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
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
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DOI：
10.1182/blood-2006-04-015693
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发表时间：
2006-10-01
期刊：
影响因子：
20.3
通讯作者：
Pende, Daniela
中科院分区：
文献类型：
--
作者：
Marcenaro, Stefania;Gallo, Federico;Pende, Daniela
自然杀伤细胞（NK）来自家族性噬血细胞性淋巴组织细胞病患者，由于PRF1 （FHL2, n = 5）或MUNC13-4 （FHL3, n = 8）突变，在IL-2中培养，然后用于各种功能测定。在这里，我们报道了CD107a表面表达作为一种新的快速识别Munc13-4缺陷患者的工具。在靶相互作用和脱颗粒过程中，与健康对照者或穿孔蛋白缺陷NK细胞相比，FHL3 NK细胞表面CD107a染色水平较低。B-EBV细胞系和树突状细胞靶点显示FHL3 nk细胞缺陷，而高度敏感的肿瘤靶点被仅表达微量Munc13-4蛋白的FHL3 INK细胞部分裂解。穿孔素缺乏的NK细胞完全缺乏任何裂解靶细胞的能力。单克隆抗体-交联触发受体诱导的细胞因子产生在患者和健康对照者中具有可比性。然而，当与721.221 B-EBV细胞共培养诱导细胞因子产生时，FHL NK细胞产生大量细胞因子，而对照细胞几乎无效。这可能反映了患者与健康对照组相比B-EBV细胞（即nk细胞刺激的来源）的存活与消除，从而模拟了FHL的病理生理情况。
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 INK 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.