Cleavage of Langerhans cell surface CD1a molecule by trypsin.
The T-cell surface differentiation antigens expressed on cortical thymocytes are composed of 3 molecules, CD1a (Mr 49,000), CD1b (Mr 45,000), and CD1c (Mr 43,000), which are non-covalently attached to beta 2-microglobulin. In the present study, differences in quantitative binding (immunogold labelling) were observed with four CD1a monoclonal antibodies (mAb), Na1/34, L544, Vit6 and OKT6, on epidermal Langerhans cells obtained through trypsinization and Ficoll-Hypaque sedimentation. These cells were surface-labelled with 125I and then lysed. Immunoprecipitation was carried out with five CD1a mAb, BL6, 10D12.2, L404, L544 and OKT6, and immunoprecipitates were electrophoretically run. All CD1a mAb except OKT6 immunoprecipitated an additional molecule with an apparent relative mass of 27,000, under reducing conditions. CD1a antigen (Mr 49,000) was borne by the same chain of Mr 49,000 on cortical thymocytes and Langerhans cells, whereas the Mr 27,000 molecule was never found on thymic cells. On two-dimensional gel analysis, the Mr 27,000 molecule showed a pattern with 3 major spots with pI of 5.6, 5.9 and 6.2. This Mr 27,000 protein was found to contain one N-linked oligosaccharide residue by endoglycosidase-F treatment. By sequential immunoprecipitation, this Mr 27,000 molecule was shown to be different from the major histocompatibility complex class II beta-chains (DR, DP). As the Mr 27,000 molecule was not precipitated with OKT6, sequential immunoprecipitation confirmed specific recognition of this low molecular weight protein by other CD1a mAb. The protein of apparent molecular mass 27,000 was considered to be a breakdown product of Mr 49,000 (CD1a) antigen. These results suggested that the CD1a molecule was sensitive to trypsin.
Dezutter-Dambuyant
C
C
INSERM U209, Laboratoire de Recherche Dermatologique et Immunologie, Hôpital Edouard-Herriot, Lyon, France.
Schmitt
D
D
Staquet
M J
MJ
Boumsell
L
L
Thivolet
J
J
D000911:Antibodies, Monoclonal; D018949:Antigens, CD1; D000943:Antigens, Differentiation / Q000378:metabolism*; D000954:Antigens, Surface / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007801:Langerhans Cells / Q000378:metabolism*; D008970:Molecular Weight; D011233:Precipitin Tests; D012867:Skin / Q000166:cytology; D013601:T-Lymphocytes; D013950:Thymus Gland / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D018949:Antigens, CD1; D000943:Antigens, Differentiation; D000954:Antigens, Surface; D014357:Trypsin