IngentaConnect In vitro expansion of human basophils by interleukin-3 from granu...

Authors: Takao K.¹; Tanimoto Y.¹; Fujii M.¹; Hamada N.¹; Yoshida I.¹; Ikeda K.¹; Imajo K.²; Takahashi K.³; Harada M.¹; Tanimoto M.¹

Source: Clinical & Experimental Allergy, Volume 33, Number 11, November 2003 , pp. 1561-1567(7)

Publisher: Blackwell Publishing

Abstract:

Summary Background

A number of studies support the belief that human basophils play an important role in allergic inflammation. The exact mechanism of basophil activation at the site of allergic inflammation, however, has not been well understood, mainly due to their low number in blood and difficulty in obtaining a sufficient number of highly purified basophils for investigation. Objective

The purpose of this study is to expand human basophils in vitro with high yield and purity by culturing peripheral blood stem cells (PBSCs). Methods

We collected PBSC-rich mononuclear cells containing CD34⁺ cells (0.15–4.9%) by leukapheresis from patients with malignant lymphoma and lung cancer during haematopoietic recovery after chemotherapy plus granulocyte colony-stimulating factor-induced mobilization. PBSC-rich mononuclear cells were cultured in the presence of IL-3. Results

When PBSC-rich mononuclear cells containing more than 1% of CD34⁺ cells were cultured, 20.0–83.3% of the cells, mostly with a yield of >10%, were metachromatic cells after 3 weeks of culture. These cells resembled mature peripheral blood basophils morphologically when examined by light and electron microscopy. Flow cytometric analysis showed that they expressed both Fc epsiv

RI and Fc

RII. Fc

RI cross-linking resulted in intracellular calcium mobilization, histamine release and synthesis of cysteinyl leukotrienes. The intracellular histamine content and the release of these chemical mediators triggered by anti-IgE antibodies were comparable to those of peripheral blood basophils. Conclusion

These findings suggest that PBSC-derived basophils expanded in vitro are morphologically and functionally mature and will be a useful tool for the analysis of basophil functions.

Keywords: Fc receptor; human basophil; interleukin-3; peripheral blood stem cell

Document Type: Research article

DOI: 10.1046/j.1365-2222.2003.01791.x

Affiliations: 1: Department of Internal Medicine II, Okayama University Medical School, 2: Department of Internal Medicine, Okayama Municipal Hospital and 3: Department of Internal Medicine, National Minami-Okayama Hospital, Okayama, Japan

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In vitro expansion of human basophils by interleukin-3 from granulocyte colony-stimulating factor-mobilized peripheral blood stem cells