The CDR-H3 Repertoire from TdT-Deficient Adult Bone Marrow Is a Close, but Not Exact, Homologue of the CDR-H3 Repertoire from Perinatal Liver
| Autor: | Harry W. Schroeder, Robert L. Schelonka, G. Larry Gartland, Andre M. Vale, Michael Zemlin, Ewa Szymanska, Ivaylo I. Ivanov |
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| Rok vydání: | 2010 |
| Předmět: |
Lineage (genetic)
B cell maturation Molecular Sequence Data Immunology Cell Cell Separation Biology Article Mice Bone Marrow DNA Nucleotidylexotransferase medicine Animals Immunology and Allergy Nucleotide Amino Acid Sequence B cell Genetics chemistry.chemical_classification B-Lymphocytes Mice Inbred BALB C Fetus Reverse Transcriptase Polymerase Chain Reaction Precursor Cells B-Lymphoid Repertoire Flow Cytometry Complementarity Determining Regions Molecular biology medicine.anatomical_structure Liver chemistry Bone marrow Immunoglobulin Heavy Chains |
| Zdroj: | The Journal of Immunology. 185:6075-6084 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.1001419 |
| Popis: | Compared with adult bone marrow (BM), the composition of the perinatal liver CDR-3 of the Ig H chain (CDR-H3) repertoire is marked by a paucity of N nucleotides and by enrichment for use of JH proximal DQ52 and DH proximal VH and JH gene segments. To test the extent to which these differences reflect limited perinatal TdT activity versus differences in the fetal/adult environment, we used the Hardy scheme to sort fractions B–F B lineage cells from TdT-deficient BALB/c adult BM. VH7183-containing VDJCμ transcripts from these cells were amplified, cloned, sequenced, and compared with transcripts from wild-type perinatal liver and adult BM. The pattern of VHDJH usage in TdT-deficient BM largely matched that of TdT-sufficient adult cells. What minor differences were detected in the pro-B cell stage tended to diminish with B cell maturation, suggesting strong environmental or Ag-driven pressure to achieve a specific range of VHDJH usage regardless of the extent of N nucleotide addition. However, although the patterns of VHDJH usage in the TdT-deficient B lineage cells paralleled that of wild-type adult cells, the length distribution, global amino acid composition, and charge distribution of the CDR-H3 repertoire proved to be a close, although not exact, homologue of the CDR-H3 repertoire first expressed by late pre-B cells in the TdT-insufficient perinatal liver. Thus, although differing in VH content, TdT-deficient mice appear to represent a good, although not perfect, model for testing the role of perinatal CDR-H3 limitations on late B cell development and Ab responses. |
| Databáze: | OpenAIRE |
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