Detection of HTLV-I and HTLV-II infection in Africans using type-specific envelope peptides

The influence of human cytomegalovirus (HCMV) strain variation on neutralizing antibody titers was investigated in sequential sera obtained from 12 organ transplant recipients (11 renal transplant recipients, 1 liver transplant patient) suffering from primary or secondary HCMV infection. Cross-neutralization assays using either the international HCMV reference strain AD169 or individual clinical isolates from patients showed congruent results. Restriction enzyme analysis of the hypervariable α-sequence of the L-S junction of AD169 and HCMV isolates amplified by polymerase chain reaction (PCR) were carried out to confirm the expected strain differences. At least 6 groups of clinical strains were differentiated. The results of this study demonstrated that despite the strain heterogenicity of HCMV, the neutralization assay using AD169 permitted a reliable and quantitative serologic detection of neutralizing antibodies against HCMV. © 1993 Wiley-Liss, Inc.     

Infection with Retroviruses Other Than HIV-1 in Spain: A Retrospective Analysis for HIV-2, HTLV-I,and/or HTLV-II

Abstract

HIV-2, human T-cell lymphotropic virus (HTLV)-I, and HTLV-II infections are currently circulating in Spain with no evidence of an increase in the number of reported cases over time. Up to June 2002, a total of 106, 53, and 460 cases of HIV-2, HTLV-I, and HTLV-II infection, respectively, have been identified in Spain. Most HIV-2-infected and HTLV-I-infected individuals are immigrants who come from endemic areas or are Spaniards with a past history of travel to or sexual contacts with persons originating in those areas. In contrast, HTLV-II infection is mainly limited to native intravenous drug users who are frequently coinfected with HIV-1.     

Infection with retroviruses other than HIV-1 in Spain: a retrospective analysis for HIV-2, HTLV-I,and/or HTLV-II

HIV-2, human T-cell lymphotropic virus (HTLV)-I, and HTLV-II infections are currently circulating in Spain with no evidence of an increase in the number of reported cases over time. Up to June 2002, a total of 106, 53, and 460 cases of HIV-2, HTLV-I, and HTLV-II infection, respectively, have been identified in Spain. Most HIV-2-infected and HTLV-I-infected individuals are immigrants who come from endemic areas or are Spaniards with a past history of travel to or sexual contacts with persons originating in those areas. In contrast, HTLV-II infection is mainly limited to native intravenous drug users who are frequently coinfected with HIV-1.     

Evaluation of a new, fully automated immunoassay for detection of HTLV-I and HTLV-II antibodies

Screening blood donations for human T-lymphotropic virus types I and II (HTLV-I/II) continues to be important in protecting the safety of blood products and controlling the global spread of these retroviruses. We have developed a fully automated, third generation chemiluminescent immunoassay, ARCHITECT rHTLV-I/II, for detection of antibodies to HTLV-I/II. The assay utilizes recombinant proteins and synthetic peptides and is configured in a double antigen sandwich assay format. Specificity of the assay was 99.98% (9,254/9,256, 95% CI = 99.92-100%) with the negative specimens from the general population including blood donors, hospital patients and pregnant women from the US, Japan and Nicaragua. The assay demonstrated 100% sensitivity by detecting 498 specimens from individuals infected with HTLV-I (n = 385) and HTLV-II (n = 113). ARCHITECT rHTLV-I/II results were in complete agreement with the Murex HTLV-I/II reference assay and 99.7% agreement with the Genelabs HTLV Blot 2.4 confirmatory assay. Analytical sensitivity of the assay was equivalent to Murex HTLV-I/II assay based on end point dilutions. Furthermore, using a panel of 397 specimens from Japan, the ARCHITECT rHTLV-I/II assay exhibited distinct discrimination between the antibody negative (Delta Value = -7.6) and positive (Delta Value = 7.6) populations. Based on the excellent specificity and sensitivity, the new ARCHITECT rHTLV-I/II assay should be an effective test for the diagnosis of HTLV-I/II infection and also for blood donor screening.     

In vivo fluctuation of HTLV-I and HTLV-II proviral load in patients receiving antiretroviral drugs

HTLV-I and HTLV-II infect T lymphocytes. A high HTLV-I proviral load in peripheral blood mononuclear cells (PBMCs) has been associated with a higher risk of neurologic disease. For HTLV-II, large numbers of infected lymphocytes might contribute to accelerate the immunodeficiency and increase the risk of neuropathy in HTLV-II/HIV-1 coinfected people. We have examined the impact of antiretroviral drugs on HTLV proviral load, testing longitudinal samples collected from 1 HTLV-I infected patient suffering HTLV-I-associated myelopathy (HAM), and two HTLV-II/ HIV-1 coinfected subjects. The HAM patient showed a reduction greater than 2 log in the peripheral proviral load after being treated with zidovudine and lamivudine. In contrast, potent antiretroviral treatment in HIV-1/HTLV-II coinfected carriers produced an initial increase in the HTLV proviral load, which was followed by a reduction greater than 1 log thereafter. In conclusion, antiretroviral drugs seem to reduce HTLV proviral load, although in HIV-1 coinfected persons a transient increase in HTLV proviral load could reflect the rapid blocking of HIV-1 replication occurring in response to therapy, thus causing an increase in the number of circulating T lymphocytes carrying HTLV proviral DNA.     

