Functional and biochemical parameters of peptide antigen presentation.
To understand the mechanism by which peptide antigens are processed and presented to T cells, we examined the T-cell response to the 13-amino-acid peptide alpha-melanocyte-stimulating hormone (alpha-MSH). To determine the fine specificity of T-cell recognition, T cells specific for alpha-MSH, and genetically restricted by I-Ab/d, were challenged with different alpha-MSH analogs and homologs. It was found that intact alpha-MSH, including the blocked amino and carboxy termini of the native molecule, was required for T-cell responsiveness. Antigen-presenting cells (APC) could be briefly pulsed with alpha-MSH and then present the alpha-MSH antigenic determinant to T cells, indicating that the relevant antigen was retained by the APC. APC stimulatory capacity was dramatically reduced by aldehyde treatment of the APC, or by pulsing the APC with alpha-MSH at low temperature. Efficient alpha-MSH pulsing was also impaired by treatment of the APC with the carboxylic ionophore, monensin, but not by the lysosomotropic agents chloroquine and methylamine. In addition, isolated APC plasma membranes added to the T cells in the presence of soluble alpha-MSH were not stimulatory. However, plasma membranes isolated from APC that had been previously pulsed with alpha-MSH retained stimulatory activity for T-cell responses. The only detectable alpha-MSH contained in these pulsed APC membranes was in an acid-stable complex of higher molecular weight than native peptide. The amount of alpha-MSH detected in the cellular membrane fraction isolated by density gradient sedimentation was also reduced by treatments that reduced the APC stimulatory capacity, such as pulsing at low temperature or in the presence of monensin. Taken together, these results suggest that processing of alpha-MSH is unlike that heretofore described for other peptide antigens and seems to involve APC handling to form the stimulatory moiety presented on the APC surface.
Thomas
D W
DW
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109.
Solvay
M J
MJ
Hadley
G
G
Betancourt
S
S
Jun
S
S
Nairn
R
R
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D002462:Cell Membrane / Q000276:immunology; D002738:Chloroquine / Q000494:pharmacology; D005976:Glutaral / Q000494:pharmacology; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007376:Interleukin-2 / Q000096:biosynthesis; D008213:Lymphocyte Activation; D008744:Methylamines / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D008985:Monensin / Q000494:pharmacology; D010455:Peptides / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D000521:alpha-MSH / Q000276:immunology* / Q000378:metabolism
D000949:Histocompatibility Antigens Class II; D007376:Interleukin-2; D008744:Methylamines; D010455:Peptides; D000521:alpha-MSH; D002738:Chloroquine; D008985:Monensin; D005976:Glutaral