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* Servicio Regional de Inmunología y Alergia, Hospital Universitario Virgen Macarena, Sevilla, Spain;
Fundación IMABIS, Laboratorio de Investigación, Hospital Civil, Málaga, Spain;
Bial-Aristegui, Research and Development Department, Bilbao, Spain; and
Departamento de Medicina, Universidad de Sevilla, Sevilla, Spain
The production of eosinophil cationic protein (ECP) in IgE-mediated diseases has been associated mainly with eosinophils, although no IgE-dependent ECP release has been observed in these cells. Because there is increasing evidence of neutrophil participation in allergic processes, we have examined whether human neutrophils from allergic patients were able to produce ECP by an IgE-dependent mechanism. After challenge with specific Ags to which the patients were sensitized, ECP release was detected in the culture medium. Furthermore, intracellular protein was detected by flow cytometry, immunofluorescence staining, and Western blotting. Expression at both mRNA and de novo protein synthesis were detected, respectively, by RT-PCR and radiolabeling with 35S. Ag effect was mimicked by cell treatment with anti-IgE Abs or Abs against Fc
RI and galectin-3 (FcRI>galectin-3), but not against FcRII. These observations represent a novel view of neutrophils as possible source of ECP in IgE-dependent diseases.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This work was supported by grants from the Junta de Andalucia (Ayudas Grupos de Investigación), Fundación de la Sociedad Española de Alergia e Inmunología Clínica, Fundación Sanitaria Virgen Macarena, and Fundación Alergol, Spain. A.V. and P.C. were supported by fellowships from the Ministerio de Ciencia y Tecnología, and Fundación Alergol, Spain. R.E. is a recipient of a postdoctoral grant (Juan de la Cierva) (SAF2003-00200) from the Ministerio de Educación y Ciencia, Spain. J.M. is under the Programa de Intensificación de la Actividad Investigadora del Sistema Nacional de Salud.
2 Address correspondence and reprint requests to Dr. Javier Monteseirín, Servicio Regional de Inmunología y Alergia, Hospital Universitario Virgen Macarena, Sevilla, Spain, c/Asunción 27, 3° Izda. E-41011, Sevilla, Spain. E-mail address: fmonteserinm{at}meditex.es
3 J.M., A.V., and P.C. contributed equally to this work.
4 Abbreviations used in this paper: ECP, eosinophil cationic protein; PVDF, polyvinylidene difluoride; CLC, Charcot-Leyden crystal protein; PAF, platelet-activating factor.
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