Gastroenterology

Gastroenterology

Volume 157, Issue 4, October 2019, Pages 1067-1080.e9
Gastroenterology

Original Research
Full Report: Clinical—Biliary
Imbalance of Genes Encoding Natural Killer Immunoglobulin-Like Receptors and Human Leukocyte Antigen in Patients With Biliary Cancer

https://doi.org/10.1053/j.gastro.2019.06.023Get rights and content

Background & Aims

Bile duct tumors are rare and have poor prognoses. Natural killer (NK) cells are frequent in human liver and infiltrate these tumors but do not control their progression. Responses of NK cells are regulated by NK immunoglobulin-like receptors (KIRs), which interact with HLA class I ligands. We aimed to characterize the features of the KIR gene loci and their ligands in patients with bile duct cancer (BDC).

Methods

We performed combined multidimensional characterization of genes that encode KIRs and their ligands in blood samples from patients with BDC from Sweden, followed for up to 8 years after diagnosis (n = 148), in 2 geographically matched cohorts of healthy individuals from Northern Europe (n = 204 and n = 900), and in healthy individuals from 6 geographically unrelated populations (n = 2917). We used real-time polymerase chain reaction, RNA sequencing, immunohistochemistry, and flow cytometry to evaluate NK-cell presence, as well as KIR and KIR-ligand expression in bile duct tumors and control tissues.

Results

Patients with bile duct tumors had multiple alterations at the KIR gene loci. KIR loci are grouped into genotypes that encode more inhibitory (group A) and more activating (group B) receptors, which can be subdivided into centromeric and telomeric fragments. Patients with BDC had a lower prevalence of KIR2DL3, which was linked to disequilibrium in centromeric A/B and B/B genotypes, compared with control individuals. The associations between KIRs and KIR ligands differed between patients with BDC and control individuals; patients had an altered balance between activating and inhibitory KIRs. KIR-positive NK cells infiltrated biliary tumors that expressed matched KIR ligands.

Conclusions

In a multidimensional analysis of DNA from blood samples of patients with BDC in Europe, we found patients to have multiple alterations at the KIR and HLA gene loci compared with control individuals. These alterations might affect NK-cell tumor surveillance. NK cells from bile duct tumors expressed KIRs and were found in tumors that expressed cognate ligands. This should be considered in development of immune-based therapies for BDC.

Section snippets

Clinical Cohorts

This study was approved by the regional ethics committee in Stockholm, Sweden, and conducted in accordance with the Declaration of Helsinki. Patients with BDC were recruited at the Department of Upper Gastrointestinal Tract Diseases, Karolinska University Hospital Huddinge, Stockholm, Sweden, a specialized tertiary center for surgical treatment of liver and biliary tract cancer. Oral and written informed consent was obtained from all patients. The Swedish healthy cohort consisted of randomly

Study Design

A total of 148 Swedish patients with BDC were recruited to the study at the Department of Upper Gastrointestinal Tract Diseases, Karolinska University Hospital, a specialized tertiary center for surgical treatment of liver and biliary tract cancer. Few patients had relevant comorbidities (Figure 1A). Because PSC patients with BDCs usually receive liver transplantation, only 7% of patients in the current cohort had underlying PSC. Roughly equal numbers of patients with GBC and CCA were included

Discussion

The importance of the KIR-HLA system in human disease is clear from a plethora of work conducted over the last decades.10, 17, 19, 20, 21 However, most studies have assessed the presence or absence of single receptors in a case-control fashion without integrating functional modules governed by clusters of genes. The KIR-KIR ligand loci are unique genetic modules that have evolved with the human species and are critical for immune cell recognition of altered self.10 Based on the current

Acknowledgments

Author contributions: All authors contributed to interpretation of data. Martin Cornillet designed the study, performed experiments, analyzed the data, and drafted the manuscript. Marie Schaffer performed KIR and HLA genotyping. Ernesto Sparrelid and Bengt Isaksson contributed to recruitment and collection of the BDC cohort. Karl-Johan Malmberg, Derek Middleton, and Faviel F. Gonzalez-Galarza contributed to collection of control populations. Lena Berglin performed immunohistochemistry

References (53)

  • J.M. Banales et al.

