CD4 is expressed by epidermal Langerhans' cells predominantly as covalent dimers.
Langerhans' cells (LC) of skin are CD4 expressing, dendritic, antigen-presenting cells, that are essential for activation of primary immune responses and are productively infected by HIV. We have shown previously that lymphocytes and monocytes express CD4 both as monomers and covalently linked homodimers. In those cells the 55-kDa monomer structure predominates. LC in un-fractionated human epidermal cell (EC) suspension also expresses both forms of CD4, but in EC the dimer form is predominant. Because isolation of LC into single cell suspension by trypsin, as is routinely used for LC isolation, degrades CD4, a systematic study for an alternate procedure for LC isolation was performed. Thus it was found that collagenase blend F treatment can efficiently release LC into suspension, under conditions of only minimal degradation of control soluble recombinant CD4 or CEM-T4 or THP-1 cell CD4, or importantly of LC surface CD4. SDS-PAGE immunoblotting of purified LC extracted from EC by collagenase confirmed CD4 structure as predominantly 110-kDa dimers, with only minimal 55-kDa monomers. The suitability of LC prepared thus for functional studies was demonstrated with binding of functional ligand HIV gp120. It remains to be determined, however, why tissue embedded LC express mainly CD4 dimers, but single-celled blood lymphocytes and monocytes mainly monomers.
Lynch
G W
GW
Centre for Virus Research, Westmead Millennium Institute, University of Sydney, Westmead Hospital, Sydney, Australia. Garry_Lynch@optusnet.com.au
Slaytor
E K
EK
Elliott
F D
FD
Saurajen
A
A
Turville
S G
SG
Sloane
A J
AJ
Cameron
P U
PU
Cunningham
A L
AL
Halliday
G M
GM
D018949:Antigens, CD1 / Q000378:metabolism; D015704:CD4 Antigens / Q000737:chemistry* / Q000378:metabolism*; D002458:Cell Fractionation; D002478:Cells, Cultured; D017364:Collagenases / Q000494:pharmacology; D019281:Dimerization; D010450:Endopeptidases / Q000494:pharmacology; D000078404:Epidermal Cells; D006801:Humans; D007801:Langerhans Cells / Q000166:cytology / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D020836:Protein Structure, Quaternary; D014357:Trypsin / Q000494:pharmacology
D018949:Antigens, CD1; C419393:CD1a antigen; D015704:CD4 Antigens; D008565:Membrane Proteins; D010450:Endopeptidases; D014357:Trypsin; D017364:Collagenases