Wastling J M, Scudamore C L, Thornton E M, Newlands G F, Miller H R
Department of Veterinary Clinical Studies, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Roslin, UK.
Immunology. 1997 Feb;90(2):308-13. doi: 10.1046/j.1365-2567.1997.00155.x.
Rodent intestinal mucosal mast cells (IMMC) store and secrete soluble granule serine proteases, the beta-chymases, which may promote epithelial permeability during intestinal hypersensitivity reactions. The beta-chymase mouse mast cell protease-1 (mMCP-1) is generally considered to be expressed late in the in vitro differentiation of mast cells. The purpose of this study was to determine the kinetics of mMCP-1 transcription and expression in vivo during nematode-induced IMMC hyperplasia. Concentrations of mMCP-1 in blood and jejunum of BALB/c mice were quantified by enzyme-linked immunosorbent assay before and at various stages after infection with the intestinal nematode Nippostronglyus brasilliensis. Mature mMCP-1 enzyme was detected in jejunal homogenate (194 ng/mg soluble protein) and in blood (8.3 ng/ml serum) from normal uninfected BALB/c mice. Maximal IMMC hyperplasia occurred 7-14 days post infection and was significantly correlated with increased levels of mMCP-1 in jejunum (r = 0.58, P < 0.001) and with raised concentrations of mMCP-1 in serum (r = 0.66, P < 0.001). Transcription of the mMCP-1 gene was detected by RNA blotting in normal, uninfected jejunum, but transcription was up-regulated after infection with maximal transcription occurring on days 7 and 14. In conclusion, mMCP-1 transcription, storage and secretion occur constitutively in normal BALB/c jejunum but this basal secretion is up-regulated during nematode infection, suggesting both a physiological and pathological function for this protease.
啮齿动物肠道黏膜肥大细胞(IMMC)储存并分泌可溶性颗粒丝氨酸蛋白酶,即β-糜蛋白酶,在肠道过敏反应期间,这些酶可能会促进上皮通透性。β-糜蛋白酶小鼠肥大细胞蛋白酶-1(mMCP-1)通常被认为在肥大细胞的体外分化后期表达。本研究的目的是确定线虫诱导的IMMC增生过程中,体内mMCP-1转录和表达的动力学变化。在用巴西日圆线虫感染BALB/c小鼠之前及感染后不同阶段,通过酶联免疫吸附测定法定量检测血液和空肠中mMCP-1的浓度。在未感染的正常BALB/c小鼠的空肠匀浆(194 ng/mg可溶性蛋白)和血液(8.3 ng/ml血清)中检测到成熟的mMCP-1酶。感染后7-14天出现最大程度的IMMC增生,且与空肠中mMCP-1水平升高显著相关(r = 0.58,P < 0.001),也与血清中mMCP-1浓度升高相关(r = 0.66,P < 0.001)。通过RNA印迹法在正常未感染的空肠中检测到mMCP-1基因的转录,但感染后转录上调,在第7天和第14天出现最大转录。总之,mMCP-1的转录、储存和分泌在正常BALB/c空肠中持续存在,但在感染线虫期间这种基础分泌会上调,提示该蛋白酶具有生理和病理功能。