TY - JOUR
T1 - Different NK cell-activating receptors preferentially recruit Rab27a or Munc13-4 to perforin-containing granules for cytotoxicity
AU - Wood, Stephanie M
AU - Meeths, Marie
AU - Chiang, Samuel C C
AU - Bechensteen, Anne Grete
AU - Boelens, Jaap J
AU - Heilmann, Carsten
AU - Horiuchi, Hisanori
AU - Rosthøj, Steen
AU - Rutynowska, Olga
AU - Winiarski, Jacek
AU - Stow, Jennifer L
AU - Nordenskjöld, Magnus
AU - Henter, Jan-Inge
AU - Ljunggren, Hans-Gustaf
AU - Bryceson, Yenan T
PY - 2009
Y1 - 2009
N2 - The autosomal recessive immunodeficiencies Griscelli syndrome type 2 (GS2) and familial hemophagocytic lymphohistiocytosis type 3 (FHL3) are associated with loss-of-function mutations in RAB27A (encoding Rab27a) and UNC13D (encoding Munc13-4). Munc13-4 deficiency abrogates NK-cell release of perforin-containing lytic granules induced by signals for natural and antibody-dependent cellular cytotoxicity. We demonstrate here that these signals fail to induce degranulation in resting NK cells from Rab27a-deficient patients. In resting NK cells from healthy subjects, endogenous Rab27a and Munc13-4 do not colocalize extensively with perforin. However, phorbol 12-myristate 13-acetate and ionomycin stimulation or conjugation to susceptible target cells induced myosin-dependent colocalization of Rab27a and Munc13-4 with perforin. Unexpectedly, individual engagement of receptors leukocyte functional antigen-1, NKG2D, or 2B4 induced colocalization of Rab27a, but not Munc13-4, with perforin. Conversely, engagement of antibody-dependent cellular cytotoxicity receptor CD16 induced colocalization of Munc13-4, but not Rab27a, with perforin. Furthermore, colocalization of Munc13-4 with perforin was Rab27a-dependent. In conclusion, Rab27a or Munc13-4 recruitment to lytic granules is preferentially regulated by different receptor signals, demonstrating that individual target cell ligands regulate discrete molecular events for lytic granule maturation. The data suggest Rab27a facilitates degranulation at an early step yet highlight a reciprocal relationship between Munc13-4 and Rab27a for degranulation.
AB - The autosomal recessive immunodeficiencies Griscelli syndrome type 2 (GS2) and familial hemophagocytic lymphohistiocytosis type 3 (FHL3) are associated with loss-of-function mutations in RAB27A (encoding Rab27a) and UNC13D (encoding Munc13-4). Munc13-4 deficiency abrogates NK-cell release of perforin-containing lytic granules induced by signals for natural and antibody-dependent cellular cytotoxicity. We demonstrate here that these signals fail to induce degranulation in resting NK cells from Rab27a-deficient patients. In resting NK cells from healthy subjects, endogenous Rab27a and Munc13-4 do not colocalize extensively with perforin. However, phorbol 12-myristate 13-acetate and ionomycin stimulation or conjugation to susceptible target cells induced myosin-dependent colocalization of Rab27a and Munc13-4 with perforin. Unexpectedly, individual engagement of receptors leukocyte functional antigen-1, NKG2D, or 2B4 induced colocalization of Rab27a, but not Munc13-4, with perforin. Conversely, engagement of antibody-dependent cellular cytotoxicity receptor CD16 induced colocalization of Munc13-4, but not Rab27a, with perforin. Furthermore, colocalization of Munc13-4 with perforin was Rab27a-dependent. In conclusion, Rab27a or Munc13-4 recruitment to lytic granules is preferentially regulated by different receptor signals, demonstrating that individual target cell ligands regulate discrete molecular events for lytic granule maturation. The data suggest Rab27a facilitates degranulation at an early step yet highlight a reciprocal relationship between Munc13-4 and Rab27a for degranulation.
KW - Case-Control Studies
KW - Cell Degranulation
KW - Child
KW - Cytoplasmic Granules
KW - Cytotoxicity, Immunologic
KW - Humans
KW - Ionomycin
KW - Killer Cells, Natural
KW - Lymphocyte Activation
KW - Lymphocyte Function-Associated Antigen-1
KW - Lymphohistiocytosis, Hemophagocytic
KW - Membrane Proteins
KW - Mutation
KW - Perforin
KW - Receptors, IgG
KW - Receptors, Immunologic
KW - Signal Transduction
KW - Tetradecanoylphorbol Acetate
KW - rab GTP-Binding Proteins
U2 - 10.1182/blood-2009-06-225359
DO - 10.1182/blood-2009-06-225359
M3 - Journal article
SN - 0006-4971
VL - 114
SP - 4117
EP - 4127
JO - Blood
JF - Blood
ER -