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IL-3-dependent growth of basophil-like cells and mastlike cells from human bone marrow
A S Kirshenbaum1, J P Goff, S C Dreskin
1Mast Cell Physiology Section, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Abstract:
Human bone marrow cultured in the presence of human rIL-3 has been reported to give rise to basophils. In contrast, mouse bone marrow, cultured in the presence of mouse IL-3, leads to the growth of mast cells. To determine if human rIL-3 might also stimulate the growth of human mast cells, we cultured human bone marrow in the presence of human rIL-3 in suspension cultures, methylcellulose, and in "interphase" cultures where cells are layered over agar. The presence of mast cells was determined using a variety of histochemical techniques. In agreement with previous reports, basophil-like cells were identified in all culture systems. Mastlike cells were identified only in interphase cultures. By 3 wk, such cultures consisted of basophil-like cells (20 to 50%) and mastlike cells (1 to 5%). Cultures supplemented with rIL-4 showed no additional increase in basophil-like and mastlike cells. Both basophil-like and mastlike cells fluoresced with o-phthaldialdehyde and exhibited IgE receptors. Unlike basophil-like cells, mastlike cells were chloroacetate esterase, amidase, and human mast cell tryptase positive. We conclude that human rIL-3 can support the growth of human mastlike cells under selected culture conditions.
Insights
Human recombinant interleukin-3 (rIL-3) can induce human mast cell growth in specific culture conditions. This finding contrasts with previous studies suggesting rIL-3 primarily generates basophils from human bone marrow.
Area of Science:
- Immunology
- Cell Biology
- Hematopoiesis
Background:
- Human recombinant interleukin-3 (rIL-3) is known to induce basophil development from human bone marrow.
- Mouse IL-3 promotes mast cell growth from mouse bone marrow, suggesting species-specific effects of IL-3.
Purpose of the Study:
- To investigate whether human rIL-3 can stimulate the growth of human mast cells.
- To compare the differentiation potential of human bone marrow cells under different culture conditions with human rIL-3.
Main Methods:
- Human bone marrow cells were cultured with human rIL-3 in suspension, methylcellulose, and interphase cultures.
- Histochemical techniques, including o-phthaldialdehyde staining and IgE receptor identification, were used to identify cell types.
- Enzyme and tryptase staining differentiated mast-like cells from basophil-like cells.
Main Results:
- Basophil-like cells were observed in all culture systems.
- Mast-like cells were identified exclusively in interphase cultures, comprising 1-5% of cells by 3 weeks.
- Both cell types expressed IgE receptors, but mast-like cells were positive for chloroacetate esterase, amidase, and human mast cell tryptase.
Conclusions:
- Human rIL-3 supports the growth of human mast-like cells, but only under specific culture conditions (interphase cultures).
- This study highlights the differential effects of human rIL-3 on human bone marrow cell differentiation, enabling mast cell development.