Cristina Crespi

Early occurrence and recurrence of hepatocellular carcinoma in HCV-related cirrhosis treated with direct-acting antivirals

BACKGROUND & AIMS Hepatocellular carcinoma (HCC) represents a serious complication of HCV-related cirrhosis. New direct-acting antivirals (DAA) cure HCV infection in over 90% of patients. The aim of this study was to evaluate the early occurrence and recurrence of HCC in cirrhotic patients treated with DAA. METHODS We analysed 344 consecutive cirrhotic patients, without HCC, who were treated with DAA, and followed for 24weeks. Fifty-nine patients had previous HCC. RESULTS DAA therapy induced sustained virological response in 91% of patients. During 24-week follow-up, HCC was detected in 26 patients (7.6%, 95% CI: 4.99-10.84): 17 of 59 patients (28.81%, 95% CI: 17.76-42.07) with previous HCC and 9 of 285 patients (3.16%, 95% CI: 1.45-5.90) without previous HCC. Child-Pugh Class B, more severe liver fibrosis, lower platelet count, and previous HCC were significantly associated with HCC development, at univariate analysis. At multivariate analysis, Child-Pugh class (p=0.03, OR: 4.18, 95% CI: 1.17-14.8) and history of HCC (p<0.0001, OR: 12.0, 95% CI: 4.02-35.74) resulted independently associated with HCC development. Among the 59 patients with previous HCC, younger age and more severe liver fibrosis were significantly associated with HCC recurrence, both at univariate and at multivariate analysis. CONCLUSIONS In patients with HCV-related cirrhosis, DAA-induced resolution of HCV infection does not seem to reduce occurrence of HCC, and patients previously treated for HCC have still a high risk of tumour recurrence, in the short term. For these reasons, all cirrhotic patients should be closely monitored and followed during and after antiviral therapy. LAY SUMMARY New direct-acting antivirals are able to eradicate HCV infection in over 90% of patients with advanced liver disease. Unfortunately, the occurrence of liver cancer is not reduced in effectively treated cirrhotic patients. In addition, patients previously treated for HCC have still a high risk of tumour recurrence in the short term, despite DAA treatment.

Autoimmunity in chronic liver disease caused by hepatitis delta virus.

Recent evidence shows that chronic liver disease induced by hepatitis delta virus is closely associated with production of autoantibodies. To verify this sera from patients with hepatitis delta virus and from those with hepatitis B virus mediated chronic liver disease were tested for a panel of autoantibodies. Although the traditional non-organ specific autoantibodies were similarly distributed in the two groups, a considerably higher prevalence of IgG basal cell layer antibodies, IgM anti-intermediate filaments, and IgG antimicrotubule antibodies was found. Although these phenomena might be secondary to hepatitis delta virus infection, they might be the serological markers of underlying immune events.

Antibodies to the Cytoskeleton Components and Other Autoantibodies in Inflammatory Bowel Disease

A high prevalence of antibodies to cytoskeleton components (anti-CYTO) and of anti-smooth muscle (SMA) and antinuclear (ANA) antibodies has been found in inflammatory bowel disease (IBD). In particular, a significant correlation has been documented between anti-CYTO and activity of the disease in ulcerative colitis and anti-fibroblast intermediate filaments (anti-vimentin) antibodies and Crohns disease. The antibodies are detectable in the three major immunoglobulin classes, including IgA. While titres of anti-CYTO did not exceed 1:40, ANA and SMA were of high titres (up to 1:1,280). Antibodies to epithelial cells intermediate filaments (anti-cytokeratin) were detected only occasionally. The significance of these findings in IBD is discussed.

Close association between antibodies to cytoskeletal intermediate filaments, and chronic graft-versus-host disease

Chronic graft-versus-host disease can mimic various autoimmune disorders, although autoantibodies are rarely detected in the sera of affected patients. Antibodies to cytoskeleton are a frequent finding in patients affected by autoimmune disorders. In all the sera of 16 patients who were submitted to allogeneic bone marrow transplantation, we have found antibodies against cytoskeletal intermediate filaments. Moreover, the titer of such antibodies is quite elevated when compared with those reported in autoimmune disorders. A statistically significant difference between the titers found in patients without and with cGVHD (median 1:40 vs. 1:256, P<0.05) has been found. This would suggest that such antibodies might be relevant in monitoring clinical course. Furthermore, since certain cytoskeleton antigens have been shown to be expressed also on cell membrane, antibodies against intermediate filaments might also play a more important role by interfering with such surface structures.

