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Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, University Hospital, Ghent, Belgium
Murine NK cells express inhibitory receptors belonging to the Ly49
and CD94/NKG2 family. Ly49E and CD94 are the only NK cell receptor
transcripts detectable in fetal NK cells. Still unproved is the surface
expression of Ly49E on NK cells. Here we generated two novel mAbs, a
mAb recognizing Ly49E with cross-reactivity to Ly49C, and a mAb against
NKG2A/C/E. Ly49E was immunoprecipitated as a disulfide-linked homodimer
with 46-kDa subunits. Removal of N-linked carbohydrates revealed a
31-kDa protein backbone. NKG2A was immunoprecipitated as a 38-kDa
protein. Although the frequency of fetal NK cells expressing Ly49E was
higher than 25%, it decreased drastically from 2 wk after birth.
Phenotypic analysis showed that 
90% of fetal NK cells and 50%
of adult NK cells express high levels of CD94/NKG2. The remaining 50%
of adult NK cells expressed low surface levels of CD94/NKG2. Expression
of Ly49E and CD94/NKG2 was not restricted to NK cells, but was also
observed on NK T and memory T cells. Functional analysis showed that
sorted Ly49E+ and CD94/NKG2+ fetal NK cells
could discriminate between MHC class I-positive and MHC class
I-negative tumor cells. We also demonstrated that Ly49E becomes
phosphorylated following pervanadate stimulation of fetal NK cells. The
expression levels of Ly49E and CD94/NKG2 were similar in wild-type
compared with 
2-microglobulin-/- mice. In
conclusion, generation of mAbs against Ly49E and NKG2 extended the
phenotypic and functional characterization of NK
cells.
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