Abstract
Src homology region 2-containing protein tyrosine phosphatase-2 (SHP-2) is required for full activation of Ras/ERK in many cytokine and growth factor receptor signaling pathways. In contrast, SHP-2 inhibits activation of human NK cells upon recruitment to killer cell Ig-like receptors (KIR). To determine how SHP-2 impacts NK cell activation in KIR-dependent or KIR-independent signaling pathways, we employed knockdown and overexpression strategies in NK-like cell lines and analyzed the consequences on functional responses. In response to stimulation with susceptible target cells, SHP-2-silenced NK cells had elevated cytolytic activity and IFN-γ production, whereas cells overexpressing wild-type or gain-of-function mutants of SHP-2 exhibited dampened activities. Increased levels of SHP-2 expression over this range significantly suppressed microtubule organizing center polarization and granzyme B release in response to target cells. Interestingly, NK-target cell conjugation was only reduced by overexpressing SHP-2, but not potentiated in SHP-2-silenced cells, indicating that conjugation is not influenced by physiological levels of SHP-2 expression. KIR-dependent inhibition of cytotoxicity was unaffected by significant reductions in SHP-2 levels, presumably because KIR were still capable of recruiting the phosphatase under these limiting conditions. In contrast, the general suppressive effect of SHP-2 on cytotoxicity and cytokine release was much more sensitive to changes in cellular SHP-2 levels. In summary, our studies have identified a new, KIR-independent role for SHP-2 in dampening NK cell activation in response to tumor target cells in a concentration-dependent manner. This suppression of activation impacts microtubule organizing center-based cytoskeletal rearrangement and granule release.
Original language | English |
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Pages (from-to) | 7234-7243 |
Number of pages | 10 |
Journal | Journal of Immunology |
Volume | 183 |
Issue number | 11 |
DOIs | |
State | Published - Dec 1 2009 |
Keywords
- Blotting, Western
- Cell Line
- Cytoplasmic Granules/immunology
- Cytoskeleton/immunology
- Cytotoxicity, Immunologic/immunology
- Gene Knockdown Techniques
- Gene Silencing
- Granzymes/biosynthesis
- Humans
- Interferon-gamma/biosynthesis
- Killer Cells, Natural/immunology
- Lymphocyte Activation/immunology
- Microtubules/immunology
- Protein Tyrosine Phosphatase, Non-Receptor Type 11/biosynthesis
- Receptors, KIR/immunology
- Signal Transduction/immunology
- Transduction, Genetic