HEPATOLOGY Vol. 22, No. 4, Pt. 2, 1995 AASLD ABSTRACTS 353A

985 HLA-B8 CONFERS SUSCEPTIBILITY TO HCV-RELATED MIXED CRYOGLOBUMNAEMIA. M Lenzi, M Frisoni*, V Mantovani °. P Ricci ̂ . L Muratori, L Baffoni*. S Fen'i* and FB Bianchi. Medicina Interna II, Servizio di Reumatologia*, Laboratorio Analisi*, Istituto di Ematologia ̂ , Policlinico S. Oraola-Malpighi, University of Bologna, Bologna, Italy.

Mixed cryoglobulinaemia (MC) has been shown to be HCV- related, but its frequency is fairly low when compared to that of HCV infection in the Mediterranean area. Since no relationship has been found between HCV genomic heterogeneity and the occurrence of HCV-related MC, it is possible that host genetic factors may be required for the onset of this condition. We have therefore performed HLA typing in 20 patients with HCV-relatsd MC and compared the results to those of 351 healthy controls and 30 patients with HCV- related chronic hepatitis (CH). Purpura, arthralgias and weakness were required for the admission of MC patients to the study; 14/25 (56%) had type II and 11/25 had type III cryoglobulinemia. One patient was anti-HCV positive/HCV RNA negative, one was anti-HCV negative/HCV RNA positive, and the remaining 23 were positive for both antibodies to HCV and serum HCV RNA. Evidence of chronic liver disease was available in 20 patients while the remaining S had persistently normal transamineses values. A significant association was found between MC and HLA BS which was detected in 10/25 (40.0%) MC patients compared with 36/351(10.2%) healthy controls (p 0.0002, pc<0.005, RR 5,83) and 2/30 (6.7%) CH patients (p 0.004, pc NS). No other HLA association was found, including Class II specificities in linkage disequilibrium with data suggest that HLA-B8 may favour the onset of MC in the presence of HCV infection

986 SEQUENCE ANALYSIS IN HYPERVARIABLE REGION OF HCV IN CRYOGLOBULINS WITH CHRONIC HCV DISEASE T Aivama. K yoshioka. A Ok~lmura. M Takavanaai. K Iwata. and S Kakurnu. Third Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan.

Backoround /A ims : Mixed cryoglobulinemia is frequently associated with hepatitis C virus (HCV) infection. HCV RNA was detected in the cryoprecipitate, and cryoglobulins (CGs) have been suggested to be consisted of HCV antigen- antibody complex. The hypervariable region (HVR) of E2/NS1 has been reported to be epitopes for the neutralizing antibodies, and it is also likely that there are escape mutants from the antibodies. To characterize the nature of HCV- associated CGs, of HCV genome in the HVR was sequenced for the supernatants and cryoprecipitates from Japanese patients' sera with chronic HCV disease. Methods : Four patients with Cgs; two had liver cirrhosis and the other two chronic active hepatitis were studied. Two patients without CGs were served as controls. The gnotype of all patients was lb. HCV RNA was detected by RT nested PCR and sequencing was performed directly by Taq dye primer method. Results : In the all patients with CGs, the amount of HCV RNA in the cryoprecipitate was larger than that in the supernatant. There was no cryoprecipitate in patients without CGs. The sequence in HVR of HCV in the cryoprecipitate revealed that amino acid (aa) differed from that in the supernatant at 6~13 sites per 27 aa (22.2%~48.1% with a mean of 38.9%) analyzed. However, no apparent genomic changes characteristic to crycprecipitate was found. Conclusions : These findings suggest that in patents with CGs, a majority of HCV in sera was found in the cryoprecipitate, and that the HCV genomes in cryoprecipitate differed from those in supernatant.

987 HEPATITIS C VIRUS INFECTION AND MONOCLONAL GAMMOPATHY (MG): POSSIBLE ROLE OF GENOTYPE 2a. P Andreone. AL Zi~,ne~,o (D. C Cursaro, A Gramenzi. C Giannini (1~. A Buzzi (2~. MG Covarelli. F Giannelli ( D . M Bemardi. P Gentilini ( lk G Gasbarrini (2~. Patologia Medica I, Bologna; (1)lstituto di Medicina Intema, Firenze; (2)Clinica Medica, Universit~ Cattolica, Roma; Ricerca in Medicina, Bologna; Italy.

