The presence of hepatitis C virus (HCV) antibody in human immunodeficiency virus-positive hemophilic men undergoing HCV "seroreversion"

Hepatitis C virus (HCV) is a major cause of transfusion-induced chronic liver disease in hemophiliacs, with 70% to 90% being anti-HCV positive. Seroreversion or loss of antibody response to HCV has been observed in a small proportion of human immunodeficiency virus-positive [HIV(+)] anti-HCV(+) hemophilic men. Despite the seroreversion to an anti-HCV- negative state, such patients continue to show serum alanine aminotransferase (ALT) elevations and biopsy evidence of cirrhosis and/or chronic active hepatitis. To determine the cause for the loss of anti-HCV antibody, we compared first- and second-generation anti-HCV enzyme immunosorbent assay (EIA 1.0 and 2.0), second-generation recombinant immunoblot (RIBA 2.0), and HCV-RNA amplification using polymerase chain reaction (PCR) in 19 "seroreverters" before and after seroreversion. There was no difference between 19 seroreverters and 59 persistently anti-HCV-positive hemophiliacs in mean ALT (1.1 +/- 0.1 XUL v 2.0 +/- 0.2 XUL; chi 2 = 1.80, P > .05), in mean CD4 (188 +/- 36/microL v 232 +/- 28/microL; t = 0.965, P > .05), or in the rate of progression to acquired immunodeficiency syndrome (13 of 19 [68.4%] v 30 of 59 [50.9%]; chi 2 = .987, P > .05, respectively). Before seroreversion, all 19 seroreverters (100%) were positive for EIA 1.0 and 2.0 and PCR, and all but 2 of 19 (89.5%) were RIBA 2.0 positive, whereas, after seroreversion, none were positive for EIA 1.0, 15 of 19 (78.9%) were positive for EIA 2.0, 8 of 18 (44.4%) were positive for RIBA 2.0, and 18 of 19 (94.7%) were positive for PCR. There was a lower CD4 lymphocyte number after seroreversion in those who were RIBA 2.0 negative as compared with those who were RIBA 2.0 positive (32 +/- 10/microL v 171 +/- 52/microL; t = 2.638, P > .05). These results indicate that HIV(+) anti-HCV(+) hemophilic men who undergo "HCV seroreversion" are truly infectious and anti-HCV positive by second- generation tests. Anti-HCV detection in immunosuppressed hosts is significantly improved by second-generation EIA and RIBA assays.     

Cellular immune responses and occult infection in seronegative heterosexual partners of chronic hepatitis C patients

It is unknown whether hepatitis C virus (HCV)-specific cellular immune responses can develop in seronegative sexual partners of chronically HCV-infected patients and whether they have occult infection. Thirty-one heterosexual partners of patients with chronic HCV were studied, fifteen of them with HCV transmission risks. Ten healthy individuals and 17 anti-HCV seropositive patients, without viremia, were used as controls. Virus-specific CD4+ and CD8+ T-cell responses were measured by flow cytometry against six HCV peptides, situated within the nonstructural (NS) proteins NS3, NS4 and NS5, through intracellular detection of gamma interferon (IFN-γ) or interleukin 4 (IL-4) production and CD69 expression. Sexual partners had a higher production of IFN-γ and IL-4 by CD4+ cells against NS3-p124 (P = 0.003), NS5b-p257 (P = 0.005) and NS5b-p294 (P = 0.012), and CD8+ cells against NS3-p124 (P = 0.002), NS4b-p177 (P = 0.001) and NS3-p294 (P = 0.004) as compared with healthy controls. We observed elevated IFN-γ production by CD4+ T cells against NS5b-p257 (P = 0.042) and NS5b-p294 (P = 0.009) in the sexual partners with HCV transmission risks (sexual, professional and familial altogether) than in those without risks. RNA was extracted from peripheral blood mononuclear cells (PBMC), and detection of HCV-RNA positive and replicative (negative) strands was performed by strand-specific real-time PCR. In four sexual partners, the presence of positive and negative HCV- RNA strands in PBMC was confirmed. Hence, we found an HCV-specific cellular immune response as well as occult HCV infection in seronegative and aviremic sexual partners of chronically HCV-infected patients.     

