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. 1994 Jul 1;180(1):67–73. doi: 10.1084/jem.180.1.67

Mouse bone marrow-derived mast cells (mBMMC) obtained in vitro from mice that are mast cell-deficient in vivo express the same panel of granule proteases as mBMMC and serosal mast cells from their normal littermates

PMCID: PMC2191541  PMID: 8006601

Abstract

The ear, skin, and purified serosal mast cells of WBB6F1/J-(+/+) (WB- (+/+)) and WCB6F1/J-(+/+) (WC-(+/+)) mice contain high steady-state levels of the transcripts that encode mouse mast cell protease (mMCP) 2, mMCP-4, mMCP-5, mMCP-6, and mouse mast cell carboxypeptidase A (mMC- CPA). In contrast, no mast cell protease transcripts are present in abundance in the ear and skin of WBB6F1/J-W/Wv (W/Wv) and WCB6F1/J- Sl/Sld (Sl/Sld) mice which are mast cell-deficient in vivo due to defects in their c-kit and c-kit ligand genes, respectively. We now report that the immature bone marrow-derived mast cells (mBMMC) obtained in vitro with recombinant interleukin 3 (rIL-3) or WEHI-3 cell conditioned medium from WB-(+/+), WC-(+/+), W/Wv, and Sl/Sld mice all contain high steady-state levels of the mMCP-2, mMCP-4, mMCP-5, mMCP-6, and mMC-CPA transcripts. As assessed immunohistochemically, mMCP-2 protein and mMCP-5 protein are also present in the granules of mBMMC from WB-(+/+), WC-(+/+), and W/Wv mice. That Sl/Sld and W/Wv mBMMC contain high steady-state levels of five granule protease transcripts expressed by the mature serosal, ear, and skin mast cells of their normal +/+ littermates suggests that c-kit-mediated signal transduction is not essential for inducing transcription of these protease genes. Because rIL-4 inhibits the rIL-10-induced expression of mMCP-1 and mMCP- 2 in BALB/cJ mBMMC, the ability of rIL-4 to influence protease mRNA levels in WC-(+/+) mBMMC and W/Wv mBMMC was investigated. Although rIL- 10 induced expression of the mMCP-1 transcript in WC-(+/+) and W/Wv mBMMC, rIL-4 was not able to suppress the steady-state levels of the mMCP-1 transcript or any other protease transcript in these cultured mast cells. Thus, not only do BALB/cJ mBMMC express fewer granule proteases than mBMMC from mast cell-deficient strains and their normal littermates but the subsequent induction of late-expressed proteases in BALB/cJ mBMMC is more tightly regulated by IL-3 and IL-4.