Recent studies suggested that HCV-infection may be associated with B-cell lymphoproliferative disorders, such as cryoglobulinemia and non-Hodgkin's lymphoma. We examined the prevalence of MG in 198 outpatients with HCV- positive chronic liver disease (mean age 51,1-+13,4 yrs; M/F: 118/80) compared to 54 outpatients with HBsAg-positive chronic liver disease (mean age 47,3-+12,8 yrs; M/F: 36/18). The two groups were comparable concerning age and sex. MG was found in 22/198 (11,1%) HCV-positive patients (pts) and in 1/54 (1,9%) HBsAg-positive pts (p=:02). In order to evaluate the role of specific HCV variants, we analyzed HCV genotypes in HCV-positive pts with MG and in a control group (matched for age, sex and histology) of HCV- positive pts without lymphoproliferative disorders. To date, genotyping of HCV (by amplification of core region with type-specific primers, according to Okamoto et al. with minor modifications) has been performed on serum samples of 13 MG pts and 38 controls. Analysis of HCV genotypes showed an higher prevalence of genotype 2a in pts with MG compared to controls as reported in the table:

HCV genotypes la lb 2a 3a la+lb lb+2a MG (13 pts) 1 (8%) 3 (23%) 8 (62%) 0 0 1 (8%)

p n.s. .02 .006 n.s. n.s. n.s.

Conlrols(38pts) 5(13%) 23(61%) 17(18%) 2(5%) 1(3%) 0 ms.=not significant Five MG pts were treated with IFN-cc and a reduction of monoclonal protein was observed in the 2 long responders. Our results show an association between HCV infection and MG. The observed prevalence of 11,1% is significantly higher compared both to HBsAg-positive pts and to the reported prevalence of MG in general population. The high prevalence of genotype 2a in pts with MG suggests that viral factors could be involved in the pathogenesis of this B-cell disorder. Further studies are needed to establish the role of HCV in inducing MG and the possible therapeutic efficacy of antiviral treatment.

988 MIXED CRYOGLOBULINEMIA AND HEPATITIS C VIRUS GENOTYPES AL Zignego. C Fen'i*, L La Civita*, M Monti, F Lombardini,* C Giaunini, G

Longombardo*, G Careccia, G Buzzelli, S Bombardieri, P Gentilini. P, heumatology and Clinical Immunology Units, University of Pisa * and Istituto Medicina Interna, University of Florence, Italy.

Mixed cryoglobulinemia (MC) is a systemic vasculitis characterized by skin parpara, palyarthralgias, chronic hepatitis (CH), nephritis, and peripheral neuropathy due to vascular deposition of immune complexes. The striking association between MC and hepatitis C virus (HCV) infection suggested that this virus is the trigger factor of B-lymphocyte expansion underlying the disease. This benign lymphoproliferation in some subjects can evolve to a non-Hedgkin's lymphoma (NHL). Hypothetically, clinical polymorphism of the MC could be correlated with some host and/or enviromental factors, and not secondarily to different HCV genotypes. The analysis ofHCV genotypes was carded out by two methods, based on type-specific primers or probes respectively, to better classify, the majority of isolated HCV. The study included 25 HCV-positive MC patients (7M, 18F, mean age 60-25 years) and 25 age and sex-roached subjects with chronic HCV infection without MC. Results are summarized in the table.

MCtotal MC+CH MC-CH MC+NIK, Controls 25pts. (%) 12pts. (%) 13pts. (%) 10pts.(%) 25pts.(%)

lb 60 92 31 50 64 2a 40 8 69 50 16

The analysis of HCV genotypes demonstrated a significanly different distribution in MC and controls. In particular, genotype 2a was higher in MC patients than in controls (p<0.05), and relatively more frequent in the MC subset with complicating NHL or without clinical signs of chronic hepatitis. In contrast genotype lb was detectable in the majority of MC with chronic elevation of liver enzymes. On the whole these data suggest that MC is not associated to a unique HCV genotype even if genotype 2a seems to be particularly involved in the pathogenesis of lymphoproliferation underlying MC.