Antibodies to recombinant and synthetic peptides derived from the hepatitis C virus genome in long-term-studied patients with posttransfusion hepatitis C.

Eight of 13 Swedish patients (62%), studied prospectively, who developed posttransfusion non-A, non-B hepatitis (PT-NANBH) had earlier been found to seroconvert for antibodies to hepatitis C virus (anti-HCV) c100-3 in the first-generation anti-HCV enzyme-linked immunosorbent assay 1-18 (mean, 8) weeks after onset of hepatitis. By using a second-generation test utilizing antigens encoded by the core NS3 and NS4 region of HCV, a further four patients non-reactive to c100-3 (NS4) were found to seroconvert. Thus 12 of 13 (92%) Swedish patients with PT-NANBH were shown to have HCV infection. In addition, the serologic reactivity for several individual synthetic peptides and/or recombinant HCV proteins was studied in seven anti-HCV c100-3 seroconverts studied long-term after onset of acute PT-HCV infection. No special patterns were found that could differentiate patients who recovered from those who developed chronic HCV infection. It was concluded that the addition of new recombinant antigens derived from the core and NS3 region to c100-3 (NS4) both improved the sensitivity of the anti-HCV test and shortened the window phase to seroconversion.     

Long-term evolution of serum and liver viral markers in patients treated for chronic hepatitis C and sustained response

Few studies have analysed the evolution of HCV markers in chronic hepatitis C (CHC)-treated patients. We have evaluated the presence or absence of serum and liver HCV-RNA, the core antigen (HCV-cAg) and the loss of specific antibodies (anti-HCV), in long-term sustained responders (SR). One hundred and seventy-six patients (132 SR and 44 nonresponders (NR) were included in the study. HCV-RNA was determined in serum and liver by a commercial PCR-kit. HCV-Ag was determined by ELISA and specific antibodies against HCV by means of a commercial line immunoblot assay (LIA) technique. Serum HCVcAg was found positive in three (4.2%) SR and in one (4%) NR (NS). Four SR (3.6%) and 44 NR (100%) were also HCV-RNA (+) in liver tissue. Two patients were HCV-cAg (+). A good correlation was found between the serum levels of HCV-cAg and HCV-RNA (r = 0.847, P < 0.001). Specific antibodies (anti-HCV) were determined by LIA in 45 patients. A decrease was found in the number of patients who presented reactivity to bands E2 and NS4 when we compared SR with a follow-up of more than 5 years with NR and SR with a follow-up <5 years (P < 0.01 and 0.005). A good correlation was found between the HCV-cAg and HCV-RNA serum levels in CHC-treated patients (P < 0.001). Few SR (3.6%) had HCV-RNA in the liver, and HCV-cAg (1.8%) in serum. In SR with more than 5 years of follow-up a clear tendency exists in the trend to clarify the bands E2 and NS4 of anti-HCV in serum.     

Preparation and application of monoclonal antibodies against hepatitis C virus nonstructural proteins

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Seroconversion to hepatitis C virus antibodies in patients with acute posttransfusion non-A, non-B hepatitis in Sweden with a second generation test.

28 patients with posttransfusion non-A, non-B (NANB) hepatitis in Stockholm, Sweden, were studied for seroconversion to hepatitis C virus antibodies (anti-HCV) and time lag to seroconversion by first and second generation tests. 15/28 patients (54%) seroconverted to anti-HCV with a first generation anti-HCV ELISA using C100-3 from the nonstructural (NS) region 4 of the HCV genome and 23 (82%) with a second generation anti-HCV ELISA including also antigens from the core and NS3 regions of the HCV genome. The mean time from onset of hepatitis to seroconversion was 6.1 weeks (0-18 weeks) with the first generation test and 2.3 weeks (0-7 weeks) with the second generation test. Development of chronic hepatitis was noticed in 14/23 (61%) patients who seroconverted to anti-HCV with the second generation ELISA and in none of 5 patients with posttransfusion NANB hepatitis who did not seroconvert. The inclusion of antigens from the core and NS3 regions of the HCV genome has increased the sensitivity of the second generation anti-HCV ELISA as compared to the first generation ELISA and also shortened the time lag to seroconversion in patients with posttransfusion hepatitis C. Patients with posttransfusion NANB hepatitis seroconverting seem more prone to develop chronic disease than patients not seroconverting.     