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Selected References

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  1. Avraham S., Stevens R. L., Nicodemus C. F., Gartner M. C., Austen K. F., Weis J. H. Molecular cloning of a cDNA that encodes the peptide core of a mouse mast cell secretory granule proteoglycan and comparison with the analogous rat and human cDNA. Proc Natl Acad Sci U S A. 1989 May;86(10):3763–3767. doi: 10.1073/pnas.86.10.3763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Crowle P. K., Reed N. D. Rejection of the intestinal parasite Nippostrongylus brasiliensis by mast cell-deficient W/Wv anemic mice. Infect Immun. 1981 Jul;33(1):54–58. doi: 10.1128/iai.33.1.54-58.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eklund K. K., Ghildyal N., Austen K. F., Stevens R. L. Induction by IL-9 and suppression by IL-3 and IL-4 of the levels of chromosome 14-derived transcripts that encode late-expressed mouse mast cell proteases. J Immunol. 1993 Oct 15;151(8):4266–4273. [PubMed] [Google Scholar]
  5. Flanagan J. G., Chan D. C., Leder P. Transmembrane form of the kit ligand growth factor is determined by alternative splicing and is missing in the Sld mutant. Cell. 1991 Mar 8;64(5):1025–1035. doi: 10.1016/0092-8674(91)90326-t. [DOI] [PubMed] [Google Scholar]
  6. Flanagan J. G., Leder P. The kit ligand: a cell surface molecule altered in steel mutant fibroblasts. Cell. 1990 Oct 5;63(1):185–194. doi: 10.1016/0092-8674(90)90299-t. [DOI] [PubMed] [Google Scholar]
  7. Galli S. J., Arizono N., Murakami T., Dvorak A. M., Fox J. G. Development of large numbers of mast cells at sites of idiopathic chronic dermatitis in genetically mast cell-deficient WBB6F1-W/Wv mice. Blood. 1987 Jun;69(6):1661–1666. [PubMed] [Google Scholar]
  8. Geissler E. N., Ryan M. A., Housman D. E. The dominant-white spotting (W) locus of the mouse encodes the c-kit proto-oncogene. Cell. 1988 Oct 7;55(1):185–192. doi: 10.1016/0092-8674(88)90020-7. [DOI] [PubMed] [Google Scholar]
  9. Ghildyal N., Friend D. S., Nicodemus C. F., Austen K. F., Stevens R. L. Reversible expression of mouse mast cell protease 2 mRNA and protein in cultured mast cells exposed to IL-10. J Immunol. 1993 Sep 15;151(6):3206–3214. [PubMed] [Google Scholar]
  10. Ghildyal N., McNeil H. P., Gurish M. F., Austen K. F., Stevens R. L. Transcriptional regulation of the mucosal mast cell-specific protease gene, MMCP-2, by interleukin 10 and interleukin 3. J Biol Chem. 1992 Apr 25;267(12):8473–8477. [PubMed] [Google Scholar]
  11. Ghildyal N., McNeil H. P., Stechschulte S., Austen K. F., Silberstein D., Gurish M. F., Somerville L. L., Stevens R. L. IL-10 induces transcription of the gene for mouse mast cell protease-1, a serine protease preferentially expressed in mucosal mast cells of Trichinella spiralis-infected mice. J Immunol. 1992 Sep 15;149(6):2123–2129. [PubMed] [Google Scholar]
  12. Gordon J. R., Galli S. J. Phorbol 12-myristate 13-acetate-induced development of functionally active mast cells in W/Wv but not Sl/Sld genetically mast cell-deficient mice. Blood. 1990 Apr 15;75(8):1637–1645. [PubMed] [Google Scholar]
  13. Gurish M. F., Ghildyal N., McNeil H. P., Austen K. F., Gillis S., Stevens R. L. Differential expression of secretory granule proteases in mouse mast cells exposed to interleukin 3 and c-kit ligand. J Exp Med. 1992 Apr 1;175(4):1003–1012. doi: 10.1084/jem.175.4.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Huang R. Y., Blom T., Hellman L. Cloning and structural analysis of MMCP-1, MMCP-4 and MMCP-5, three mouse mast cell-specific serine proteases. Eur J Immunol. 1991 Jul;21(7):1611–1621. doi: 10.1002/eji.1830210706. [DOI] [PubMed] [Google Scholar]
  15. Kitamura Y., Go S. Decreased production of mast cells in S1/S1d anemic mice. Blood. 1979 Mar;53(3):492–497. [PubMed] [Google Scholar]
  16. Kitamura Y., Go S., Hatanaka K. Decrease of mast cells in W/Wv mice and their increase by bone marrow transplantation. Blood. 1978 Aug;52(2):447–452. [PubMed] [Google Scholar]
  17. McBride P., Bradley D., Kaliner M. Evaluation of a radioimmunoassay for histamine measurement in biologic fluids. J Allergy Clin Immunol. 1988 Oct;82(4):638–646. doi: 10.1016/0091-6749(88)90977-3. [DOI] [PubMed] [Google Scholar]
  18. McNeil H. P., Austen K. F., Somerville L. L., Gurish M. F., Stevens R. L. Molecular cloning of the mouse mast cell protease-5 gene. A novel secretory granule protease expressed early in the differentiation of serosal mast cells. J Biol Chem. 1991 Oct 25;266(30):20316–20322. [PubMed] [Google Scholar]
  19. McNeil H. P., Frenkel D. P., Austen K. F., Friend D. S., Stevens R. L. Translation and granule localization of mouse mast cell protease-5. Immunodetection with specific antipeptide Ig. J Immunol. 1992 Oct 1;149(7):2466–2472. [PubMed] [Google Scholar]
