Bcl-2 expression by eosinophils in a patient with hypereosinophilia☆,☆☆,★,★★,♢,♢♢
Section snippets
CASE REPORT
A 61-year-old woman had recurrent bilateral periorbital swelling and intermittent urticaria with annular erythematous skin lesions for more than 8 years. The laboratory findings included a marked persistent blood eosinophilia. The absolute eosinophil numbers in blood during the time of these investigations ranged between 2079/mm3 (without glucocorticosteroid therapy) and 600/mm3 (with glucocorticosteroid therapy). Recently, diarrhea, nausea, and vomiting occurred sporadically, and the patient
Antibodies
Anti-Bcl-2 and control IgG1 mAb were obtained from Dako (Zurich, Switzerland); FITC-conjugated anti-Bcl-2 mAb from Ancell Corp (Bayport, Minn); FITC-conjugated control IgG1 mAb from Coulter (Hialeah, Fla); anti-Fas receptor mAb (clone CH-11, IgM) from Immunotech (Marseille, Fance); and anti-CD16 mAb microbeads from Miltenyi Biotec (Bergisch-Gladbach, Germany).
Eosinophil purification and cultures
Eosinophils were purified as previously described.3, 4 Eosinophils were cultured at 1 × 106 /mL for the indicated times with complete
RT-PCR
mRNA expression of Bcl-2 and Fas receptor was studied by using RT-PCR. Primers for Bcl-2 (5 ́-ACA ACA TCG CCC TGT GGA TGA C-3 ́ and 5 ́-ATA GCT GAT TCG ACG TTT TGC C-3 ́) and Fas receptor (5 ́-GTG GGA TCC CAC TTC GGA GGA TTG CTC AAC AAC C-3 ́ and 5 ́-GTG CTG CAG TAT GTT GGC TCT TCA GCG CTA ATA-3 ́) amplifications were synthesized (HSC Biotechnology Service Centre, Toronto, Ontario) according to previously published sequences.3, 4, 5 Primers for β-actin control amplification were obtained from
RESULTS AND DISCUSSION
As demonstrated in Fig 1 (left panel), the purified eosinophils of the presented patient with HES demonstrated, compared with normal control eosinophils, a marked delay of eosinophil death in vitro.
Acknowledgements
We thank Martina Weber for excellent technical assistance.
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Cited by (0)
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From a the Department of Dermatology, Technical University Munich, Munich; and b Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos.
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*S. G. P. and B. D. contributed equally to this work.
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Supported by grants from the Swiss National Science Foundation (32-49210.96); the OPO-Foundation, Zurich; and the Foundation for Scientific Research, University of Zurich.
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Reprint requests: Hans-Uwe Simon, MD, Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Obere Strasse 22, CH-7270 Davos, Switzerland.
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J Allergy Clin Immunol 1998;102:1037-40.
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