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J Clin Immunol. 2017 Jan;37(1):92-99. doi: 10.1007/s10875-016-0357-3. Epub 2016 Nov 28.

A CD57+ CTL Degranulation Assay Effectively Identifies Familial Hemophagocytic Lymphohistiocytosis Type 3 Patients.

Author information

1
Department of Pediatrics, Kyoto University Graduate School of Medicine, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan.
2
Department of Pediatrics, Kyoto University Graduate School of Medicine, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. yasumi@kuhp.kyoto-u.ac.jp.
3
Laboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan.
4
Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
5
Department of Pediatrics, Ehime University Graduate School of Medicine, Toon, Japan.
6
Department of Molecular and Cellular Biology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.
7
Department of Biomedical Statistics and Bioinformatics, Kyoto University Graduate School of Medicine, Kyoto, Japan.
8
KAZUSA DNA Research Institute, Kisarazu, Japan.

Abstract

PURPOSE:

Familial hemophagocytic lymphohistiocytosis type 3 (FHL3) is a genetic disorder that results in immune dysregulation. It requires prompt and accurate diagnosis. A natural killer (NK) cell degranulation assay is often used to screen for FHL3 patients. However, we recently encountered two cases of late-onset FHL3 carrying novel UNC13D missense mutations: in these cases, the degranulation assays using freshly isolated and interleukin (IL)-2-activated NK cells yielded contradictory results. Since the defective degranulation of CD57+ cytotoxic T lymphocytes (CTLs) in these cases was helpful for making the diagnosis, we assessed whether the CD57+ CTL degranulation assay more effectively identified FHL3 patients than the NK cell assays.

METHODS:

Forty additional patients with hemophagocytic lymphohistiocytosis were prospectively screened for FHL3 by measuring the perforin expression in NK cells and the expression of Munc13-4, syntaxin-11, and Munc18-2 in platelets and by performing NK cell and CTL degranulation assays. The results were confirmed by genetic analysis.

RESULTS:

The freshly isolated NK cell degranulation assay detected FHL3 patients with high sensitivity (100%) but low specificity (71%). The IL-2-stimulated NK cell assay had improved specificity, but 3 out of the 31 non-FHL3 patients still showed degranulation below the threshold level. The CD57+ CTL degranulation assay identified FHL3 patients with high sensitivity and specificity (both 100%).

CONCLUSIONS:

The CD57+ CTL degranulation assay more effectively identified FHL3 patients than the NK cell-based assays.

KEYWORDS:

Familial hemophagocytic lymphohistiocytosis type 3; UNC13D; functional screening assay; lysosomal degranulation defect

PMID:
27896523
DOI:
10.1007/s10875-016-0357-3
[Indexed for MEDLINE]

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