The influence of induction therapy on graft and patient survival in patients with and without hepatitis C after liver transplantation.

We used the United Network for Organ Sharing Database to determine the influence of antibody-based induction therapy on patient and graft survival in orthotopic liver transplant (OLT) recipients with and without hepatitis C (HCV). We identified all initial OLT patients with HCV serology. Patients were divided into four groups: HCV positive without induction (17 362), HCV positive with induction (3479), HCV negative without induction (20 417) and HCV negative with induction (4357). Both HCV positive and negative patients who received induction did better than those who did not. For HCV positive patients, 5-year patient survival was 70.8% versus 68.7% (p = 0.004) and graft survival was 65.2% versus 62.1% (p < 0.001). For HCV negative patients, 5-year patient survival was 78.8% versus 76.7% (p < 0.001) and graft survival was 74.0% versus 70.8% (p < 0.001). On multivariate analysis, induction was associated with improved patient (HR = 0.91: p = 0.024) and graft (HR = 0.88: p < 0.001) survival in HCV positive patients and improved patient (HR = 0.87: p = 0.003) and graft survival (HR = 0.87: p < 0.001) in HCV negative patients. The benefit of induction occurred early and largely dissipated when patients with death within a year were censored. The benefit of induction therapy appeared most pronounced in patients with renal insufficiency or on organ-perfusion support at transplant.

Human T cell leukemia virus-I-associated T-suppressor cell inhibition of erythropoiesis in a patient with pure red cell aplasia and chronic T gamma-lymphoproliferative disease.

Human retroviruses have recently been linked with T cell lymphoproliferative disorders and with the acquired immune deficiency syndrome. We investigated the mechanisms for acquired pure red cell aplasia and cutaneous anergy in a patient with the chronic T gamma-lymphoproliferative disease (T gamma-LPD) syndrome. Patient marrow erythroid progenitors (BFU-E) were 17 +/- 9% of control and were selectively increased to 88-102% of control after marrow T cell depletion. Patient Leu 2+ suppressor T cells spontaneously produced high titers of human gamma-interferon and resulted in a concentration-dependent selective inhibition (74-91%) of BFU-E when co-cultured with autologous or allogeneic marrow. Conditioned media (CM) derived from patient Leu 2+ T cells similarly inhibited growth of autologous or allogeneic marrow BFU-E. The inhibitory factor derived from patient CM was acid-labile (pH 2) and sensitive to trypsin; prior treatment of patient T cells with anti-HLA-DR monoclonal antibody plus complement abrogated the suppressive effect of T cell-derived CM. Patient peripheral blood mononuclear cells (PBMC) were unable to support growth of cultured interleukin 2 (IL 2)-dependent T cells, but responded to exogenous IL 2 in vitro with a 16-21-fold augmentation, relative to control, in mitogen-induced proliferation. Antibodies to HTLV-I core proteins p19 and p24 but not to HTLV-III proteins were detected in patient serum by Western blotting; patient cultured PBMC stained (7-11%) with antibodies to p19 and p24. Patient cultured PBMC demonstrated integrated HTLV-I genomic sequences by the Southern technique and expressed both specific HTLV-I genomic sequences by RNA dot blot plus reverse transcriptase activity. Utilizing a cloned DNA probe for the beta chain of the T cell receptor gene, patient PMBC demonstrated gene rearrangements providing presumptive evidence for clonality. The presence in serum of HTLV-I p19 and p24 antibodies, the expression of p19 and p24 core antigens on patient mononuclear cells, the evidence of HTLV-I proviral integration sequences and the expression of HTLV-I genomic sequences in patient cells, indicates infection with HTLV-I and raises the possibility of an etiologic link between human retrovirus infection and some instances of large granular lymphocytic leukemia (T gamma-LPD).

Chronic occlusion of the monkey fallopian tube with silicone polymer.

Investigators have previously reported good retention, antifertility efficacy, and oviduct polymer compatibility in rabbit oviducts in studies of both short-term and chronic occlusion with high viscosity silicone polymer. The present study attempts to determine the chronic effects of the silicone polymer installed into the oviducts of 10 rhesus monkeys. Unlike former studies with rabbits, the polymer was installed into the fimbrial end of the monkey's oviducts because of the complexity of the uterotubal junction's musculature. Studies with light and scanning electron microscopy were conducted to examine tissue reaction to plug retention. Despite our major modification of the installation technique, we obtained a retention rate of 50%. In addition, there was no evidence of inflammation or tissue distortion, and the polymer plugs were not affected by menstrual bleeding.

The effect of selected amino acids on gelatin-induced inflammation in adult male mice.

