Lipid antigens in bile from patients with liver diseases activate NKT cells — ASN Events
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  3. Lipid antigens in bile from patients with liver diseases activate NKT cells

Lipid antigens in bile from patients with liver diseases activate NKT cells (#189)

Laura Valestrand 1 2 3 , Natalie L. Berntsen 1 2 3 , Elisabeth Schrumpf 1 2 3 , Tom H. Karlsen 1 2 3 4 , Richard S. Blumberg 5 , Espen Melum 1 2 4
  1. Norwegian PSC Research Center, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital, Rikshospitalet, Oslo, Norway
  2. Research Institute of Internal Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital, Rikshospitalet, Oslo, Norway
  3. Institute of Clinical Medicine, University of Oslo, Oslo, Norway
  4. Section of Gastroenterology, Division of Surgery, Inflammatory Medicine and Transplantation, Surgery, and Transplantation, Oslo University Hospital, Rikshospitalet, Oslo, Norway
  5. Division of Gastroenterology, Hepatology and Endoscopy, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA

Background

Natural Killer T (NKT) cells are abundant in the liver of mice and humans. We have previously demonstrated that CD1d on the biliary epithelium can present antigens to NKT cells1. We hypothesized that lipids dissolved in bile could act as antigens and activate NKT cells. 

 

Methods

Bile was collected from ten patients undergoing liver transplantation due to primary sclerosing cholangitis (PSC) (n=5), cryptogenic cirrhosis (n=3), autoimmune hepatitis (n=1), or hepatocellular carcinoma (n=1). Murine invariant (24.7, 24.8 and DN32.D3) and non-invariant (14S.6, 14S.7, 14S.10 and 14S.15) NKT cell hybridomas covering a range of different antigen specificities were used to test bile samples for reactivity. α-Galactosylceramide was used as a positive antigen control. Cell-culture plates were coated with a murine CD1d-monomer and incubated with the different bile samples over night. NKT hybridomas were added to the plates and IL-2 was analyzed in the culture supernatants by ELISA.

 

Results

Eight out of ten bile samples activated at least one of the NKT hybridomas. The NKT hybridoma that demonstrated the clearest activation by the bile samples was 14S.10, which is a non-invariant NKT hybridoma14S.10 was further used in the subsequent experiments along with four bile samples that demonstrated distinct activation. The bile samples were then tested in dilutions from 1:100 to 1:1.000.000. For all of the samples there was an evident dose-response relationship. Of note there was clear activation even at the highest dilutions.

 

Conclusions

Bile from patients with a diverse set of liver diseases contains antigens that activate both invariant and non-invariant NKT hybridomas. The biliary antigens that activate these cells demonstrate a dose-response relationship and do not seem to be present in bile from all patients. These findings indicate that activation of NKT cells by lipid antigens in bile can be of relevance in the disease development of inflammatory biliary diseases.

  1. Schrumpf E, Tan C, Karlsen TH, et al. The biliary epithelium presents antigens to and activates natural killer T cells. Hepatology. April 2015. doi:10.1002/hep.27840.