Quantifying lymphocyte vacuolization serves as a measure of CLN3 disease severity.

Autor: Kuper WFE; Department of Metabolic Diseases, Wilhelmina Children's Hospital University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., Oostendorp M; Department of Clinical Chemistry University Medical Center Utrecht Utrecht the Netherlands., van den Broek BTA; Sylvia Toth Center for the Multidisciplinary Follow up of Lysosomal Storage Disorders University Medical Center Utrecht, Utrecht University Utrecht the Netherlands.; Laboratory of Translational Immunology University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., van Veghel K; Laboratory of Translational Immunology University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., Nonkes LJP; Department of Clinical Chemistry University Medical Center Utrecht Utrecht the Netherlands., Nieuwenhuis EES; Department of Gastroenterology Wilhelmina Children's Hospital, University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., Fuchs SA; Department of Metabolic Diseases, Wilhelmina Children's Hospital University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., Veenendaal T; Section Cell Biology, Center for Molecular Medicine University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., Klumperman J; Section Cell Biology, Center for Molecular Medicine University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., Huisman A; Department of Clinical Chemistry University Medical Center Utrecht Utrecht the Netherlands., Nierkens S; Laboratory of Translational Immunology University Medical Center Utrecht, Utrecht University Utrecht the Netherlands., van Hasselt PM; Department of Metabolic Diseases, Wilhelmina Children's Hospital University Medical Center Utrecht, Utrecht University Utrecht the Netherlands.; Sylvia Toth Center for the Multidisciplinary Follow up of Lysosomal Storage Disorders University Medical Center Utrecht, Utrecht University Utrecht the Netherlands.
Jazyk: angličtina
Zdroj: JIMD reports [JIMD Rep] 2020 Jun 02; Vol. 54 (1), pp. 87-97. Date of Electronic Publication: 2020 Jun 02 (Print Publication: 2020).
DOI: 10.1002/jmd2.12128
Abstrakt: Background: The CLN3 disease spectrum ranges from a childhood-onset neurodegenerative disorder to a retina-only disease. Given the lack of metabolic disease severity markers, it may be difficult to provide adequate counseling, particularly when novel genetic variants are identified. In this study, we assessed whether lymphocyte vacuolization, a well-known yet poorly explored characteristic of CLN3 disease, could serve as a measure of disease severity.
Methods: Peripheral blood obtained from healthy controls and CLN3 disease patients was used to assess lymphocyte vacuolization by (a) calculating the degree of vacuolization using light microscopy and (b) quantifying expression of lysosomal-associated membrane protein 1 (LAMP-1), using flow cytometry in lymphocyte subsets as well as a qualitative analysis using electron microscopy and ImageStream analysis.
Results: Quantifying lymphocyte vacuolization allowed to differentiate between CLN3 disease phenotypes ( P = .0001). On immunofluorescence, classical CLN3 disease lymphocytes exhibited abundant vacuole-shaped LAMP-1 expression, suggesting the use of LAMP-1 as a proxy for lymphocyte vacuolization. Using flow cytometry in lymphocyte subsets, quantifying intracellular LAMP-1 expression additionally allowed to differentiate between infection and storage and to differentiate between CLN3 phenotypes even more in-depth revealing that intracellular LAMP-1 expression was most pronounced in T-cells of classical-protracted CLN3 disease while it was most pronounced in B-cells of "retina-only" CLN3 disease.
Conclusion: Lymphocyte vacuolization serves as a proxy for CLN3 disease severity. Quantifying vacuolization may help interpretation of novel genetic variants and provide an individualized readout for upcoming therapies.
Competing Interests: The authors declare no potential conflicts of interest.
(© 2020 The Authors. Journal of Inherited Metabolic Disease published by John Wiley & Sons Ltd on behalf of SSIEM.)
Databáze: MEDLINE
Externí odkaz: