Imbalance in X-chromosome expression: evidence for a human X-linked gene affecting growth of hemopoietic cells.

In each of six family members who were heterozygous at the X-linked locus for glucose-6-phosphate dehydrogenase, only one or the other of the two alleles at that locus was almost exclusively expressed. The data are consistent with evidence that X-chromosome inactivation is a random process that may be followed by selection for one of the two resulting cell types on the basis of an unknown gene, which is located on the X chromosome and which can affect the rate of proliferation of hemopoietic cells in humans.

Immunization of liver and renal transplant recipients: a seroepidemiological and sociodemographic survey.

Several life-threatening infections, a major risk to adult solid organ transplant (SOT) recipients on immunosuppressive therapy, can be prevented by immunization. We analyzed sociodemographic parameters and the immunization status of adult liver transplant recipients (LTX-R, n=267) and renal transplant recipients (RTX-R, n=197) SOT recipients at the Transplantation Center, Berlin, Germany. Date, number, and provider of recommended vaccines were recorded and seroprotection rates determined. The social status in both groups was similar. Most patients (89%) were not adequately informed about immunizations; and if informed, main sources were physicians (47%) and the media (40%). Vaccinations were predominantly provided by family doctors (LTX-R, 66%; RTX-R, 31%) or hemodialysis centers (RTX-R, 37%). Before transplantation, RTX-R had significantly more often received booster vaccinations against tetanus and diphtheria (P<0.005), and a primary hepatitis B immunization (55%); whereas in LTX-R, post-transplant vaccinations against hepatitis A (16%) and pneumococcal disease (13%) were more frequent. Seroprotection rates against tetanus were fairly high in LTX-R (85.3%) and RTX-R (86.8%), and considerably lower for diphtheria, hepatitis A, and influenza. Immunization rates are too low in SOT recipients. Improvement will depend on a more active role of health care providers.

A fast and reliable single-run method for genotyping of the human cytochrome P450 2C8 gene for different ethnic groups.

Patients vary widely in their response to drug therapy according to their genetic background of drug metabolizing enzymes. The cytochrome P450 enzyme CYP2C8 is one of the major metabolizing enzymes involved in drug metabolism and thus a candidate for routine pharmacogenetic screening. The aim of this work was establishing a fast and reliable method to detect the three CYP2C8 genotypes CYP2C8*2, CYP2C8*3, CYP2C8*4, and the wildtype allele. An established real-time polymerase chain reaction (PCR) to detect two CYP2C8 genotypes was extended by introduction of a third hybridization probe. After optimization of running conditions, the new triplex method was evaluated using 200 DNAs of African origin as templates. Standard methods were performed as controls. The new triplex real-time PCR was fast, reliable and reproducible. The obtained results showed no deviation from the results of the established technique. The polymorphism of the CYP2C8 gene among an African population showed the expected distribution (68% wildtype gene, 32% at least one CYP2C8*2 allele). Pharmacogenetics gain increasing interest in routine medical care to prevent severe adverse effects or the application of ineffective drugs. We here provide a fast, reliable and reproducible method in one single assay run to detect three relevant CYP2C8 alleles independent of the patient's ethnic origin.

Toll-like receptor (TLR) polymorphisms in African children: common TLR-4 variants predispose to severe malaria.

