scholarly journals Restoration of arylsulphatase A activity in human-metachromatic-leucodystrophy fibroblasts via retroviral-vector-mediated gene transfer

1991 ◽  
Vol 280 (2) ◽  
pp. 459-461 ◽  
Author(s):  
W Rommerskirch ◽  
A L Fluharty ◽  
C Peters ◽  
K von Figura ◽  
V Gieselmann
Keyword(s):  
Storage Disease ◽  
Metachromatic Leukodystrophy ◽  
Normal Individual ◽  
Human Fibroblasts ◽  
Retroviral Vector ◽  
Lysosomal Storage ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Metachromatic Leucodystrophy ◽  
Arylsulphatase A

Metachromatic leukodystrophy is a lysosomal storage disease caused by the deficiency of arylsulphatase A (ASA). A human ASA cDNA was subcloned into the retroviral vector pXT1. Replication-defective retrovirus was generated by transfection of the vector into the amphotropic packaging cell line PA317. Human fibroblasts from a patient suffering from metachromatic leucodystrophy was infected with the recombinant retrovirus. Infected fibroblasts expressed ten times more ASA compared with control fibroblasts from a normal individual. The ASA encoded by the integrated provirus was shown to be correctly transported into the lysosomes and to normalize the impaired degradation of cerebroside sulphate.

scholarly journals Restoration of arylsulphatase B activity in human mucopolysaccharidosis-type-VI fibroblasts by retroviral-vector-mediated gene transfer

1991 ◽  
Vol 276 (2) ◽  
pp. 499-504 ◽  
Author(s):  
C Peters ◽  
W Rommerskirch ◽  
S Modaressi ◽  
K von Figura
Keyword(s):  
Gene Transfer ◽  
Retroviral Vector ◽  
Mucopolysaccharidosis Type ◽  
Lysosomal Storage ◽  
Packaging Cell Line ◽  
Retroviral Gene Transfer ◽  
Gene Replication ◽  
Packaging Cell ◽  
Mucopolysaccharidosis Type Vi ◽  
Mps Vi

The Maroteaux-Lamy syndrome (mucopolysaccharidosis type VI; MPS VI) is a lysosomal storage disease caused by deficiency of the enzyme arylsulphatase B (ASB). A human ASB cDNA has been subcloned into the retroviral vector pXT1 containing the bacterial neomycin-resistance gene and an internal thymidine kinase promoter for transcription of the inserted gene. Replication defective retrovirus was generated by transfecting the construct into the amphotropic packaging cell line PA317. Human MPS VI fibroblasts infected with recombinant retrovirus integrated the provirus into their genome and expressed retrovirus-encoded ASB mRNAs. In infected fibroblasts the level of ASB was up to 36-fold higher than in normal fibroblasts. Biosynthesis and processing of ASB in infected MPS VI fibroblasts was accomplished as in normal fibroblasts, and mature, enzymically active, ASB accumulated in dense lysosomes, indicating that the ASB deficiency in MPS VI fibroblasts was corrected by the retroviral gene transfer.

Efficient transduction and selection of human T-lymphocytes with bicistronic Thy1/HSV1-TK retroviral vector produced by a human packaging cell line

2004 ◽  
Vol 6 (4) ◽  
pp. 374-386 ◽  
Author(s):  
François M. Lemoine ◽  
Mariana Mesel-Lemoine ◽  
Mustapha Cherai ◽  
Géraldine Gallot ◽  
Henri Vié ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cell Line ◽  
Retroviral Vector ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Human T Lymphocytes ◽  
Selection Of

scholarly journals Optimization of a retroviral vector for transduction of human CD34 positive cells.