  20. Nakano T., Sonoda T., Hayashi C., Yamatodani A., Kanayama Y., Yamamura T., Asai H., Yonezawa T., Kitamura Y., Galli S. J. Fate of bone marrow-derived cultured mast cells after intracutaneous, intraperitoneal, and intravenous transfer into genetically mast cell-deficient W/Wv mice. Evidence that cultured mast cells can give rise to both connective tissue type and mucosal mast cells. J Exp Med. 1985 Sep 1;162(3):1025–1043. doi: 10.1084/jem.162.3.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ody C., Kindler V., Vassalli P. Interleukin 3 perfusion in W/Wv mice allows the development of macroscopic hematopoietic spleen colonies and restores cutaneous mast cell number. J Exp Med. 1990 Jul 1;172(1):403–406. doi: 10.1084/jem.172.1.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Razin E., Ihle J. N., Seldin D., Mencia-Huerta J. M., Katz H. R., LeBlanc P. A., Hein A., Caulfield J. P., Austen K. F., Stevens R. L. Interleukin 3: A differentiation and growth factor for the mouse mast cell that contains chondroitin sulfate E proteoglycan. J Immunol. 1984 Mar;132(3):1479–1486. [PubMed] [Google Scholar]
  23. Reynolds D. S., Gurley D. S., Austen K. F., Serafin W. E. Cloning of the cDNA and gene of mouse mast cell protease-6. Transcription by progenitor mast cells and mast cells of the connective tissue subclass. J Biol Chem. 1991 Feb 25;266(6):3847–3853. [PubMed] [Google Scholar]
  24. Reynolds D. S., Stevens R. L., Gurley D. S., Lane W. S., Austen K. F., Serafin W. E. Isolation and molecular cloning of mast cell carboxypeptidase A. A novel member of the carboxypeptidase gene family. J Biol Chem. 1989 Nov 25;264(33):20094–20099. [PubMed] [Google Scholar]
  25. Reynolds D. S., Stevens R. L., Lane W. S., Carr M. H., Austen K. F., Serafin W. E. Different mouse mast cell populations express various combinations of at least six distinct mast cell serine proteases. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3230–3234. doi: 10.1073/pnas.87.8.3230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Serafin W. E., Reynolds D. S., Rogelj S., Lane W. S., Conder G. A., Johnson S. S., Austen K. F., Stevens R. L. Identification and molecular cloning of a novel mouse mucosal mast cell serine protease. J Biol Chem. 1990 Jan 5;265(1):423–429. [PubMed] [Google Scholar]
  27. Serafin W. E., Sullivan T. P., Conder G. A., Ebrahimi A., Marcham P., Johnson S. S., Austen K. F., Reynolds D. S. Cloning of the cDNA and gene for mouse mast cell protease 4. Demonstration of its late transcription in mast cell subclasses and analysis of its homology to subclass-specific neutral proteases of the mouse and rat. J Biol Chem. 1991 Jan 25;266(3):1934–1941. [PubMed] [Google Scholar]
  28. Spiegelman B. M., Frank M., Green H. Molecular cloning of mRNA from 3T3 adipocytes. Regulation of mRNA content for glycerophosphate dehydrogenase and other differentiation-dependent proteins during adipocyte development. J Biol Chem. 1983 Aug 25;258(16):10083–10089. [PubMed] [Google Scholar]
  29. Stevens R. L., Friend D. S., McNeil H. P., Schiller V., Ghildyal N., Austen K. F. Strain-specific and tissue-specific expression of mouse mast cell secretory granule proteases. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):128–132. doi: 10.1073/pnas.91.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tertian G., Yung Y. P., Guy-Grand D., Moore M. A. Long-term in vitro culture of murine mast cells. I. Description of a growth factor-dependent culture technique. J Immunol. 1981 Aug;127(2):788–794. [PubMed] [Google Scholar]
  31. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Trong H. L., Newlands G. F., Miller H. R., Charbonneau H., Neurath H., Woodbury R. G. Amino acid sequence of a mouse mucosal mast cell protease. Biochemistry. 1989 Jan 10;28(1):391–395. doi: 10.1021/bi00427a054. [DOI] [PubMed] [Google Scholar]
  33. Tsai M., Shih L. S., Newlands G. F., Takeishi T., Langley K. E., Zsebo K. M., Miller H. R., Geissler E. N., Galli S. J. The rat c-kit ligand, stem cell factor, induces the development of connective tissue-type and mucosal mast cells in vivo. Analysis by anatomical distribution, histochemistry, and protease phenotype. J Exp Med. 1991 Jul 1;174(1):125–131. doi: 10.1084/jem.174.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tsai M., Takeishi T., Thompson H., Langley K. E., Zsebo K. M., Metcalfe D. D., Geissler E. N., Galli S. J. Induction of mast cell proliferation, maturation, and heparin synthesis by the rat c-kit ligand, stem cell factor. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6382–6386. doi: 10.1073/pnas.88.14.6382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yung Y. P., Moore M. A. Long-term in vitro culture of murine mast cells. III. Discrimination of mast cells growth factor and granulocyte-CSF. J Immunol. 1982 Sep;129(3):1256–1261. [PubMed] [Google Scholar]

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