Genetic host factors play a substantial role in susceptibility to and severity of malaria, which continues to cause at least one million deaths per year. Recently, members of the toll-like receptor (TLR) family have been shown to be involved in recognition of the etiologic organism Plasmodium falciparum: The glycosylphosphatidylinisitol anchor induces signaling in host cells via TLR-2 and -4, while hemozoin-induced immune activation involves TLR-9. Binding of microbial ligands to the respective TLRs triggers the release of pro-inflammatory cytokines via the TLR/IL-1 receptor (TIR) domain and may contribute to the host response, including pro-inflammatory cytokine induction and malarial fever. In a case-control study among 870 Ghanaian children, we examined the influence of TLR-2, -4, and -9 polymorphisms in susceptibility to severe malaria. TLR-2 variants common in Caucasians and Asians were completely absent. However, we found a new, rare mutation (Leu658Pro), which impairs signaling via TLR-2. We failed to detect any polymorphisms within the TLR-9/interleukin-1 receptor domain. Two frequent TLR-9 promoter polymorphisms did not show a clear association with malaria severity. In contrast, the TLR-4-Asp299Gly variant occurred at a high rate of 17.6% in healthy controls, and was even more frequent in severe malaria patients (24.1%, p<0.05). Likewise, TLR-4-Thr399Ile was seen in 2.4% of healthy children and in 6.2% of patients (p=0.02). TLR-4-Asp299Gly and TLR-4-Thr399Ile conferred an 1.5- and 2.6-fold increased risk of severe malaria, respectively. These findings suggest TLR4-mediated responses to malaria in vivo and TLR-4 polymorphisms to be associated with disease manifestation. However some gray areas also suggest the scope for further improvements.

Imprisonment: a risk factor for HIV infection counteracting education and prevention programmes for intravenous drug users.

OBJECTIVES: To examine changes of risk behaviour and its determinants as well as risk factors for HIV infection in intravenous drug users (IVDU) with particular attention to imprisonment and its risk patterns. SETTING: In 1993 a multisite cross-sectional study was carried out by standardized questionnaires and blood/saliva samples in which 612 IVDU from Berlin were enrolled. RESULTS: Multifactorial analysis revealed that the most important risk factor for HIV infection was needle-sharing in prison. In total, 353 IVDU (58%) reported reduced risk behaviour; changes related more to injection behaviour than sexual practices (91 versus 68%). Important determinants for needle-sharing during the last 6 months were intravenous drug use in prison, duration of drug-taking history, and knowledge of a negative HIV test. The most frequently reported reasons for current needle-sharing were having shared needles with only one regular partner (45%) and imprisonment (26%). CONCLUSION: Information campaigns and other prevention measures appear to have produced risk awareness in IVDU, and as a consequence, a reduction in risk behaviour. The situation in prisons (no sterile injecting equipment, no effective disinfectants), however, is counteractive to prevention measures implemented outside prisons. An important task for future strategies should be to enable IVDU to avoid HIV transmission while in prisons.

Frontloading: a risk factor for HIV and hepatitis C virus infection among injecting drug users in Berlin.

OBJECTIVE: To determine whether frontloading (i.e., syringe-mediated drug-sharing) is a risk factor for HIV, hepatitis B virus (HBV) and hepatitis C virus (HCV) infection among injecting drug users (IDU). DESIGN: Cross-sectional study. Data on sociodemographic and behavioural characteristics were obtained by a standardized questionnaire. Serum samples were tested for seromarkers for HIV, HBV and HCV. SETTING AND PARTICIPANTS: IDU were recruited at 'low-threshold' storefront agencies (out-of-treatment sample), and at a centre for long-term drug use treatment (in-treatment sample). Individuals were included in the study if they had injected drugs within the previous 3 months. MAIN OUTCOME MEASURES: Serological evidence for HIV, HBV, HCV exposure. RESULTS: Of all IDU (n = 324), 84% had ever practised frontloading with non-sterile injecting equipment, and 46% had done so more than 100 times; 32% had front-loaded during the 6 months prior to the interview. The crude seroprevalence rates for HIV, HBV and HCV increased with the overall frequency of frontloading, and reached 22, 71 and 94%, respectively, among IDU who had frontloaded more than 100 times. After controlling for confounding effects by logistic regression, having practised frontloading more than 100 times was significantly associated with HIV infection [adjusted prevalence odds ratio (POR) 3.5; 95% confidence interval (CI), 1.4-9], and HCV infection (adjusted POR, 5.4; 95% CI, 2.3-12), but not with HBV infection. Another independent risk factor for all three virus infections was needle-sharing in prison. CONCLUSIONS: In communities where sterile injection equipment is readily available, and IDU have substantially reduced their overall levels of needle-sharing, the practice of frontloading appears to be a major risk factor for infections by blood-borne viruses among IDU. Prevention activities should specifically address this risk behaviour.

