Macrophage chemotactic factor (MCF) produced by a human T cell hybridoma clone

Yoshizuka, Naonobu; Yoshimura, Masaaki; Tsuchiya, Satoshi; Okamoto, Kikuhiko; Kobayashi, Yoshiro; Osawa, Toshiaki

doi:10.1016/0008-8749(89)90281-5

Cited by 8 publications

(2 citation statements)

References 12 publications

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“…(21)(22)(23)(24)(25)(26)(27)(28)(29)(30) This cell selection method was used to omit HAT cytotoxicity. The F3B6 heteromyeloma cells were cultured at 10 6 streptomycin, 0.1 mM hypoxanthine, 0.016 mM thymidine (all from Sigma, St. Louis, MO), and 10% fetal bovine serum (FBS, Gibco, BRL Life Technologies) (complete RPMI medium) in the presence of 5Â10 À5 M emetine-Cl and 0.25 mg=mL actinomycin D (both from Sigma) for 2 h in a humidified 378C, 5% CO 2 incubator prior to fusion.…”

Section: Pretreatment Of F3b6 Cells With Actinomycin D and Emetinementioning

confidence: 99%

An Extreme Strategy for the Production of Hybridoma

2009

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The ethical issues surrounding human immunization hamper the production of human monoclonal antibody through scarcity of immunized B cells in peripheral blood. This defect can be compensated in part by improvement of hybridoma production techniques. We have developed a new strategy to bypass the toxic effects of polyethylene glycol (PEG) as fusogenic reagent and hypoxanthine aminoptrin thymidine (HAT) as selective medium on newly fused cells. The Epstein-Barr virus (EBV) transformed peripheral blood mononuclear cells (PBMC) of accidentally Rh antigen sensitized persons were fused using cephalin as fugenic reagent, with emetine and actinomycin D pretreated heteromyeloma cells. Our results showed that 19-34% of EBV-transformed B cells were grown as hybridoma clones following selection. This extreme improvement in hybridoma production rate may end the fusion efficiency problem and make hybridoma production a plug-and-play technology.

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Section: Pretreatment Of F3b6 Cells With Actinomycin D and Emetinementioning

confidence: 99%

An Extreme Strategy for the Production of Hybridoma

2009

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“…Macrophages exert various functions in response to a variety of lymphokines (4,5,9,16). Macrophage migration inhibitory factor (MIF) is a lymphokine produced by stimulated lymphocytes and is thought to play an important role in cellular immunity, especially delayed-type hypersensitivity (1,2).…”

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Macrophage Migration Inhibitory Factor (MIF) Produced by a Human T Cell Hybridoma Clone

Hirose¹,

Ooki

Higuchi

et al. 1991

Microbiology and Immunology

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A human T cell hybridoma clone, F5, producing high levels of macrophage migration inhibitory factor (MIF) was established by the emetine-actinomycin D selection method. This clone produced two species of MIF which were separated on a Phenyl Sepharose column. We purified MIF-2 (the more hydrophobic species of the two) to homogeneity from the conditioned medium of stimulated F5 cells by a series of steps that included hydrophobic chromatography, ion-exchange chromatography. Ricinus communis lectin affinity chromatography, and high-performance liquid chromatography on anion exchange and reverse-phase columns. Purified MIF was digested with endoproteinase Lys-C and Asp-N. The amino acid sequences of the generated peptides were determined. No sequence similarity with any other protein was found. The molecular weight of MIF-2 was estimated to be 45 kDa from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of immunoprecipitates with anti-peptide antibodies. These results show that F5MIF-2 is a novel cytokine.

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Estrogen regulation of JE/MCP-1 mRNA expression in fibroblasts

Kovacs

Faunce

Ramer-Quinn

et al. 1996

Journal of Leukocyte Biology

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We have recently demonstrated that 17 beta-estradiol (E2) inhibits peritoneal adhesion formation. Because macrophages play a central role in inflammation and wound healing, we chose to investigate whether the E2 could inhibit the expression of JE, the murine monocyte chemoattractant protein-1 (MCP-1). To accomplish this, murine fibroblasts were cultured with physiological concentrations of E2 (3-300 pg/ml) with or without inducers of JE/MCP-1 mRNA expression. Untreated cells failed to express the message, but, following stimulation, a marked increase in JE/MCP-1 mRNA expression was observed. At 10-30 pg/ml, E2 had no effect on JE/MCP-1 mRNA expression in stimulated fibroblasts. In contrast, lower and higher doses of E2 inhibited the expression of JE/MCP-1 mRNA in stimulated fibroblasts. Treatment with tamoxifen reversed the E2-inhibition of expression of the message. These data demonstrate that JE/MCP-1 mRNA expression is controlled, in part, by estrogen and suggest that macrophage recruitment may be affected by circulating levels of E2.

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Macrophage chemotactic factor (MCF) produced by a human T cell hybridoma clone

Cited by 8 publications

References 12 publications

An Extreme Strategy for the Production of Hybridoma

An Extreme Strategy for the Production of Hybridoma

Macrophage Migration Inhibitory Factor (MIF) Produced by a Human T Cell Hybridoma Clone

Estrogen regulation of JE/MCP-1 mRNA expression in fibroblasts

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