CD24 is a glycosyl-phosphatidyl-inositol linked glycoprotein expressed in a broad range of cell types and is heavily glycosylated. It has been found to be over expressed in cancers and tumors and is also a costimulatory molecule. Therefore, this study was carried out to define the structures of the carbohydrates associated with the CD24 recombinant protein. The CD24 glycoprotein's oligosaccharides were released by chemical and enzymatic means prior to being analyzed by MALDI-TOF-MS. The results obtained showed that CD24 is both N-and O-glycosylated. The major oligosaccharides were found to be Neu5Acα-2,3/6Galβ-1,3GalNAc, NeuAc 2 Gal β-1,3GalNAc 1 (O-glycans), GalNAc 2 GlcNAc 2 Man 3 Fuc 1 , Gal 1 GalNAc 2 GlcNAc 2 Man 3 Fuc 1 , and Gal 2 GalNAc 2 GlcNAc 2 Man 3 Fuc 1 (N-glycans). The results showed that Neu5Acα-2,3/6Galβ-1,3GalNAc (sialyl-tumor antigen, sT), a cancer-associated carbohydrate, was the most abundant glycan associated with CD24. This result raised the intriguing possibility that CD24 may be a major carrier of the sialyl-T abundantly found in cancer cells.
Keywords
CD24; Glycans; MALDI-TOF-MS; T antigen
IntroductionCD24 is a glycosyl-phosphatidyl-inositol (GPI)-anchored glycoprotein with widespread expression in both hematopoietic and non-hematopoietic cells. The protein is composed of 27-35 amino acids with nearly half of the amino acids being serine and threonine [1,2]. Both serine and threonine are potential sites for O-glycosylation and asparagine gives potential for N-glycosylation. The human form of CD24 has an unusual structure with the core having [3]. The CD24 molecule is expressed in developing or regenerating tissue and in pre-B-cells, granulocytes, keratinocytes, and renal tubular epithelial cells [1,4,5]. CD24 is over-expressed in both hematological malignancies and a variety of solid tumors such as gastric, renal, nasopharyngeal, hepatocellular, colonic, and small cell lung carcinomas [6,7].The CD24 gene has been implicated in immunity, cancer and autoimmune diseases. Thus, expressing of CD24 in non-professional antigen-presenting cells, such as activated B cells [8,9], astrocytes and oligodendritic cells [10,11] conveys a potent costimulatory activity. In T cells, CD24 promotes homeostatic proliferation [12]. In contrast, CD24 expression in dendritic cells is a negative regulator that modulates the pace of homeostatic proliferation [13], a process that is important for development of autoimmune diseases and cancer immunotherapy [14,15]. Expression of CD24 in pre-B cells on the other hand, modulates fibronectin/VLA-4:VCAM-1 interactions [16,17]. Outside the immune system, CD24 expressed in the central nervous system (CNS) has been shown to inhibit neurite growth [18]. In humans, CD24 polymorphism controls risk and progression of autoimmune diseases, such as multiple sclerosis [19]. Concerning cancerous cells, CD24 has been found to be prevalent on many tumors and carcinomas [20][21][22]. Carbohydrates of CD24 on tumor cells have been shown to play an important role in metastasi...