Human keratinocyte membrane lectins: characterization and modulation of their expression by cytokines.
In an attempt to identify cell surface molecules involved in recognition phenomena between cells such as keratinocytes and melanocytes and putatively target biological responses modifiers to keratinocytes, we undertook the detection of cell surface sugar specific receptors: membrane lectins. Keratinocyte membrane lectins were found to bind synthetic glycoproteins (neoglycoproteins) carrying either alpha-L-fucosyl or alpha-L-rhamnosyl residues. Fluorescence microscopy observations indicate that cultured keratinocytes are able to bind these two neoglycoproteins while frozen sections of human skin labelled with neoglycoprotein-coated covaspheres show that the selectivity of the binding to keratinocytes is restricted to alpha-L-rhamnosyl-BSA. Keratinocytes were adapted to grow on collagen; harvesting conditions allowing the analysis of keratinocytes by flow cytometry are described. This technique allows the quantification of the binding at 4 degrees C, and the estimation of the endocytosis of F-, neoglycoproteins: F-, alpha-L-Rha-BSA and F-, alpha-L-Fuc-BSA were efficiently internalized. Thereafter, alpha-L-rhamnose-substituted liposomes containing 5-(6)carboxyfluorescein were prepared in order to follow the delivery of the fluorescent dye into cells. This was measured both by flow cytometry and by spectrofluorimetry. The expression of surface lectins was checked upon action of cytokines (IL1 alpha, IL1 beta, IL2 and TNF) which are known as biological response modifiers of keratinocytes.
Cerdan
D
D
Département de biochimie des glycoconjugués, Centre de biophysique moléculaire, CNRS, Orléans, France.
Grillon
C
C
Monsigny
M
M
Redziniak
G
G
Kieda
C
C
D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D046508:Culture Techniques / Q000379:methods; D016207:Cytokines / Q000494:pharmacology*; D004705:Endocytosis; D005643:Fucose / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes; D008856:Microscopy, Fluorescence; D012210:Rhamnose / Q000378:metabolism
D016207:Cytokines; D006023:Glycoproteins; D037102:Lectins; D008081:Liposomes; D005643:Fucose; D012210:Rhamnose