Absence of Leishmania infantum in cave bats in an endemic area in Spain

Though dogs have been historically considered the main reservoir of Leishmania infantum, the role of wildlife in its epidemiology is attracting increasing attention. Rodents, wild carnivores and, recently, hares (Lepus spp.) have been proposed as sylvatic reservoirs for this parasite. Bats have never been tested for L. infantum infection in Europe. Nevertheless, bats have a widespread distribution, they live in abundant colonies, and some species inhabit caves, where constant temperatures and humidity provide ideal habitat for the sand fly vector. We tested blood samples from 35 Schreibers’ bats (Miniopterus schreibersii), abundant cave bats in NE Spain, which is an enzootic area of leishmaniasis. A PCR-amplifying fragment of the high copy of Leishmania donovani group kDNA minicircles was used. None of the analyzed samples were positive (maximum possible prevalence?=?8.20 %). Though the susceptibility of this bat to parasitization by L. infantum cannot be ruled out, our survey indicates that this species may not be a relevant sylvatic reservoir of L. infantum in the Mediterranean area. Nevertheless, even if the prevalence of infection in bats is low, such an abundant taxonomic group would still provide a significant maintenance population for the parasite.     

High frequencies of human T-lymphotropic virus type I (HTLV-I) infection and presence of HTLV-II proviral DNA in blood donors with anti-thyroid antibodies

To investigate the relationship between human T-lymphotropic virus (HTLV) types I and II and the pathogenesis of autoimmune thyroid diseases, we examined serum anti-thyroid antibodies in 1019 blood donors with or without serum anti-HTLV-I antibody as well as proviral DNA for HTLV-II in leukocyte DNA by the polymerase chain reaction in 395 blood donors with or without anti-thyroid antibodies. The frequency of donors with anti-HTLV-I antibody who also showed anti-thyroid antibodies (7.9%) tended to be higher than that (6.3%) among donors who did not have the anti-HTLV-I antibody. The frequency of anti-thyroid antibodies in 125 young male donors aged 16–39 years with anti-HTLV-I antibody (4.8%) was significantly higher (P<0.05) than that (0.6%) in 164 control donors without the antibody. In blood donors with anti-thyroid antibody, 25.0% of those with anti-HTLV-I antibody and 14.3% of those without the antibody had HTLV-II proviral DNA. In contrast, in donors without anti-thyroid antibody HTLV-II proviral DNA was detected in 2.3% of those with anti-HTLV-I antibody and in 0.6% of those without the anti body. Thus the detection rates in donors with anti-thyroid antibody were significantly higher (P<0.001) than those in donors without the antibody, regardless of HTLV-I infection. These results suggest that HTLV-I infection and the presence of HTLV-II proviral DNA may be independently related to the pathogenesis of autoimmune thyroid diseases.Abbreviations HTLV Human T-lymphotropic virus - PCR Polymerase chain reaction     

Absence of p53 gene mutations in hepatocarcinomas from a Mediterranean area of Spain

 The incidence of p53 gene abnormalities in human hepatocellular carcinoma (HCC) varies in different geographical areas, being higher in regions where hepatitis virus infection and dietary exposure to aflatoxin B1 are the most common aetiological agents. These mutations are less frequently encountered in Europe, although some studies have reported p53 protein overexpression in up to 45% of cases analysed. We have analysed 129 tumour samples of primary malignant hepatic neoplasms recovered from paraffin blocks processed in two pathology laboratories in a Mediterranean area of Spain (Valencia and Gerona). Among 14 cases in which p53 immunohistochemistry expression proved positive, 5 stained in more than 50% of the cell nuclei. By PCR-SSCP analysis we could detect the complete sequence from exon 5 through 8 in 70 cases and part of this region in the remaining cases, but no mutations were found. We found no relationship with the clinical stage, tumour stage or clinical outcome. We conclude that p53 gene alterations are not a major event in the malignant transformation of hepatic cells in this region of the Mediterranean. The variable incidence of p53 gene alterations in other geographical areas may reflect a different genetic background for the aetiology of HCC. Received: 14 September 1998 / Accepted: 28 January 1999     

Discrimination between human T-cell lymphotropic virus type I and II (HTLV-I and HTLV-II) infections by using synthetic peptides representing an immunodominant region of the core protein (p19) of HTLV-I and HTLV-II.

