and “type”:”entrez-nucleotide”,”attrs”:”text”:”GM065230″,”term_id”:”221621386″GM065230 to N.R.J.G. DNAM-1? and DNAM-1+ Ly49H+ degranulated and produced IFN- when co-cultured with RMA target cells transduced to express m157 (Number 1E). Co-expression of CD155, a ligand of DNAM-1, within the m157+ RMA cells did not increase the rate of recurrence of Ly49H+ NK cells degranulating or generating IFN-. In a similar fashion we adoptively transferred wildtype (WT) Ly49H+ NK cells into syngeneic DAP12-deficient recipient mice, which lack functionally proficient Ly49H+ NK cells and are unable to control early replication of MCMV (Sj?lin et al., 2002; Sun et al., 2009). After illness with MCMV, both DNAM-1? and DNAM-1+ Ly49H+ NK cells produced IFN- on day time 1.5 post-infection (pi) (Figure 1F). These findings demonstrate that DNAM-1 is not required for m157-induced degranulation or cytokine production by Ly49H+ NK cells. DNAM-1 antibody blockade suppresses the NK cell response to MCMV We tackled the part of DNAM-1 in the control of MCMV by treating C57BL/6 mice having a nondepleting, neutralizing anti-DNAM-1 mAb one day prior to illness and measured viral titers on day time 3, when NK cells are mainly responsible for antiviral immunity. Blocking DNAM-1 resulted in a significant increase in viral weight in the spleen and liver (Number 2A). Open in a CD117 separate window Number 2 DNAM-1 antibody blockade suppresses the NK cell response to MCMV(A) WT mice were inoculated with control Ig or a neutralizing mAb against DNAM-1 on TAK-063 the day before illness with 5 105 pfu MCMV. Viral burden in the liver and spleen TAK-063 was measured on day time 3 pi. (B) One hundred thousand WT Ly49H+ NK cells were transferred into DAP12-deficient mice and infected with 1 105 pfu MCMV. Mice were inoculated with 100 g control Ig or anti-DNAM-1 mAb on the day before illness and day time 3 pi. The complete quantity of Ly49H+ NK cells per ml blood. (C) DAP12-deficient mice receiving 1 105 WT Ly49H+ NK cells were infected TAK-063 with MCMV, and treated with 100 g control Ig or anti-DNAM-1 mAb on days 7, 14, and 21 pi. The complete quantity of Ly49H+ NK cells per TAK-063 ml blood and percentages of Ly49H+ memory space NK cells in the spleen on day time 28 pi. Data were pooled from 2 experiments (= 6 mice per mAb group). Error bars display s.e.m. *<0.05, **<0.01. See also Figure S1. Ly49H+ NK cells preferentially increase after MCMV illness and are required for early control of viral replication (Brown et al., 2001; Dokun et al., 2001; Lee et al., 2001). When a limiting quantity of Ly49H+ NK cells are adoptively transferred into Ly49H-deficient mice and infected with MCMV, these Ly49H+ NK cells undergo extensive development and give rise to memory space Ly49H+ NK cells (Sun et al., 2009). Monitoring of donor Ly49H+ NK cells in the blood displays the reactions in the spleen and liver, as previously shown (Sun et al., 2010). Memory space Ly49H+ NK cells can be TAK-063 operationally defined as KLRG1high, CD11b+, CD27?, Ly6Chigh Ly49H+ NK cells that persist for more than 25 days after illness with MCMV. To determine the effect of DNAM-1 blockade within the clonal development of Ly49H+ NK cells and generation of memory space NK cells, we enriched NK cells and adoptively transferred wildtype (WT) Ly49H+ NK cells into syngeneic DAP12-deficient recipient mice (Number S1). These mice were injected having a neutralizing anti-DNAM-1 mAb on the day before illness and on day time 3 pi. Development of donor Ly49H+ NK cells in the peak of the NK cell response during MCMV illness was suppressed by anti-DNAM-1 antibody (Number 2B); however, Ly49H+ NK.