title: Feasibility of using cryopreserved lymphoblastoid cells to diagnose some lysosomal storage diseases.
abstract: OBJECTIVE
  The Epstein-Barr virus (EBV) is utilized as a tool in the study of cellular biology because of its capacity to transform B-lymphocytes. For this reason, EBV is used in conservation of human B-lymphocytes for long periods for subsequent evaluation of lysosomal hydrolase activity. Lymphoblastoid cell lines have several advantages for use over other cell types, such as prompt availability and possibility to develop, characterize and standardize cell banks, to test effects of promising pharmaceutical reagents. The study below presents biochemical data that demonstrate validity of lymphoblastoid cell lines for diagnosis of GM1-gangliosidosis, Gaucher, Fabry and Pompe diseases and mucopolysaccharidosis type I.
  
  
  METHODS
  Cultures were prepared from peripheral blood, collected from 25 normal subjects and 13 affected individuals. Enzyme activities and immunohistochemistry (IHC) were measured. Activities of enzymes beta-galactosidase, beta-glucosidase, alpha-iduronidase, alpha-galactosidase and alpha-glucosidase were measured before and after cryopreservation for 180 days. Enzymatic activity was measured when transformation was confirmed by IHC.
  
  
  RESULTS
  We observed some significant alterations in enzymatic activity of non-cultured cells when compared to others that had been cultured for 12 days and kept frozen for 180 days.
  
  
  CONCLUSIONS
  However, these alterations did not invalidate use of the technology of transformation of lymphoblastoid cell lines with EBV, to diagnose the diseases mentioned above, in view of the fact that the cultured cells, before and after freezing, demonstrated similar enzymatic activities.
authors.affiliation: Post Graduate Program in Biological Sciences - Biochemistry, Department of Biochemistry, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: A S
authors.initials: AS
authors.lastname: de Mello
authors.forename: F
authors.initials: F
authors.lastname: Provin
authors.forename: K
authors.initials: K
authors.lastname: Michelin-Tireli
authors.forename: M V
authors.initials: MV
authors.lastname: Camelier
authors.forename: J C
authors.initials: JC
authors.lastname: Coelho
mesh_terms: D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000821:virology; D016022:Case-Control Studies; D002460:Cell Line; D015925:Cryopreservation; D000795:Fabry Disease / Q000175:diagnosis / Q000201:enzymology; D005240:Feasibility Studies; D016537:Gangliosidosis, GM1 / Q000175:diagnosis / Q000201:enzymology; D005776:Gaucher Disease / Q000175:diagnosis / Q000201:enzymology; D006009:Glycogen Storage Disease Type II / Q000175:diagnosis / Q000201:enzymology; D004854:Herpesvirus 4, Human / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007068:Iduronidase / Q000276:immunology / Q000378:metabolism; D007150:Immunohistochemistry; D008213:Lymphocyte Activation / Q000276:immunology; D016464:Lysosomal Storage Diseases / Q000175:diagnosis* / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology / Q000276:immunology / Q000821:virology; D008059:Mucopolysaccharidosis I / Q000175:diagnosis / Q000201:enzymology; D000519:alpha-Galactosidase / Q000276:immunology / Q000378:metabolism; D000520:alpha-Glucosidases / Q000276:immunology / Q000378:metabolism; D001616:beta-Galactosidase / Q000276:immunology / Q000378:metabolism; D001617:beta-Glucosidase / Q000276:immunology / Q000378:metabolism
chemical_list: D000520:alpha-Glucosidases; D001617:beta-Glucosidase; D000519:alpha-Galactosidase; D001616:beta-Galactosidase; D007068:Iduronidase
references.citation: Clin Chim Acta. 1979 Mar 1;92(2):257-65
references.pmid: 114339
references.citation: Biochim Biophys Acta. 1984 Jun 6;794(1):89-95
references.pmid: 6329310
references.citation: Biomedicine. 1980 May;33(3):78-80
references.pmid: 7448326
references.citation: J Inherit Metab Dis. 1986;9(2):193-201
references.pmid: 3018358
references.citation: Cell Prolif. 2003 Aug;36(4):191-7
references.pmid: 12950388
references.citation: Biochim Biophys Acta. 1998 Jul 23;1381(2):123-30
references.pmid: 9685606
references.citation: Biol Cell. 1985;53(3):293-6
references.pmid: 2990625
references.citation: Cell Prolif. 2006 Feb;39(1):29-36
references.pmid: 16426420
references.citation: Biochim Biophys Acta. 1981 Jun 15;659(2):445-56
references.pmid: 6266491
references.citation: Arch Biochem Biophys. 1976 Aug;175(2):569-82
references.pmid: 958319
references.citation: Clin Chem. 2004 Oct;50(10):1785-96
references.pmid: 15292070
references.citation: Hum Genet. 1978 Oct 19;44(1):79-87
references.pmid: 213379
references.citation: Eur J Pediatr. 1997 Aug;156(8):650-4
references.pmid: 9266201
references.citation: J Lab Clin Med. 1973 Feb;81(2):157-71
references.pmid: 4683418
references.citation: Ann Biol Clin (Paris). 1983;41(5):299-309
references.pmid: 6320695
references.citation: Biol Cell. 1987;59(1):101-4
references.pmid: 3038233
references.citation: Tohoku J Exp Med. 1977 Aug;122(4):393-6
references.pmid: 199964
references.citation: J Immunol. 1982 May;128(5):1986-91
references.pmid: 6278023
references.citation: J Inherit Metab Dis. 2004;27(3):385-410
references.pmid: 15190196
references.citation: Hum Genet. 1986 Aug;73(4):320-6
references.pmid: 3017841
references.citation: J Clin Pathol. 1995 May;48(5):438-42
references.pmid: 7543118
references.citation: J Biol Chem. 1951 Nov;193(1):265-75
references.pmid: 14907713
references.citation: Nat Rev Genet. 2002 Dec;3(12):954-66
references.pmid: 12459725