CD4 T lymphocyte proliferative responses to hepatitis C virus (HCV) antigens in patients coinfected with HCV and human immunodeficiency virus who responded to anti-HCV treatment

CD4 T lymphocyte proliferative responses to hepatitis C virus (HCV) antigens were evaluated before and during an anti-HCV regimen (interferon-alpha2a and ribavirin) in 36 patients coinfected with HCV and human immunodeficiency virus (HIV), to determine whether immune responses against HCV antigens are present in such patients, whether these responses are modified by anti-HCV treatment, and whether they are correlated with treatment efficacy. The CD4 responses against HCV antigens (primarily core antigens) detected at study entry in one-half of the patients did not correlate with anti-HCV treatment efficacy. Of 36 patients, 8 had patterns of persistent immune response to infection by genotypes 3 or 4 that were significantly correlated with sustained virologic response. Persistent immunologic reactivity and sustained virologic response coexisted only in patients infected with genotype 3. These findings suggest that HCV genotype may influence specific immune response, which, in turn, is implicated in virologic control.     

Retracted: outcome and immune reconstitution of HBV-specific immunity in patients with reactivation of occult HBV infection after alemtuzumab-containing chemotherapy regimen

Whether preemptive anti- hepatitis B virus (HBV) therapy should be considered in all hepatitis B surface antigen (HBsAg)-negative patients with occult HBV infection receiving alemtuzumab containing chemotherapy is uncertain. We determined the outcome and effect on HBV-specific immunity of an alemtuzumab-containing chemotherapy regimen in occult HBV-infected patients. Twenty-one consecutive occult HBV-infected patients treated with an alemtuzumab containing chemotherapy regimen were studied. T cell reactivity to HBV antigens and -peptides were quantified by ELISpot and the T cell subset by flow cytometry. Six of the 21 patients (28.6%) developed HBsAg seroreversion. The median (range) time to development of HBsAg seroreversion after the end of chemotherapy was 1.8 months (0.2-2.3 months). Direct sequencing showed that the occult HBV infection of all six patients (100%) was reactivated. These six patients developed severe HBV-related hepatitis. At the end of follow-up, four of these six patients (66.7%) had become negative for HBsAg again. Recovery of CD4+ T cell count and CD4+T cell reactivity against hepatitis B core antigen (HBcAg) occurred 9 months after the end of chemotherapy. Loss of HBsAg occurred after recovery of the CD4+T cell count and increased CD4+T cell reactivity against HBcAg 9 months after the end of chemotherapy. CONCLUSION: An alemtuzumab-containing chemotherapy regimen is associated with a high risk of reactivation of occult HBV infection. Suppression of HBV immunity by an alemtuzumab-containing chemotherapy regimen would persist until 9 months after the end of chemotherapy. In occult HBV-infected patients receiving an alemtuzumab-containing chemotherapy regimen, preemptive anti-HBV therapy should be continued until 9 months after the end of chemotherapy, when recovery of HBV immunity has occurred.     

