OKT3 monoclonal antibody induces production of colony-stimulating factor(s) for granulocytes and macrophages in cultures of human T lymphocytes and adherent cells

E. Platzer, B. Y. Rubin, L. Lu, K. Welte, Hal Broxmeyer, M. A. Moore

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

OKT3 monoclonal antibody (mab) recognizes a membrane antigen asociated with the T cell antigen recognition receptor, and is known to be mitogenic and to induce lymphokine production. Our studies demonstrate the ability of OKT3 mab to induce from cultures of human T lymphocytes supplemented with adherent cells the production of colony-stimulating factor(s) for granulocytes and macrophages (GM-CSF) and interferon-γ (IFN-γ), an inhibitor of clonal growth of hematopoietic progenitor cells. As has been shown for the mitogenic and IFN-γ-inducing activity of OKT3 mab, the induction of GM-CSF release in cultures of T cells is strictly dependent on the presence of adherent cells. However, the concentrations of OKT3 mab required for optimal GM-CSF production (50 ng/ml) were found to be 80-fold higher than those sufficient for maximal IFN-γ production, proliferation, and interleukin 2 production. IFN-γ activity induced by OKT3 mab partially inhibited colony and cluster formation from progenitor cells of granulocytes and macrophages in vitro. Therefore, neutralization of the IFN-γ by monoclonal anti-human-IFN-γ antibody before assay of conditioned medium in bone marrow cultures significantly enhanced the detection of GM-CSF production in response to optimal OKT3 mab concentrations on days 4 through 6 in cultures of T cells supplemented with 15% adherent cells. Highly enriched OKT4+ and OKT8+ T cell subsets co-cultured with adherent cells in the presence of OKT3 mab both produced GM-CSF and IFN-γ and showed similar dose-response curves to OKT3 mab. The requirement for the presence of adherent cells could not be overcome by the addition of purified interleukin 1 or macrophage supernatants. Studies using irreversible inhibitors of DNA (mitomycin C) or protein biosynthesis (emetine-HC1) revealed the necessity of intact DNA synthesis and translation in mononuclear cells to produce GM-CSF in response to OKT3 mab. Loss of GM-CSF production was observed when either adherent cells or T lymphocytes were treated with emetine before co-culture with untreated cells of the other population in the presence of OKT3 mab. In contrast, mitomycin C reduced GM-CSF production significantly when T cells, but not adherent cells, were pretreated. These results suggest that T lymphocytes and adherent cells closely cooperate in the production of GM-CSF induced by OKT3 mab.

Original languageEnglish (US)
Pages (from-to)265-271
Number of pages7
JournalJournal of Immunology
Volume134
Issue number1
StatePublished - 1985
Externally publishedYes

Fingerprint

Muromonab-CD3
Granulocyte-Macrophage Colony-Stimulating Factor
Antibody Formation
T-Lymphocytes
Interferons
Emetine
Mitomycin
Growth Inhibitors
Granulocyte-Macrophage Progenitor Cells
Lymphokines
DNA
Protein Biosynthesis
T-Lymphocyte Subsets
Conditioned Culture Medium
Hematopoietic Stem Cells
Coculture Techniques
T-Cell Antigen Receptor
Interleukin-1
Interleukin-2
Bone Marrow

ASJC Scopus subject areas

  • Immunology

Cite this

OKT3 monoclonal antibody induces production of colony-stimulating factor(s) for granulocytes and macrophages in cultures of human T lymphocytes and adherent cells. / Platzer, E.; Rubin, B. Y.; Lu, L.; Welte, K.; Broxmeyer, Hal; Moore, M. A.

In: Journal of Immunology, Vol. 134, No. 1, 1985, p. 265-271.

