Tracking the kinetics of intrahepatic immune responses by repeated fine needle aspiration of the liver

Tom Pembroke,Awen Gallimore,Andrew Godkin

doi:10.1016/j.jim.2015.04.011

Tom Pembroke, Awen Gallimore + Show 1 more

Open Access

https://doi.org/10.1016/j.jim.2015.04.011

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Abstract

Liver disease is an increasing global health burden. The final sequalae of cirrhosis, liver failure and hepatocellular carcinoma are often the result of inflammation driven by intrahepatic lymphocytes. Accurate assessment of organ-specific diseases ideally employs tissue sampling though this is rarely performed. Here we report our experiences of utilising repeated fine needle aspirations (FNAs) to assess liver-derived leukocytes. In 88 patient samples, we obtained a mean of 36,959 lymphocytes from each FNA-derived biopsy (SD 22,319 cells, range 5034–91,242 cells) measured by flow cytometry. This quick technique required minimal analgesia compared to liver biopsy (p=0.03); was well tolerated and safe, and hence repeated sampling up to 3 times within a week was feasible. We detail the technique to rapidly derive a single cell suspension suitable for multiparameter flow cytometry analysis. Finally we illustrate the importance of organ-derived sampling by showing that natural killer (NK) cells from FNA samples have a markedly altered phenotype compared to those assessed in peripheral blood. In combination these data validate FNA as a powerful and well-tolerated method of sampling intrahepatic lymphocytes to study the immunology of acute and chronic liver diseases.

Highlights

Inflammatory liver diseases, including chronic viral hepatitis, autoimmune hepatitis and steatohepatitis, represent a major global health burden
Nine patients had fine needle aspirations (FNAs) samples taken at multiple time points (Table 1) resulting in a total of 81 intrahepatic samples obtained for lymphocyte analysis
When patients proceeded to liver biopsies following FNA there was a marked increase in the need for analgesia compared to FNA alone, with 16 patients requiring cocodamol and 2 patients required intramuscular pethidine (χ2 p = 0.0004, Fig. 1)

Summary

Introduction

Inflammatory liver diseases, including chronic viral hepatitis, autoimmune hepatitis and steatohepatitis, represent a major global health burden. It is well recognised that, easy to sample, peripheral blood lymphocytes may not reflect the function and phenotype of cells within the liver. Sampling cells from within the liver has remained problematic. Liver biopsy, using 16–18 gauge sheathed (Tru Cut) needles or suction (Menghini) needles, remains the clinical gold standard to assess liver fibrosis and inflammation and can provide valuable diagnostic information. The invasive nature of the liver biopsy, coupled with significant morbidity (and very rarely mortality) has driven interest in development of biochemical and non-invasive markers (i.e. liver elastography) to assess chronic liver disease (Castera, 2012). Percutaneous liver biopsy has a mortality rate of 1 in 10,000 and the most common complication is abdominal or right shoulder pain in up to 25% of patients (Bravo et al, 2001).

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