Hepatitis C virus‐specific CD8+ T cell frequencies are associated with the responses of pegylated interferon‐α and ribavirin combination therapy in patients with chronic hepatitis C virus infection

Aim: Hepatitis C virus (HCV)‐specific cytotoxic T lymphocytes (CTLs) play critical roles in elimination of the HCV‐infected hepatocytes. However, the mechanism of HCV elimination by pegylated interferon‐α (peg‐IFNα) plus ribavirin is not fully understood. We examined HCV‐specific CTL responses during this combination therapy. Methods: CD8+ T cells were isolated from 16 HCV infected patients treated by this combination therapy and were subjected to IFN‐γ enzyme‐linked immunospot (ELISPOT) assay. Results: The numbers of IFN‐γ spots against HCV Core or NS3 protein‐derived peptides in HCV patients before treatment were similar to those in healthy donors, and those in HCV patients significantly increased 4 weeks after the initiation of combination therapy. All HCV Core or NS3 proteins‐derived peptides specific CD8+ T cells responses in pre‐treated patients were not associated with ALT levels and HCV viral loads of HCV patients before treatment. And those in pre‐treated patients were similar between sustained virologic responder (SVR) patients and non‐SVR patients. Significant increase of HCV Core or NS3 proteins‐derived peptides specific CD8+ T cells responses between before and 4 weeks after this combination therapy were observed in SVR patients, but not in non‐SVR patients. Conclusions: These results demonstrated that significant increase of HCV‐specific CD8+ T cells at 4 weeks after the initiation of IFN treatment might be associated with the elimination of HCV. Our findings suggest that the reactivity against HCV Core and NS3 proteins‐derived peptides might be useful in predicting the clinical outcome of the combination therapy of peg‐IFNα and ribavirin.     

Induction of IgA and sustained deficiency of cell proliferative response in chronic hepatitis C

AIM： In the present study, antibody and peripheral blood mononuclear cells （PBMC） proliferative responses against hepatitis C virus （HCV） antigens were evaluated in HCV chronically infected patients. METHODS： Paired serum and PBMC samples were taken six months apart from 34 individuals, either treated or not, and tested by enzyme-linked immunosorbent assay （ELISA） and carboxyfluorescein succinimidyl ester staining. RSULTS： Over 70% of the patients showed specific IgG and IgM against capsid, E1 and NS3, while HVR-1 was recognized by half of the patients. An increase in the levels of the anti-capsid IgM （P = 0.027） and IgG （P = 0.0006） was observed in six-month samples, compared to baseline. Similarly, a significantly higher percent of patients had detectable IgA reactivity to capsid （P = 0.017） and NS3 （P = 0.005） after six months, compared to baseline. Particularly, IgA against structural antigens positively correlated with hepatic damage （P = 0.036）. IgG subclasses evaluation against capsid and NS3 revealed a positive recognition mediated by IgG1 in more than 80% of the individuals. On the contrary, less than 30% of the patients showed a positive proliferative response either of CD4＋ or CD8＋ T cells, being the capsid poorly recognized. CONCLUSION： These results confirm that while the cellular immune response is narrow and weak, a broad and vigorous humoral response occurs in HCV chronic infection. The observed correlation between IgA and hepatic damage may have diagnostic significance, although it warrants further confirmation.     

Epidemiologic study of anti-HCV antibody reactivity with immunoblot assay

To investigate the relationship between the reactivity of anti-HCV antibody and detection of HCV RNA, we evaluated the anti-core, NS3 and NS4 antibodies by immunoblot assay. The subjects were 79 inhabitants of H. Town who were anti-HCV positive. HCV RNA-positive ratios were significantly higher in high titer group (10 >S /CO) than in low titer group (10 ≤ S/CO) (P < 0.001). Subjects were classified into four groups according to the antibodies reactivity: A, anti-core reactive only (nine subjects); B, anti-core and anti-NS3 reactive (19 subjects); C, reactive to all epitopes (47 subjects); and D, others (four subjects). In group A, seven (77.8%) were negative for HCV RNA. In groups B, C and D, 17 (24.3%) were negative for HCV RNA. Anti-HCV positive, but HCV RNA-negative, subjects were frequently observed in group A (P < 0.01) and in female (P < 0.05). We concluded that (1) the high titer of antibody against each epitope may reflect the presence of HCV RNA; (2) the natural elimination of HCV may have occurred in group A subjects and in females.     

