Preferential utilization of specific immunoglobulin heavy chain diversity and joining segments in adult human peripheral blood B lymphocytes
Autor: | Sara Shane, Masao Yamada, Betty Anne Reichard, Giovanni Rovera, Robert Wasserman, Andrew J. Caton |
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Jazyk: | angličtina |
Rok vydání: | 1991 |
Předmět: |
Immunology
Molecular Sequence Data Gene Rearrangement B-Lymphocyte Heavy Chain Immunoglobulin Variable Region Biology Polymerase Chain Reaction Homology (biology) law.invention Plasmid law Sequence Homology Nucleic Acid Immunology and Allergy Humans Bacteriophages Amino Acid Sequence Codon Gene Polymerase chain reaction Genetics Immunoglobulin Joining Region B-Lymphocytes Polymorphism Genetic Base Sequence Nucleic acid sequence Gene rearrangement Articles Molecular biology Immunoglobulin heavy chain Chromosome Deletion Immunoglobulin Heavy Chains Oligonucleotide Probes Plasmids |
Zdroj: | The Journal of Experimental Medicine |
ISSN: | 1540-9538 0022-1007 |
Popis: | We have examined at the molecular level the CDR3 and adjacent regions in peripheral blood B lymphocytes of normal individuals. A total of 111 sequences (12-28 sequences from six individuals) were obtained after cloning of the polymerase chain reaction-amplified segments into plasmids or phage. The average length of the VDJ joining was 109 nucleotides, with a range from 79 to 151. Approximately 75% of the sequences were in frame when translated into amino acids. Among the JH segments, JH4 was found most frequently (in 52.5% of the sequences), and JH1 and JH2 segments the least frequently (approximately 1% of the clones). A polymorphic JH6 gene with a one-codon deletion accompanied by a base change was present in two of six patients. Preferential breakpoints were found for JH2, JH3, JH4, and JH5, although the breakpoints of JH6 were distributed more heterogenously. In approximately 90% of the cases, significant homology of the D regions with published D sequences was found. Preferential usage of a particular coding frame was observed in in-frame sequences utilizing DA, D21/9, and DM1 segments. However, in general, all coding frames of germline D genes were used to generate CDR3s. Eight sequences that have a DN1-like D sequence with two base changes at the same positions were identified, suggesting the likely existence of a new germ line D gene belonging to the DN families. Using probes specific for a particular CDR3, the frequency of a specific B cell clone in the peripheral blood of normal individuals was estimated to be at most as high as 1/20,000. |
Databáze: | OpenAIRE |
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