Research output: Contribution to journalArticle

@article{872c24bd6db849dc8ddfa0d051d4b83a,
title = "OKT3 monoclonal antibody induces production of colony-stimulating factor(s) for granulocytes and macrophages in cultures of human T lymphocytes and adherent cells",
abstract = "OKT3 monoclonal antibody (mab) recognizes a membrane antigen asociated with the T cell antigen recognition receptor, and is known to be mitogenic and to induce lymphokine production. Our studies demonstrate the ability of OKT3 mab to induce from cultures of human T lymphocytes supplemented with adherent cells the production of colony-stimulating factor(s) for granulocytes and macrophages (GM-CSF) and interferon-γ (IFN-γ), an inhibitor of clonal growth of hematopoietic progenitor cells. As has been shown for the mitogenic and IFN-γ-inducing activity of OKT3 mab, the induction of GM-CSF release in cultures of T cells is strictly dependent on the presence of adherent cells. However, the concentrations of OKT3 mab required for optimal GM-CSF production (50 ng/ml) were found to be 80-fold higher than those sufficient for maximal IFN-γ production, proliferation, and interleukin 2 production. IFN-γ activity induced by OKT3 mab partially inhibited colony and cluster formation from progenitor cells of granulocytes and macrophages in vitro. Therefore, neutralization of the IFN-γ by monoclonal anti-human-IFN-γ antibody before assay of conditioned medium in bone marrow cultures significantly enhanced the detection of GM-CSF production in response to optimal OKT3 mab concentrations on days 4 through 6 in cultures of T cells supplemented with 15{\%} adherent cells. Highly enriched OKT4+ and OKT8+ T cell subsets co-cultured with adherent cells in the presence of OKT3 mab both produced GM-CSF and IFN-γ and showed similar dose-response curves to OKT3 mab. The requirement for the presence of adherent cells could not be overcome by the addition of purified interleukin 1 or macrophage supernatants. Studies using irreversible inhibitors of DNA (mitomycin C) or protein biosynthesis (emetine-HC1) revealed the necessity of intact DNA synthesis and translation in mononuclear cells to produce GM-CSF in response to OKT3 mab. Loss of GM-CSF production was observed when either adherent cells or T lymphocytes were treated with emetine before co-culture with untreated cells of the other population in the presence of OKT3 mab. In contrast, mitomycin C reduced GM-CSF production significantly when T cells, but not adherent cells, were pretreated. These results suggest that T lymphocytes and adherent cells closely cooperate in the production of GM-CSF induced by OKT3 mab.",
author = "E. Platzer and Rubin, {B. Y.} and L. Lu and K. Welte and Hal Broxmeyer and Moore, {M. A.}",
year = "1985",
language = "English (US)",
volume = "134",
pages = "265--271",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "1",

}

TY - JOUR

T1 - OKT3 monoclonal antibody induces production of colony-stimulating factor(s) for granulocytes and macrophages in cultures of human T lymphocytes and adherent cells

AU - Platzer, E.

AU - Rubin, B. Y.

AU - Lu, L.

AU - Welte, K.

AU - Broxmeyer, Hal

AU - Moore, M. A.

