Recipient HLA-DR3, tumour necrosis factor-alpha promoter allele-2 (tumour necrosis factor-2) and cytomegalovirus infection are interrelated risk factors for chronic rejection of liver grafts.

BACKGROUND/AIMS: The tumour necrosis factor (TNF)-2 promoter allele, which elicits elevated expression of TNF-alpha, is in linkage disequilibrium with the extended haplotype HLA-A1-B8-DR3-DQ2. TNF-2 and HLA-DR3 have been implicated in renal and cardiac graft rejection and loss. Cytomegalovirus (CMV) infection has been associated with chronic allograft rejection. We examined the relationship between HLA-DR3, promoter allele TNF-2 and cytomegalovirus in relation to chronic rejection following liver transplantation. METHODS: (i) Retrospective analysis of HLA-DR3 was performed in 307 liver transplant recipients and 283 donors. (ii) Prospective analysis of TNF-alpha promoter allele status, HLA-DR3 status and cytomegalovirus infection was assessed in 123 recipients. RESULTS: (i) Retrospective analysis. Recipient HLA-DR3 (relative risk 1.9; 95% C.I. 1.01-3.58) was a risk factor for chronic rejection. (ii) Prospective analysis. Recipient HLA-DR3 was a risk factor for chronic rejection (relative risk 3.41; 95% C.I. 1.66-7.03) which was elevated further by superimposed CMV infection (relative risk 5.01; 95% C.I. 2-12.55). Recipient TNF-2 was associated with chronic rejection (relative risk 2.29; 95% C.I. 0.9-5.83) through linkage to HLA-DR3. CONCLUSIONS: Recipient HLA-DR3, TNF-2 status and CMV infection were inter-related risk factors for chronic rejection of liver grafts.

An association between cytomegalovirus infection and chronic rejection after liver transplantation.

BACKGROUND: Previous studies suggest a link between cytomegalovirus (CMV) infection and chronic rejection. Since these studies, more sophisticated diagnostic methods with high sensitivity and specificity for CMV have been developed and effective therapy/prophylaxis for CMV is now available. We sought CMV prospectively by polymerase chain reaction of serum and urine and by conventional methods in a group of 33 patients undergoing 57 transplants during 1993 or 1994, selected from a larger series. There were 13 grafts lost to chronic rejection. The remaining 44 grafts that did not develop chronic rejection served as controls and comprised 15 successful primary grafts, 15 second transplants, 8 third transplants, and 6 primary grafts that were lost for reasons other than chronic rejection. RESULTS: The combination donor CMV antibody negative with recipient antibody positive and the duration of CMV infection >30 days were associated with an increased relative risk of chronic rejection. In contrast, the presence of CMV infection alone, symptomatic CMV infection, the detection of CMV by PCR of serum or urine, and the peak/cumulative viral load were not predictive. CMV infection occurred earlier in those undergoing a second transplant for chronic rejection than for those undergoing a second transplant for other reasons. In addition, a human leukocyte antigen B mismatch was associated with prolonged CMV infection. CONCLUSION: These data are consistent with the hypothesis that prolonged subclinical cytomegalovirus infection is associated with an increased risk of chronic rejection.

Cytomegalovirus infection of bile duct epithelial cells, hepatic artery and portal venous endothelium in relation to chronic rejection of liver grafts.

BACKGROUND/AIM: Chronic rejection is an important cause of graft loss following liver transplantation. A number of risk factors for chronic rejection have been identified previously, albeit inconsistently. These include cytomegalovirus infection detected by a number of different techniques, including immunohistochemical staining and in situ hybridisation of liver grafts. However, tissue-based techniques for the detection of CMV have not been applied to grafts lost to conditions other than chronic rejection. The purpose of this study was to investigate the relationship between the presence of cytomegalovirus infection detected by in situ hybridisation and immunohistochemistry with respect to graft outcome, the presence of cytomegalovirus infection detected by other techniques and in addition, the type of infected cell. METHODS: The 29 patients studied included 15 patients who lost their primary liver graft to chronic rejection in 8 cases, to hepatic artery thrombosis in 4 cases and to causes other than chronic rejection or hepatic artery thrombosis in 3 further cases. In each case, sections containing septal or large ducts and vessels were selected (usually blocks) since these may be more representative. Needle biopsies from 14 further patients who ultimately achieved satisfactory graft function served as control tissue. Of these, ten had evidence of cytomegalovirus infection at the time of study by serum/urine PCR, DEAFF testing or seroconversion, while 4 patients had no evidence of cytomegalovirus infection according to these techniques. RESULTS: Cytomegalovirus infection was detected in the liver of 12 of the 29 patients. These included 8/15 grafts lost, which comprised 3/8 with chronic rejection, 2/3 with hepatic artery thrombosis and 3/4 with grafts lost to other causes, as well as 4/14 who retained grafts. CMV was detected most commonly in association with symptomatic infection and notably was detected only by in situ hybridisation in two cases. Predominant cell types that contained cytomegalovirus were hepatocytes and mononuclear cells. However, bile duct epithelial cells, hepatic artery endothelial cells and portal venous endothelial cells also contained cytomegalovirus in some cases. CONCLUSIONS: These data support previous studies that cytomegalovirus infection is detectable in patients with chronic rejection and are consistent with the theory that CMV is involved in chronic rejection. However, cytomegalovirus infection was detected in explanted grafts lost to conditions other than chronic rejection, and the association may not be causal but a consequence of graft injury.

