&partnerID=8YFLogxK

M3 - Article

C2 - 2995486

AN - SCOPUS:0022202935

VL - 135

SP - 3087

EP - 3093

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 5

ER -