PY - 1985

Y1 - 1985

N2 - OKT3 monoclonal antibody (mab) recognizes a membrane antigen asociated with the T cell antigen recognition receptor, and is known to be mitogenic and to induce lymphokine production. Our studies demonstrate the ability of OKT3 mab to induce from cultures of human T lymphocytes supplemented with adherent cells the production of colony-stimulating factor(s) for granulocytes and macrophages (GM-CSF) and interferon-γ (IFN-γ), an inhibitor of clonal growth of hematopoietic progenitor cells. As has been shown for the mitogenic and IFN-γ-inducing activity of OKT3 mab, the induction of GM-CSF release in cultures of T cells is strictly dependent on the presence of adherent cells. However, the concentrations of OKT3 mab required for optimal GM-CSF production (50 ng/ml) were found to be 80-fold higher than those sufficient for maximal IFN-γ production, proliferation, and interleukin 2 production. IFN-γ activity induced by OKT3 mab partially inhibited colony and cluster formation from progenitor cells of granulocytes and macrophages in vitro. Therefore, neutralization of the IFN-γ by monoclonal anti-human-IFN-γ antibody before assay of conditioned medium in bone marrow cultures significantly enhanced the detection of GM-CSF production in response to optimal OKT3 mab concentrations on days 4 through 6 in cultures of T cells supplemented with 15% adherent cells. Highly enriched OKT4+ and OKT8+ T cell subsets co-cultured with adherent cells in the presence of OKT3 mab both produced GM-CSF and IFN-γ and showed similar dose-response curves to OKT3 mab. The requirement for the presence of adherent cells could not be overcome by the addition of purified interleukin 1 or macrophage supernatants. Studies using irreversible inhibitors of DNA (mitomycin C) or protein biosynthesis (emetine-HC1) revealed the necessity of intact DNA synthesis and translation in mononuclear cells to produce GM-CSF in response to OKT3 mab. Loss of GM-CSF production was observed when either adherent cells or T lymphocytes were treated with emetine before co-culture with untreated cells of the other population in the presence of OKT3 mab. In contrast, mitomycin C reduced GM-CSF production significantly when T cells, but not adherent cells, were pretreated. These results suggest that T lymphocytes and adherent cells closely cooperate in the production of GM-CSF induced by OKT3 mab.

AB - OKT3 monoclonal antibody (mab) recognizes a membrane antigen asociated with the T cell antigen recognition receptor, and is known to be mitogenic and to induce lymphokine production. Our studies demonstrate the ability of OKT3 mab to induce from cultures of human T lymphocytes supplemented with adherent cells the production of colony-stimulating factor(s) for granulocytes and macrophages (GM-CSF) and interferon-γ (IFN-γ), an inhibitor of clonal growth of hematopoietic progenitor cells. As has been shown for the mitogenic and IFN-γ-inducing activity of OKT3 mab, the induction of GM-CSF release in cultures of T cells is strictly dependent on the presence of adherent cells. However, the concentrations of OKT3 mab required for optimal GM-CSF production (50 ng/ml) were found to be 80-fold higher than those sufficient for maximal IFN-γ production, proliferation, and interleukin 2 production. IFN-γ activity induced by OKT3 mab partially inhibited colony and cluster formation from progenitor cells of granulocytes and macrophages in vitro. Therefore, neutralization of the IFN-γ by monoclonal anti-human-IFN-γ antibody before assay of conditioned medium in bone marrow cultures significantly enhanced the detection of GM-CSF production in response to optimal OKT3 mab concentrations on days 4 through 6 in cultures of T cells supplemented with 15% adherent cells. Highly enriched OKT4+ and OKT8+ T cell subsets co-cultured with adherent cells in the presence of OKT3 mab both produced GM-CSF and IFN-γ and showed similar dose-response curves to OKT3 mab. The requirement for the presence of adherent cells could not be overcome by the addition of purified interleukin 1 or macrophage supernatants. Studies using irreversible inhibitors of DNA (mitomycin C) or protein biosynthesis (emetine-HC1) revealed the necessity of intact DNA synthesis and translation in mononuclear cells to produce GM-CSF in response to OKT3 mab. Loss of GM-CSF production was observed when either adherent cells or T lymphocytes were treated with emetine before co-culture with untreated cells of the other population in the presence of OKT3 mab. In contrast, mitomycin C reduced GM-CSF production significantly when T cells, but not adherent cells, were pretreated. These results suggest that T lymphocytes and adherent cells closely cooperate in the production of GM-CSF induced by OKT3 mab.

UR - http://www.scopus.com/inward/record.url?scp=0021997402&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021997402&partnerID=8YFLogxK

M3 - Article

C2 - 3917276

AN - SCOPUS:0021997402

VL - 134

SP - 265

EP - 271

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 1

ER -