Routine immunizations in adult renal transplant recipients.

BACKGROUND: Vaccination guidelines for transplant recipients include regular boosters of tetanus, diphtheria, and inactivated polio vaccine, but there are few published data on the efficacy of these vaccines in patients receiving immunosuppressive therapy. METHODS: Serum antibody values were evaluated before and 4 weeks after tetanus, diphtheria, and inactivated polio vaccination in 164 renal transplant recipients compared with healthy controls. Twelve months later, antibody levels were evaluated in 55 patients. RESULTS: Prebooster tetanus antitoxin values were lower in transplant recipients than in controls. All patients developed protective tetanus antibody levels (> or = 0.01 IU/ml) after vaccination. After 12 months, serum antibodies had decreased, but all patients maintained protective values. Diphtheria antitoxin titers before and after booster vaccination were lower in patients than in controls: 88.5% of patients and 96.2% of controls developed protective diphtheria antibody values. Twelve months after vaccination, diphtheria antitoxin values were below the protective level (0.1 IU/ml) in 38% of patients. Prebooster antibody values to poliovirus types 1 and 3 were comparable in patients and controls, whereas antibodies to poliovirus type 2 were lower in transplant recipients. Seroprotection rates and geometric mean antibody titers after vaccination were equivalent between the two groups for all three poliovirus types. No difference was observed in antibody levels between patients on different immunosuppressive drug regimens. Adverse reactions were significantly less often reported by transplant recipients. CONCLUSIONS: In transplant recipients, tetanus and inactivated polio vaccinations are well tolerated and induce protective antibody levels; diphtheria vaccination as currently recommended is less effective and protective antitoxin values decrease rapidly in these patients within 1 year after vaccination.

Rapid decline of antibodies after hepatitis A immunization in liver and renal transplant recipients.

BACKGROUND: Hepatitis A vaccine is safe and achieves good seroconversion rates in liver (LTX) and renal (RTX) transplant recipients. METHODS: A study was performed to determine the anti-hepatitis A virus (HAV) antibody decline in LTX and RTX patients, and in healthy controls who have been immunized with two doses of hepatitis A vaccine. RESULTS: LTX and RTX patients had a satisfactory seroconversion rate after complete immunisation. However, 2 years later they had experienced a much more rapid antibody decline than controls, and only 59% of LTX and 26% of RTX seroconverters showed titres above the cut-off level defined as protective. CONCLUSIONS: Patients on immunosuppressive therapy may not be adequately protected against hepatitis A a few years after vaccination and alternative vaccination schemes may have to be considered.

Intravenous or intramuscular anti-HBs immunoglobulin for the prevention of hepatitis B reinfection after orthotopic liver transplantation.

To prevent reinfection with hepatitis B virus after orthotopic liver transplantation, patients receive long-term intravenous anti-HBs immunoprophylaxis. We compared the pharmacokinetics of intravenously and intramuscularly administered commercially available hepatitis B virus immunoglobulins. The study group consisted of 12 patients on immunoprophylaxis after orthotopic liver transplantation, who were Hbs antigen negative; 11 were anti-HBe positive and one was HBe positive. The patients first received intravenous immunoglobulin, and six of them were then transferred to intramuscular immunoglobulin. Our findings show that with fortnightly intramuscular application of 1000 IU of anti-HBs, reproducible and stable antibody titers above 100 IU of anti-HBs can be achieved. Side effects of intramuscular immunoprophylaxis are minimal and the method is safe. The switch from intravenous (1500 IU of anti-HBs) to intramuscular (1000 IU of anti-HBs) reduced the cost of immunoprophylaxis by more than 50%.

Influenza vaccination in liver transplant recipients.