    Expert consensus document: cholangiocarcinoma: current knowledge and future perspectives consensus statement from the European Network for the Study of Cholangiocarcinoma (ENS-CCA)

    Nat Rev Gastroenterol Hepatol

    (2016)
  • T.H. Karlsen et al.

    Primary sclerosing cholangitis—a comprehensive review

    J Hepatol

    (2017)
  • V. Thorsson et al.

    The immune landscape of cancer

    Immunity

    (2018)
  • B. Goeppert et al.

    Prognostic impact of tumour-infiltrating immune cells on biliary tract cancer

    Br J Cancer

    (2013)
  • D. Walter et al.

    PD-L1 expression in extrahepatic cholangiocarcinoma

    Histopathology

    (2017)
  • G. Liu et al.

    Distribution of natural killer cells and T-lymphocyte subsets in peripheral blood, gallbladder cancer and surrounding tissue

    Hepatobiliary Pancreat Dis Int

    (2007)
  • Y. Nakakubo et al.

    Clinical significance of immune cell infiltration within gallbladder cancer

    Br J Cancer

    (2003)
  • N.K. Björkström et al.

    Emerging insights into natural killer cells in human peripheral tissues

    Nat Rev Immunol

    (2016)
  • M.G. Morvan et al.

    NK cells and cancer: you can teach innate cells new tricks

    Nat Rev Cancer

    (2015)
  • P. Parham et al.

    Variable NK cell receptors and their MHC class I ligands in immunity, reproduction and human evolution

    Nat Rev Immunol

    (2013)
  • A. Horowitz et al.

    Class I HLA haplotypes form two schools that educate NK cells in different ways

    Sci Immunol

    (2016)
  • S. Kim et al.

    Licensing of natural killer cells by host major histocompatibility complex class I molecules

    Nature

    (2005)
  • N. Anfossi et al.

    Human NK cell education by inhibitory receptors for MHC class I

    Immunity

    (2006)
  • J.P. Goodridge et al.

    Newtonian cell interactions shape natural killer cell education

    Immunol Rev

    (2015)
  • P. Parham et al.

    Genetics of natural killer cells in human health, disease, and survival

    Annu Rev Immunol

    (2018)
  • A.R. Manser et al.

    Human KIR repertoires: shaped by genetic diversity and evolution

    Immunol Rev

    (2015)
  • S.I. Khakoo et al.

    HLA and NK cell inhibitory receptor genes in resolving hepatitis C virus infection

    Science

    (2004)
  • V. Ramsuran et al.

    Elevated HLA-A expression impairs HIV control through inhibition of NKG2A-expressing cells

    Science

    (2018)
  • V. Varbanova et al.

    Killer-cell immunoglobulin-like receptor genes and ligands and their role in hematologic malignancies

    Cancer Immunol Immunother

    (2016)
  • M.P. Martin et al.

    Epistatic interaction between KIR3DS1 and HLA-B delays the progression to AIDS

    Nat Genet

    (2002)
  • F. Colucci

    The role of KIR and HLA interactions in pregnancy complications

    Immunogenetics

    (2017)
  • C. Guillerey et al.

    Targeting natural killer cells in cancer immunotherapy

    Nat Immunol

    (2016)
  • T.H. Karlsen et al.

    Particular genetic variants of ligands for natural killer cell receptors may contribute to the HLA associated risk of primary sclerosing cholangitis

    J Hepatol

    (2007)
  • F.F. González-Galarza et al.

    Allele frequency net 2015 update: new features for HLA epitopes, KIR and disease and HLA adverse drug reaction associations

    Nucleic Acids Res

    (2015)
  • S. Cooley et al.

    Donor selection for natural killer cell receptor genes leads to superior survival after unrelated transplantation for acute myelogenous leukemia

    Blood

    (2010)
  • The Genotype-Tissue Expression (GTEx) project

    Nat Genet

    (2013)
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    Conflicts of interest The authors disclose no conflicts.

    Funding This work was funded by the Swedish Research Council, Swedish Cancer Society, Swedish Foundation for Strategic Research, Swedish Society for Medical Research, Cancer Research Foundations of Radiumhemmet, Knut and Alice Wallenberg Foundation, Novo Nordisk Foundation, Center for Innovative Medicine at Karolinska Institutet, Stockholm County Council, Karolinska Institutet, Research Council of Norway, Norwegian Cancer Foundation, and K.G. Jebsen Foundation.

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