Serological response to infection with different isolates of hepatitis C virus

Different isolates of hepatitis C virus (HCV) show nucleotide sequence variability throughout the genome. Detection of antibodies to recombinant proteins derived from hepatitis C virus genotype 1, the prototype HCV clone HCV-PT, constitutes the main method for screening HCV infection. The influence of the genomic variability on the serological diagnosis of HCV by enzyme immunoassay remains poorly defined. The aim of this study was to assess the serological reactivity of a panel of well characterized French HCV isolates typed by sequence analysis from patients with chronic hepatitis. The 73 sera samples were tested in three third generation EIA tests and three confirmatory assays. HCV isolates were determined by RT-PCR and sequencing in NS5B region of the genome. The 73 sera were positive in the three EIA tests. The three confirmatory tests showed a weaker reactivity with NS5 protein whatever the genotype, and a lower reactivity in NS4 antigens of non-type 1 sequences, particularly for genotype 3. Even though the reactivity of the antigens differed among the HCV isolates, the 73 isolates from genotype 1-6 were reactive with the three commercial screening assays. These results demonstrate that using a single test is adequate in the routine diagnosis of HCV infection in clinical laboratory, as recommended by the last French and European consensus conference.     

Quantitative analysis of anti-hepatitis C virus antibody-secreting B cells in patients with chronic hepatitis C

To investigate the quantitative characteristics of humoral immunity in patients with hepatitis C, we established an enzyme-linked immunosorbent spot (ELISpot) assay for detection of anti-hepatitis C virus (HCV)-secreting B cells. Receiver operating characteristic curve analysis demonstrated 100% specificity and 58% to 92% sensitivity for detecting B-cell responses to NS5b, NS3, E2, and core antigens. The median sum of anti-HCV-secreting B cells to all HCV antigens tested was significantly higher in 39 patients with chronic hepatitis C (47.3 spot forming cells [SFCs]/10(6) peripheral blood mononuclear cells [PBMCs]) than in 9 recovered subjects (15.3 SFCs/10(6) PBMCs; P = .05) or 11 uninfected controls (5.3 SFCs/10(6) PBMCs; P < .001); the significant difference (P = .018) in chronic versus recovered patients was in reactivity to nonstructural antigens NS3 and NS5b. Anti-HCV immunoglubulin M (IgM)-secreting B cells were also readily detected and persisted decades into HCV infection; there was no difference in IgM-positive cells between chronic and recovered patients. ELISpot reactivity to genotype 1-derived antigens was equivalent in patients of genotypes 1, 2, and 3. There was significant correlation between the numbers of anti-HCV IgG-secreting B cells and serum aminotransferase and to the level of circulating antibody. In conclusion, ELISpot assays can be adapted to study B-cell as well as T-cell responses to HCV. Measurement at the single-cell level suggests that humoral immunity plays a minor role in recovery from HCV infection and that B-cell immunity is strongest in those with persistent infection.     

Long-term outcome of vertically acquired and post-transfusion hepatitis C infection in children

BACKGROUND AND AIM: To determine the natural history of perinatally acquired hepatitis C virus (HCV) infection, clinical and laboratory outcomes among 31 children with HCV infection were retrospectively reviewed. Fifteen children had acquired HCV by blood transfusion (BT) prior to 6 months of age and 16 had vertically acquired (VT) HCV. METHODS: Demographic data, clinical symptoms and signs, liver biochemistry, HCV antibody, HCV-RNA and liver histology were evaluated. RESULTS: Mean age at last visit was 13.0 years (range 9.0-16.8 years) in the BT group and 8.6 years (range 0.5-18.1 years) in the VT group. There were no abnormal clinical findings of chronic liver disease in either group. Estimated HCV-RNA clearance rate was 19%, with no significant difference between the groups. In HCV-RNA-negative children (n = 6), two lost anti-HCV antibody and two developed indeterminate anti-HCV antibody results, while all HCV-RNA-positive children (n = 25) remained both anti-HCV antibody positive and HCV-RNA positive throughout follow up. The alanine aminotransferase level was significantly higher in the VT group than in the BT group during the first 5 years of life. Liver biopsy, which was carried out in four children, revealed mild to moderate fibrosis and/or necroinflammatory activity, but no cirrhosis. CONCLUSIONS: Outcomes among children with HCV acquired in infancy demonstrate asymptomatic and slowly progressive disease, at least for the initial decade of infection. Mode of acquisition appears to have a limited impact on outcomes, with similar viral clearance and anti-HCV antibody seroreversion rates in vertical and transfusion acquired infection.     

