A new germline VH36-60 gene is used in the neonatal primary and adult memory response to (T,G)-A--L.
Our previous studies of the neonatal primary response to (T,G)-A--L showed that the majority of anti-(T,G)-A--L antibodies bind the copolymer L-Glu:L-Tyr (GT), share idiotypy (Id), and use the H10 germline VH gene from the VHJ558 family and a V kappa 1 gene. We also identified two hybridomas from different neonatal donors that produced GT+, Id+ antibodies using a V kappa 1 gene with a VH gene from the VH36-60 family. In the study reported here, we show that both neonatal hybridomas use the same germline VH gene from the VH36-60 gene family. However, the VH gene sequence is different from previously identified germline genes of the VH36-60 gene family. To determine whether the expressed heavy chain gene had undergone somatic mutation, we isolated the corresponding germline gene from kidney DNA. Sequence analysis of this gene shows that it is a new member of the VH36-60 family which is not mutated in the neonatal antibodies. Furthermore, the deduced amino acid sequences of the two neonatal antibodies are identical not only in the VH region but also in the VH-D-JH joins, suggesting that there is a strong selection for CDRIII among neonatal anti-(T,G)-A--L antibodies using this germline gene (designated here as VH3A1) with a V kappa 1 gene. Also, the VH gene from the VH36-60 family that we showed previously was used by an adult memory B cell clone specific for (T,G)-A--L, can now be identified as a rearrangement of the VH3A1 germline gene. Elucidation of the germline variable region genes that are used in the antigen-specific neonatal response will help us understand the mechanisms that shape the preimmune B cell repertoire during B cell development.
Department of Biology and Rosenstiel Research Center, Brandeis University, Waltham, MA 02254-9110.
J L
JL
Press
C A
CA
Giorgetti
W F
WF
Busby
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D015321:Gene Rearrangement; D005803:Genes, Immunoglobulin / Q000235:genetics*; D006825:Hybridomas / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007156:Immunologic Memory / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D010455:Peptides / Q000276:immunology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012689:Sequence Homology, Nucleic Acid; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*; D014158:Transcription, Genetic
D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D015345:Oligonucleotide Probes; D010455:Peptides; C021759:(T,G)-A-L