title: Heterozygosity for the common perforin mutation, p.A91V, impairs the cytotoxicity of primary natural killer cells from healthy individuals.
abstract: The production and delivery of functional perforin (PRF; PRF1 gene) by cytotoxic lymphocytes maintains immune homeostasis and tumour immune surveillance. In humans, inheritance of the common PRF1 polymorphism, p.A91V, (c.272C>T) found in 8-9% of the Caucasian population, with another mutated allele resulting in reduced PRF function or trafficking, has been shown to result in hyperinflammatory diseases and/or haematological cancers. In this study, we sought to investigate the function of p.A91V on a wild-type (WT) perforin background. We first developed an assay that distinguishes the relative levels of transcription of individual PRF1 alleles, including p.A91V. The p.A91V allele was seen to be expressed at similar levels as the WT allele in primary human natural killer (NK) cells, ruling out that allelic expression imbalance influenced their function. We then demonstrated that the p.A91V mutation results in protein misfolding and an appreciable reduction in NK-cell cytotoxicity in healthy carriers of p.A91V. We propose that this level of cytotoxic dysfunction may readily account for the predisposition to immune-mediated disease in individuals homozygous for p.A91V. Also, the fact that monoallelic mutations of PRF1 decrease NK-cell cytotoxicity should be considered in individuals presenting with the manifestations of immune deficiency states that impinge on NK-cell cytotoxicity.
authors.affiliation: 1] Cancer Immunology Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia [2] Sir Peter MacCallum Department of Oncology, Parkville, Victoria, Australia [3] Department of Pathology, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Imran G
authors.initials: IG
authors.lastname: House
authors.affiliation: Cancer Immunology Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.
authors.forename: Kevin
authors.initials: K
authors.lastname: Thia
authors.affiliation: Cancer Immunology Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.
authors.forename: Amelia J
authors.initials: AJ
authors.lastname: Brennan
authors.affiliation: 1] Cancer Immunology Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia [2] Department of Pathology, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Richard
authors.initials: R
authors.lastname: Tothill
authors.affiliation: Ludwig Institute for Cancer Research, Olivia Newton-John Cancer and Wellness Centre, Heidelberg (Melbourne), Victoria, Australia.
authors.forename: Alexander
authors.initials: A
authors.lastname: Dobrovic
authors.affiliation: Cancer Immunology Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.
authors.forename: Wei Z
authors.initials: WZ
authors.lastname: Yeh
authors.affiliation: Murdoch Children's Research Institute; Department of Paediatrics; The University of Melbourne; Royal Children's Hospital, Melbourne, Victoria, Australia.
authors.forename: Richard
authors.initials: R
authors.lastname: Saffery
authors.affiliation: Murdoch Children's Research Institute; Department of Paediatrics; The University of Melbourne; Royal Children's Hospital, Melbourne, Victoria, Australia.
authors.forename: Zac
authors.initials: Z
authors.lastname: Chatterton
authors.affiliation: 1] Cancer Immunology Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia [2] Sir Peter MacCallum Department of Oncology, Parkville, Victoria, Australia [3] Department of Pathology, University of Melbourne, Parkville, Victoria, Australia [4] Department of Immunology and Microbiology, Parkville, Victoria, Australia.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Trapani
authors.affiliation: 1] Cancer Immunology Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia [2] Sir Peter MacCallum Department of Oncology, Parkville, Victoria, Australia [3] Department of Pathology, University of Melbourne, Parkville, Victoria, Australia [4] Department of Genetics, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Ilia
authors.initials: I
authors.lastname: Voskoboinik
mesh_terms: D000483:Alleles; D019943:Amino Acid Substitution; D003062:Codon; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D015870:Gene Expression; D005799:Genes, Dominant; D064368:Healthy Volunteers; D006579:Heterozygote; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism*; D009154:Mutation; D054353:Perforin / Q000737:chemistry / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D017510:Protein Folding; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D003062:Codon; D012333:RNA, Messenger; D054353:Perforin