Neutrophil chemotactic factors produced by malignant fibrous histiocytoma cell lines

Abstract

(Tani et al., 1990).The infiltration of monocytes-macrophage into the tumour is a typical feature of MFH (Weiss & Enzinger, 1978). Among the several active cytokines secreted by infiltrating macrophages, IL-1 and tumour necrosis factor (TNF) are likely to be an important in inflammation and tissue damage (Oppenheim et al., 1986;Le & Vilcek, 1987). Stimulation by exogenious IL-1 and TNF can induce promotion and modification of production of GM-CSF and IL-8/NAP-1 in various cells (Munker et al., 1986;Zucali et al., 1986;Matsushima et al., 1988;Strieter et al., 1989;Brennan et al., 1990;Seitz et al., 1991;Zoja et al., 1991; Meir et al., 1992).In this paper, we examined neutrophil chemotactic factor production by MFH cell lines and their expression of GM-CSF and IL-8/NAP-1 mRNA with or without IL-1p stimulation. Materials and methods Cell linesThe three human MFH cell lines (MF-1, MF-3, MF-4) were established by H. Okabe (Okabe et al., 1987;Takeya et al., 1991) and MF-SH was the generous gift of Dr K. Shirasuna (Shirasuna et al., 1985). The Human thyroid carcinoma cell line, KHM-5M (Yoshida et al., 1992) was used as a positive control for GM-CSF and IL-8/NAP-1 expression, and a human multiple myeloma cell line, KHM-7 established in our laboratory was used as the negative control.Culture conditions Cells were grown to confluence in 25 mm2 flasks at 37'C in humidified 95% air/5% C02, using RPMI 1640 medium containing 10% fetal bovine serum (FBS) as the culture medium. On the day of use cells were washed with RPMI1640 medium and incubated with IL-1p in RPMI1640 medium containing 0.1% bovine serum albumin (BSA) for the specified times and doses. Cell-free supernatants were harvested, dialysed with Dulbecco's calcium-and magnesium-free phosphate-buffered saline (CMF-PBS) (pH. 7.4), and then tested for neutrophil chemotactic activity. Total cellular RNA was extracted from 1-2 x 107 cells and analysed as described below. Reagent preparationHuman IL-1p, with a specific activity of 2 x 107 UMmg', was a generous gift of Otsuka Parmaceutical Co. Ltd. (Tokyo, Japan). Dilutions of this cytokine were prepared in CMF-PBS with 0.1% BSA. Chemotaxis assayHeparinised human venous blood from healthy volunteers was layered on to Ficoll-Conray (specific gravity 1.078) and centrifuged at 400 g for 30 min. Polymorphonuclear cell (PMN)-rich pellets were collected and the contaminating erythrocytes were lysed by treatment with 0.85% (w/v) NH4Cl (pH 7.0) for 5 min at room temperature. The PMN were then washed three times and suspended in RPMI1640 medium at 1 x 106 cellsml-'. PMN contained more than 92% neutrophils and 5% eosinophils, and the viability was confirmed to be more than 98% by trypan blue dye exclusion.Neutrophil chemotaxis activity experiments were set up in multiwell chemotasis chambers (Neuro Probe. Inc., Bethesda, MD) (Harvath et al., 1980). Briefly, 25 gl aliquots of specimens in CMF-PBS with 0.1% BSA were placed into triplet wells. A polycarbonate filter with 3 ytm pores (polyvinylpyrrolidone-free, Nucleopore Corp., Ple...

Origin of histiocyte‐like cells and multinucleated giant cells in malignant fibrous histiocytoma: Neoplastic or reactive?

“…Previous studies have shown that MFH produce various bioactive factors, such as monocyte chemoattractant protein‐1, CSF‐1, granulocyte–macrophage CSF, granulocyte CSF, tumor necrosis factor‐α and interleukin‐8 40 –42 . We identified the expression of the human CSF‐1 mRNA and intense positive signals for mouse c‐fms (receptor of CSF‐1) mRNA on the histiocyte‐like cells using ISH.…”

“…Previous studies have shown that MFH produce various bioactive factors, such as monocyte chemoattractant protein-1, CSF-1, granulocyte-macrophage CSF, granulocyte CSF, tumor necrosis factor-α and interleukin-8. [40][41][42] We identified the expression of the human CSF-1 mRNA and intense positive signals for mouse c-fms (receptor of CSF-1) mRNA on the histiocyte-like cells using ISH. Among several macrophagic growth factors, CSF-1 plays an important role in the development, differentiation, proliferation and function of macrophages.…”

Interleukin-6/Soluble Interleukin-6 Receptor Signaling Attenuates Proliferation and Invasion, and Induces Morphological Changes of a Newly Established Pleomorphic Malignant Fibrous Histiocytoma Cell Line

“…Indeed, MIPS-2 cells were also found to overproduce hematopoietic growth factor IL-8 (data not shown). Yoshida and colleagues 44 considered chemoat- tractants and/or differentiation factors such as CSF-1 produced by MFH to stimulate proliferation and infiltration of the host monocyte/macrophage lineage. Takeya and colleagues 45 suggested that histiocyte-like cells comprising MFH tumors are the infiltrated macrophages that originate from blood monocytes attracted by tumor-derived MCP-1.…”

Malignant Fibrous Histiocytoma and Macrophage Differentiation.

“…We also demonstrated the production by these cells of monocyte chemoattractant protein-1, a potent monocyte-specific chemotactic factor (32). In addition, we recently detected the production of granulocyte-macrophage colony-stimulating factor (MG-CSF) which also possesses the chemotactic acitivity for monocytes (58) by MFH cell lines (56). Since GM-CSF can induce proliferation, survival, and differentiation of monocyte/macrophages (7,12,13,35), it may sustain and induce differentiation of infiltrated macrophages.…”