A randomized trial of solvent/detergent-treated and standard fresh-frozen plasma in the coagulopathy of liver disease and liver transplantation.

BACKGROUND: Virus inactivation of pooled fresh-frozen plasma (FFP) by the solvent/detergent (SD) method results in a loss of approximately 20 percent of factor VIII. This study aimed to assess the efficacy of SD-treated plasma in correcting the coagulopathy associated with liver disease and liver transplantation. STUDY DESIGN AND METHODS: Forty-nine patients with coagulation deficits due to liver disease, who required FFP for invasive procedures or liver transplantation, were randomly assigned to receive either FFP or SD-treated plasma. Patients were assessed for side effects, correction of coagulopathy over 24 hours, and seroconversion for viral markers 6 to 18 months after treatment. RESULTS: In the liver disease group, equal correction of clotting factors and partial thromboplastin time was seen with FFP and SD-treated plasma, with a similar return to baseline values over 24 hours. There was greater correction of the International Normalised Ratio in patients receiving SD-treated plasma (p = 0.037), but this patient group had higher baseline values than recipients of FFP (p = 0.024). Liver transplant patients also showed equivalent correction of coagulopathy with the same dose of FFP and SD-treated plasma. The use of other blood components during transplantation was identical in the two treatment groups. No seroconversions were seen for HIV or hepatitis B or C virus. One patient who had received FFP seroconverted for human parvovirus B19. Apparent seroconversion for hepatitis A virus seen at 9 to 13 months in four other patients was probably due to detection of passively transferred antibodies, as later testing of these patients gave negative results. Minor side effects were rare in both groups. CONCLUSION: SD-treated plasma is an efficacious source of coagulation factors for patients with liver disease who are undergoing biopsy or transplantation. Assessment of seroconversion for viral markers in recipients of plasma-derived products and plasma components should include consideration of the possibility that passively transferred antibodies were detected.

Comparison of three PCR techniques for detecting cytomegalovirus (CMV) DNA in serum, detection of early antigen fluorescent foci and culture for the diagnosis of CMV infection.

Three PCR assays were developed for detection of cytomegalovirus (CMV) DNA in serum and were evaluated with samples from organ transplant recipients. The Qiamp Blood Kit was efficient for extraction of DNA from sera. Single-round PCR of a 293-bp region of CMV DNA was sensitive and highly specific for CMV targets and was more sensitive than detection of early antigen fluorescent foci (DEAFF) testing or isolation of CMV from buffy coat by cell culture. The results of a significant proportion of buffy coat samples were not interpretable because of toxicity in conventional culture or DEAFF tests. A non-competitive quantitative PCR test and semi-quantitative PCR test for the detection of CMV DNA in serum yielded comparable results for samples taken serially from three bone marrow transplant recipients. Single-round PCR was superior to conventional techniques for the diagnosis of CMV infection, was simple to perform and was completed rapidly. The semi-quantitative technique has added advantages where quantification is important.

Serological reactivity against cyst and tachyzoite antigens of Toxoplasma gondii determined by FAST-ELISA.

AIMS: To obtain quantitative data on the human serological response to Toxoplasma gondii tachyzoite and bradyzoite antigens. METHODS: Serum samples from 30 patients who had positive antibody titres against T gondii and from 14 who were seronegative, together with sequential serum samples from four infected individuals, were screened by FAST-ELISA. RESULTS: Serum samples from the 30 seropositive patients showed high IgG and IgM titres against the T gondii tachyzoite antigen but very low responses to cyst antigen. This result was borne out in sequential serum samples from patients with toxoplasmosis. CONCLUSION: Antibody recognition of the cystic stage of T gondii is low, implying that either this stage is poorly immunogenic or that the antigen load is low.