Immunofluorescent detection of anti-cytoskeleton antibodies using vinblastine-treated mononuclear cells

A reliable and reproducible immunofluorescence method is described for the detection of anti-cytoskeleton antibodies in human sera, based on the use of vinblastine-treated peripheral blood mononuclear cells as substrate. Three immunofluorescence patterns associated with antibodies to microfilaments, intermediate filaments and microtubules are readily identified.

Vimentin and keratin intermediate filaments expression by K562 leukemic cell line

The expression of intermediate filaments (IMF) by K562 leukemic cell line has been studied by the use of polyclonal and monoclonal antibodies. These cells exhibited an extremely rich network of IMF reacting with both anti-vimentin and anti-keratin antibodies. Although these results need to be confirmed by different techniques, such as immunoblotting, they suggest the possibility that leukemic cell lines can express different types of IMF similarly to other cultured cells.

Cytoskeleton organization of normal and neoplastic lymphocytes and lymphoid cell lines of T and B origin

Summary An anomalous organization of the cytoskeleton has been described in lymphocytes from chronic lymphatic leukaemia and in only few cell lines. We have now studied normal and neoplastic lymphocytes and lymphoid cell lines of both T and B lineage in order to detect morphological differences in the expression of microfilaments and intermediate filaments. Microfilaments appear to be well expressed by all the B cells, whereas a rich network of intermediate filaments is present in T cells and plasma cells. Most prominent changes occur in the latter system, which is almost lacking in cells of B chronic lymphatic and hairy cell leukaemia. Although the significance of the present findings is not yet clear, one might speculate that such alterations account for some of the aberrant functions and peculiar biologic properties of neoplastic lymphocytes.

Role of target and effector cell structures in natural killer-mediated cytotoxicity

SummaryAn analysis of target and effector cell structures involved in thein vitro natural killer (NK)-mediated cytotoxicity has been performed. The degree of surface expression of transferrin receptor (TR) was only in part correlated with that of cell lysis. Moreover, the lysis could not be blocked by treating target cells with two anti-TR monoclonal antibodies. Finally, cell lines poorly affected by NK cells express TR only at the cytoplasmic level. As to the effector cells, the integrity of cytoskeleton components (especially microtubules) was found to be essential for the occurrence of cell lysis. In fact, vinblastine, an anti-microtubule agent, was able to significantly reduce the percentage cell lysis. This effect was not due to a selective depletion in NK cells induced by the drug. It is concluded that the mechanisms underlying NK activity are complex and involve both target and effector cell structures.

Relationship between smooth muscle and cytoskeleton antibodies in neuroblastoma.

The previously reported high prevalence of smooth muscle antibodies in neuroblastoma has been found to be associated with a similarly elevated prevalence of anti-cytoskeleton antibodies. The most relevant finding is related to anti-microfilaments (anti-actin) and anti-microtubules antibodies, which were detected with highly significantly different prevalences when compared with a disease control group (p < 0.001 and p < 0.000001, respectively). The correspondence between smooth muscle antibodies and anti-cytoskeleton antibodies is incomplete, and it is more relevant for anti-microfilaments. It is concluded that antigen specificities of smooth muscle antibodies in neuroblastoma are as complex as documented in other diseases. Possible pathogenetic and clinical implications emerging from these data are discussed.

Expression of Cytoskeleton Components in Various Epstein-Barr Virus Infected Cell Lines

&NA; It has been shown that viruses can induce alterations in the content and distribution of cytoskeleton structures, particularly actin microfilaments and microtubules. An immunomorphologic study of the cytoskeleton components of various EBV‐infected cell lines with expression of different functions of EBV genome has been performed using antibodies to each of its three major components. Intermediate filaments and microtubules were similarly represented in all examined lymphoblastoid cell lines. The distribution of actin microfilaments, on the contrary, differed significantly from cell line to cell line. It is concluded that the morphologic expression of actin might depend on the expression of EBV genome. Furthermore, some of these cell lines might represent a useful substrate for the identification of anticytoskeleton antibodies, mainly anti‐actin antibodies, in human sera.