title: The monoclonal antibody HCM31 specifically recognises the Sd(a) tetrasaccharide in goblet cell mucin.
abstract: Rat small intestinal goblet cell mucins reacting with monoclonal antibody HCM31 increase significantly during regeneration from experimental mucosal damage and at the period of expulsion of parasitic nematode, Nippostrongylus brasiliensis (N.b). The reduction in reactivity of HCM31 with mucin upon neuraminidase treatment, suggested that HCM31 recognizes sialylated oligosaccharide on mucin. HCM31-reactive sialomucins are therefore considered to play an important role in the physiological and pathological changes in the gastrointestinal mucosa. To determine the epitope for HCM31, oligosaccharide-alditols reacted with HCM31 were obtained from the small intestinal mucins of N.b-infected rats and purified by ion-exchange chromatography followed by normal-phase HPLC. Two HCM31-reactive oligosaccharide-alditols were obtained. Analyses using tandem mass spectrometry and NMR spectroscopy showed that these oligosaccharides were core 4 mucin-type oligosaccharides having a common tetrasaccharide sequence, NeuAcα2-3(GalNAcβ1-4)Galβ1-4GlcNAcβ- (Sd(a) blood group antigen). These structures were not found in the small intestinal mucin oligosaccharides from uninfected rats. This epitope specificity of HCM31 was also confirmed using previously established anti-GM2 and anti-Sd(a) antibodies. Taken together, these results strongly suggest that HCM31 specifically recognizes mucin-type oligosaccharides with the Sd(a) tetrasaccharide sequence. Immunohistochemical examination of human gastrointestinal tracts showed that HCM31 site-specifically stained the goblet cells in normal sigmoid colon and normal rectum, but the goblet cells stained with HCM31 were reduced in the corresponding cancer tissues. HCM31 seems to be useful for diagnosis of colonic cancer and for examining the function of secretory-type mucin with Sd(a) antigen.
authors: Department of Biochemistry, Kitasato University Graduate School of Medical Sciences, Sagamihara, Japan ; Department of Parasitology, Kitasato University School of Medicine, Sagamihara, Japan.
authors: Daigo
authors: D
authors: Tsubokawa
authors: Yukinobu
authors: Y
authors: Goso
authors: Rei
authors: R
authors: Kawashima
authors: Hiroyoshi
authors: H
authors: Ota
authors: Takeshi
authors: T
authors: Nakamura
authors: Kazuo
authors: K
authors: Nakamura
authors: Noriko
authors: N
authors: Sato
authors: Makoto
authors: M
authors: Kurihara
authors: Taeko
authors: T
authors: Dohi
authors: Yuki I
authors: YI
authors: Kawamura
authors: Takafumi
authors: T
authors: Ichikawa
authors: Kazuhiko
authors: K
authors: Ishihara
keywords: Colon cancer; GalNAc-ol, N-acetylgalactosaminitol; MALDI-TOF/MS; MALDI-TOF/MS, matrix-assisted laser desorption/ionization-time of flight mass spectrometry; MS/MS, tandem mass spectrometry; Monoclonal antibody; Mucin; N.b, Nippostrongylus brasiliensis; NMR; Nippostrongylus brasiliensis; Sda antigen; Sialylated oligosaccharide; mAb, monoclonal antibody
references: J Clin Invest. 2012 Mar;122(3):923-34
references: 22307328
references: Biochem J. 2000 Sep 15;350 Pt 3:805-14
references: 10970796
references: J Biol Chem. 1999 Feb 19;274(8):4504-12
references: 9988682
references: J Exp Med. 1988 Jan 1;167(1):119-31
references: 2447219
references: Annu Rev Immunol. 2004;22:129-56
references: 15032576
references: Gastroenterology. 1995 Sep;109(3):735-47
references: 7657101
references: Rapid Commun Mass Spectrom. 1993 Jul;7(7):614-9
references: 8347867
references: Scand J Gastroenterol. 2008 Jan;43(1):59-65
references: 18938749
references: Histochem J. 1991 Jan;23(1):22-8
references: 1718927
references: Biochim Biophys Acta. 1992 Jul 21;1117(1):97-103
references: 1627599
references: Biochem J. 2001 Sep 15;358(Pt 3):657-64
references: 11577689
references: Science. 2004 Aug 13;305(5686):1003-6
references: 15310903
references: Biochim Biophys Acta. 1999 Dec 23;1489(2-3):336-44
references: 10673034
references: Comp Biochem Physiol B. 1993 Apr;104(4):781-6
references: 8472545
references: J Biol Chem. 1968 Feb 10;243(3):616-26
references: 5637714
references: Exp Parasitol. 2009 Dec;123(4):319-25
references: 19703448
references: J Biol Chem. 1990 May 15;265(14):7880-5
references: 2139874
references: Glycoconj J. 1996 Oct;13(5):857-64
references: 8910013
references: Biochem J. 2004 Dec 1;384(Pt 2):307-16
references: 15361072
references: Rapid Commun Mass Spectrom. 1996;10(9):1027-32
references: 8755235
references: Histochem J. 1992 Feb;24(2):86-92
references: 1374372
references: J Gastroenterol. 2009;44(4):277-84
references: 19280111
references: Biochem J. 1996 Sep 1;318 ( Pt 2):409-16
references: 8809027
references: FEBS J. 2007 Apr;274(7):1833-48
references: 17381513
references: Cancer Res. 2005 Jul 15;65(14):6220-7
references: 16024623
references: Nat Rev Cancer. 2004 Jan;4(1):45-60
references: 14681689
references: Hybridoma. 1993 Oct;12(5):609-20
references: 7507882
references: Clin Chim Acta. 1993 Nov 30;221(1-2):219-25
references: 8149639
references: Biochim Biophys Acta. 2008 Mar;1780(3):467-71
references: 17961927
references: Methods Enzymol. 2003;362:160-95
references: 12968363
references: Cancer Res. 1989 Dec 1;49(23):6466-70
references: 2510926
references: Gastroenterology. 1995 Sep;109(3):723-34
references: 7657100
references: Glycoconj J. 1998 Aug;15(8):737-47
references: 9870349