Epstein-Barr virus infection in heart and heart-lung transplant recipients: incidence and clinical impact.

BACKGROUND: A retrospective serologic study was made of 67 heart-lung and 295 heart transplant recipients (with transplantations at Papworth Hospital, Cambridge, England) to determine the incidence and clinical impact of Epstein-Barr virus infection. METHODS: Epstein-Barr virus capsid antigen immunofluorescence tests were performed, and the antibody avidity was determined by modifying the washing procedure to include a mild reducing agent (8M urea). RESULTS: This testing showed that 6.0% of the patients had primary Epstein-Barr virus infections, whereas 17.4% had the reactivation of a past infection. Primary infections were only detected in patients who were Epstein-Barr virus antibody-negative before transplantation, who had received an organ from an Epstein-Barr virus antibody-positive donor. Of the patients with serologically proven Epstein-Barr virus infections, 52.9% had symptoms. Although these were generally mild, five heart and two heart-lung transplant recipients had malignant lymphoma and one heart and one heart-lung transplant recipient had lymphoproliferative disease after Epstein-Barr virus infection. Additional four heart transplant recipients had lymphoma after transplantation. None of these four patients had evidence of active Epstein-Barr virus infection; one was Epstein-Barr virus antibody-negative during the study period and three had stable Epstein-Barr virus virus capsid antigen immunoglobulin G titers throughout. CONCLUSIONS: Epstein-Barr virus infection in organ transplant recipients may lead on to life-threatening lymphoproliferative disease or lymphoma. For this reason it may be beneficial to monitor patients after transplantation for evidence of Epstein-Barr virus infection and to follow the progress of those affected.

Testing for HIV antibody: a comparison of two services offered in a genitourinary medicine clinic.

Rates of screening for sexually transmissible infections in patients using different services provided by a genitourinary medicine clinic for testing for HIV antibodies are presented. Those patients whose primary reason for attending the clinic was HIV antibody testing and used the same day result (SDR) service were significantly less likely to be screened for other infections than those using the normal waiting time (NWT) service, (P < 0.00001). This was true for both males and females. Of those patients screened for other infections in the SDR and NWT groups 29% and 35% respectively were found to have a sexually transmitted infection. It would appear that an SDR service offers little benefit for the majority of patients as only a few patients would not have had an HIV antibody test had the SDR not been available.

Serum folate and chronic fatigue syndrome.

We assayed serum folate levels of 60 patients with chronic fatigue syndrome (CFS) and found that 50% had values below 3.0 micrograms/l. Some patients with CFS are deficient in folic acid.

A case of transfusion-acquired hepatitis C.

The Blood Transfusion Service introduced screening for Hepatitis C antibody (HCV) in September 1991. This is done by second generation enzyme linked immunosorbent assay (ELISA) tests. We present a case of post-transfusion hepatitis C hepatitis in a patient with myeloma. Infection was acquired before screening was introduced. Both the patient and the infected blood donor were diagnosed using ELISA assays and the polymerase chain reaction (PCR). In this way we prevented the blood donor from spreading the virus via subsequent blood donations. There were some interesting discrepancies in the HCV assays. Blood samples, when tested by different methods, gave both positive and negative results. The results also varied according to when the blood samples to be tested were taken. The case illustrates the importance of confirming positive results and that no single laboratory test is entirely satisfactory in diagnosing HCV infection.

Herpes simplex virus infection in heart-lung transplant recipients.

We report our experience of herpes simplex virus infection in a series of 51 recipients of heart lung transplantation (HLT). Nine patients, all of whom were seropositive for the virus preoperatively, developed HSV infection. Seven episodes of culture-proved mucocutaneous HSV infection without evidence of pulmonary involvement occurred in four patients. Six episodes of HSV pneumonia were seen in a further five patients, one of whom died. Diagnosis of HSV pneumonia was by histological appearances on transbronchial biopsy, together with culture of lung tissue or bronchoalveolar lavage. Concomitant cytomegalovirus infection occurred in four patients. All patients who developed HSV pneumonia did so within the first two postoperative months; in four patients following augmented immunosuppression. We now suggest that HLT recipients who are HSV antibody-positive should receive prophylactic acyclovir for the first two months after surgery and at times of augmented immunosuppression.

Viral and toxoplasma gondii infections in children after liver transplantation.