Localization of hepatitis c virus antigens in liver and skin tissues of chronic hepatitis c virus—Infected patients with mixed cryoglobulinemia

Skin and/or liver biopsy specimens were obtained from the following patients: 15 anti-hepatitis C virus (HCV), HCV RNA-positive patients and 3 anti-HCV, HCV RNA-negative patients with type II mixed cryoglobulinemia (MC); 7 anti-HCV, HCV RNA-positive patients with chronic active liver disease (CALD); 5 anti-HCV, HCV RNA-negative patients with noncryoglobulinemic vasculitis; and 7 anti-HCV, HCV RNA-negative patients with lichen ruber planus. A pool of marine monoclonal antibodies (MAbs) developed against c22-3, c33c, and c100-3 proteins was used to detect HCV-related antigens (Ags) by indirect immunohistochemistry. Acid electroelution (AEE) of tissue sections was performed to enhance the sensitivity of the immunohistochemical method. In anti-HCV-positive MC patients, specific HCV-related Ags were detected in the small vessels of the skin and in the cytoplasm of hepatocytes. Prior AEE of biopsy sections allowed detection of HCV Ags in 6 of 15 (40%) skin biopsy and in 9 of 14 (64.3%) liver biopsy specimens. HCV immunoreactive deposits in the skin displayed two immunohistochemical patterns: (1) coarse intraluminal material associated with dermal inflammatory infiltrates and intravascular deposition of eosinophilic hyaline material; and (2) reactivity confined to the vessel wall in the context of an apparently normal tissue. Immunoglobulin (Ig) G and IgM deposition in the skin showed immunohistochemical features comparable with those found for HCV Ag deposits. In addition, tissue complement reactivity was detected in 6 (40%) of them and was strictly associated with histological and clinical signs of active vasculitis. Five of 7 (71.4%) liver biopsy specimens, but none of 7 skin biopsy specimens, from anti-HCV, HCV RNA-positive patients without circulating cryoglobulins displayed immune reactivity after AEE procedure. Consistently negative results were obtained with skin and liver sections from all anti-HCV, HCV RNA-negative patients. These findings indicate that, under the experimental conditions used, in 40% of anti-HCV, HCV RNA-positive patients with MC, skin tissue deposits consist of HCV-containing immune complexes. In addition, the occurrence of HCV reactivity in apparently normal blood vessels suggests that deposition of viral Ags precedes and possibly initiates tissue damage. Whether in the remaining patients HCV Ags cannot be detected because of the insufficient sensitivity of the method or the involvement of Ags other than those assayed, remains to be determined.     

The systemic and local immune responses in patients with alcoholic liver cirrhosis depending on hepatitis C viral infection (HCV)

