Localization, quantitation, and in situ detection of specific peptide-MHC class I complexes using a monoclonal antibody.
CD8+ T lymphocytes recognize antigens as short peptides bound to MHC class I molecules. Available methods cannot determine the number and distribution of these ligands on individual cells or detect antigen-presenting cells in tissues. Here we describe a method for eliciting and identifying monoclonal antibodies specific for a particular peptide-MHC class I combination. One such antibody can identify antigen complexes with a limit of detection approaching that of T cells. We used this antibody to determine the number of peptide-class I complexes generated upon viral infection, to identify antigen-presenting cells in cell mixtures, to determine the site of peptide-MHC class I interaction inside cells, and to visualize cells bearing specific peptide-MHC class I complexes after in vivo infection. Similar antibodies may prove useful for diagnostic or therapeutic purposes in cancer, infectious diseases, and autoimmune disorders.
Porgador
A
A
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-1892, USA.
Yewdell
J W
JW
Deng
Y
Y
Bennink
J R
JR
Germain
R N
RN
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000938:Antigen-Presenting Cells / Q000276:immunology; D002451:Cell Compartmentation; D002469:Cell Separation / Q000379:methods; D019084:Fluorescent Antibody Technique, Indirect; D006183:H-2 Antigens / Q000276:immunology* / Q000378:metabolism; D007150:Immunohistochemistry; D046911:Macromolecular Substances; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D011485:Protein Binding; D014616:Vaccinia virus; D014777:Virus Diseases / Q000276:immunology
D000911:Antibodies, Monoclonal; D006183:H-2 Antigens; D046911:Macromolecular Substances; D010455:Peptides; D010047:Ovalbumin