TY - JOUR
T1 - HMC-1 human mast cells synthesize neurotensin (NT) precursor, secrete bioactive NT-like peptide(s) and express NT receptor NTS1
AU - Cochrane, David E.
AU - Carraway, Robert E.
AU - Harrington, Kimberly
AU - Laudano, Melissa
AU - Rawlings, Stephen
AU - Feldberg, Ross S.
N1 - Funding Information:
Acknowledgments We thank students Alyssa Lillo, Katherine Moon, David Buzanoski and Richard Scarpa for technical help, and Liz Palmer for secretarial help. We thank Sanofi Recherche for the gift of SR48692. Supported by a Tufts Faculty Research Award to D.E.C., by the Department of Biology Carpenter Fellowship program to S.R., by the Arts and Sciences Dean’s Office, by the Tufts University Summer Scholar’s Program and N.I.H. grant # AR47652-06 to Dr TC Theohardides, Department of Pharmacology, Tufts University Medical School (D.E.C., co-investigator).
PY - 2011/12
Y1 - 2011/12
N2 - Objective and design: To determine if mast cells synthesize the inflammatory peptide, neurotensin (NT), secrete immunoreactive and bioactive NT, and express the NT receptor NTS1. Materials: HMC-1 cells, pleural mast cells from Sprague-Dawley rats, LAD2 mast cells, and human cord blood mast cells were used. Treatment: HMC-1 cells were stimulated with NT, C48/80, mastoparan, or PGE 2. For changes in cutaneous vascular permeability, anesthetized rats were injected intravenously with Evans Blue dye and intradermally with saline, NT, histamine, diphenhydramine, and C48/80. Methods: RT-PCR was used to identify RNA transcripts. Histamine was measured by fluorometric assay. In vivo cutaneous vascular permeability assays, radio-immunoassays for NT, Western blotting for the NT precursor protein and NTS1 protein from HMC-1 cells and tissues from rats were used. Immunohistochemistry was used to identify NT precursor-like proteins in HMC-1 mast cells. Results: HMC-1 cells express mRNAs for NT precursor, PC5A processing enzyme and NTS1 receptor. Human cord blood mast cells and LAD2 mast cells express mRNA transcripts for NT precursor and NTS1. Western blotting showed NT precursor and NTS1 receptor in HMC1. Rat tissues with high numbers of mast cells contained NT precursor proteins. NT-like peptides from HMC-1 displayed NT-like bioactivity. Conclusions: HMC-1 mast cells synthesize and secrete immunoreactive and bioactive NT-like peptide(s) and express the NT receptor, suggesting that NT from mast cells might serve autocrine and paracrine roles.
AB - Objective and design: To determine if mast cells synthesize the inflammatory peptide, neurotensin (NT), secrete immunoreactive and bioactive NT, and express the NT receptor NTS1. Materials: HMC-1 cells, pleural mast cells from Sprague-Dawley rats, LAD2 mast cells, and human cord blood mast cells were used. Treatment: HMC-1 cells were stimulated with NT, C48/80, mastoparan, or PGE 2. For changes in cutaneous vascular permeability, anesthetized rats were injected intravenously with Evans Blue dye and intradermally with saline, NT, histamine, diphenhydramine, and C48/80. Methods: RT-PCR was used to identify RNA transcripts. Histamine was measured by fluorometric assay. In vivo cutaneous vascular permeability assays, radio-immunoassays for NT, Western blotting for the NT precursor protein and NTS1 protein from HMC-1 cells and tissues from rats were used. Immunohistochemistry was used to identify NT precursor-like proteins in HMC-1 mast cells. Results: HMC-1 cells express mRNAs for NT precursor, PC5A processing enzyme and NTS1 receptor. Human cord blood mast cells and LAD2 mast cells express mRNA transcripts for NT precursor and NTS1. Western blotting showed NT precursor and NTS1 receptor in HMC1. Rat tissues with high numbers of mast cells contained NT precursor proteins. NT-like peptides from HMC-1 displayed NT-like bioactivity. Conclusions: HMC-1 mast cells synthesize and secrete immunoreactive and bioactive NT-like peptide(s) and express the NT receptor, suggesting that NT from mast cells might serve autocrine and paracrine roles.
KW - HMC-1 mast cell
KW - Neurotensin
KW - Neurotensin receptor
KW - Tumor growth
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U2 - 10.1007/s00011-011-0378-6
DO - 10.1007/s00011-011-0378-6
M3 - Article
C2 - 21927981
AN - SCOPUS:84855248584
SN - 1023-3830
VL - 60
SP - 1139
EP - 1151
JO - Inflammation Research
JF - Inflammation Research
IS - 12
ER -