A platform for complementation and characterization of familial haemophagocytic lymphohistiocytosis 3 mutations

作者：Elstak Edo; de Jong Arjan; van der Sluijs Peter^*

来源：Journal of Immunological Methods, 2011, 365(1-2): 58-66.

DOI：10.1016/j.jim.2010.12.009

• Haemophagocytic lymphohistiocytosis
• Munc13-4
• Degranulation
• RBL-2H3

Mutations in UNC13D cause the severe immune disorder familial haemophagocytic lymphohistiocytosis type 3 (FHL3). The gene product munc13-4 is expressed in hematopoietic cells and is essential for degranulation. Little information is available on genotype-phenotype relationships of UNC13D mutations. Some mutants may have residual functionality which qualifies them as promising targets for attempts to enhance function pharmacologically. A problem for such analysis is the scarcity of patient material. We established assays in the RBL-2H3 cell line to assess functionality of lentivirally transduced munc13-4 mutants. The basic principle of which is to silence endogenous rat munc13-4 and replace it with siRNA resistant YFP-tagged human variants. Localization, degranulation, and membrane binding kinetics can now easily be analyzed quantitatively. Such a system might also be, useful to screen small molecular weight compounds for their ability to rescue degranulation in cells with reduced functional munc13-4.

• 出版日期2011-2-28