The order and orientation of mouse lambda-genes explain lambda-rearrangement patterns.

A cDNA library derived from spleen cells of an unimmunized rabbit expressing the c7 phenotype of Ig lambda chains (c7+, c21-) was screened with V lambda or C lambda probes of a lambda light chain bearing c21 epitopes. The nucleotide sequences of three hybridizing clones were found to be identical within the V lambda, J lambda and C lambda regions. The V lambda region was 97% similar to that of the functional germ-line gene V lambda 2, and the C lambda region was identical to that of gene C lambda 6, recently identified. Gene C lambda 6 exhibited four codon differences when compared with gene C lambda 5, the latter encoding c21 epitopes. The data presented here and in the accompanying report (Jaton, J.-C. et al., Eur. J. Immunol. 1990, 20:2713) support the view that gene C lambda 6 encodes the C region of c7 lambda chains and that c7 and c21 markers designate two distinct isotypic forms of lambda chains. On the basis of comparative Southern blotting analyses and restriction maps of cloned genomic regions containing V lambda and C lambda genes, a scheme is proposed to account for the c7- and c21- phenotypes.

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cDNA clones encoding immunoglobulin λ chains from rabbit expressing the phenotype c7

Hayzer

Young-Cooper

Mage

et al. 1990

Eur J Immunol

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Mapping of the duplicated rabbit immunoglobulin χ light chain locus

1991

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The rabbit has two isotypic forms of the immunoglobulin kappa light chain, K1 and K2, which probably arose by duplication. In the normal rabbit, only traces of K2 light chains are produced. However, K2 levels are elevated in allotype-suppressed rabbits and in the Basilea strain which does not produce K1 because of a K1 mRNA splice site mutation. Previous cloning and sequencing showed that each isotype has its own set of J kappa genes but it was not known whether the two isotypes utilize shared or separate sets of V kappa genes. In addition, although genetic linkage of allotypes associated with the K1 and K2 genes has been demonstrated, physical linkage had not been previously demonstrated by overlapping cosmid or phage clones. We used pulsed field and transverse alternating field electrophoresis to obtain megabase maps and to estimate the size of the duplication of the rabbit kappa light chain locus. We found that the two C kappa genes are about 1 megabase apart. One explanation for the poor expression of K2, could be great physical distance from V kappa genes. However, we found that there are V kappa, J kappa and C kappa 2 genes within a approximately 105-kb fragment. Thus, physical distance of V kappa from C kappa 2 may not be the basis for poor K2 expression.

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A λ1 transgene under the control of a heavy chain promoter and enhancer does not undergo somatic hypermutation

Hengstschläger¹,

Williams²,

Maizels³

1994

Eur J Immunol

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To identify cis-acting elements responsible for targeting somatic hypermutation to immunoglobulin variable regions, we generated transgenic mice which carry a rearranged lambda 1 gene regulated by the heavy chain intron enhancer, E mu, and the heavy chain promoter PH186.2 from the VH186.2 variable region. C57BL/6 x SJL founders were bred with C57BL/6 mice to establish a line carrying a single copy of the transgene. Somatic hypermutation was studied by generating hybridoma cell lines from mice immunized with the hapten (4-hydroxy-3-nitrophenyl)acetyl (NP) coupled to chicken gamma globulin. The immune response in this transgenic line was dominated by the endogenous VH186.2 heavy chain variable region and the transgenic lambda 1 light chain, and the transgene was actively expressed in all hybridomas analyzed. In this work we show that the transgenic V lambda 1 regions do not undergo hypermutation, despite high levels of expression, while the expressed heavy chain V regions accumulate mutations at a rate typical of the NP response in C57BL/6 mice. Thus, within the same B cell, the PH186.2 promoter in connection with E mu drives efficient expression of both a VH and a V lambda region, but only the VH is a target for somatic hypermutation. Our observations show that cis-acting sequences that activate immunoglobulin gene transcription are not sufficient to target somatic hypermutation.

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The order and orientation of mouse lambda-genes explain lambda-rearrangement patterns.

Cited by 26 publications

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cDNA clones encoding immunoglobulin λ chains from rabbit expressing the phenotype c7

cDNA clones encoding immunoglobulin λ chains from rabbit expressing the phenotype c7

Mapping of the duplicated rabbit immunoglobulin χ light chain locus

A λ1 transgene under the control of a heavy chain promoter and enhancer does not undergo somatic hypermutation

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