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Cited by 669Immunotherapy and Immune Responsesvaccines and immunoinformatics approachesT-cell and B-cell Immunology

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Localization, Quantitation, and In Situ Detection of Specific Peptide–MHC Class I Complexes Using a Monoclonal Antibody

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Porgador, Angel, Yewdell, Jonathan W, Deng, Yuping, Bennink, Jack R, Germain, Ronald N·1997·Immunity
DOI 10.1016/s1074-7613(00)80447-1
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Abstract

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.

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  • The amount of SIINFEKL/MHC complexes required for induction of robust IFN-γ secretion of specific T cells in this particular system is 56 pmol.

    “(having been reported as the amount required for induction of robust IFN-␥ secretion of specific T cells in this particular system 17 )”

    From Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells (2006) · cites this paper as [17]

    In “Sensitivity of Ligand Detection” (page unavailable), the paper compares SIINFEKL concentrations with IFN-γ secretion by a specific CD8+ T-cell line. Our copy discusses minimal concentrations for measurable secretion, but does not establish 56 pmol as an amount required for robust secretion; it reports peptide concentrations in pM, not amounts in pmol. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

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Metadata

Authors
Porgador, A., Yewdell, J. W., Deng, Y., Bennink, J. R., & Germain, R. N.
Affiliations
National Institutes of Health (US)
Corresponding author
Germain, Ronald N
ORCID
Venue
Immunity
Publisher
Elsevier BV
Published
1997-06
Volume
6
Issue
6
Pages
715-726
ISSN
1074-7613 (ISSN-L)
DOI
10.1016/s1074-7613(00)80447-1
OpenAlex
W2001656273
Cited by
669
References
0
Citations by year
2026: 8 · 2025: 13 · 2024: 5 · 2023: 11 · 2022: 9 · 2021: 11 · 2020: 11 · 2019: 28 · 2018: 10 · 2017: 17 · 2016: 19 · 2015: 16 · 2014: 12 · 2013: 25 · 2012: 34
FWCI
9.57
Type
journal article
Language
en
Open access
Licence
Locations
  • Immunity (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › vaccines and immunoinformatics approaches
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
Keywords
peptide-MHC class I complexes, monoclonal antibody production, antigen-presenting cells, viral infection, in situ detection, detection limit, cancer immunotherapy
MeSH terms
Animals, Antibodies, Monoclonal, Antibody Affinity, Antibody Specificity, Antigen-Presenting Cells, Cell Compartmentation, Cell Separation, H-2 Antigens, Immunohistochemistry, Mice, Inbred BALB C, Mice, Inbred C57BL, Ovalbumin, Peptides, Protein Binding, Vaccinia virus, Virus Diseases, Fluorescent Antibody Technique, Indirect, Macromolecular Substances, Mice

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