The systemic and local immune response was studied in patients with alcoholic liver cirrhosis and the significance of the combined infection with HCV. To investigation were submitted 23 patients (16 males and 7 females) aged between 29 and 61 years with alcoholic liver cirrhosis. Of them 14 were anti-HCV(+) and 9 anti-HCV(-). As controls were used 36 clinically healthy individuals, matched by sex and age to the patients. The flow cytometric analysis of the lymphocyte (Ly) populations from the peripheral venous blood and of cells from liver aspirate obtained by blind liver biopsy according to Menghini, was performed with FacsTAR (Becton Dickinson). In the anti-HCV(-) patients, as compared to the controls (patients/controls) the Ly subpopulations were increased: CD3+/mm3:2010 +/- 738/1440 +/- 388; CD4+/mm3:1350 +/- 441/991 +/- 442; IL-2R+/mm3:133 +/- 78.5/31 +/- 20. In the anti-HVC(+) patients we established increased IL-2R+/mm3: 170 +/- 126 as compared with the controls and anti-HCV(-) patients. The suppressor/cytotoxic (CD8+) Ly with their suppressor (CD8+CD11b+) and cytotoxic (CD8+CD11b-) subpopulations and natural killers (CD16+) had a tendency to diminution in the anti-HCV(+) patients. In both examined groups the B (CD19+) Ly were non-significantly increased. The flow cytometric analysis of the cells from the liver specimen in 9 patients of whom 3 anti-HCV(-) and 6 anti-HCV(+) revealed that CD3+ on the average were 32.8% +/- 20.4% (from 9.2% to 65.1%); CD4+ were 21.1% +/- 7.4% (from 12.0% to 34.5%); CD8+ 22.6% +/- 11.8% (from 4.7% to 39.8%) and their values were higher in the anti-HCV(+) patients; the correlation CD4+/CD8+ = 1/1.09 +/- 0.6; CD16+ were 12.9% +/- 10.1% (from 1.9% to 34.8%); CD19+ varied from 3.2% to 27.8%; monocytes (CD14+) were 7.69% +/- 5.65 (from 2.0% to 15.8%) from the cells of the aspirate and their percentage contents was higher in the anti-HCV(+) patients. The results of out study revealed that in patients with alcoholic liver cirrhosis changes in the cell immune response were also observed and that they were more marked in infection with HCV.     

Identification of a hepatitis C virus-reactive T cell receptor that does not require CD8 for target cell recognition

Hepatitis C virus (HCV) has been reported to elicit B and T cell immunity in infected patients. Despite the presence of antiviral immunity, many patients develop chronic infections leading to cirrhosis, hepatocellular carcinoma, and liver failure that can require transplantation. We have previously described the presence of HLA-A2-restricted, HCV NS3-reactive cytotoxic T lymphocytes (CTL) in the blood of HLA-A2- liver transplantation patients that received an HLA-A2+ liver allograft. These T cells are analogous to the "allospecific" T cells that have been described in hematopoietic stem cell transplantation patients. It has been speculated that allospecific T cells express high-affinity T cell receptors (TCRs). To determine if our HCV-reactive T cells expressed TCRs with relatively high affinity for antigen, we identified and cloned a TCR from an allospecific HLA-A2-restricted, HCV:NS3:1406-1415-reactive CD8+ T cell clone and expressed this HCV TCR in Jurkat cells. Tetramer binding to HCV TCR-transduced Jurkat cells required CD8 expression, whereas antigen recognition did not. In conclusion, based on the reactivity of the TCR-transduced Jurkat cells, we have identified a TCR that transfers anti-HCV reactivity to alternate effectors. These data suggest this high affinity HCV-specific TCR might have potential new immunotherapic implications.     

人类免疫缺陷病毒和丙型肝炎病毒共感染患者肝硬化发病情况

目的探讨人类免疫缺陷病毒(HIV)和丙型肝炎病毒(HCV)共感染患者发展到肝硬化的情况及原因。方法将观察对象分为HIV和HCV共感染组、单独HCV感染组。回顾性比较HIV和HCV共感染患者、单独HCV感染患者15年内发展到肝硬化的情况。结果HIV和HCV共感染患者15年内肝硬化发生率为16.4％(23／140)明显高于单独HCV感染组3.0％(1／33),差异有统计学意义,x2=4.012,P=0.045。CD4+、CD8+T淋巴细胞计数在HIV和HCV共感染组分别为(200.0±134.1)个／μl及(880.6±444.2)个／μl,而单独HCV感染组分别为(752.3±251.7)个／μl及(529.0±170.7)个／μl,两组比较差异有统计学意义,t值分别为12.020和7.600,P值均<0.01;HCVRNA(+)、抗HCV(+)数与HCVRNA(+)、抗-HCV(-)数,在HIV和HCV共感染组为89例和15例,在HCV感染组为25例和0例,两组间差异有统计学意义,x2=4.080,P=0.043。结论HIV和HCV共感染可加速肝硬化的进展,可能与HIV对机体的细胞免疫、体液免疫有关。     

