LAMP2 as a marker of EBV-mediated B lymphocyte transformation in the study of lysosomal storage diseases
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Following the degradative pathway, vesicles loaded with extracellular material, eventually, dock and fuse with lysosomes, acquiring specific membrane markers of these organelles and acid hydrolases responsible for digest their content. The lysosomal-associated membrane protein 2 (LAMP-2), the best characterized lysosomal membrane protein, is found in late stages of endosome maturation and may be used as a marker of lysosome-associated membranes. Lysosomal storage disorders (LSDs) are described by the absence or deficiency in hydrolase activity leading to substrate accumulation within lysosomal components and to the onset of several diseases. It is known that lymphocytes infected by Epstein–Barr virus (EBV) are able to form cytoplasmic vacuoles, which work as a storage compartment for lysosomal acidic hydrolases. At the present study, we validate the EBV as a transforming agent of B lymphocytes in stability studies of long-term stored samples, since the methods used to keep samples in liquid nitrogen and thaw them have all proven to be efficient in samples frozen for up to 2 years. To confirm and investigate some of the most prevalent LSDs in the South of Brazil—Pompe, Fabry and Gaucher diseases—we first measured the enzymatic activity of α-glicosidase, α-galactosidase, and β-glicosidase in those cytoplasmic-formed vacuoles and then looked to LAMP-2 immunoreactivity by employing confocal microscopy techniques.
KeywordsLymphocyte transformation Inborn errors of metabolism Epstein–Barr virus Lysosomal storage disorders Pompe disease Fabry disease Gaucher disease
- 1.Brooks DA (2009) The endosomal network. Int J Clin Pharmacol Ther 47:9–17Google Scholar
- 10.Elliot J, Coulter-Mackie MB, Jung JH, Rodenhiser DI, Singh SM (1991) A method for transforming lymphocytes from very small blood volumes suitable for paediatric samples. Hum Genet 86(6):615–616Google Scholar
- 20.Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275Google Scholar