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Blood, Vol. 107, Issue 6, 2364-2372, March 15, 2006
Multiplicity and plasticity of natural killer cell signaling pathways
Blood Chiesa et al.
107: 2364
Supplemental materials for: Chiesa et al
Files in this Data Supplement:
- Figure S1 (43.4 KB) -
(A) Indicated mice were weighed every two weeks. Groups of age- and sex-matched 5 mice have been tested. Data have been analyzed by statistical unpaired t Student test. As shown by comparable growth of RAGKO and ZGR mice, no sign of IBD can be detected in ZGR mice. CTR : control +/+ littermates. (B) NK cell (CD3—MHC class II—NK1.1+) percentages in splenocytes isolated from indicated mice have been represented. Dots indicate individual mice. An important augmentation of total splenocyte number was detected in ZGR and ZGKR mice, supporting that reduced splenic NK cell percentages are the consequence of the expansion of a non-NK cell population in these mice. Statistical analysis was performed as indicated in Figure 1. (C) Upper panel: expression of NK1.1 and 2B4 on CD3—MHC class II—DX5+ splenocytes isolated from RAGKO (thick line) and ZGR (thin line) mice. In ZGR mice, 2B4 NK cell receptor is not detectable because the anti-2B4 mAb is selectively reacting with C57BL/6 2B4 protein, but cannot recognize 129-derived 2b4 gene product encoded in these mutant mice (data not shown). In addition, NK1.1 expression is down-regulated in ZGR mice, consistent with the reported association of NKRP1-C with FcR .1 Lower panel: mixed bone marrow chimeras reconstituted with bone marrow cell precursors isolated from RAGKO and ZGR mice were analyzed at least four weeks after reconstitution. Splenic NK cells derived from RAGKO precursors (R1=NK1.1bright2B4+) and from ZGR precursors (R2=NK1.1dim2B4—) are present in comparable percentages, indicating that the reduced NK cell percentages in ZGR mice are not due to an intrinsic NK cell defect, but to an alteration in NK cell microenvironment. Since these experiments were performed using RAG-1-deficient bone marrow cells, the alteration in NK cell percentages is also T- and B-independent, and is thus most likely the consequence of FcR-deficiency in myeloid cells that functionally interact with NK cells, such as dendritic cells and/or macrophages.2 Data are representative of 8 MBMC analyzed. 1. Arase N, Arase H, Park SY, Ohno H, Ra C, Saito T. Association with FcRgamma Is Essential for Activation Signal through NKR-P1 (CD161) in Natural Killer (NK) Cells and NK1.1(+) T Cells. J Exp Med. 1997;186:1957-1963.
2. Degli-Esposti MA, Smyth MJ. Close encounters of different kinds: dendritic cells and NK cells take centre stage. Nat Rev Immunol. 2005;5:112-124.
- Figure S2. Anti-NKG2D mAb inhibits YAC-1 lysis mediated by control and DAP12 deficient LAK cells at similar extent (PDF, 87.6 KB) -
Purified LAK cells prepared from indicated mice were assayed for their cytolytic activity against YAC-1 cells in the presence or absence of anti-NKG2D mAb or of respective isotype control mAb. Of note, a residual lytic activity is still detectable after NKG2D blocking with both types of NK cells.
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