BACKGROUND: The immunogenicity of the trivalent inactivated influenza split virus vaccine (Infusplit SSW 97/98) containing A/Bayern/07/95 (H1N1)-like (A/Johannesburg/82/96 [NIB-39]), A/Wuhan/359/95 (H3N2)-like (A/Nanchang/933/95 [Resvir-0]), and B/Beijing/184/93-like (B/Harbin/7/94) hemagglutinin antigens was tested in liver transplant recipients (TXL-R). SUBJECTS AND METHODS: Serum antibody titers were determined 21+/-2 days after a single vaccination in 62 adult TXL-R and 59 adult volunteers. RESULTS: Protective postimmunization antibody titers for the three antigens were similar in TXL-R (protection rates 92%, 92%, and 95%) and the comparison group (97%, 100%, and 100%). Adverse reactions were mild and less frequent in TXL-R. A significant decrease of CD8+CD38+ lymphocytes after vaccination was found in TXL-R. No association between antibody response and age, gender, time interval since transplantation, anti-hepatitis B surface antigen immunoprophylaxis, or immunosuppressive medication was detected. CONCLUSION: Our results show that the vaccine is safe and effective and should be recommended to TXL-R.

Hepatitis G virus RNA and hepatitis G virus antibodies in renal transplant recipients: prevalence and risk factors.

BACKGROUND: Hepatitis G virus (HGV/GBV-C) RNA indicating current infection has been frequently isolated from the sera of transplant recipients and other multitransfused individuals. Lifetime exposure to the virus, however, is unknown. We carried out a study to determine the prevalence and risk factors of HGV antibodies and of HGV RNA among renal transplant recipients, and to investigate possible associations between HGV RNA and immunosuppressive treatment. METHODS: HGV RNA was detected by reverse transcriptase-polymerase chain reaction, and HGV antibodies (anti-E2) by a newly developed immunoassay. To assess risk factors for HGV exposure, univariate and multivariate analysis was performed. RESULTS: Of the 221 patients, 14% were HGV RNA positive and 40% had HGV antibodies. Both HGV RNA and anti-HGV were present in only two individuals. Thus, the overall HGV exposure prevalence was 53%. It increased significantly with the number of blood transfusions. In logistic regression, the adjusted HGV exposure prevalence odds ratio was 5.7 (95% confidence interval [CI]: 2.2-15) among patients with > or =10 transfusions (baseline: no transfusions). Other independent risk factors were a longer duration of hemodialysis and a longer time interval since transplantation. HGV viremia was not associated with the type of immunosuppressive treatment. Alanine aminotransferase levels were not significantly increased among HGV RNA-positive patients. CONCLUSIONS: Much higher proportions of renal transplant recipients were exposed to HGV than is suggested by HGV RNA detection alone. The majority of infected individuals apparently eliminate the virus over time. Contaminated blood transfusions have to be regarded as a main risk factor for HGV infection.

Neutralizing antibodies and the course of HIV-induced disease.

The capacity to neutralize the human immunodeficiency virus (HIV) in vitro was examined in 52 sera obtained from 23 seropositive individuals in addition to 7 negative control sera. Neutralization was measured as the activity of a serum to protect MT-4 cells against the cytopathic effect of HTLV-IIIB. Virus neutralization depended on HIV antibodies. Some sera had HIV neutralizing antibody titers of several thousands. All serum samples had been titrated in two ELISAs based either on disrupted HTLV-IIIB or on a bacterially synthesized polypeptide (ENV-80) of gp41 as a test antigen. The correlation of neutralizing activity of the sera with ELISA titers was low. A correlation of serum neutralizing titers with the stage of the disease could not be observed. However, in a longitudinal study with 6 patients over up to 22 months an increase in neutralizing antibodies seemed to protect against progression of the disease. The implications of these findings for antibody treatment and vaccine development are discussed.

T cell phenotype alterations in hepatosplenic schistosomiasis mansoni normalize after chemotherapy.

