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Biomedical subjects

J P Allain

Publications and source records attributed to J P Allain.

At least 19 recordsLinked to original sources
PubMed2001

The dynamics of hepatitis C virus binding to platelets and 2 mononuclear cell lines.

Hepatitis C virus (HCV) binds to platelets in chronically infected patients where free HCV constitutes only about 5% of total circulating virus. Free HCV preferentially binds to human mononuclear cell lines but free and complexed virus binds equally to platelets. The extent of free HCV binding to human Molt-4 T cells (which express CD81) and to human promonocytic U937 cells or to platelets (which do not express CD81) was similar. The binding of free HCV to the cell lines was saturated at a virus dose of 1 IU HCV RNA per cell but binding to platelets was not saturable. Human anti-HCV IgG, but not anti-CD81, markedly inhibited HCV binding to target cells in a dose-dependent manner. Human antibodies to HCV hypervariable region 1 of E2 glycoprotein partially inhibited viral binding to target cells. Recombinant E2 also inhibited viral binding to target cells in a dose-dependent manner, with the efficacy of this decreasing in the rank order of Molt-4 cells more than U937 cells more than platelets. In contrast to HCV, recombinant E2 bound to Molt-4 cells to an extent markedly greater than that apparent with U937 cells or platelets. These results suggest that the binding of HCV to blood cells is mediated by multiple cell surface receptors and that recombinant E2 binding may not be representative of the interaction of the intact virus with target cells.

Antibodies, Viral↗
PubMed2001

Emerging viruses and transfusion.

The development of new technologies leads to the discovery of new viruses. For each of these new infectious agents, their possible relevance to blood transfusion needs to be assessed. The questions to be answered are transmissibility by transfusion, pathogenicity, prevalence in blood donors, persistence, and the availability of screening assays. Since 1995, three new viruses have been identified and extensively studied.

Blood Transfusion↗
PubMed2001

Residual risk of transfusion in Ghana.

UNLABELLED: Plasma samples from replacement and volunteer blood donors in Kumasi, Ghana were pooled and tested using a duplex human immunodeficiency virus (HIV) and hepatitis C virus (HCV) RNA detection METHOD: Individual plasmas constitutive of reactive pools were confirmed using reverse transcription-polymerase chain reaction. HIV and HCV infections were significantly higher in 1569 replacement donors than in 1169 volunteers; 2.4 and 1.7 versus 0.3 and 0.7% respectively (P < 0.01). Two duplex RNA-positive plasma pools contained a confirmed/seronegative HIV or HCV RNA individual plasma. The residual post-transfusion risk of HIV and HCV infection of blood collected from replacement blood donors ranged between 1:260 and 1:16 393 after screening for anti-HIV, p24 antigen and anti-HCV. These data indicate that in high-prevalence HIV and HCV blood donor populations, a substantial residual post-transfusion risk of infection remains. This risk might be reduced by collecting blood in younger volunteer donors or by genomic screening.

Adult↗
PubMed2001

Screening for viral markers in volunteer and replacement blood donors in West Africa.

BACKGROUND AND OBJECTIVES: West Africa is a highly endemic area for viral infections. The prevalence of five viral markers was determined in Ghanaian blood donors. MATERIALS AND METHODS: Replacement and volunteer blood donors were screened using enzyme immunoassays (EIAs) for hepatitis B surface antigen (HBsAg), human immunodeficiency virus antibodies (anti-HIV), HIV p24 antigen, human T-cell lymphocytotrophic virus-I and -II antibodies (anti-HTLV-I/II) and hepatitis C virus antibodies (anti-HCV). RESULTS: HBsAg was present at an equally high frequency (15%) in young volunteer (median age 18 years) and older replacement (median age 33 years) blood donors. In contrast, the prevalence of anti-HIV and anti-HCV was significantly higher in replacement blood donors (2.4 and 0.3%, respectively, P < 0.001). HCV RNA was detected in 74 or 55% of seropositive donors, depending on the confirmatory criteria used. No p24 antigen-positive/anti-HIV-negative donations were found. The prevalence of HTLV-I/II was generally low (0.5%). CONCLUSION: All blood donations should be screened for hepatitis B virus (HBV), HIV and HCV markers.

