OBJECTIVE AND DESIGN: To determine whether RBL-2H3 cells have endogenous substance P (SP) expression under immunoglobulin E (IgE)-activated and inactivated conditions, and to ascertain the function of endogenous SP in the antigen-induced degranulation of RBL-2H3 cells. MATERIALS AND METHODS: SP mRNA and protein expression in both inactivated and 2,4-dinitrophenol (DNP)-specific IgE-activated RBL-2H3 cells were assessed by real-time PCR and immunofluorescence, respectively. Following activation with DNP-specific IgE, the degranulation of RBL-2H3 cells in response to DNP-bovine serum albumin (BSA), with and without endogenous SP expression, was assessed by monitoring the release of the granular enzyme β-hexosaminidase. RESULTS: Endogenous SP mRNA and peptide expression increased in activated RBL-2H3 cells, compared with inactivated RBL-2H3 cells. The small hairpin RNA (shRNA)-mediated knockdown of endogenous SP reduced the degranulation ability of RBL-2H3 cells. CONCLUSIONS: Activated RBL-2H3 cells express endogenous SP which plays a role in antigen-induced degranulation.
OBJECTIVE AND DESIGN: To determine whether RBL-2H3 cells have endogenous substance P (SP) expression under immunoglobulin E (IgE)-activated and inactivated conditions, and to ascertain the function of endogenous SP in the antigen-induced degranulation of RBL-2H3 cells. MATERIALS AND METHODS: SP mRNA and protein expression in both inactivated and 2,4-dinitrophenol (DNP)-specific IgE-activated RBL-2H3 cells were assessed by real-time PCR and immunofluorescence, respectively. Following activation with DNP-specific IgE, the degranulation of RBL-2H3 cells in response to DNP-bovine serum albumin (BSA), with and without endogenous SP expression, was assessed by monitoring the release of the granular enzyme β-hexosaminidase. RESULTS: Endogenous SP mRNA and peptide expression increased in activated RBL-2H3 cells, compared with inactivated RBL-2H3 cells. The small hairpin RNA (shRNA)-mediated knockdown of endogenous SP reduced the degranulation ability of RBL-2H3 cells. CONCLUSIONS: Activated RBL-2H3 cells express endogenous SP which plays a role in antigen-induced degranulation.
Authors: Terence M O'Connor; Joseph O'Connell; Darren I O'Brien; Triona Goode; Charles P Bredin; Fergus Shanahan Journal: J Cell Physiol Date: 2004-11 Impact factor: 6.384
Authors: A Leon; A Buriani; R Dal Toso; M Fabris; S Romanello; L Aloe; R Levi-Montalcini Journal: Proc Natl Acad Sci U S A Date: 1994-04-26 Impact factor: 11.205
Authors: D Lorenz; B Wiesner; J Zipper; A Winkler; E Krause; M Beyermann; M Lindau; M Bienert Journal: J Gen Physiol Date: 1998-11 Impact factor: 4.086
Authors: M Swieter; R J Midura; H Nishikata; C Oliver; E H Berenstein; S E Mergenhagen; V C Hascall; R P Siraganian Journal: J Immunol Date: 1993-01-15 Impact factor: 5.422