Several studies have indicated to us that certain amino acids may exhibit antiinflammatory activity. In the present study we attempted to evaluate the inhibitory effect of various amino acids on gelatin-induced abdominal inflammation in mice using peritoneal fluid cytology as the diagnostic tool. The L-amino acids tested were tryptophan, phenylalanine, alanine, cystine, hydroxyproline, tyrosine, citrulline, leucine, and valine. Hydrocortisone was used as an antiphlogistic steroid control. Tryptophan, phenylalanine, alanine, cystine, hydroxyproline, and tyrosine all significantly decreased the inflammation. Citrulline and valine, however, both exhibited strong antiiflammatory responses. Based on these results, three related dipeptides were also screened: L-valy-L-alanine, L-valyl-L-tryptophan, and L-tyrosl-L-valine. Valyl alanine was found to produce a strong antiinflammatory effect. In a final test, the combination of the steroid, hydrocortisone, and the amino acid, cystine, was screened for a synergistic effect. The combined treatment inhibited the gelatin-induced inflammation more than either the amino acid or the steroid administered alone.

Survival after liver transplantation in patients who develop renal insufficiency.

BACKGROUND: Renal insufficiency (RI) after liver transplantation (OLT) is associated with worse outcomes but the actual survival after RI ensues is not well described. We examined the survival of OLT recipients who developed moderate or severe RI or end-stage renal disease (ESRD), seeking to identify variables associated with these outcomes. METHODS: Between 1993 and 2007, 731 patients underwent OLT. After excluding patients undergoing retransplantation, combined kidney-liver grafts, and those who died within 1 year, we had a cohort of 527 subjects whose basic demographic data were obtained. Glomerular filtration rate (GFR) calculated (by MDRD4-Modification of Diet in Renal Disease 4-formula) at 3-month intervals in the first year and then at 6-month intervals. Moderate RI was defined as a GFR < 60 mL/min/1.73 m(2); severe RI, GFR < 30; and ESRD by need for dialysis or renal transplantation. We determined survival from the point of developing RI. An analysis determined factors associated with survival. RESULTS: Among 527 patients, 251 developed moderate (47.6%) and 40 (7.6%) severe RI as well as 40 (7.6%) with ESRD. Once RI ensued, the 5-year survivals for patients with moderate RI, severe RI or ESRD were 84.0%, 67.7%, and 48.5%, respectively. Five-year survival, for patients receiving a renal transplant was 100%. On multivariate Cox regression analysis, the only variables associated with time to death for patients with any RI were higher age at transplant (hazard ratio [HR] = 1.04, P = .02), higher creatinine at transplant (HR = 1.25, P = .01), pretransplant diabetes (HR = 2.34, P = .008), and transplantation in the Model for End-stage Liver Disease (MELD) era (HR = 0.15, P = .002). CONCLUSION: Development of severe RI or ESRD correlated with diminished survival. For patients with RI, age and creatinine at transplant, pretransplant diabetes, and transplantation in the pre-MELD era were associated with lower survival rates. Five-year survival for dialysis patients was somewhat higher than that previously reported but worse than that of subjects treated by renal transplantation.

Liver transplantation.

Survival after liver transplantation has steadily improved, in part because of newer immunosuppression, which may offer decreased long-term side effects. Reduction of steroids early in the course of transplant continues to be a goal, with satisfactory results in terms of both risk of rejection and reduction of side effects. Dominating the literature and the press in 1999 was the controversy surrounding the way in which livers are allocated. Regulation by the federal government was proposed to change the way the United Network of Organ Sharing distributes and allocates livers. Prompted by the shortage of organs, living-donor liver transplantation has blossomed. Continued experience in pediatric patients has shown excellent survival rate and quality of life. In adults, further experience is being gained with respect to the use of right lobes for transplantation. Early data suggest that this is a potential alternative to cadaveric transplantation in adults, with acceptable risk to the donor. Despite advances made in improving the technical aspects of transplantation, recurrent disease remains a significant issue. Lamivudine appears to be a potent inhibitor of hepatitis B virus DNA replication after liver transplantation, although resistance remains a significant problem. Further review of transplantation for hepatitis C virus is encouraging, with excellent five-year survival rate. However, studies evaluating the evolution of fibrosis in these patients throw caution on those results, showing increased progression to cirrhosis over time. Further follow-up of these patients is needed to more accurately assess long-term impact of hepatitis C on morbidity and mortality rates after liver transplantation.

Liver transplantation.