Peripheral blood T cell phenotypes, CD3-induced mitogenesis and soluble IL 2 receptor and CD8 in sera were studied in intestinal and hepatosplenic Schistosomiasis mansoni before and three to six months after therapy with praziquantel. Fifteen pairs matched for intensity of infection were analyzed and compared with local, non-infected age-matched controls. CD3+ cell counts were lower in untreated hepatosplenic schistosomiasis (median 1040 cells/microliters; 95% confidence interval 608-1269) compared to controls (1534; 1264-1620). This difference was largely accounted for by immature CD1+/CD3-cells circulating in these patients (median 388/microliters, 252-474). The frequency of CD1+ T cells in circulation decreased drastically after chemotherapy. Similar, but less marked, alterations were seen in intestinal schistosomiasis. Lymphocyte proliferation initiated by agonistic anti-CD3 monoclonal antibody was severely impaired in hepatosplenic patients, who had suffered haemorrhagic complications, but not in the cases of incipient hepatomegaly. Soluble CD8 antigen circulated in increased amounts in hepatosplenic schistosomiasis. Remarkably, a negative correlation between CD3-induced mitogenesis and circulating levels of CD8 was noted in these patients. Whereas CD3-induced mitogenesis in hepatosplenic schistosomiasis normalized after therapy, circulating IL 2R and CD8 antigen in hepatosplenic patients still exceeded control levels. The results demonstrate disturbances of CD3 and CD8 expression and/or T cell maturation in hepatosplenic schistosomiasis. Imbalanced CD4/CD8 ratios and an increased IL 2R/CD8 turnover may reflect an inhibitory circuit within the T cell compartment.

Plasmodium falciparum malaria and human red cells. II. Red cell genetic traits and resistance against malaria.

In 702 Nigerian children under 6 years of age the incidence and the severity of malarial infection was studied with respect to haemoglobin types and red cell glucose-6-phosphate dehydrogenase variants. The results suggest that Hb AS as well as the female genotype GdA-/GdB offer selective advantage against the disease. Parasite densities in carriers of these genotypes were significantly lower than in other subjects. Whereas protection by Hb AS was found mainly in children between 2 and 4 years of age, the advantage afforded by GdA-/GdB was similar in all age groups. Possible mechanisms are discussed.

Plasmodium falciparum malaria and human red cells. I. A genetic and clinical study in children.

In a holoendemic malaria region of Nigeria children of both sexes aged from 9 months to 6 years with an acute febrile illness were examined for malaria. In 461 children with malaria, predominantly P. falciparum, and in 241 children without malaria, haemoglobin levels, haemoglobin types, red cell glucose-6-phosphate dehydrogenase variants and parasite densities were recorded. The G6PD status was determined by a combination of spectrophotometric enzyme quantitation, electrophoresis and the cytochemical methaemoglobin elution technique. Malaria morbidity and parasitaemia decreased with increasing age. Frequencies of haemoglobin types and G6PD variants were not significantly different in the malaria and the non-malaria series. Haemoglobin values were significantly lower in children with malaria to about the same extent in HbAA and HbAS subjects, but no close correlation existed between haemoglobin level and parasite density. Details of the G6PD classification and the effect of malaria on enzyme activity are discussed.

History of syringe sharing in prison and risk of hepatitis B virus, hepatitis C virus, and human immunodeficiency virus infection among injecting drug users in Berlin.

BACKGROUND: Injecting drug users (IDU) are at risk of parenterally transmitted diseases such as hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) infection. We investigated whether a history of syringe sharing in prison is a risk factor for these infections. In the longitudinal part of the study, HBV, HCV, and HIV seroincidence rates were determined. METHODS: The participants were recruited by multisite-sampling at different agencies for IDU. Data on risk behaviour were obtained by a standardized questionnaire. Serological markers for HBV, HCV, and HIV were determined. Logistic regression analysis was performed to adjust for confounding effects. RESULTS: A history of syringe sharing in prison was significantly associated with HBV (adjusted prevalence odds ratio [POR] = 3.9, 95% confidence interval [CI]: 2-10), HCV (POR = 9.7, 95% CI: 3-33), and HIV infection (POR = 10.4, 95% CI: 4-29). The HIV seroincidence rate was 5.9 per 100 person-years. None of the IDU receiving methadone maintenance treatment (MMT) seroconverted whereas the HIV incidence was 8.5 among IDU not in MMT (P = 0.01). CONCLUSIONS: The increased risk of HBV, HCV, and HIV infection among IDU who had shared syringes in prison warrants specific preventive action. The longitudinal data suggest that IDU in MMT have a lower risk of HIV infection.

