title: Analysis of natural killer-cell function in familial hemophagocytic lymphohistiocytosis (FHL): defective CD107a surface expression heralds Munc13-4 defect and discriminates between genetic subtypes of the disease.
abstract: Natural killer (NK) cells from patients with familial hemophagocytic lymphohistiocytosis because of PRF1 (FHL2, n = 5) or MUNC13-4 (FHL3, n = 8) mutations were cultured in IL-2 prior to their use in various functional assays. Here, we report on the surface CD107a expression as a novel rapid tool for identification of patients with Munc13-4 defect. On target interaction and degranulation, FHL3 NK cells displayed low levels of surface CD107a staining, in contrast to healthy control subjects or perforin-deficient NK cells. B-EBV cell lines and dendritic cell targets reveal the FHL3 NK-cell defect, whereas highly susceptible tumor targets were partially lysed by FHL3 NK cells expressing only trace amounts of Munc13-4 protein. Perforin-deficient NK cells were completely devoid of any ability to lyse target cells. Cytokine production induced by mAb-crosslinking of triggering receptors was comparable in patients and healthy control subjects. However, when cytokine production was induced by coculture with 721.221 B-EBV cells, FHL NK cells resulted in high producers, whereas control cells were almost ineffective. This could reflect survival versus elimination of B-EBV cells (ie, the source of NK-cell stimulation) in patients versus healthy control subjects, thus mimicking the pathophysiologic scenario of FHL.
authors.affiliation: Istituto Giannina Gaslini, Genoa, Italy; the Sir William Dunn School of Pathology, Oxford, United Kingdom.
authors.forename: Stefania
authors.initials: S
authors.lastname: Marcenaro
authors.forename: Federico
authors.initials: F
authors.lastname: Gallo
authors.forename: Stefania
authors.initials: S
authors.lastname: Martini
authors.forename: Alessandra
authors.initials: A
authors.lastname: Santoro
authors.forename: Gillian M
authors.initials: GM
authors.lastname: Griffiths
authors.forename: Maurizio
authors.initials: M
authors.lastname: Aricó
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Moretta
authors.forename: Daniela
authors.initials: D
authors.lastname: Pende
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D002648:Child; D002675:Child, Preschool; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007223:Infant; D007694:Killer Cells, Natural / Q000166:cytology* / Q000378:metabolism; D051359:Lymphohistiocytosis, Hemophagocytic / Q000097:blood* / Q000235:genetics*; D052118:Lysosomal-Associated Membrane Protein 1 / Q000096:biosynthesis*; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics*; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins
chemical_list: D016207:Cytokines; D052118:Lysosomal-Associated Membrane Protein 1; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D052899:Pore Forming Cytotoxic Proteins; C507969:UNC13D protein, human; D054353:Perforin