Molecular and physiological properties of murine CD14.
We have previously cloned the murine homolog of cDNA for the human myelomonocytic differentiation antigen, CD14. We synthesized three hydrophilic peptides derived from the predicted amino acid sequence of murine CD14 (mCD14), designated MS7.1, MS7.2, and MS7.3, respectively, and raised antisera against them. Each antiserum showed specific reactivity to the same peptide used for immunization. One of the anti-mCD14 antisera directed against MS7.3 peptide (AMS7.3) demonstrated the highest titer and definitively reacted with monocytic cell lines, inflammatory polymorphonuclear cells, and macrophages. Significant cross-reactivity of AMS7.3 was observed in the human monocytic cell line, THP-1. COS-1 cells transfected with MS7 cDNA expressed an antigen recognized by AMS7.3. Resident peritoneal and alveolar macrophages both expressed mCD14. mCD14 expression in peritoneal but not alveolar macrophages increased after treatment with lipopolysaccharide. Expression of mCD14 varied among monocytic cell lines and roughly paralleled the mRNA levels except in MI cells. SDS-PAGE and isoelectric focusing analysis of immunoprecipitated mCD14 showed that mCD14 was a 53 kd disulfide-linked protein with a pI of 4.5-5.1. Reduction of molecular weight by endo F treatment demonstrated that mCD14 was an N-linked glycoprotein. Since mCD14 is shed from the cell surface membrane by phosphatidylinositol-specific phospholipase C treatment, the indication is that mCD14 is a phosphatidylinositol-linked protein. The soluble form of mCD14 was detectable. Treatment with anti-mCD14 before interferon gamma (IFN gamma) stimulation significantly enhanced IFN gamma-induced H-2 antigen expression in the macrophage cell line.
Nasu
N
N
Department of Pathology, Medical College of Oita, Japan.
Yoshida
S
S
Akizuki
S
S
Higuchi
Y
Y
Setoguchi
M
M
Yamamoto
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D015214:Antigens, Differentiation, Myelomonocytic / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D003429:Cross Reactions / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D015870:Gene Expression; D006183:H-2 Antigens / Q000276:immunology; D018950:Lipopolysaccharide Receptors; D008264:Macrophages / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D014162:Transfection; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000494:pharmacology
D015214:Antigens, Differentiation, Myelomonocytic; D000939:Epitopes; D006183:H-2 Antigens; D018950:Lipopolysaccharide Receptors; D010455:Peptides; D012333:RNA, Messenger; D010738:Type C Phospholipases