Adolescent↗
PubMed2001

A complex human immunodeficiency virus type 1 A/G/J recombinant virus isolated from a seronegative patient with AIDS from Benin, West Africa.

A human immunodeficiency virus type 1 (HIV-1(B76)) originating from Benin (West Africa) was isolated and characterized. The patient had severe clinical AIDS and presented an unusual serological profile. Only one out of five different detection assays was able to demonstrate the presence of antibodies to HIV, whereas confirmatory assays remained indeterminate. In contrast, both plasma viral load and p24 antigen level were unusually high. HIV-1 infection was proved by viral RNA and proviral DNA amplification. HIV-1(B76) partially purified lysate reacted strongly with all anti-HIV-1-positive sera from the region but B76 plasma did not react with subtype A control viral antigen. This patient is likely to have had severe acquired immune dysfunction explaining her lack of immunological reactivity. Phylogenetic analysis of the genome identified a complex HIV-1 A/G/J recombinant. The gag and pol genes, and the majority of nef,are characteristic of subtype A; the gag/pol junction, the 3' end of pol, vpu and env genes were characteristic of subtype G; vif, vpr and the 5' end of nef were subtype J. In addition, part of the HIV-1(B76) genome had considerable sequence similarity with the previously described CRF06 cpx (BFP90) isolate. HIV-1(B76) did not exhibit any remarkable replication properties or cell tropism in vitro.

Acquired Immunodeficiency Syndrome↗
PubMed2001

Production and characterization of monoclonal antibodies specific for a conserved epitope within hepatitis C virus hypervariable region 1.

Frequent mutations in hypervariable region 1 (HVR1) of the main envelope protein of hepatitis C virus (HCV) is a major mechanism of persistence by escaping the host immune recognition. HVR1 contains an epitope eliciting neutralizing antibodies. This study was aimed to prepare broadly cross-reacting, high-affinity, monoclonal antibodies (MAb) to the HVR1 C terminus of HCV with potential therapeutic neutralizing capacity. A conserved amino residue group of glycine (G) at position 23 and glutamic acid (Q) at position 26 in HVR1 was confirmed as a key epitope against which two MAbs were selected and characterized. MAbs 2P24 and 15H4 were immunoglobulin G1 kappa chain [IgG1(kappa)], cross-reacted with 32 and 30 of 39 random C-terminal HVR1 peptides, respectively, and did not react with other HCV peptides. The V(H) of 2P24 and 15H4 heavy chains originated from Igh germ line v gene family 1 and 8, respectively. In contrast, the V(L) kappa sequences were highly homologous. The affinity (K(d)) of 2P24 and 15H4 (10(-9) or 10(-8) M with two immunizing peptides and 10(-8) M with two nonimmunizing HVR1 peptides) paralleled the reactivity obtained with peptide enzyme immunoassay. MAbs 2P24 and 15H4 captured 25 of 31 (81%) HCV in unselected patients' plasmas. These antibodies also blocked HCV binding to Molt-4 cells in a dose-dependent fashion. The data presented suggest that broadly cross-reactive MAbs to a conserved epitope within HCV HVR1 can be produced. Clinical application for passive immunization in HCV-related chronic liver disease and after liver transplantation is considered.

Amino Acid Sequence↗
PubMed2001

Serological and molecular screening for viruses in blood donors from Ntcheu, Malawi: high prevalence of HIV-1 subtype C and of markers of hepatitis B and C viruses.

The prevalence of antibodies to human immunodeficiency virus type 1 (HIV-1), hepatitis C virus (HCV), human T lymphotropic virus I (HTLV-I), and hepatitis B (HBV) surface antigen (HBsAg) was determined in blood donors from Ntcheu, Malawi. Each donation was also screened for HIV-1 RNA and HCV RNA. Among 159 blood donations, the prevalence of HIV-1 infection was 10.7%, 8.1% for HBV carriage, 6.8% for anti-HCV, and 2.5% for anti-HTLV-I. HIV-1/HTLV-I and HIV-1/HCV dual infections were observed in 1.2% of the donations. Consequently, 13% of blood donors from Ntcheu should be deferred for retroviral infections and 15% for hepatitis viral infections. Sequence analyses of the HIV-1 strains revealed a relatively homogeneous circulation of subtype C viruses in Malawi. These findings confirm the high endemicity of blood-borne viruses in Malawi and the need for a sensitive viral screening of blood donations to improve blood safety.

