Serum autoantibody measurement for the detection of hepatocellular carcinoma.

BACKGROUND: Individuals with liver disease, and especially those with Hepatitis B or C, are at an increased risk of developing hepatocellular carcinoma (HCC) which is the third most common cause of cancer-related death worldwide. Inadequate screening tests largely account for presentation of advanced tumours and high mortality rates. Early detection of HCC amongst high-risk groups is paramount in improving prognosis. This research aimed to further characterise the previously described humoral immune response raised to tumour-associated antigens (TAAs) in the serum of patients with HCC. METHODS: Serum from 96 patients with confirmed HCC, 96 healthy controls matched for age and sex, 78 patients with confirmed liver cirrhosis and 91 patients with confirmed chronic liver disease were analysed for the presence of IgG autoantibodies raised to 41 recombinant TAAs/antigen fragments by ELISA. RESULTS: Varying autoantibody specificities (97-100%) and sensitivities (0-10%) were observed to individual TAAs. A 21-antigen panel achieved a specificity of 92% and sensitivity of 45% for the detection of HCC. This same panel identified 21% of 169 high-risk controls as having elevated autoantibody levels. A reproducible panel of 10 antigens achieved a specificity of 91% and sensitivity of 41% in HCC. 15% of 152 high-risk controls gave positive results with this panel. CONCLUSIONS: This minimally invasive blood test has the potential to offer advantages over currently available tools for the identification of HCC amongst pre-disposed patients. Results are comparable to current gold standards in HCC (Ultrasonography) and to similar tests in other cancers (EarlyCDT-Lung).

Accumulation and dissemination of prion protein in experimental sheep scrapie in the natural host.

BACKGROUND: In order to study the sites of uptake and mechanisms of dissemination of scrapie prions in the natural host under controlled conditions, lambs aged 14 days and homozygous for the VRQ allele of the PrP gene were infected by the oral route. Infection occurred in all lambs with a remarkably short and highly consistent incubation period of approximately 6 months. Challenge of lambs at approximately eight months of age resulted in disease in all animals, but with more variable incubation periods averaging significantly longer than those challenged at 14 days. This model provides an excellent system in which to study the disease in the natural host by virtue of the relatively short incubation period and close resemblance to natural infection. RESULTS: Multiple sites of prion uptake were identified, of which the most important was the Peyer's patch of the distal ileum. Neuroinvasion was detected initially in the enteric nervous system prior to infection of the central nervous system. At end stage disease prion accumulation was widespread throughout the entire neuraxis, but vacuolar pathology was absent in most animals that developed disease at 6-7 months of age. CONCLUSION: Initial spread of detectable PrP was consistent with drainage in afferent lymph to dependent lymph nodes. Subsequent accumulation of prions in lymphoid tissue not associated with the gut is consistent with haematogenous spread. In addition to macrophages and follicular dendritic cells, prion containing cells consistent with afferent lymph dendritic cells were identified and are suggested as a likely vehicle for carriage of prions from initial site of uptake to the lymphoreticular system, and as potential carriers of prion protein in blood. It is apparent that spongiform change, the characteristic lesion of scrapie and other prion diseases, is not responsible for the clinical signs in sheep, but may develop in an age dependent manner.

Modification of blood cell PrP epitope exposure during prion disease.

PrPC [normal cellular PrP (prion-related protein)] is a glycosylphosphatidylinositol-linked cell-surface glycoprotein that is expressed primarily by cells of the central and peripheral nervous system and the lymphoreticular system. During prion disease, PrPC undergoes structural modification to PrPSc (abnormal disease-specific conformation of PrP). The appearance of prion infectivity and PrPSc within different peripheral lymphoid tissue sites during natural scrapie infection in sheep is suggestive of haematogenic dissemination. For this to occur, blood cells may harbour or carry disease-associated PrP and in doing so present altered conformations of PrP on their cell-surface. In the present study, we show that changes in PrP epitope expression, or accessibility, can be detected on peripheral blood mononuclear cells during the course of experimental scrapie in susceptible sheep. Peripheral blood mononuclear cells isolated from VRQ homozygous lambs inoculated orally with scrapie were probed with either N- or C-terminal-specific anti-PrP monoclonal antibodies and analysed by flow cytometry. During the progression of scrapie, significant alterations were seen in the exposure of particular cell-surface PrP epitopes. These modifications included increased accessibility to N-terminal regions of the PrP molecule, to the region between beta-strand-2 and residue 171, and to the C-terminal region of helix-3. Increased accessibility in the globular C-terminal domain of PrP occurred in the vicinity of tyrosine dimers, which are believed to have increased solvent exposure in disease-associated PrP. We suggest that the alterations in anti-PrP monoclonal antibody recognition of cell-surface PrP on blood cells from scrapie-infected sheep are indicative of structural changes within this molecule that may be relevant to prion disease.

Studies of the transmissibility of the agent of bovine spongiform encephalopathy to pigs.

Studies to test the transmissibility of the bovine spongiform encephalopathy (BSE) agent to pigs began in 1989. Parenteral inoculation of the agent by three routes simultaneously (intracranially, intravenously and intraperitoneally) produced disease with an incubation period range of 69-150 weeks. Pre-clinical pathological changes were detected in two pigs killed electively at 105 and 106 weeks post-inoculation. Infectivity was detected by bioassay in inbred mice in the CNS of those pigs that developed spongiform encephalopathy. Infectivity was also found in the stomach, jejunum, distal ileum and pancreas of terminally affected pigs. These findings show that pigs are susceptible to BSE. In contrast, disease failed to occur in pigs retained for 7 years after exposure by feeding BSE-affected brain on three separate days, at 1-2 week intervals. The amounts fed each day were equivalent to the maximum daily intake of meat and bone meal in rations for pigs aged 8 weeks. No infectivity was found in tissues assayed from the pigs exposed orally. This included tissues of the alimentary tract. It is suggested that these pigs did not become infected. The relatively high oral exposure used in these experiments compared with feed-borne exposure in the field may explain the absence of an epidemic of spongiform encephalopathy in domestic pigs concurrent with the BSE epidemic in the UK.