The Journal of Immunology, Vol 146, Issue 3 981-987, Copyright © 1991 by American Association of Immunologists

ARTICLES

Connective tissue mast cells in contact with fibroblasts express IL-3 mRNA. Analysis of single cells by polymerase chain reaction

E Razin, KB Leslie and JW Schrader
Institute of Biochemistry, Hebrew University, Hadassah Medical School, Jerusalem, Israel.

Mast cell-fibroblast interactions have been extensively investigated in the last few years. Fibroblasts support the in vitro survival but not proliferation of mouse connective-tissue type mast cells. However, the factor(s) that allow their survival on fibroblast monolayers has not been identified. We have investigated the presence of mRNA for IL-3 and granulocyte-macrophage-CSF in single mouse mast cells, before and after co-culture with 3T3 fibroblasts, using the polymerase chain reaction technique. The system was calibrated first by using in vitro generated population of mouse bone-marrow derived mast cells (BMMC). Significant differences in the amplification of IL-3 cDNA were observed in each of the BMMC cells examined, whereas the amplification of cDNA for the alpha-subunit of the Fc epsilon RI were similar. Inasmuch as murine cultured IL-3-dependent mast cells differentiate into connective tissue- like mast cells when co-cultured with 3T3 fibroblasts without any exogenous supply of growth factors, it was of interest to determine whether these connective tissue-like mast cells produce IL-3 message. Separation of the differentiated BMMC from the fibroblast monolayer, by either trypsinization or by single cell manipulation revealed the synthesis of a detectable amount of IL-3 mRNA in these mast cells. Whether this IL-3 mRNA was induced by fibroblasts was further investigated using connective tissue mast cells freshly purified from the mouse peritoneal cavity. Only about 20% of these connective tissue mast cells produced detectable amount of granulocyte-macrophage-CSF mRNA whereas in less than 10% of the cells IL-3 mRNA was detected. However, when these connective tissue mast cells were co-cultured with 3T3 fibroblasts for 18 hours and then separated, IL-3 mRNA were detected in most of the cells whereas no such mRNA was detected in tissue mast cells incubated for 18 h with medium derived from 3T3 fibroblasts. Therefore we conclude that fibroblasts induce the accumulation of IL-3 mRNA in connective tissue mast cells. The production of IL-3 may play a role in the survival of this type of mast cells on the fibroblast monolayer.

This article has been cited by other articles:

				H. Oren, A. R. Erbay, M. Balci, and S. Cehreli Role of Novel Biomarkers of Inflammation in Patients With Stable Coronary Heart Disease Angiology, April 1, 2007; 58(2): 148 - 155. [Abstract] [PDF]

				H. Oren, A. R. Erbay, M. Balc, and S. Cehreli Role of Novel Mediators of Inflammation in Left Ventricular Remodeling in Patients With Acute Myocardial Infarction: Do They Affect the Outcome of Patients? Angiology, February 1, 2007; 58(1): 45 - 54. [Abstract] [PDF]

				K. Roundy, A. Kollhoff, E. J. Eichwald, J. J. Weis, and J. H. Weis Microphthalmic Mice Display a B Cell Deficiency Similar to that Seen for Mast and NK Cells J. Immunol., December 15, 1999; 163(12): 6671 - 6678. [Abstract] [Full Text] [PDF]

				E. D. Bullock and E. M. Johnson Jr. Nerve Growth Factor Induces the Expression of Certain Cytokine Genes and bcl-2 in Mast Cells. POTENTIAL ROLE IN SURVIVAL PROMOTION J. Biol. Chem., November 1, 1996; 271(44): 27500 - 27508. [Abstract] [Full Text] [PDF]

				W C Gause and J Adamovicz The use of the PCR to quantitate gene expression. Genome Res., June 1, 1994; 3(6): S123 - S135. [PDF]