Nerve growth factor is preformed in and activates human peripheral blood eosinophils,☆☆,,★★

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Abstract

Background: Recent studies have shown that nerve growth factor (NGF) is produced by and can act on several immune-inflammatory cells.

Objectives: The objective of this study was to study the effects of NGF on human peripheral blood eosinophils and assess whether these cells produce and store NGF.

Methods: Eosinophils were purified by negative immunoselection (magnetic cell sorting systems, purity 98% to 100%) from 13 subjects (9 to 26 years old) with mild blood eosinophilia, mainly of allergic origin. Eosinophils were incubated with NGF (50 to 1000 ng/mL), and supernatants were collected for measurement of eosinophil peroxidase (EPO, 20 minutes, colorimetric enzymatic assay) and IL-6 (12 hours, ELISA). Eosinophil viability was evaluated by Trypan blue test (days 2, 3, and 4). NGF content in freshly isolated eosinophils, after ultrasound disruption, was determined with a 2-site immunoenzymatic assay. The presence of mRNA for NGF was evaluated by reverse transcription PCR.

Results: NGF caused EPO release (highly significant at 1000 ng/mL NGF). IL-6 release from eosinophils was not higher than IL-6 spontaneously released into culture medium alone. NGF did not significantly affect the number of viable eosinophils. NGF was found in the eosinophil sonicates (1.5 to 17.8 pg/mL per 106 cells). Similarly, mRNA for NGF was detected by reverse transcription PCR in the freshly isolated eosinophils.

Conclusions: NGF activates human peripheral blood eosinophils from subjects with mild eosinophilia to selectively release inflammatory mediators. Eosinophils store and produce NGF. Therefore the capability of NGF to induce a secretory response and its production and storage by circulating human eosinophils suggest a possible role for NGF in conditions associated with eosinophilia, including allergic disease. (J Allergy Clin Immunol 1998;102:454-60.)

Section snippets

Subjects

Thirteen subjects (mean age, 18 years; range, 9 to 26 years) were studied. All subjects had mild blood eosinophilia (range, 4% to 10%). The majority of subjects had signs and symptoms of allergic disease: 5 patients had vernal keratoconjunctivitis, 3 patients had allergic rhinitis, 2 patients had allergic conjunctivitis, and 1 patient had mild bronchial asthma. Two subjects were asymptomatic, and the cause for their eosinophilia could not be attributed to any specific morbid condition by common

Effect of NGF on EPO release

Twenty-minute incubation of eosinophils with various concentrations of NGF caused a dose-dependent EPO release compared with incubation of eosinophils in medium alone (Fig 1).

. EPO release from eosinophils cultured with NGF. EPO levels were assessed in supernatants of isolated eosinophils cultured for 20 minutes with different NGF concentrations by enzymatic colorimetric assay. Data are mean ± SEM of experiments performed with eosinophils isolated from 13 subjects with mild eosinophilia. Assays

DISCUSSION

Attention and interest have recently focused on the role NGF plays in allergic and immunologic diseases.10, 11, 12 Several studies indicate that NGF can affect various immune and inflammatory cells through specific receptors present on their membranes.7, 8, 9 On the other hand, NGF is also produced and stored by immune cells such as mast cells and CD4+ cells.8, 9, 10, 13

In this report we show that NGF can induce circulating human eosinophils to release EPO, an important proinflammatory

Acknowledgements

We thank Mrs Dalia Pickholz for skillful technical assistance.

References (35)

  • M Yanagida et al.

    Effects of T-helper 2-type cytokines interleukin 3 (IL-3), IL-4, IL-5 and IL-6 on the survival of cultured human mast cells

    Blood

    (1995)
  • K Kawamoto et al.

    Nerve growth factor prevents apoptosis of rat peritoneal mast cells through the Trk proto-oncogene receptor

    Blood

    (1995)
  • P Callissano et al.

    The nerve growth factor (NGF).

  • H Thoenen et al.

    Neurotrophic factors

    Science

    (1995)
  • L Aloe et al.

    The expanding role of nerve growth factor: from neurotrophic activity to immunologic diseases

    Allergy

    (1997)
  • JS Marshal et al.

    The role of mast cell degranulation products in mast cell hyperplasia, I: mechanism of action of nerve growth factor

    J Immunol

    (1990)
  • H Matsuda et al.

    Nerve growth factor induces development of connective tissue-type mast cells in vitro from murine bone marrow cells

    J Exp Med

    (1991)
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    From athe Department of Ophthalmology, Hadassah University Hospital, and bthe Department of Pharmacology, School of Pharmacy, Hebrew University–Hadassah Medical School, Jerusalem; cInstitute of Neurobiology, Consiglio Nazionale delle Ricerche (CNR); dthe Department of Ophthalmology, University Tor Vergata–G.B. Bietti Eye Foundation, and eInstitute of Experimental Medicine CNR, Rome; and fClinical Immunology and Allergology, Second University of Naples.

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    Supported by a grant from The Joint Research Fund of The Hebrew University and Hadassah University Hospital, Jerusalem, Israel.

    Reprint requests: Francesca Levi-Schaffer, PhD, Department of Pharmacology, School of Pharmacy, The Hebrew University–Hadassah Medical School, POB 12065, Jerusalem 91120, Israel.

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