The growing disparity between available organs for liver transplantation and the number of waiting recipients has prompted significant debate over organ allocation and distribution. In light of this debate, recipient selection and prediction of factors relating to outcome have become increasingly important. Current immunosuppressive regimens provide excellent short-and long-term survival for patients and grafts. Increasingly, efforts are being made to decrease or withdraw immunosuppression late after transplantation to minimize long-term side effects. Viral disease, particularly cytomegalovirus infection, results in significant morbidity and mortality in patients. However, strategies for targeting high-risk patients with prophylactic antiviral therapy have been successful in reducing the incidence of cytomegalovirus disease. Recurrent viral hepatitis following liver transplantation may limit long-term graft success. Lamivudine appears to limit recurrent infection with hepatitis B virus in a significant number of patients who develop this condition following liver transplantation and may represent a cost savings over hepatitis B immunoglobulin. Although the overall survival of patients with chronic hepatitis C virus infection after orthotopic liver transplantation is excellent, significant morbidity and mortality occur in the subset of patients with severe recurrent disease. Interferon may delay the onset of disease in patients infected with hepatitis C virus following orthotopic liver transplantation, and investigation continues into antiviral therapy in this group of patients.

Liver transplantation.

Continued discussion over organ allocation and distribution remained a focal point in the field of liver transplantation in the year 2000. Despite the ongoing debate, no significant changes were implemented in the current allocation system. By far, the most widely discussed topic in liver transplantation this year was live donor adult-to-adult liver transplantation. Several authors reported on their initial experience, with both recipient and donor outcomes appearing excellent. As the number of transplant centers performing this procedure increases we look forward to further studies regarding the safety and long-term outcome of this innovative procedure. Studies on viral hepatitis after liver transplantation again focused on the problem of recurrent hepatitis B and hepatitis C. Several small studies found benefit in patients with hepatitis B treated with intramuscular hepatitis B immunoglobulin and lamivudine after transplantation. Although breakthrough replication remains a problem in some patients, these studies offer hope that combination therapy for hepatitis B may provide improved long-term graft survival in these patients. In patients with hepatitis C, several studies focused on identifying risk factors to predict graft recurrence of the virus after liver transplantation. Both cellular rejection and level of viral replication may be important predictors of recurrent hepatitis C virus in the graft. Early treatment reports using interferon and ribavirin suggest that some patients may have a viral response during therapy; however, it is short lived, and tolerance of medication is difficult. Certainly, we look forward to further studies looking at means of prevention and treatment of viral hepatitis in patients undergoing liver transplantation.

A human alloreactive inducer T cell clone that selectively activates antigen-specific suppressor T cells.

We showed previously that T cells with the phenotype Leu-3+,8+ are required for the induction of antigen-specific Leu-2+ suppressor cells. Furthermore, when mixed lymphocyte reactions are carried out in the presence of 1 microgram/ml cyclosporin A (CsA), such cultures lead preferentially to the activation of alloantigen-specific suppressor-inducer Leu-3+,8+ cells. In an attempt to generate a clone of T cells with such specific suppressor-inducer properties, we activated Leu-3+,8+ T cells with allogeneic (HLA-DR4+) lymphocytes in the presence of CsA. Clone SP-21, derived by propagating such activated T cells with conditioned medium containing IL 2, is a noncytotoxic, nonsuppressor clone that specifically proliferates to allogeneic cells bearing HLA-DR4 antigen. When cultured with fresh autologous Leu-2+ cells in the absence of HLA-DR4+ cells, clone SP-21 selectively activates Leu-2+ suppressor cells, which inhibit the response of fresh Leu-3+ cells to DR4+ stimulator cells. On the other hand, clone SP-21 fails to induce cytolytic T cells or to help B cell differentiation. These results demonstrate that a T cell clone with a remarkably narrow functional repertoire nonetheless contains and transmits all of the signals necessary for the activation of antigen-specific suppressor cells.

Quantitative assessment of hepatitis C virus RNA in peripheral blood mononuclear cells during therapy with interferon-alpha2a.

A significant number of patients with hepatitis C (HCV) treated with interferon (IFN) will initially clear their serum of HCV RNA, but will then have recurrence of viraemia either during or after therapy. One proposed mechanism for relapse is that HCV may persist in peripheral blood mononuclear cells (PBMCs) and that the PBMCs serve as a 'viral reservoir' that is resistant to IFN. To address this hypothesis, we performed serial, quantitative polymerase chain reaction (PCR) of HCV RNA in serum and PBMCs from 26 consecutive patients treated with IFN-alpha2a. Of the 26 patients, 11 (42%) did not clear virus from their serum during therapy and were termed non-responders. Five patients (19%) had sustained clearance of virus from serum and were termed complete responders. The remaining 10 patients (39%) initially eliminated HCV RNA from their serum, but had relapse of viraemia. They were termed partial responders. In all 10 partial responders HCV RNA was undetectable in PBMCs at the same time that it was undetectable in serum. When virus recurred in serum, it was preceded by or occurred at the same time as the return of virus in PBMCs. The results of our study indicate that PBMCs did not serve as an IFN-resistant 'viral reservoir' during therapy. Partial responders who transiently cleared virus from serum also cleared virus from PBMCs and the presence or titre of HCV RNA in PBMCs at the initiation of therapy did not predict response to therapy.

