A receptor for the lipocalin placental protein 14 on human monocytes.
Human placental protein 14 (PP14), a member of the lipocalin structural superfamily, is an abundant amniotic fluid glycoprotein with documented immunoinhibitory activities. While receptors have been characterized for several other lipocalins, none have been reported to date for PP14. In the present study, two-color immunofluorescence and flow cytometry was used to screen peripheral blood mononuclear cell subpopulations for their capacity to engage fluoresceinated recombinant PP14. The tagged PP14 bound strongly in a specific and saturable fashion to CD14+ (monocyte lineage) cells, but not to CD20+ (B cell lineage) or CD3+ (T cell lineage) cells. This binding was both pH- and temperature-sensitive, and was reduced by proteolytic pre-digestion of the cells with trypsin or proteinase K. Scatchard analysis demonstrated a single class of receptors on CD14+ cells, with a K(D) of approximately 1 x 10(-8) and approximately 10-35,000 receptors per cell. These findings constitute the first report of a cell surface-associated binding protein for PP14 and set the stage for exploring the molecular mechanisms of PP14-mediated signaling and immunomodulation.
Miller
R E
RE
Department of Pediatrics, Case Western Reserve University, Cleveland, OH 44106, USA.
Fayen
J D
JD
Chakraborty
S
S
Weber
M C
MC
Tykocinski
M L
ML
D015703:Antigens, CD / Q000097:blood*; D018951:Antigens, CD20 / Q000097:blood; D017252:CD3 Complex / Q000097:blood; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D000071816:Glycodelin; D006023:Glycoproteins / Q000097:blood*; D006801:Humans; D007700:Kinetics; D018950:Lipopolysaccharide Receptors / Q000097:blood; D008214:Lymphocytes / Q000145:classification / Q000276:immunology / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism*; D011257:Pregnancy Proteins / Q000097:blood*; D011994:Recombinant Proteins / Q000097:blood
D015703:Antigens, CD; D018951:Antigens, CD20; D017252:CD3 Complex; D000071816:Glycodelin; D006023:Glycoproteins; D018950:Lipopolysaccharide Receptors; C045246:PAEP protein, human; D011257:Pregnancy Proteins; D011994:Recombinant Proteins