Assessment of copy-number variation in the NKG2C receptor gene in a single-tube and characterization of a reference cell panel, using standard polymerase chain reaction.
Natural killer (NK) and T-lymphocytes monitor human leukocyte antigen (HLA)-E expression through CD94:NKG2 heterodimers. Structural polymorphism is not a hallmark for NK-complex genes on chromosome 12, except for complete NKG2C deletion in some humans. We present a method for fast, simple and accurate assessment of NKG2C copy-number variation - presence or absence in the genome of an NKG2C gene, in homo- or heterozygosis, is detected by a single conventional polymerase chain reaction that yields amplicons of different lengths in each genotype. We have also determined the NKG2C genotypes of a reference cell panel comprising 13 NK- and tumour-cell lines and 39 Epstein-Barr virus transformed cells from the International Histocompatibility Workshop. Our results should facilitate research on the importance of NKG2C and its deletion for immunity.
Laboratorio de Inmunogenética-HLA, Servicio de Inmunología, Hospital Universitario Puerta de Hierro, Majadahonda, Madrid, Spain.
M
M
Moraru
M
M
Cañizares
A
A
Muntasell
R
R
de Pablo
M
M
López-Botet
C
C
Vilches
D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002881:Chromosomes, Human, Pair 12 / Q000235:genetics; D056915:DNA Copy Number Variations / Q000235:genetics*; D017931:DNA Primers; D018628:Gene Dosage; D004854:Herpesvirus 4, Human / Q000235:genetics; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D055654:NK Cell Lectin-Like Receptor Subfamily C / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*
D017931:DNA Primers; C527059:KLRC2 protein, human; D055654:NK Cell Lectin-Like Receptor Subfamily C