Expressed antibody repertoires in human cord blood cells: 454 sequencing and IMGT/HighV-QUEST analysis of germline gene usage, junctional diversity, and somatic mutations.
Human cord blood cell-derived IgM antibodies are important for the neonate immune responses and construction of germline-based immunoglobulin libraries. Several previous studies of a relatively small number of sequences found that they exhibit restrictions in the usage of germline genes and in the diversity of the variable heavy chain complementarity determining region 3 compared to adults. To further characterize such restrictions on a larger scale and to compare the early B-cell diversity to adult IgM repertoires, we performed 454 sequencing and IMGT/HighV-QUEST analysis of cord blood IG libraries from two babies and determined germline gene usage, V-D-J rearrangement, VHCDR3 diversity, and somatic mutations to characterize human neonate repertoire. Most of the germline subgroups were identified with frequencies comparable to those present in the adult IgM repertoire except for the IGHV1-2 gene that was preferentially expressed in the cord blood cells. The gene usage diversity contributed to 1,430 unique IGH V-D-J rearrangement patterns while the exonuclease trimming and N region addition at the V-D-J junctions along with gene diversity created a wide range of VHCDR3 with different lengths and sequence variability. We observed a lower degree of somatic mutations in the CDR and framework regions of antibodies from cord blood cells compared to adults. These results provide insights into the characteristics of human cord blood antibody repertoires, which have gene usage diversity and VHCDR3 lengths similar to that of the adult IgM repertoire but differ significantly in some of the gene usages, V-D-J rearrangements, junctional diversity, and somatic mutations.
Protein Interactions Group, Center for Cancer Research Nanobiology Program, National Cancer Institute (NCI)-Frederick, National Institutes of Health (NIH), Bldg 469, Rm 150B, Frederick, MD 21702, USA.
Ponraj
P
Prabakaran
Weizao
W
Chen
Maria G
MG
Singarayan
Claudia C
CC
Stewart
Emily
E
Streaker
Yang
Y
Feng
Dimiter S
DS
Dimitrov
D000328:Adult; D000916:Antibody Diversity / Q000235:genetics*; D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D022801:Complementarity Determining Regions / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology*; D015322:Gene Rearrangement, B-Lymphocyte; D018095:Germ-Line Mutation; D006801:Humans; D055261:Immunogenetic Phenomena; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007075:Immunoglobulin M / Q000097:blood* / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007231:Infant, Newborn; D008297:Male; D060152:V(D)J Recombination
D022801:Complementarity Determining Regions; D017931:DNA Primers; D007143:Immunoglobulin Heavy Chains; D007075:Immunoglobulin M; D007135:Immunoglobulin Variable Region