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 β2-microglobulin−/− mice. In conclusion, generation of mAbs against Ly49E and NKG2 extended the phenotypic and functional characterization of NK cells.

The expression of the neu oncogene product was investigated in invasive and non-invasive ductal carcinomas of the breast, non-neoplastic lesions of the breast, fragments of normal adult and fetal breasts and in several other normal and fetal tissues at different weeks of pregnancy by means of an immunohistochemical study with monoclonal antibodies. The staining pattern along the cytoplasmic membrane was specific for malignancy and occurred in 29% of the breast carcinomas. It was observed in invasive carcinomas as well as in ductal carcinoma in situ and it showed a significantly higher expression in premenopausal women than in postmenopausal women. This higher expression was also present in oestrogen receptor-negative tumours. The tubules of the fetal and adult kidney, the absorption cells of the fetal and adult small and large intestine, the sebaceous glands of the fetal and adult small and large intestine, the sebaceous glands of the fetal and adult skin, the adult endocervix, the endometrium, the C-cells of the thyroid, hepatocytes and all ductal cells of the fetal breast showed a constant diffuse intracytoplasmic granular staining. staining. The same granular intracytoplasmic staining pattern was focally observed in rare cases of normal breast tissue in adults and in some cases of epitheliosis, aprocrine metaplasia and some breast carcinoma cells, which did not express neu oncogene product on their membrane. Western blot experiments showed that the cytoplasmic protein had a molecular weight of 155 kD (kilodaltons); the membrane protein is the known 185 kD neu protein.

No abstract

No abstract

No abstract