Decreased frequency of rearrangement due to the synergistic effect of nucleotide changes in the heptamer and nonamer of the recombination signal sequence of the V kappa gene A2b, which is associated with increased susceptibility of Navajos to Haemophilus influenzae type b disease.
Navajos and genetically related populations have a 10-fold increased incidence of Haemophilus influenzae type b (Hib) disease compared with control populations. The Vkappa gene A2 is used to encode the majority of anti-Hib Abs, and these are the highest affinity anti-Hib Abs. Navajos carry a different allele of the A2 gene segment (A2b) that is defective in its ability to undergo V-J recombination. The A2b allele has only three nucleotide changes from the commonly occurring A2a allele, two of which could potentially affect its ability to recombine. In this study we used two independent in vitro assays to test whether the nucleotide change found in the A2b promoter and/or in the A2b recombination signal sequence (RSS) might be responsible for the decrease in recombination frequency observed in vivo. Using a luciferase reporter gene assay, we found no significant difference between A2a and A2b promoter activities. However, the competition recombination substrate assay showed a 4.5-fold reduction in the relative frequency of recombination of the A2b RSS compared with A2a. We show that this decreased frequency is due to a synergistic effect of the unique nucleotide change present in the heptamer of the A2b RSS and the shared nucleotide change present in the nonamer of both A2b and A2a. This in vitro relative frequency of rearrangement is not significantly different from that observed in vivo; therefore, the A2b RSS is probably the factor associated with the increased susceptibility to Hib disease among individuals carrying the A2b allele.
Nadel
B
B
Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Tang
A
A
Lugo
G
G
Love
V
V
Escuro
G
G
Feeney
A J
AJ
D000483:Alleles; D001483:Base Sequence; D002461:Cell Line, Transformed; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain; D005803:Genes, Immunoglobulin; D020022:Genetic Predisposition to Disease / Q000276:immunology*; D019741:Haemophilus influenzae type b / Q000276:immunology*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics*; D007198:Indians, North American / Q000235:genetics; D008583:Meningitis, Haemophilus / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000276:immunology; D011995:Recombination, Genetic / Q000276:immunology*; D012045:Regulatory Sequences, Nucleic Acid / Q000276:immunology; D014162:Transfection / Q000276:immunology
D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D009711:Nucleotides