Relationship between early HCV kinetics and T-cell reactivity in chronic hepatitis C genotype 1 during peginterferon and ribavirin therapy

BACKGROUND/AIMS: To gain understanding of inter-individual differences of treatment response in hepatitis C virus genotype 1 (HCV-G1) patients, we investigated simultaneously the early HCV kinetics and virus-specific T-cell reactivity. METHODS: Thirty, treatment-na?ve HCV-G1 patients received peginterferon-alfa2a 180 microg/week plus ribavirin 1000-1200 mg/day, with blood samples collected prospectively at protocol time-points. HCV RNA was quantitated with a TaqMan assay with mathematical modelling of HCV decay. Virus-specific CD4+/CD8+ T-cells were enumerated by Elispot assays. RESULTS: HCV kinetic analysis identified two subgroups: fast (18/30) and slow (12/30) treatment-responders. Although these subgroups did not differ in any baseline characteristics, fast responders (FR) showed greater antiviral efficacy (epsilon) than slow responders (SR) (84.5+/-3.2 vs. 65.2+/-7.0%, P=0.002), and a higher rate of infected cell loss (delta) (0.56+/-0.2 vs. 0.04+/-0.02, P=0.038). The viral load drop (baseline to treatment week 4) was higher in FR vs. SR group (3.5+/-1.1 vs. 1.4+/-0.6 log10IU/mL, P<0.001). T-cell reactivity to HCV increased only in FR (after the loss of viraemia), but not in SR patients. CONCLUSIONS: Assessment of early viral and T-cell kinetics during treatment reveals marked differences amongst HCV-G1 patients and may provide a basis for treatment individualization. Enhancement of antiviral T-cell reactivity requires rapid viraemia clearance, rather than immunostimulation alone.     

Antibody Responses to Hepatitis C Virus by Second-Generation Immunoassays in a Cohort of Patients with Bleeding Disorders

The antibody responses and the prevalence patterns of antibodies to hepatitis C virus (anti-HCV) in a cohort of patients (n = 210) with bleeding disorders were studied using a first-generation and a second-generation enzyme immunoassays (EIA-1, EIA-2) as well as a second-generation recombinant immunoblot assay (RIBA-2). The anti-HCV prevalence as determined by EIA-1 and EIA-2 was 183/210 (87.1%) and 197/210 (93.8%), respectively (p = 0.0026). None of the 17 EIA-2(+)/EIA-1(-) samples was scored nonreactive by RIBA-2. At follow-up, samples of 123 patients were tested. Twenty-nine out of 111 patients reactive by EIA-1 seroreverted according to EIA-1 while the seroreversion rate with EIA-2 was 0 out of the 121 (p < 10(-8)). The anti-HCV prevalence by EIA-2 was 150/154 (97.4%) in anti-HIV-1-positive individuals and 47/56 (83.9%) in the anti-HIV-1-negative ones (p = 0.001). However, high assay signals (OD 492 nm > 2.0) were observed in 94/150 (62.7%) and 45/47 (95.7%) of the anti-HIV-1-positive and -negative patients, respectively (p = 10(-5)). The decreasing anti-HCV reactivity among anti-HIV-1-positive individuals was mainly due to diminishing c33c reactivity. Seroconversion to anti-HCV was observed in 3/7 (42.9%) cases with acute icteric non-A, non-B hepatitis by both EIA-1 and EIA-2, while the remaining 4 cases had detectable levels of anti-HCV 1-18 months before the acute episode.