A Fibrogenic Cytokine, Platelet-Derived Growth Factor (PDGF), Enhances Mast Cell Growth Indirectly Via a SCF- and Fibroblast-Dependent Pathway Takaaki.

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A Fibrogenic Cytokine, Platelet-Derived Growth Factor (PDGF), Enhances Mast Cell Growth Indirectly Via a SCF- and Fibroblast-Dependent Pathway Takaaki Hiragun, Eishin Morita, Toshihiko Tanaka, Yoshikazu Kameyoshi, Shoso Yamamoto Journal of Investigative Dermatology Volume 111, Issue 2, Pages 213-217 (August 1998) DOI: 10.1046/j.1523-1747.1998.00260.x Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 rhPDGF-AA, -AB, and -BB induced mast cell growth. BMMC obtained from +/+ mice were co-cultured with NIH/3T3 fibroblasts in the presence of a series of concentrations of PDGF, and histamine contents (A) and the number of mast cells (B) were determined on day 6. Data are mean ± SEM of three experiments done in duplicate (*p < 0.05, **p < 0.01). Journal of Investigative Dermatology 1998 111, 213-217DOI: (10.1046/j.1523-1747.1998.00260.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Time-dependent mast cell growth induced by rhPDGF-AB and -BB. BMMC obtained from +/+ mice were co-cultured with NIH/3T3 fibroblasts in the presence of 10 ng rhPDGF-AB or rhPDGF-BB per ml, and histamine content in the co-culture was determined on the indicated days. One typical experiment from three experiments performed independently is shown. Journal of Investigative Dermatology 1998 111, 213-217DOI: (10.1046/j.1523-1747.1998.00260.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Expression of PDGF receptor (α- and β-subunit) mRNA in BMMC and 3T3 fibroblasts analyzed by reverse transcriptase polymerase chain reaction. Total RNA was extracted from NIH/3T3 fibroblasts and BMMC obtained from +/+ mice, reverse transcribed, and analyzed by polymerase chain reaction with specific primers for PDGF α-receptor and PDGF β-receptor. The amplified polymerase chain reaction products were 544 bp for PDGF α-receptor and 381 bp for PDGF β-receptor. Journal of Investigative Dermatology 1998 111, 213-217DOI: (10.1046/j.1523-1747.1998.00260.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Separation of mast cells from fibroblasts by using 0.2 μm pore membrane abrogated the mast cell growth induced by PDGF. BMMC obtained from +/+ mice were cultured with NIH/3T3 fibroblasts either with or without tissue culture inserts attached with 0.2 μm pore membrane, and stimulated with 10 ng rhPDGF-AB per ml or 10 ng rhPDGF-BB per ml. PDGF-containing complete α-MEM was added into both the inner and the outer sides of the inserts at the starting of the co-culture. Culture medium of the outer side was replaced on days 2 and 4, then BMMC in the inserts were harvested on day 6. Data are mean ± SEM of three experiments done in duplicate. Journal of Investigative Dermatology 1998 111, 213-217DOI: (10.1046/j.1523-1747.1998.00260.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 SCF expression in 3T3 fibroblasts was enhanced by PDGF. Confluent NIH/3T3 fibroblasts were either untreated or stimulated with 10 ng rhPDGF-AB per ml, and the membrane fraction was extracted after 18 h or 36 h. SCF associated with membrane fraction was visualized by western blotting with anti-SCF monoclonal antibody. Journal of Investigative Dermatology 1998 111, 213-217DOI: (10.1046/j.1523-1747.1998.00260.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions