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KIR gene content diversity and haplotype analysis in Chinese Han population in Shanghai area
Abstracts
S167
6.05 #241
MOLECULAR INVESTIGATION OF THE KILLER IMMUNOGLOBULIN LIKE RECEPTOR, KIR2DS4. Lynn D. Maxwell, Paula M. Gilmore, Fionnuala Williams, Derek Middleton. Histocompatibility & Immunogenetics Laboratory, City Hospital, Belfast, Northern Ireland, United Kingdom. The killer immunoglobulin-like receptor (KIR) genes, located on chromosome 19, are a highly polymorphic multigene family. In addition to variable gene content, significant allelic diversity within the known KIR genes is now apparent. The activating 2DS4 gene exists in 2 known allelic formats. Of the 4 named 2DS4 alleles *00101, *00102 *002 encode full-length membrane bound receptors. 2DS4*003 encodes a variant allele, containing a 22 base pair deletion in exon 5, which is predicted to encode a protein lacking the transmembrane domain. This finding implies the identification of a soluble version of the 2DS4 protein that is secreted by NK cells. A PCR-SSOP method to identify 2DS4 allele variation has been developed. Typing of a Northern Irish control group (n⫽100), identified the presence of 2DS4*00101 and 003 within this group. Additionally, 2 further allelic variants of the deleted form of 2DS4 (*003) were detected. Further investigation of these variants is currently underway. The method has also been applied to the 13th IHW NK cell line panel (n⫽84), 12 CEPH families (n⫽48), and 40 Northern Irish families (n⫽220). The same allelic variation was identified throughout all samples investigated. Allele typing of the Northern Irish families reveals 2DS4 segregation on B haplotypes at a higher than expected frequency, as compared to studies based on KIR gene segregation alone (14.5%, 17/120 v 1.7%, 2/120). The potential functional consequences of the 2DS4*003 deletion are undergoing investigation. Expression of V5-tagged constructs of full-length and deleted 2DS4 is being evaluated following their transfection into COS7 cells and subsequent western blotting and immunohistochemical staining of the V5-tagged proteins.
6.05 #242
KIR GENE CONTENT DIVERSITY AND HAPLOTYPE ANALYSIS IN CHINESE HAN POPULATION IN SHANGHAI AREA Lei Zhang,1 Katharine C. Hsu,2 Xiao-Rong Liu,2 Bo Dupont,2 Lian Fan.1 1 Immunogenetics Laboratory, Shanghai Institute of Immunology, Shanghai, Shanghai, China; 2Immunology Program, Sloan-Kettering Institute for Cancer Research, New York, NY Natural killer immunoglobulin-like receptors(KIRs) are one of the Ig-like receptor families that recognize HLA class I ligands and are expressed on NK cells and a subset of T cells. The number and distribution of KIR genes vary among individuals and populations. Also, the combination of haplotypes and genotypes may vary in different races and populations . We detect the diversity of KIR gene content and the combination of haplotypes in 87 randomly unrelated healthy individuals in Shanghai Han population with SSP/PCR method. Results show that :(1) frequencies of KIR genes: All of 18 known KIRs genes, such as KIR1D, 2DL1⬃5, 2DS1⬃5, 3DL1⬃3, 3DS1 and the pseudogenes KIR3DP1 (X, Xv) and KIR2DP1 (KIR-Z) are observed in Shanghai Hans. All individuals contain 3DL3, 2DL4, 3DL2 and 3DL1; the most common genes are 2DL3, Z, 2DL1 and X with gene frequency 0.85; the following are 2DS4, 1D, 2DL5, 2DS1, 3DS1 and 2DS5 with gene frequency 0.51, 0.27, 0.21, 0.20,0.19, 0.13 respectively; the next are 2DS2, 2DL2, 2DS3 and Xv with gene frequency below 0.10. (2) frequencies of KIR gene haplotypes: There are 12 haplotypes detected in 87 Han individuals, among them, the most frequent is the type 2 (haplotypeA-2DS4). (3) frequencies of KIR genotypes: The 18 kinds of KIR genotypes are observed in our study, and the most frequent is the AJ (2,2), AF (1,2) with 26.4%, 23% respectively. Also, we find five new genotypes FZ1 (2,9), FZ2 (1,16), FZ3 (6,17), FZ4 (4,13), and FZ5 (2,6), which are not observed in Caucasians so far. Our findings suggest that, there are distinctive frequencies of KIR gene content and haplotype, as well as the genotype in Chinese Han population in Shanghai area. This work is supported by the Science and Technology Committee of Shanghai municipality 02DJ14054 and the National Natural Science Foundation of China (30070784).
S167
6.05 #241
MOLECULAR INVESTIGATION OF THE KILLER IMMUNOGLOBULIN LIKE RECEPTOR, KIR2DS4. Lynn D. Maxwell, Paula M. Gilmore, Fionnuala Williams, Derek Middleton. Histocompatibility & Immunogenetics Laboratory, City Hospital, Belfast, Northern Ireland, United Kingdom. The killer immunoglobulin-like receptor (KIR) genes, located on chromosome 19, are a highly polymorphic multigene family. In addition to variable gene content, significant allelic diversity within the known KIR genes is now apparent. The activating 2DS4 gene exists in 2 known allelic formats. Of the 4 named 2DS4 alleles *00101, *00102 *002 encode full-length membrane bound receptors. 2DS4*003 encodes a variant allele, containing a 22 base pair deletion in exon 5, which is predicted to encode a protein lacking the transmembrane domain. This finding implies the identification of a soluble version of the 2DS4 protein that is secreted by NK cells. A PCR-SSOP method to identify 2DS4 allele variation has been developed. Typing of a Northern Irish control group (n⫽100), identified the presence of 2DS4*00101 and 003 within this group. Additionally, 2 further allelic variants of the deleted form of 2DS4 (*003) were detected. Further investigation of these variants is currently underway. The method has also been applied to the 13th IHW NK cell line panel (n⫽84), 12 CEPH families (n⫽48), and 40 Northern Irish families (n⫽220). The same allelic variation was identified throughout all samples investigated. Allele typing of the Northern Irish families reveals 2DS4 segregation on B haplotypes at a higher than expected frequency, as compared to studies based on KIR gene segregation alone (14.5%, 17/120 v 1.7%, 2/120). The potential functional consequences of the 2DS4*003 deletion are undergoing investigation. Expression of V5-tagged constructs of full-length and deleted 2DS4 is being evaluated following their transfection into COS7 cells and subsequent western blotting and immunohistochemical staining of the V5-tagged proteins.
6.05 #242
KIR GENE CONTENT DIVERSITY AND HAPLOTYPE ANALYSIS IN CHINESE HAN POPULATION IN SHANGHAI AREA Lei Zhang,1 Katharine C. Hsu,2 Xiao-Rong Liu,2 Bo Dupont,2 Lian Fan.1 1 Immunogenetics Laboratory, Shanghai Institute of Immunology, Shanghai, Shanghai, China; 2Immunology Program, Sloan-Kettering Institute for Cancer Research, New York, NY Natural killer immunoglobulin-like receptors(KIRs) are one of the Ig-like receptor families that recognize HLA class I ligands and are expressed on NK cells and a subset of T cells. The number and distribution of KIR genes vary among individuals and populations. Also, the combination of haplotypes and genotypes may vary in different races and populations . We detect the diversity of KIR gene content and the combination of haplotypes in 87 randomly unrelated healthy individuals in Shanghai Han population with SSP/PCR method. Results show that :(1) frequencies of KIR genes: All of 18 known KIRs genes, such as KIR1D, 2DL1⬃5, 2DS1⬃5, 3DL1⬃3, 3DS1 and the pseudogenes KIR3DP1 (X, Xv) and KIR2DP1 (KIR-Z) are observed in Shanghai Hans. All individuals contain 3DL3, 2DL4, 3DL2 and 3DL1; the most common genes are 2DL3, Z, 2DL1 and X with gene frequency 0.85; the following are 2DS4, 1D, 2DL5, 2DS1, 3DS1 and 2DS5 with gene frequency 0.51, 0.27, 0.21, 0.20,0.19, 0.13 respectively; the next are 2DS2, 2DL2, 2DS3 and Xv with gene frequency below 0.10. (2) frequencies of KIR gene haplotypes: There are 12 haplotypes detected in 87 Han individuals, among them, the most frequent is the type 2 (haplotypeA-2DS4). (3) frequencies of KIR genotypes: The 18 kinds of KIR genotypes are observed in our study, and the most frequent is the AJ (2,2), AF (1,2) with 26.4%, 23% respectively. Also, we find five new genotypes FZ1 (2,9), FZ2 (1,16), FZ3 (6,17), FZ4 (4,13), and FZ5 (2,6), which are not observed in Caucasians so far. Our findings suggest that, there are distinctive frequencies of KIR gene content and haplotype, as well as the genotype in Chinese Han population in Shanghai area. This work is supported by the Science and Technology Committee of Shanghai municipality 02DJ14054 and the National Natural Science Foundation of China (30070784).