2006 ◽  
Vol 53 (4) ◽  
pp. 815-823 ◽  
Author(s):  
Anna Szyda ◽  
Maria Paprocka ◽  
Agnieszka Krawczenko ◽  
Katarzyna Lenart ◽  
Jerzy Heimrath ◽  
...  
Keyword(s):  
Cord Blood ◽  
Cell Lines ◽  
Ex Vivo ◽  
Retroviral Vectors ◽  
Retroviral Vector ◽  
Transduction Efficiency ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Human Cd34 ◽  
Stem And Progenitor Cells

Human stem and progenitor cells have recently become objects of intensive studies as an important target for gene therapy and regenerative medicine. Retroviral vectors are among the most effective tools for genetic modification of these cells. However, their transduction efficiency strongly depends on the choice of the ex vivo transduction system. The aim of this study was to elaborate a system for retroviral vector transduction of human CD34 positive cells isolated from cord blood. The retroviral vector pMINV EGFP was chosen for transduction of two human erythroblastoid cell lines: KG-1a (CD34 positive) and K562 (CD34 negative). For vector construction, three promoters and two retroviral vector packaging cell lines were used. To optimize the physicochemical conditions of the transduction process, different temperatures of supernatant harvesting, the influence of centrifugation and the presence of transduction enhancing agents were tested. The conditions elaborated with KG-1a cells were further applied for transduction of CD34 positive cells isolated from cord blood. The optimal efficiency of transduction of CD34 positive cells with pMINV EGFP retroviral vector (26% of EGFP positive cells), was obtained using infective vector with LTR retroviral promoter, produced by TE FLY GA MINV EGFP packaging cell line. The transduction was performed in the presence of serum, at 37 degrees C, with co-centrifugation of cells with viral supernatants and the use of transduction enhancing agents. This study confirmed that for gene transfer into CD34 positive cells, the detailed optimization of each element of the transduction process is of great importance.

scholarly journals Epidemiology and Genotyping of Patients with Lysosomal Storage Disease in Malaysia.

2021 ◽  
Author(s):  
Affandi Omar ◽  
Rosnani Mohamed ◽  
Fatimah Diana Amin Nordin ◽  
Norashareena Mohamed Shakrin ◽  
Sofwatul Mukhtaroh Nasohah ◽  
...  
Keyword(s):  
Storage Disease ◽  
Metachromatic Leukodystrophy ◽  
Neuronal Ceroid Lipofuscinoses ◽  
Type Ii ◽  
Lysosomal Storage ◽  
Asian Countries ◽  
Birth Prevalence ◽  
Mps Ii ◽  
Storage Disorders ◽  
Lysosomal Transport

Abstract Background: Lysosomal storage disorders (LSD) are storage disorders involving malfunction of degradation enzymes in lysosome. More than 50 types of LSD have been discovered, which includes the group of mucopolysaccharidoses (MPS), sphingolipidoses, oligosaccharidoses, mucolipidoses, lipoprotein storage disorders, lysosomal transport defects and neuronal ceroid lipofuscinoses and others. The aims of this study were to calculate the birth prevalence and carrier frequency of LSDs in the Malaysian population; to compare our results with reported epidemiologic data from other populations, and to describe the mutation spectrum in Malaysia. From 2008 to 2017, 2.1% (92/4338) suspected patients were diagnosed with LSD. Results: The prevalence of LSD in Malaysia was 1/231,904 live births. The combined prevalence of MPS was 1/292,401 with its subtype of MPS II presented the highest calculated birth prevalence of 1/221,425. Within the group of sphingolipidoses, the combine prevalence was 1/770,777 with Fabry as the most common disorder with calculated prevalence of 1/193,203 followed by metachromatic leukodystrophy (MLD) (1/494,514). MLD is more common among people of Iban ethnicity with the prevalence of 1/6,981. Pompe and mucolipidoses type II are the less common subtypes of LSD with a prevalence of 1/1,694,634 and 1/2,229,516, respectively.Conclusion: Overall, although the prevalence of LSD in Malaysia may be underestimated, the prevalence of MPS is consistent with other reported in East Asian countries.

scholarly journals Successful Reconstitution of Human Hematopoiesis in the SCID-hu Mouse by Genetically Modified, Highly Enriched Progenitors Isolated From Fetal Liver

Blood ◽  
1997 ◽  
Vol 90 (9) ◽  
pp. 3496-3506 ◽  
Author(s):  
Laurent Humeau ◽  
Christian Chabannon ◽  
Meri T. Firpo ◽  
Patrice Mannoni ◽  
Claude Bagnis ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fetal Liver ◽  
Genetically Modified ◽  
Retroviral Vector ◽  
Hematopoietic Progenitors ◽  
Packaging Cell Line ◽  
Retroviral Gene Transfer ◽  
Packaging Cell ◽  
Hematopoietic Reconstitution ◽  
B Lineage