Extracorporeal photopheresis alone and with interferon-alpha2a in chronic hepatitis C patients who failed previous interferon therapy.

Extracorporeal photopheresis (ECP) is approved for treatment of cutaneous, T-cell lymphoma. Evidence suggests that ECP can induce an immune response against tumor antigens expressed by malignant T lymphocytes. We theorized that if HCV-infected PBMCs express viral antigens, ECP could demonstrate antiviral activity by eliciting an immune response against these antigens. Fifteen cirrhotic patients with genotype-1 HCV, who had previously relapsed or not responded to interferon-alpha (IFN-alpha) therapy were stratified by their HCV RNA titer into one of three treatment groups: (1) ECP alone, (2) ECP + 3 MIU IFN-alpha2a subcutaneously three times a week and (3) ECP + 6 MIU IFN-alpha2a subcutaneously three times a week. All patients received treatment for 24 weeks. Group 1 had no significant decrease in HCV RNA. Two patients in group 2 had undetectable HCV RNA at the end of treatment. One patient in group 3 had undetectable HCV RNA at the end of treatment. However, HCV RNA was detected in all three patients during follow-up. ECP alone or with IFN-alpha was well tolerated. ECP alone demonstrated no clear antiviral activity. The combination of ECP and IFN-alpha resulted in an end-of-treatment response (ETR) in three of 10 patients. All responders had elimination of serum HCV RNA by three months, although no patient had a sustained response. More intensive therapy for a longer duration may result in sustained responses. A multicenter trial is now underway.

Chronic occlusion of the rabbit Fallopian tube with silicone polymer.

Investigators have previously reported good retention, antifertility efficacy, and good retrievability with no adverse tissue reactions in rabbit oviducts occluded with silicone rubber for 55 days. The present study attempts to determine the long-term effects of this silicone polymer system installed into the oviducts of 12 New Zealand White rabbits for 7 months. 9 rabbits had both oviducts occluded while 3 had only one oviduct treated. All animals were visualized by X-rays and were mated an average of six times over the experimental period. Oviduct tissue was examined with light and scanning electron microscopy. There was no evidence of inflammation, distortion, or tissue irritation. Good retention and antifertility efficacy were exhibited.

A pilot trial of recombinant interleukin-12 in patients with chronic hepatitis C who previously failed treatment with interferon-alpha.

OBJECTIVE: Interleukin-12 is a cytokine with a multitude of immunomodulatory actions. Currently, interferon-alpha (IFN-alpha) monotherapy and combination treatment with IFN and ribavirin are the only therapies with proven efficacy against chronic hepatitis C infection. The purpose of this study was to assess the safety and antiviral activity of recombinant interleukin-12 (rhIL-12) in adults with chronic hepatitis C who did not achieve a sustained response to previous IFN-alpha therapy. METHODS: This was a randomized, placebo-controlled, double-blind trial. We randomized 24 patients to one of three dose groups: 30 ng/kg, 100 ng/kg, and 300 ng/kg. Within each group, six patients received rhIL-12, and two patients received placebo administered s.c. twice a week for 12 wk. RESULTS: Three of six patients treated with rhIL-12 at a dose of 300 ng/kg had loss of detectable hepatitis C RNA by reverse transcription-polymerase chain reaction compared with the placebo group (p = 0.05). All patients relapsed at the end of the 3-month treatment period. No other dose group demonstrated a loss of detectable hepatitis C RNA. CONCLUSIONS: RhIL-12 at 300 ng/kg can suppress hepatitis C RNA to undetectable levels by reverse transcription-polymerase chain reaction, although relapse occurred when treatment was stopped. RhIL-12 was well tolerated with the most common side effects being flu-like symptoms and headaches.

High levels of hepatitis C virus RNA in native livers correlate with the development of cholestatic hepatitis in liver allografts and a poor outcome.

