Divergent and convergent evolution of NK-cell receptors.

Natural killer (NK)-cell receptors specific for major histocompatibility complex (MHC) class I molecules have been identified in humans and mice. Some of the most important receptors are structurally unrelated in the two species: the murine Ly-49 receptors are C-type lectins, while human killer-cell inhibitory receptors (KIRs) belong to the immunoglobulin superfamily. Here, Roland Barten and colleagues describe the divergent and convergent evolution of NK-cell receptors.

Plasticity in the organization and sequences of human KIR/ILT gene families.

The approximately 1-Mb leukocyte receptor complex at 19q13.4 is a key polymorphic immunoregion containing all of the natural killer-receptor KIR and related ILT genes. When the organization of the leukocyte receptor complex was compared from two haplotypes, the gene content in the KIR region varied dramatically, with framework loci flanking regions of widely variable gene content. The ILT genes were more stable in number except for ILT6, which was present only in one haplotype. Analysis of Alu repeats and comparison of KIR gene sequences, which are over 90% identical, are consistent with a recent origin. KIR genesis was followed by extensive duplication/deletion as well as intergenic sequence exchange, reminiscent of MHC class I genes, which provide KIR ligands.

(1)H-(17)O nuclear-quadrupole double-resonance study of hydrogen disorder in 2-nitrobenzoic acid.

Temperature dependence of (17)O nuclear quadrupole resonance frequencies was measured in solid 2-nitrobenzoic acid by a (1)H-(17)O nuclear quadrupole double resonance technique. The experimental results show the presence of a fast exchange of hydrogen atoms between two nonequivalent positions within the O-H ellipsis O hydrogen bonds. The hydrogen disorder is ascribed to concerted jumps of two hydrogen atoms within the hydrogen bonds connecting two molecules in a dimer. The energy difference DeltaE of the two hydrogen configurations is equal to DeltaE = 60 meV = 5.8 kJ/mol. The dipole structure of the (17)O NQR lines from the C-O-H oxygen positions was also measured at -100 degrees C and at room temperature. The orientation of the principal axes of the electric field gradient tensor with respect to the O-H bond and the sign of the quadrupole coupling constant were determined. The oxygen-hydrogen distance R(O-H), as determined from the dipole structure of the (17)O NQR lines is at -100 degrees C equal to 0.099 nm. At room temperature we observe a longer distance, R(O-H) = 0.101 nm, in agreement with the hydrogen intrabond exchange.

Arrangement of the ILT gene cluster: a common null allele of the ILT6 gene results from a 6.7-kbp deletion.

The leukocyte receptor cluster (LRC) is a highly polymorphic region of human chromosome 19q13.4 that encompasses at least 24 members of the immunoglobulin superfamily (Ig-SF). The centromeric end of the LRC contains eight Ig-SF loci, namely LAIR1 and seven ILT genes. All ILT genes conform to prototypic ILT gene structures. ILT6 is the only member of the ILT family that lacks a transmembrane and cytoplasmic domain. Close examination of the ILT6 genomic sequence reveals high similarity of this locus with the organization of activating ILT genes. However, the ILT6 transcript runs through the putative splice site of exon 8 that encodes for an extracellular stalk region, leading to a premature in-frame stop codon. Downstream of exon 8 are three pseudo exons that are not included in any of the known ILT6 transcripts, but share high homology to the equivalent region in activating ILT loci, suggesting that these genes have evolved from a common ancestral sequence. Comparison of two haplotypes over this region revealed a remarkable polymorphism with respect to the ILT6 gene which lacks exons 1-7 in one allele, reminiscent of the presence/absence variation displayed by the closely related and genetically linked KIR loci. Detailed sequence analysis of the two LAIR/ILT clusters suggests that the two complexes may have evolved from an inverted duplication.

Isotypic variation of novel immunoglobulin-like transcript/killer cell inhibitory receptor loci in the leukocyte receptor complex.

The leukocyte receptor complex (LRC) on human chromosome 19q13.4 encompasses at least four families of related genes: immunoglobulin-like transcripts (ILT), killer cell inhibitory receptors (KIR), the leukocyte-associated inhibitory receptors (LAIR) and the Fcalpha receptor (Fc(alpha)R). We determined the genomic organization of a region of DNA spanning the junction of the ILT and KIR gene complexes. Extensive sequence data were collected for ILT3, two novel genes, ILT9 and ILT10, and one novel KIR locus (KIRCI). These loci, along with other reported sequences from the region, encoded a leader sequence split into two exons, upstream of two to four immunoglobulin (Ig) domains, each on a separate exon. Downstream of the Ig domains, however, the organization differs markedly between inhibitory and activating ILT. These data are consistent with a highly conserved gene arrangement for all superfamily members suggesting duplication of primordial sequences. ILT3 and KIRCI were in the same head-to-tail orientation as has been described for other KIR loci which may facilitate addition or loss of genes between different haplotypes.

Genomic organization of a human killer cell inhibitory receptor gene.

We have cloned a region of human chromosome 19q13.4 which contains multiple killer cell inhibitory receptor (KIR) loci. By random and directed sequence analysis of these KIR-specific clones, we deduced the genomic structure of KIR genes. A locus encoding a member of the NKAT-2 family of KIRs is presented here. The structure of the gene is reminiscent of loci of the Fc receptor gene family, and the two sets of genes may derive from a common ancestor. The KIR gene contains potentially nine exons. The first two exons encode the leader sequence, as in Fc receptor genes. The third exon encodes an untranslated pseudo exon specifying an immunoglobulin domain with an in-frame stop codon. Expressed cDNAs do not contain this exon. This finding is consistent with the hypothesis that certain KIR genes may have been derived from the duplication of a primordial three immunoglobulin domain structure with subsequent skipping of one exon to derive genes with two expressed immunoglobulin domains. Variation in numbers of immunoglobulin domains in different KIR genes is facilitated by conservation of splicing frame in respect to the codon triplet for each immunoglobulin domain.

Transgene-encoded human CD2 acts in a dominant negative fashion to modify thymocyte selection signals in mice.

CD2 is a cell surface glycoprotein present on all T cells which has been shown to function as an adhesion and signaling molecule. Expressed early in T cell development, human CD2 (HCD2) has been suggested to play a role during thymopoiesis. However, the relevance of CD2 in T cell development has been called into question recently, as neither disruption of the CD2 gene nor anti-CD2 antibody treatment of fetal thymic organ cultures in mouse were shown to have any discernible consequences. We have expressed HCD2 at high levels in transgenic mice and found a profound effect of the transgene on thymocyte differentiation. Transgenic thymuses are considerably reduced in cell number as a consequence of increased apoptosis of double-positive (DP) thymocytes in the cortex. The remaining DP cells have up-regulated levels of T cell receptor (TCR) and are resistant to apoptosis mediated by administration of antigen. These effects are dependent on the cytoplasmic domain of HCD2, as mice expressing comparable levels of a tailless HCD2 transgene have a normal phenotype. The HCD2 cytoplasmic domain contains several regions of identity with mouse CD2 and can interact effciently with mouse intracellular signaling machinery. These results suggest there is considerable cross-talk between CD2 and TCR on developing thymocytes with consequences for the stimulation threshold of mature T cells.