Abstract Highly purified CD34++CD38−Lin− hematopoietic progenitors isolated from human fetal liver were infected with the murine retroviral vector, MFG nls-LacZ, which encodes a modified version of the Escherichia coli β-galactosidase gene. Progenitors that were cocultured with the packaging cell line could reconstitute human bone marrow or thymus implanted in SCID-hu mice. Expression of the β-galactosidase gene was observed in primitive and committed clonogenic progenitors, mature myeloid, B-lineage cells, and T-lineage cells for up to 4 months after injection into SCID-hu mice. Furthermore, hematopoietic reconstitution by genetically modified progenitor cells could be achieved by the injection of the cells generated from as few as 500 CD34++CD38−Lin− cells, suggesting efficient retroviral gene transfer into fetal liver progenitors.

scholarly journals The effect of chloroquine on the metabolism of [35S]cystine in normal and cystinotic human skin fibroblasts

1981 ◽  
Vol 200 (3) ◽  
pp. 555-563 ◽  
Author(s):  
C J Danpure
Keyword(s):  
Storage Disease ◽  
Human Fibroblasts ◽  
Total Cell ◽  
Optimum Concentration ◽  
Cell Uptake ◽  
Lysosomal Storage ◽  
Normal Cells ◽  
Normal Human ◽  
Uptake And Metabolism

The present study concerns the effect of the lysosomotropic drug chloroquine on the uptake and metabolism of [35S]cystine in vitro by normal human fibroblasts and those from patients suffering from the lysosomal storage disease cystinosis. When the cells were cultured with [35S]cystine for periods in excess of 4 h, it was found that chloroquine considerably increased (up to 30-fold) the labelling of the intracellular cystine pool in cystinotic cells, with no increase or a much smaller increase in normal cells. For this effect chloroquine had an optimum concentration of 20 microM, with a small effect still being noticeable at 1 microM. A quinoline analogue, 4-(dimethylaminoethylamino)-7-iodoquinoline, had a similar effect to chloroquine. However, NH4Cl at concentrations of between 100 microM and 50 mM showed either no effect (at the lower concentrations) or a depression of intracellular cystine labelling (at the higher concentrations). The differences between the effects of the quinolines on cystinotic acid normal cells were not due to differences in total cell uptake of drug.

scholarly journals Defective Retroviral Endogenous RNA Is Efficiently Transmitted by Infectious Particles Produced on an Avian Retroviral Vector Packaging Cell Line

Virology ◽  
1995 ◽  
Vol 207 (1) ◽  
pp. 271-275 ◽  
Author(s):  
Corinne Ronfort ◽  
Anne Girod ◽  
François-Loı̈c Cosset ◽  
Catherine Legras ◽  
Victor Marc Nigon ◽  
...  
Keyword(s):  
Cell Line ◽  
Retroviral Vector ◽  
Packaging Cell Line ◽  
Packaging Cell

Is there a role for scintigraphic imaging of bone manifestations in Gaucher disease?

2008 ◽  
Vol 47 (06) ◽  
pp. 239-247 ◽  
Author(s):  
S. Kohlfürst ◽  
H. J. Gallowitsch ◽  
E. Kresnik ◽  
P. Lind ◽  
A. B. Mehta ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Lysosomal Storage Disease ◽  
Gaucher Disease ◽  
Storage Disease ◽  
Lysosomal Storage ◽  
Scintigraphic Imaging ◽  
Imaging Methods ◽  
X Ray ◽  
Deficient Activity

SummaryGaucher disease is the most prevalent inherited, lysosomal storage disease and is caused by deficient activity of the enzyme β-glucocerebrosidase. Bone and bone marrow alterations are frequent in the most prevalent non-neuronopathic form of Gaucher disease. Imaging of bone manifestations in Gaucher disease is performed by a variety of imaging methods, conventional X-ray and MRI as the most frequently and most important ones. However, different modalities of scintigraphic imaging have also been used. This article gives an overview on scintigraphic imaging with respect to bone manifestations in Gaucher disease discussing the advantages and limitations of scintigraphic imaging in comparison to other imaging methods.

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