Amino Acid Sequence↗
PubMed2000

Detection of TT virus DNA in patients with liver disease and recipients of liver transplant.

TT virus (TTV) is transfusion-transmissible but its involvement in post-transfusion hepatitis is uncertain. To investigate the potential association of TTV with liver diseases, the prevalence of TTV DNA was tested by semi-nested PCR in 113 carriers of hepatitis C virus (HCV), 10 patients with acute liver failure, 11 patients with cryptogenic cirrhosis and 200 control blood donors. Thirty-seven of these patients underwent liver transplantation and were tested pre- and post-transplantation. TTV DNA was semi-quantified in serial samples from seven patients with unexplained post-transplant hepatitis. TTV genotyping was performed on samples from 28 patients by sequence analysis. The prevalence of TTV DNA in blood donors was 1.5% and 17% in HCV infected haemophiliacs. In patients with acute or chronic liver disease or hepatitis, 6 to 27% prevalence was observed. After liver transplantation, the prevalence of TTV DNA increased from 16 to 46% (P < 0.01). In patients who developed unexplained hepatitis post-transplantation, TTV viraemia did not parallel ALT levels. TTV DNA either increased in titre or became detectable shortly after transplantation, suggesting that either TTV was transfusion-transmitted, or, more likely, that immunosuppression caused a recurrence of low level or undetectable TTV viraemia. TTV had considerable genomic diversity in the N22 region, corresponding to at least 4 genotypes. Genotype 2 was found in 14/28 patients.

Adult↗
PubMed2000

AIDS in an HIV-seronegative Ghanaian woman with intersubtype A/G recombinant HIV-1 infection.

A 29-year-old Ghanaian woman who developed AIDS while being HIV-antibody seronegative was investigated during a collaborative study aimed at the identification of viral causes of a HIV-seronegative AIDS syndrome in West Africa. Plasma was screened with a panel of EIA tests for antibodies to HIV and HIV-1 p24 antigen. Retroviral infection was investigated by detection of reverse transcriptase (RT) activity in plasma, viral RNA amplification and quantification, and virus isolation. Positive amplification products were sequenced and phylogenetic analyses were carried out. Most EIA tests were unable to demonstrate the presence of anti-HIV anti-bodies, whereas confirmatory assays yielded inconclusive results. Retroviral infection was documented by detection of RT activity, HIV-1-specific genomic amplification and virus isolation. This virus was HIV-1 subtype A with an unusual six amino acid insertion in the gp120 V4 loop and with the nef gene of subtype G. The patient's plasma did not react with either autologous or heterologous viral lysates or HIV-1 peptides, whereas antibodies to other viral antigens were present. In conclusion, the Ghanaian patient exhibited a rare subtype A/G recombinant HIV-1 infection with a near absence of a HIV-specific humoral response. The lack of detectable antibody response might be due to either a highly pathogenic, rapidly fatal, HIV-1 infection preventing the development of the typical humoral immune response or to a host-related dysfunction of the immune system. Direct antigenemia or genomic detection of the virus should be undertaken when clinical or biological data suggests an HIV infection in the absence of serological evidence.

Acquired Immunodeficiency Syndrome↗
PubMed2000

Genomic screening for blood-borne viruses in transfusion settings.

The residual risk of post-transfusion human immunodeficiency virus (HIV) infection is low but slightly higher for hepatitis B virus (HBV) and hepatitis C virus (HCV), the main reason being viraemia during the window period preceding antibody or antigen detection by enzyme immunoassays. Immunosilent-infected individuals and carriers of distant viral variants also play an unquantifiable role. Multiple techniques, e.g. reverse transcription-polymerase chain reaction (RT-PCR), PCR, ligase-chain reaction, nucleic acid sequence-based amplification (NASBA) and transcription-mediated amplification (TMA) have been developed to amplify and detect viral genomes as single or multiplex assays. Equipment providing various degrees of automation has been adapted to these techniques. Applying nucleic acid amplification techniques (NAT) to blood screening, two main approaches have been advocated: plasma pool and single-donation testing. Pool testing presents the advantage of lower cost and readily available equipment although it is prone to false negative and positive reactions. The time required to identify infected donations is incompatible with blood component release, and may lead to product waste. Single-unit testing, although appealing, is not yet fully automated and potentially very costly unless a systematic multiplex approach is taken. Although technically feasible, NAT applied to the blood supply needs to be clinically evaluated and its cost efficiency assessed in the general public health context. However, pool NAT is currently implemented in continental Europe and the USA.

