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

K. Takao, Y. Tanimoto, M. Fujii, N. Hamada, I. Yoshida, K. Ikeda, K. Imajo, K. Takahashi, M. Harada, M. Tanimoto

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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 FcεRI and FcγRII. FcεRI 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.

Original languageEnglish
Pages (from-to)1561-1567
Number of pages7
JournalClinical and Experimental Allergy
Volume33
Issue number11
DOIs
Publication statusPublished - Nov 2003

Fingerprint

Basophils
Interleukin-3
Granulocyte Colony-Stimulating Factor
Histamine Release
Cultured Cells
Inflammation
Leukapheresis
Peripheral Blood Stem Cells
In Vitro Techniques
Lymphoma
Lung Neoplasms
Electron Microscopy
Calcium
Light
Drug Therapy

Keywords

  • Fc receptor
  • Human basophil
  • Interleukin-3
  • Peripheral blood stem cell

ASJC Scopus subject areas

  • Immunology

Cite this

In vitro expansion of human basophils by interleukin-3 from granulocyte colony-stimulating factor-mobilized peripheral blood stem cells. / Takao, K.; Tanimoto, Y.; Fujii, M.; Hamada, N.; Yoshida, I.; Ikeda, K.; Imajo, K.; Takahashi, K.; Harada, M.; Tanimoto, M.

In: Clinical and Experimental Allergy, Vol. 33, No. 11, 11.2003, p. 1561-1567.

Research output: Contribution to journalArticle

Takao, K, Tanimoto, Y, Fujii, M, Hamada, N, Yoshida, I, Ikeda, K, Imajo, K, Takahashi, K, Harada, M & Tanimoto, M 2003, 'In vitro expansion of human basophils by interleukin-3 from granulocyte colony-stimulating factor-mobilized peripheral blood stem cells', Clinical and Experimental Allergy, vol. 33, no. 11, pp. 1561-1567. https://doi.org/10.1046/j.1365-2222.2003.01791.x
Takao, K. ; Tanimoto, Y. ; Fujii, M. ; Hamada, N. ; Yoshida, I. ; Ikeda, K. ; Imajo, K. ; Takahashi, K. ; Harada, M. ; Tanimoto, M. / In vitro expansion of human basophils by interleukin-3 from granulocyte colony-stimulating factor-mobilized peripheral blood stem cells. In: Clinical and Experimental Allergy. 2003 ; Vol. 33, No. 11. pp. 1561-1567.
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AU - Takao, K.

AU - Tanimoto, Y.

AU - Fujii, M.

AU - Hamada, N.

AU - Yoshida, I.

AU - Ikeda, K.

AU - Imajo, K.

AU - Takahashi, K.

AU - Harada, M.

AU - Tanimoto, M.

PY - 2003/11

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N2 - 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 FcεRI and FcγRII. FcεRI 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.

AB - 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 FcεRI and FcγRII. FcεRI 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.

KW - Fc receptor

KW - Human basophil

KW - Interleukin-3

KW - Peripheral blood stem cell

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