Analysis of the requirement for beta 2-microglobulin for expression and formation of human CD1 antigens.
Human CD1 form a group of nonpolymorphic leukocyte surface molecules with homology to major histocompatibility complex (MHC) proteins. Recent findings in human and in mouse demonstrate the capacity of CD1 molecules to present nonpeptide components like lipids or lipoglycans as well as peptides. We studied the involvement of beta 2-microglobulin (beta 2m) in expression of the classic human CD1 proteins CD1a, CD1b, and CD1c. The beta 2m-deficient human melanoma cell line FO-1 was transiently transfected with either CD1a, CD1b, or CD1c DNA alone, or in combination with beta 2m using the adenovirus-enhanced receptor-mediated transfer infection system. Only co-transfection of FO-1 cells with CD1+ beta 2m resulted in the detection of CD1 Ag by monoclonal antibodies (mAb). This indicated that CD1 mAb recognized determinants are dependent on beta 2m and raised the question whether beta 2m-free forms of CD1 can be expressed. Therefore, to visualize CD1 molecule expression independently of beta 2m, we expressed tagged recombinant forms. A full-length CD1b construct tagged at the very C terminus with a small peptide was transported to the plasma membrane only when beta 2m was co-transfected. beta 2m involvement in the transport of CD1 was confirmed by expression of soluble forms of CD1a, CD1b, and CD1c in three different cell types. Analogous to tagged full-length CD1b, secretion of the soluble CD1 constructs was strictly dependent on beta 2m. The soluble CD1 chimeras were secreted as complexes with endogenous beta 2m. Thus, similar to its role for MHC class I expression, beta 2m is essential for processing and surface transport of the classic human CD1 molecules CD1a, CD1b, and CD1c.
Bauer
A
A
Institute of Immunology, Vienna International Research Cooperation Center, University of Vienna, Austria.
Hüttinger
R
R
Staffler
G
G
Hansmann
C
C
Schmidt
W
W
Majdic
O
O
Knapp
W
W
Stockinger
H
H
D000911:Antibodies, Monoclonal / Q000378:metabolism; D018949:Antigens, CD1 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D001692:Biological Transport / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D006801:Humans; D007668:Kidney / Q000196:embryology; D008545:Melanoma; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D012995:Solubility; D014407:Tumor Cells, Cultured; D001613:beta 2-Microglobulin / Q000276:immunology / Q000502:physiology*
D000911:Antibodies, Monoclonal; D018949:Antigens, CD1; D011993:Recombinant Fusion Proteins; D001613:beta 2-Microglobulin