Phenotype of the accessory cell necessary for mitogen-stimulated T and B cell responses in human peripheral blood: Delineation by its sensitivity to the lysosomotropic agent, L-leucine methyl ester

Dwain L Thiele, M. Kurosaka, P. E. Lipsky

Research output: Contribution to journalArticle

308 Citations (Scopus)

Abstract

The lysosomotropic compound L-leucine methyl ester (Leu-OMe) was utilized to delineate the phenotype of the accessory cells involved in human B and T cell activation in vitro. Leu-OMe was shown to cause lysosomal disruption and selective death of human monocytes (Mφ). After 30-45 minute incubations with this agent, human peripheral blood mononuclear cells (PBM) were nearly completely depleted of Mφ. Associated with this Mφ depletion, PBM were rendered unresponsive to a variety of T and B cell mitogens including the plant lectins phytohemagglutinin, concanavalin A, and pokeweed mitogen as well as the oxidative mitogens sodium periodate and neuraminidase plus galactose oxidase. Leu-OMe mediated loss of responsiveness was the result of a selective loss of an accessory cell necessary for each of these responses since reconstitution was accomplished by the addition of an Mφ-enriched adherent cell population. While intact adherent cells could reconstitute responsiveness, crude Mφ supernatants or highly purified human IL 1 alone were ineffective. Further identification of the Leu-OMe sensitive accessory cell indicated that it was entirely contained within the fraction of the adherent population identified by the monoclonal anti-Mφ antibody, 63D3. The mechanism by which Leu-OMe killed Mφ was dependent on the lysosomal content of these cells, since agents that altered lysosomal enzyme activity such as chloroquine or NH4Cl protected Mφ from Leu-OMe. Thus, the selective killing of Mφ by Leu-OMe appeared to relate to the characteristically rich endowment of lysosomes within these cells. These results support the conclusion that a lysosome-rich, leucine methyl ester-sensitive, intact Mφ identified by the monoclonal anti-Mφ antibody 63D3 is the circulating accessory cell required for mitogen-triggered human B and T cell activation.

Original languageEnglish (US)
Pages (from-to)2282-2290
Number of pages9
JournalJournal of Immunology
Volume131
Issue number5
StatePublished - 1983

Fingerprint

Mitogens
Leucine
B-Lymphocytes
T-Lymphocytes
Phenotype
Lysosomes
Anti-Idiotypic Antibodies
Blood Cells
Galactose Oxidase
Monoclonal Antibodies
Plant Lectins
Pokeweed Mitogens
leucine methyl ester
Chloroquine
Neuraminidase
Phytohemagglutinins
Financial Management
Concanavalin A
Interleukin-1
Population

ASJC Scopus subject areas

  • Immunology

Cite this

@article{652dd3b094824d7e8ce1cf81a9fa3e32,
title = "Phenotype of the accessory cell necessary for mitogen-stimulated T and B cell responses in human peripheral blood: Delineation by its sensitivity to the lysosomotropic agent, L-leucine methyl ester",
abstract = "The lysosomotropic compound L-leucine methyl ester (Leu-OMe) was utilized to delineate the phenotype of the accessory cells involved in human B and T cell activation in vitro. Leu-OMe was shown to cause lysosomal disruption and selective death of human monocytes (Mφ). After 30-45 minute incubations with this agent, human peripheral blood mononuclear cells (PBM) were nearly completely depleted of Mφ. Associated with this Mφ depletion, PBM were rendered unresponsive to a variety of T and B cell mitogens including the plant lectins phytohemagglutinin, concanavalin A, and pokeweed mitogen as well as the oxidative mitogens sodium periodate and neuraminidase plus galactose oxidase. Leu-OMe mediated loss of responsiveness was the result of a selective loss of an accessory cell necessary for each of these responses since reconstitution was accomplished by the addition of an Mφ-enriched adherent cell population. While intact adherent cells could reconstitute responsiveness, crude Mφ supernatants or highly purified human IL 1 alone were ineffective. Further identification of the Leu-OMe sensitive accessory cell indicated that it was entirely contained within the fraction of the adherent population identified by the monoclonal anti-Mφ antibody, 63D3. The mechanism by which Leu-OMe killed Mφ was dependent on the lysosomal content of these cells, since agents that altered lysosomal enzyme activity such as chloroquine or NH4Cl protected Mφ from Leu-OMe. Thus, the selective killing of Mφ by Leu-OMe appeared to relate to the characteristically rich endowment of lysosomes within these cells. These results support the conclusion that a lysosome-rich, leucine methyl ester-sensitive, intact Mφ identified by the monoclonal anti-Mφ antibody 63D3 is the circulating accessory cell required for mitogen-triggered human B and T cell activation.",
author = "Thiele, {Dwain L} and M. Kurosaka and Lipsky, {P. E.}",
year = "1983",
language = "English (US)",
volume = "131",
pages = "2282--2290",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "5",