The incidence and morbidity of viral and Toxoplasma gondii infections were studied in 40 children who underwent liver transplantation between December 1983 and February 1988. The incidence of primary and reactivated cytomegalovirus (CMV) infection was 19% and 47%, respectively; primary infection caused clinical disease in all five cases affected and was fatal in one. Primary Epstein-Barr virus (EBV) infection occurred in 10 (26%) recipients but caused only mild disease. No reactivated EBV infection was recorded and no lymphoproliferative disorders associated with EBV were found after a maximum of four years' follow up. Adenovirus infection occurred in seven (18%) patients; this was associated in one case with fatal pneumonia and fulminant hepatitis, but otherwise with only mild respiratory disease. Primary T gondii infection was detected in one patient who remained asymptomatic. Other viruses causing infection included herpes simplex, varicella zoster, and respiratory syncytial virus. Surveillance for these infections and the long term sequelae should be included in the follow up of all children who undergo transplantation.

Antibiotic resistant fever associated with herpes simplex virus infection in neutropenic patients with haematological malignancy.

The incidence of mucocutaneous herpes simplex virus infection confirmed by culture and occurring during febrile neutropenic episodes was determined in 43 patients with haematological malignancy. The outcome of 72 episodes of neutropenic fever was determined and correlated with the presence or absence of herpes simplex virus (HSV) infection. Twenty four patients had mucocutaneous HSV infection during at least one episode. In 24 episodes in which HSV was isolated only 12.5% of fevers responded to antibiotics and 75% of fevers were otherwise unexplained. Conversely, in 48 episodes of neutropenic fever in which HSV was not isolated 67% of fevers responded to antibiotics and only 8.3% were unexplained. The difference in incidence of antibiotic resistant fever in the two groups was significant. There was, therefore, a strong association between mucocutaneous HSV infection and antibiotic resistant fever in immunosuppressed neutropenic patients. As most HSV infections are the result of virus reactivation, establishing the HSV serological state of patients would identify those at risk of infection and hence those in whom the prophylactic use of acyclovir would be indicated.

Susceptibility to acyclovir of herpes simplex virus: emergence of resistance in patients with lymphoid and myeloid neoplasia.

We have examined the susceptibility to acyclovir (ACV) of herpes simplex virus isolated from immunocompetent and immunocompromised patients. Susceptibility to ACV was determined in a dye-uptake assay with vero cells grown in 96-well microtitre plates. Resistant strains were found in two out of 35 (5.7%) patients and partially resistant strains in four out of 35 (11.4%) patients immunodeficient as a result of treatment for lymphoid or myeloid malignancy. Strains resistant to ACV were not found in organ transplant recipients or in immunocompetent patients.

Lack of association between cytomegalovirus infection of heart and rejection-like inflammation.

Serial myocardial biopsy specimens, taken up to the time of serological evidence of primary cytomegalovirus (CMV) infection in 22 heart transplant patients, were examined and compared with those taken over similar times after transplantation in 21 patients who did not develop CMV infection. None of these 43 patients had serological evidence of CMV infection before their heart transplantation. There was no evidence of an increased cellular infiltrate in the myocardium at the time of the active CMV infection, even though the donor heart is the likeliest source of infection, nor was there any change in myocyte, interstitial cell, or vascular endothelial cell nuclei to identify active CMV infection.

Cytomegalovirus infection in cardiac transplant recipients associated with chronic T cell subset ratio inversion with expansion of a Leu-7+ TS-C+ subset.

Lymphocyte subsets were analysed in 18 patients during the first 3 years after cardiac transplantation. The patients received Cyclosporin A and prednisolone for maintenance immunosuppression. Serological evidence of active cytomegalovirus (CMV) infection was found in 13 cases (72%), and in 12 of these an inversion usually of the T helper/T suppressor-cytotoxic ratio (TH/TS-C) was detected. T subset inversion usually preceded the diagnostic rise in CMV antibody titre. In 69% of patients with CMV the TH/TS-C ratio remained inverted throughout follow-up (245-951 days). Persistent T subset inversion was not found in all five patients who lacked serological evidence of active CMV. Chronic inversion consisted of an average increase in TS-C of 152% and an average decline in TH cells of 31% as compared to CMV negative patients. The proportion of lymphoid cells reacting with a phenotypic marker for natural killer (NK) cells (Leu-7) was increased by 83%. These alterations were also reflected in the absolute numbers of cells with these markers. Two-colour immunofluorescence analysis revealed that the expanded TS-C population present during chronic inversion was predominantly Leu-7+. As TS-C+ Leu-7+ cells in healthy persons may be hyporesponsive NK cells, a sustained increase in this cell type in allograft recipients could further reduce immunocompetence, thereby predisposing to superinfection or malignancy.