Analysis of antigen-specific T-cell responses with synthetic peptides--what kind of peptide for which purpose?
The analysis of T-cell responses to peptides has recently become a busy area of immunologic research. Peptides may be used as single stimulants, pools or libraries, or as part of peptide/major histocompatibility complexes (MHC) for direct T-cell receptor staining. For stimulating T cells, peptides must be bound to MHC molecules. In this study we have used 9- or 10-amino acid peptides, 15-amino acid peptides containing stimulating shorter sequences, and peptides with modified C-terminal function. On average 67% of the T cells from healthy cytomegalovirus-positive donors that bound a frequently used cytomegalovirus pp65/HLA-A*0201 tetramer were able to produce interferon-gamma on stimulation with the respective 9-amino acid peptide. Peptides of 15 amino acids length used at the same concentration (in microg/ml) stimulated CD8 T cells somewhat less efficiently (on average 77% of the frequencies induced with the respective shorter peptides). Modifications of 9-amino acid peptides such as addition of amino acids or functional groups often resulted in a decreased ability to stimulate. However, based on our own results, published data, and theoretic considerations, we conclude that sets of peptides of 15 amino acids length with 11 amino acids overlap represent a good compromise for stimulating both CD8 and CD4 T cells in a number of applications. These parameters may be modified subject to the purpose of a study.
Kiecker
Felix
F
Abteilung Klinische Immunologie, Institut für Medizinische Immunologie der Charité, Humboldt-Universität zu Berlin (Charité), Campus Charité Mitte, Berlin, Germany.
Streitz
Mathias
M
Ay
Bernhard
B
Cherepnev
Georgy
G
Volk
Hans-Dieter
HD
Volkmer-Engert
Rudolf
R
Kern
Florian
F
D000577:Amides / Q000737:chemistry / Q000276:immunology; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000276:immunology; D020126:Brefeldin A / Q000494:pharmacology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D004306:Dose-Response Relationship, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000276:immunology; D005434:Flow Cytometry; D005998:Glycine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology; D015234:HLA-A Antigens / Q000276:immunology; D006801:Humans; D017874:Immediate-Early Proteins / Q000276:immunology; D007371:Interferon-gamma / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D010750:Phosphoproteins / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology
D000577:Amides; D000596:Amino Acids; D018984:Epitopes, T-Lymphocyte; D015234:HLA-A Antigens; C088645:IE1 protein, cytomegalovirus; D017874:Immediate-Early Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C065418:cytomegalovirus matrix protein 65kDa; D020126:Brefeldin A; C018556:glycine amide; D007371:Interferon-gamma; D005998:Glycine