Malaria and erythrocyte glucose-6-phosphate dehydrogenase variants in West Africa.

In a clinical study, 702 Nigerian children aged 1-6 years were examined for malaria. Comparison of morbidity rates and parasitemia of patients with different glucose-6-phosphate dehydrogenase (G6PD) status provided evidence that in heterozygous females the gene for G6PD deficiency (GdA-/GdB) confers an advantage against malaria.

Curing of chloroquine-resistant malaria with clindamycin.

A randomized comparative trial for treating adult patients with Plasmodium falciparum malaria was performed in Lambarene, Gabon. Forty-two patients received chloroquine (25 mg/kg for 48 hr) and 38 patients received clindamycin (5 mg/kg twice a day, for five days). Chloroquine treatment cured 15 patients (36%). Twenty patients (48%) showed recrudescent malaria by day 28 of follow-up (RI resistance) and seven patients (17%) showed persistent parasitemia after chloroquine treatment (RII/III resistance). In contrast, clindamycin treatment cured 37 of 38 patients (97%) and only one (3%) showed a recrudescence by day 28 (P < 0.001). Although the parasite clearance time was significantly longer after clindamycin treatment (median five days, range 3-6) than after chloroquine treatment (median four days, range 2-8) (P < 0.01), no differences were seen in the duration of symptoms after chemotherapy. In both treatment groups, no severe side effects occurred. Clindamycin can be used as a safe alternative to achieve radical cure in semi-immune adult patients with chloroquine-resistant P. falciparum malaria in Central Africa.

Comparison of micronized halofantrine with chloroquine-antibiotic combinations for treating Plasmodium falciparum malaria in adults from Gabon.

Multidrug resistance of Plasmodium falciparum is spreading throughout Africa. In Lambarene, Gabon where chloroquine-resistant malaria is prevalent, a randomized comparative trial with three regimens for treating P. falciparum malaria in adults was performed. One hundred two patients evaluated received either a new micronized formulation of halofantrine (8 mg/kg every 6 hr in three doses) (group H) or chloroquine (25 mg/kg for a 48-hr period) plus clindamycin (5 mg/kg every 12 hr in six doses) (group CC1), or chloroquine (as above) plus doxycycline (2 mg/kg every 12 hr in six doses) (group CD). All treatment regimens were well-tolerated. In group H, 100% of the patients were cured, and in group CC1, 97% of the patients were cured by day 28 of follow-up. In group CD, a significantly lower cure rate of 75% (P < 0.01) and a slower parasite clearance was observed, but only low grade (RI) resistance occurred.

Effects of Plasmodium vinckei hemozoin on the production of oxygen radicals and nitrogen oxides in murine macrophages.

When murine peritoneal macrophages were loaded in vitro with Plasmodium vinckei hemozoin and stimulated with opsonized zymosan for 90 min or with lipopolysaccharide and/or murine interferon-gamma for 24 hr, significant decreases in the production of oxygen radicals and nitrogen oxides, respectively, could be detected by comparison with macrophages without hemozoin. Moreover, nonradioactive in situ hybridization and immunohistologic analysis in liver sections of P. vinckei-infected mice with more than 60% parasitemia showed that liver cells were still expressing considerable levels of inducible nitric oxide synthase in the late phase of murine malaria, but most of the liver macrophages presenting accumulation of malaria pigment were negative in this analysis. These results further indicate that malaria pigment accumulation may be responsible for toxicity and impairment of macrophage functions during murine malaria.