Intestinal epithelial cells express and secrete the CD43 glycoform that contains core 2 O-glycans.
Several glycoforms of CD43 are known to regulate cellular interactions in the immune system. One such glycoform, the CD43 that bears core 2 O-glycans, is also known to be expressed on T lymphocytes and natural killer cells, but only after their activation. Previous studies have also shown that when Caco-2 cells, which are derived from human colon carcinoma, differentiate into enterocytes, they also express core 2 O-glycans, though proteins bearing this glycan are unknown. To examine whether CD43 glycosylation is altered during enterocytic differentiation of Caco-2 cells, we conducted immunocytochemical studies with a monoclonal antibody, 1D4, that recognizes a glycoform of CD43 bearing core 2 O-glycans. We found that 1D4 could bind to intracellular granules but not the cell surface of differentiated Caco-2 cells, whereas hematopoietic cells expressed 1D4 epitope on the cell surface as previously shown. The reactivity with this antibody increased as the degree of cell differentiation progressed as shown by the activity of the apical enzyme marker, dipeptidyl peptidase IV. 1D4-reactive CD43 was also found in the culture medium of differentiated Caco-2 cells, suggesting this molecule may be stored and secreted. The production and secretion of this CD43 glycoform by enterocyte-like Caco-2 cells was enhanced, and most 1D4 epitope converted to a soluble form when bacterial lipopolysaccharide was present. These observations strongly support the possibility that core 2 O-glycans on mucins such as CD43 are important to primary defense on the intestinal epithelium against infection.
Amano
J
J
Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. amano@mol.f.u-tokyo.ac.jp
Morimoto
C
C
Irimura
T
T
D015703:Antigens, CD; D018938:Caco-2 Cells; D002454:Cell Differentiation / Q000187:drug effects; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D051917:Leukosialin; D008070:Lipopolysaccharides / Q000494:pharmacology; D011134:Polysaccharides / Q000737:chemistry*; D012795:Sialoglycoproteins / Q000737:chemistry / Q000378:metabolism*
D015703:Antigens, CD; D051917:Leukosialin; D008070:Lipopolysaccharides; D011134:Polysaccharides; C465014:SPN protein, human; D012795:Sialoglycoproteins