Use of cyclic peptide phage display library for the identification of a CD45RC epitope expressed on murine B cells and their precursors.
The alternative splicing and variable expression of the exons near to the N-terminus of the leukocyte common antigen (L-CA, CD45) result in distinct extracellular isoforms expressed by cells with different functional and developmental properties. Here we report the tissue reactivity pattern and epitope specificity of a novel rat monoclonal antibody (IBL-8) against a restricted epitope of mouse CD45. We found that this mAb reacts with an epitope displayed by B cells and their precursors (both in newborn spleen and adult bone marrow). Moreover, peripheral CD8-positive T cells were also recognised at an intermediate intensity, whereas the CD4 T cell subset was weakly reactive. The epitope of this mAb was determined with M13 filamentous phages that display cysteine constrained nonapeptides on their coat proteins. The isolated bacteriophages expressing the putative epitope showed an isoform-specific inhibition of the binding of exon-specific mAbs. Deduced amino acid sequence data of these phages indicate that the epitope recognised by the IBL-8 mAb lies at the 136-144 region of the mouse CD45 molecule within its C exon, with a TAFP consensus sequence at its centre.
Czömpöly
Tamás
T
Department of Immunology and Biotechnology, Faculty of Medicine, University of Pécs, Szigeti út 12, Pécs H-7634, Hungary.
Lábadi
Arpád
A
Balázs
Mercedesz
M
Németh
Péter
P
Balogh
Péter
P
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000201:enzymology*; D002454:Cell Differentiation; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D005091:Exons; D005260:Female; D007527:Isoenzymes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D016415:Sequence Alignment; D013234:Stem Cells / Q000201:enzymology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007527:Isoenzymes; D019151:Peptide Library; D010456:Peptides, Cyclic; D017493:Leukocyte Common Antigens