Mechanisms of asthma and allergic inflammation
Surface CD88 functionally distinguishes the MCTC from the MCT type of human lung mast cell

https://doi.org/10.1016/j.jaci.2005.02.022Get rights and content

Background

MCT and MCTC types of human mast cells (MCs) are distinguished from one another on the basis of the protease compositions of their secretory granules, but their functional and developmental relationships have been uncertain.

Objective

These studies better define the functional properties and developmental relationship of MCT and MCTC cells.

Methods

Mast cells were dispersed from human skin and lung, purified with anti-Kit antibody, and separated into CD88+ and CD88 populations by cell sorting. These cells were evaluated by immunocytochemistry with antitryptase and antichymase mAbs; for chymase and tryptase mRNA by real-time RT-PCR; for conversion of MCT to MCTC cells during cell culture with recombinant human stem cell factor and recombinant human IL-6; and for degranulation and leukotriene C4 (LTC4) secretion when stimulated with anti-FcεRI, substance P, C5a, and compound 48/80.

Results

Mature MCT and MCTC cells were separated from one another on the basis of selective expression of CD88, the C5aR, on MCTC cells. Lung MCT cells had negligible levels of chymase mRNA and retained their MCT phenotype in culture. Mature MCTC cells from skin and lung degranulated in response to FcεRI cross-linking, C5a, compound 48/80, and substance P. Lung MCTC cells released LTC4 on activation, but no LTC4 was detected when skin-derived MCTC cells were activated. MCT cells from lung degranulated and released LTC4 in response to anti-FcεRI and substance P, but not to C5a and compound 48/80.

Conclusion

These observations functionally distinguish MCT from MCTC types of human mast cells and suggest important differences that may affect their participation in diseases such as asthma and urticaria.

Section snippets

Culture of Lu-MC and Sk-MC

All experimental protocols involving human tissues were approved by the Human Studies Committee at Virginia Commonwealth University. Surgical lung or skin tissue samples were obtained from consented patients through the Cooperative Human Tissue Network (Columbus, Ohio), the National Disease Research Interchange Center (Philadelphia, Pa), or the Departments of Surgery or Pathology at Virginia Commonwealth University. Lung specimens were typically lobectomies from patients with lung cancer,

CD88-dependent sorting of MCT and MCTC cells from lung

Because MCTC cells from skin express the C5aR, CD88, experiments were conducted to see whether CD88 also was expressed on MCTC cells from lung and could enable MCTC cells to be separated from MCT cells by cell sorting. As shown in Fig 1, after sorting Lu-MC, <4% of the cells in the Kit+/CD88 gate were chymase+, whereas greater than 98% of the cells in the Kit+/CD88+ gate were chymase+. This result was representative of 3 independent experiments. By real-time/RT-PCR, the 18S-rRNA–normalized

Discussion

The current study makes 3 important observations. First, the mature MCTC type of mast cell in lung and skin expresses CD88, the receptor for C5a, whereas mature MCT cells in lung do not. Accordingly, cell sorting of Kithi lung-derived mast cells with anti-CD88 antibody separates CD88 MCT from CD88+ MCTC cells. Thus, surface expression of CD88 corresponds to the MCTC phenotype, regardless whether the MCTC cell originates from the skin or lung. Lu-MCT cells express negligible levels, if any, of

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    Supported by grants R01-AI27517 and R01-AI20487 to Dr. Schwartz and K08-AI057357 to Dr Zhao from the National Institutes of Health.

    Disclosure of potential conflict of interest: C. A. Oskertizian: none disclosed; W. Zhao: none disclosed; H.-K. Min: none disclosed; H.-Z. Xia: none disclosed; A. Pozez: none disclosed; J. Kiev: none disclosed; L. B. Schwartz: none disclosed.

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