}

TY - JOUR

T1 - Phenotype of the accessory cell necessary for mitogen-stimulated T and B cell responses in human peripheral blood

T2 - Delineation by its sensitivity to the lysosomotropic agent, L-leucine methyl ester

AU - Thiele, Dwain L

AU - Kurosaka, M.

AU - Lipsky, P. E.

PY - 1983

Y1 - 1983

N2 - The lysosomotropic compound L-leucine methyl ester (Leu-OMe) was utilized to delineate the phenotype of the accessory cells involved in human B and T cell activation in vitro. Leu-OMe was shown to cause lysosomal disruption and selective death of human monocytes (Mφ). After 30-45 minute incubations with this agent, human peripheral blood mononuclear cells (PBM) were nearly completely depleted of Mφ. Associated with this Mφ depletion, PBM were rendered unresponsive to a variety of T and B cell mitogens including the plant lectins phytohemagglutinin, concanavalin A, and pokeweed mitogen as well as the oxidative mitogens sodium periodate and neuraminidase plus galactose oxidase. Leu-OMe mediated loss of responsiveness was the result of a selective loss of an accessory cell necessary for each of these responses since reconstitution was accomplished by the addition of an Mφ-enriched adherent cell population. While intact adherent cells could reconstitute responsiveness, crude Mφ supernatants or highly purified human IL 1 alone were ineffective. Further identification of the Leu-OMe sensitive accessory cell indicated that it was entirely contained within the fraction of the adherent population identified by the monoclonal anti-Mφ antibody, 63D3. The mechanism by which Leu-OMe killed Mφ was dependent on the lysosomal content of these cells, since agents that altered lysosomal enzyme activity such as chloroquine or NH4Cl protected Mφ from Leu-OMe. Thus, the selective killing of Mφ by Leu-OMe appeared to relate to the characteristically rich endowment of lysosomes within these cells. These results support the conclusion that a lysosome-rich, leucine methyl ester-sensitive, intact Mφ identified by the monoclonal anti-Mφ antibody 63D3 is the circulating accessory cell required for mitogen-triggered human B and T cell activation.

AB - The lysosomotropic compound L-leucine methyl ester (Leu-OMe) was utilized to delineate the phenotype of the accessory cells involved in human B and T cell activation in vitro. Leu-OMe was shown to cause lysosomal disruption and selective death of human monocytes (Mφ). After 30-45 minute incubations with this agent, human peripheral blood mononuclear cells (PBM) were nearly completely depleted of Mφ. Associated with this Mφ depletion, PBM were rendered unresponsive to a variety of T and B cell mitogens including the plant lectins phytohemagglutinin, concanavalin A, and pokeweed mitogen as well as the oxidative mitogens sodium periodate and neuraminidase plus galactose oxidase. Leu-OMe mediated loss of responsiveness was the result of a selective loss of an accessory cell necessary for each of these responses since reconstitution was accomplished by the addition of an Mφ-enriched adherent cell population. While intact adherent cells could reconstitute responsiveness, crude Mφ supernatants or highly purified human IL 1 alone were ineffective. Further identification of the Leu-OMe sensitive accessory cell indicated that it was entirely contained within the fraction of the adherent population identified by the monoclonal anti-Mφ antibody, 63D3. The mechanism by which Leu-OMe killed Mφ was dependent on the lysosomal content of these cells, since agents that altered lysosomal enzyme activity such as chloroquine or NH4Cl protected Mφ from Leu-OMe. Thus, the selective killing of Mφ by Leu-OMe appeared to relate to the characteristically rich endowment of lysosomes within these cells. These results support the conclusion that a lysosome-rich, leucine methyl ester-sensitive, intact Mφ identified by the monoclonal anti-Mφ antibody 63D3 is the circulating accessory cell required for mitogen-triggered human B and T cell activation.

UR - http://www.scopus.com/inward/record.url?scp=0021039984&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021039984&partnerID=8YFLogxK

M3 - Article

C2 - 6605384

AN - SCOPUS:0021039984

VL - 131

SP - 2282

EP - 2290

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 5

ER -