A subset of hepatitis C virus (HCV)-positive liver transplant recipients develop cholestatic hepatitis (CH). We investigated the role of pretransplantation disease activity (estimated by Knodell score and HCV RNA quantitation) in the native liver explant on the development of CH and graft and patient outcome. Eight patients with CH were identified among HCV-positive liver transplants and were compared with 20 consecutive patients with recurrent HCV hepatitis of noncholestatic type in liver transplants. We evaluated all 28 explanted native livers histologically using the Knodell scoring system. HCV viral load was measured in the native explant and 5 allograft explants from the CH group using Amplicor HCV RNA Monitor test. Six of 8 patients with CH had HCV RNA levels of 5,000 copies/microg of DNA or greater in the native liver explant, whereas only 1 of the control group had viral loads greater than this level. Greater HCV RNA levels correlated with worse graft and patient survival (P <.001). The 3-year survival rate in the CH group was 18% compared with 77% in the control group (P <.001). There was no difference in the primary immunosuppressive regimens used in the 2 groups. We conclude that (1) CH has a uniformly poor prognosis, (2) type of immunosuppressive therapy appears to have little influence on the development of CH, (3) high pretransplantation HCV RNA levels in the native explant may predict the development of CH, and (4) patients with high HCV RNA levels in the explanted native liver may be appropriate candidates for antiviral therapy to prevent the development of CH.

Tissue quantification of hepatitis C virus RNA with morphologic correlation in the diagnosis of recurrent hepatitis C virus in human liver transplants.

BACKGROUND: Histologic findings and liver enzymes in liver transplants are often non-diagnostic of recurrent hepatitis C virus (HCV) disease. In addition, the relationship between HCV replication and the presence of recurrent HCV hepatitis after liver transplantation remains unclear. We studied liver transplant recipients to determine if quantitation of HCV RNA in liver tissue by reverse transcriptase-polymerase chain reaction (RT-PCR) correlates with histopathologic disease and/or liver enzymes. METHODS: Twenty-six patients who received liver transplants for HCV infection were evaluated. Four sequential biopsies were analyzed for each patient. HCV RNA was extracted and quantified using the Amplicor HCV Monitor Test. Histologic examination and RNA quantitation were blinded. All available liver enzymes on the day of liver biopsy were analyzed. RESULTS: HCV RNA quantity in liver tissue was significantly increased at the time of clinically-suspected recurrence (P < .0001). HCV RNA levels were highest in biopsies with lobular hepatitis and nonspecific inflammation, followed by biopsies with cytomegalovirus infection, chronic hepatitis, and acute cellular rejection. HCV RNA quantity had a significant correlation with increasing portal inflammation (P = .0002), decreasing amount of interface hepatitis (P = .0333), and presence of acidophilic bodies (P = .0316). Increasing HCV RNA levels significantly correlated with decreasing number of episodes of treated rejection. HCV RNA quantity did not correlate with other histologic features or liver enzymes. CONCLUSIONS: HCV RNA levels are highest at the time of active hepatocellular destruction. Elevated HCV RNA indicates recurrence. HCV RNA quantitation may be a useful diagnostic test for determining recurrent disease and distinguishing it from other causes of inflammation, such as rejection.

The use of hepatitis C viral RNA levels in liver tissue to distinguish rejection from recurrent hepatitis C.

Persistence of hepatitis C virus (HCV) after orthotopic liver transplantation is almost universal in HCV-infected patients. Histological examination of liver biopsy specimens can be variable in distinguishing between recurrent hepatitis C and acute cellular rejection. The purpose of this study is to determine whether hepatic HCV RNA levels can be used to distinguish rejection from recurrent HCV by determining whether hepatic HCV RNA levels correlate with histological characteristics and clinical course. Seventy-two biopsy specimens were evaluated from 36 liver transplant recipients with HCV and elevated liver-related enzyme levels. Based on histological findings and clinical response to therapy, patients were defined as belonging to 1 of 5 groups: (1) definite rejection, (2) probable rejection, (3) indeterminate findings, (4) probable HCV, and (5) definite HCV. Hepatic HCV RNA was quantified using the Amplicor Monitor assay (Roche Diagnostic Systems Inc, Branchburg, NJ). There was a difference across groups in HCV RNA levels (P =.046). The median HCV RNA level was 10,695 copies/mg of tissue DNA in the definite-HCV group compared with 1,024 copies/mg of tissue DNA in the definite-rejection group. Using pairwise comparisons, significant differences were found between definite HCV and definite rejection, probable HCV and definite rejection, probable HCV and probable rejection, and probable HCV and indeterminate. Our findings support the following conclusions. (1) In liver transplant recipients, hepatic HCV RNA levels are statistically greater in patients with recurrent HCV than rejection, although there is considerable overlap between groups. (2) Patients with low HCV RNA levels were unlikely to have recurrent HCV. (3) Patients with minimal and indeterminate findings on biopsy (group 3) had low HCV RNA levels.