Blood Donors↗
PubMed2000

Will genome detection replace serology in blood screening for microbial agents?

The residual risk of transfusion-transmitted viral infection in developed countries is considered minimal or negligible. However, zero risk remains a strong political objective. Genomic screening for HCV, HIV and HBV represents a major advance, eliminating infectious blood donations collected during the pre-seroconversion window period, rare cases of immunosilent infections and, possibly, a large spectrum of viral variants. In Western countries, HCV RNA genomic screening started on pools of 16-400 plasma samples from individual donations. Pooling may produce false-positive and false-negative results. Individual donation testing is more suitable to blood screening but requires multiplexing, automation, and affordable cost. Because donations from individuals who are HBV DNA-negative/serologically positive, or those apparently recovered from HCV infection, may remain infectious, it is unlikely that HBsAg, anti-HCV, and anti-HIV will be discontinued when genomic screening is extended to all three viruses. HIV-1 p24 antigen may prove redundant with HIV RNA screening. Anti-HTLV-I and HTLV-II will remain more effective than genomic testing.

Antibodies, Viral↗
PubMed2000

Emerging viral infections relevant to transfusion medicine.

The development of new technologies leads to the discovery of new viruses. For each of these new infectious agents relevance to transfusion needs to be assessed. The questions to be answered are transmissibility by transfusion, pathogenicity, prevalence in blood donors, persistence and the availability of screening assays. Since 1995, three new viruses have been identified and extensively studied. GB virus-C/hepatitis G virus (GBV-C/HGV), a relatively rare virus with some homology with and epidemiological features of HCV, was thought to be related to post-transfusion hepatitis but was proven to be unrelated to hepatitis and is still in search of a disease. Human herpes virus-8 (HHV-8) is a major factor in the pathogenesis of Kaposi's sarcoma and other tumours related to immunodeficiency. HHV-8 transmission by organ transplantation, but not by transfusion, has been demonstrated. The TT virus (TTV) is a ubiquitous virus infecting a very high proportion of humans in infancy. No clinical symptoms or pathogenicity is attached to TTV. To date, none of the emerging viruses have been proven relevant to transfusion.

Blood Donors↗
PubMed2000

Evolutionary rate and genetic drift of hepatitis C virus are not correlated with the host immune response: studies of infected donor-recipient clusters.

Six donor-recipient clusters of hepatitis C virus (HCV)-infected individuals were studied. For five clusters the period of infection of the donor could be estimated, and for all six clusters the time of infection of the recipients from the donor via blood transfusion was also precisely known. Detailed phylogenetic analyses were carried out to investigate the genomic evolution of the viral quasispecies within infected individuals in each cluster. The molecular clock analysis showed that HCV quasispecies within a patient are evolving at the same rate and that donors that have been infected for longer time tend to have a lower evolutionary rate. Phylogenetic analysis based on the split decomposition method revealed different evolutionary patterns in different donor-recipient clusters. Reactivity of antibody against the first hypervariable region (HVR1) of HCV in donor and recipient sera was evaluated and correlated to the calculated evolutionary rate. Results indicate that anti-HVR1 reactivity was related more to the overall level of humoral immune response of the host than to the HVR1 sequence itself, suggesting that the particular sequence of the HVR1 peptides is not the determinant of reactivity. Moreover, no correlation was found between the evolutionary rate or the heterogeneity of the viral quasispecies in the patients and the strength of the immune response to HVR1 epitopes. Rather, the results seem to imply that genetic drift is less dependent on immune pressure than on the rate of evolution and that the genetic drift of HCV is independent of the host immune pressure.

Blood Donors↗