Expression of a neutrophil chemotactic protein LECT2 in human hepatocytes revealed by immunochemical studies using polyclonal and monoclonal antibodies to a recombinant LECT2.
A recombinant human neutrophil chemotactic protein LECT2 (rhLECT2) was purified as a 16-kDa protein from the culture fluids of stable transfectants derived from CHO cells (clone C1D8-1) and L929 cells (clone L2E4-1). The N-terminal amino acid sequence of the protein secreted by both clones were homologous to the previously described bovine LECT2. We produced polyclonal and monoclonal antibodies against rhLECT2 and investigated secretion of LECT2 protein in six human hepatoma cell lines, which express LECT2 mRNA, and in hepatocytes of normal human livers by a sandwich enzyme-linked immunosorbent assay and by immunostaining using the antibodies, respectively. We revealed that five of six hepatoma cell lines secreted LECT2 into culture fluids at concentrations of 30-135 ng/mg. We also demonstrated that the cytoplasm of human hepatocytes was diffusely stained, although periportal hepatocytes tended to be weakly and granularly stained by immunostaining. These results indicated that the novel protein was expressed in hepatocytes and suggested an important role of LECT2 in the cells in addition to the activation of neutrophils.
Yamagoe
S
S
Department of Bioactive Molecules, National Institute of Infectious Diseases, Tokyo, Japan.
Akasaka
T
T
Uchida
T
T
Hachiya
T
T
Okabe
T
T
Yamakawa
Y
Y
Arai
T
T
Mizuno
S
S
Suzuki
K
K
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D016466:CHO Cells; D006528:Carcinoma, Hepatocellular; D002417:Cattle; D002630:Chemotactic Factors / Q000096:biosynthesis*; D006224:Cricetinae; D006801:Humans; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins; D007739:L Cells; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008113:Liver Neoplasms; D051379:Mice; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000911:Antibodies, Monoclonal; D002630:Chemotactic Factors; D036341:Intercellular Signaling Peptides and Proteins; C491214:LECT2 protein, Bos taurus; C103468:LECT2 protein, human; C491213:Lect2 protein, mouse; D011506:Proteins; D011994:Recombinant Proteins