We describe enzyme immunoassays that use synthetic oligopeptides to discriminate serologically between human T-cell lymphotropic virus type I and II (HTLV-I and HTLV-II) infections. The peptides represented 20-amino acid segments between residues 111 and 130 (MA1) and residues 116 and 135 (MA2) of the p19 gag proteins of HTLV-I and HTLV-II, respectively. The assays were sensitive since 69 of 74 HTLV-positive sera were reactive to at least one of the two matrix (MA) peptides (sensitivity, 93.2%). By using the ratio of the optical density of MA1 to the optical density of MA2, which represents for every serum sample the ratio between the absorbance value obtained in the MA1 assay and the absorbance value obtained in the MA2 assay, 59 of the 69 reactive serum samples were clearly and easily typed as positive for either antibody to HTLV-I or antibody to HTLV-II. Eight of the 10 remaining reactive serum samples were analyzed further by an inhibition procedure, and their type specificities were then clearly identifiable. Therefore, the results indicate that all MA-reactive sera were serologically distinguished by our peptide assays.     

Differential diagnosis of HTLV-I and HTLV-II infections by restriction enzyme analysis of 'nested' PCR products.

A 'nested' polymerase chain reaction (PCR) assay is described which is capable of detecting single copies of human T-cell lymphotropic virus (HTLV) in genomic DNA extracted from peripheral blood mononuclear cells (PBMCs). A single set of 'nested' oligonucleotide primers, based on the highly conserved tax/rex region of the viral genome, was able to detect both HTLV-I and HTLV-II proviral sequences in clinical samples of diverse geographical origins, from the United States, Great Britain, Japan, the Caribbean, Italy, Greece, Iraq and West Africa. Rapid discrimination between HTLV-I and HTLV-II infections was achieved by restriction enzyme analysis of unpurified second-round PCR products, even in those cases in which serological assays had failed to provide a definitive result. Over a 2-year period, a total of 53 HTLV infections (37 HTLV-I and 16 HTLV-II) were identified by this technique and complete concordance with serological typing, available in 41 cases, was observed.     

Protection of rabbits against HTLV-II infection with a synthetic peptide corresponding to HTLV-II neutralization region

Summary Rabbit immune sera raised against synthetic peptides of the HTLV-II envelope gp46 region were examined for HTLV-II neutralization ability by HTLV-vesicular stomatitis virus (VSV) pseudotype assay and syncytium inhibition assay. HTLV-II neutralization activity was detected in the sera against HTLV-II Env gp46, 80–103 but not in those to HTLV-II Env gp46, 171–196. Three rabbits immunized with the synthetic peptide of HTLV-II Env gp46, 80–103 and three non-immunized rabbits were challenged with intravenous inoculation of an HTLV-II-producing human cell line (MOT, 1×10⁷ cells). The non-immunized rabbits showed seroconversion for HTLV-II after 2 weeks and maintained persistent infection but the immunized rabbits were protected from HTLV-II infection. Nested or repeated polymerase chain reaction revealed the presence of HTLV-II provirus sequences in the non-immunized rabbits but not in the immunized rabbits. These results suggest that peptide vaccination with a synthetic peptide corresponding to the HTLV-II neutralization region is useful for preventing HTLV-II infection.     

Prevalence and risk factor analysis of TTV infection in prostitutes

TTV, a DNA virus, has been isolated from patients with non-A to non-E post-transfusion hepatitis. In the past it was assumed that TTV was transmitted parenterally. It is unclear whether sexual contact leads to transmission of this virus. In this study, two sets of TTV-specific polymerase chain reaction primers were used to detect serum TTV DNA in 140 prostitutes and 136 controls. The prevalence of TTV DNA in prostitutes was significantly higher than in the control group (46/140 [32.9%] vs. 29/136 [21.3%]; P = 0.043). There was no significant difference in the prevalence of positive antibody to hepatitis A virus (anti-HAV) in either group (87.8% for prostitutes, 85.3% for controls). No particular risk factor was significantly associated with positive TTV DNA in prostitutes. In summary, TTV is highly prevalent in prostitutes. Transmission of TTV via sexual contact is not as efficient as transmission of hepatitis C and D viruses and GB virus-C hepatitis G virus. The high prevalence of TTV in controls indicates that there are diverse routes of transmission of this virus.     

Sensitive and specific polymerase chain reaction assays for diagnosis of human T-cell lymphotropic virus type I (HTLV-I) and HTLV-II infections in HTLV-I/II-seropositive individuals.

To confirm and differentiate between human T-cell lymphotropic virus type I (HTLV-I) and HTLV-II infections, we analyzed by polymerase chain reaction (PCR) samples of peripheral blood lymphocytes from 98 individuals seropositive for HTLV-I/II using pol (SK110/111) and tax (SK43/44) consensus primer pairs. A total of 96 samples (97.9%) were positive by the tax generic probe, while 95 were typed by the HTLV-I and HTLV-II pol probes. The three pol-negative samples were successfully amplified and typed by nested PCR with primers internal to SK110 and SK111. Results of PCR with a lysate of leukocyte nuclei obtained by whole blood lysis were comparable to those obtained with peripheral blood lymphocytes from 16 HTLV-seropositive subjects.     

HIV-1, HIV-2, and HTLV-I infection in high-risk groups in Brazil

We conducted a serologic survey for antibodies to human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2) and human T-cell lymphotropic virus Type I (HTLV-I) in 704 Brazilians with the acquired immunodeficiency syndrome (AIDS) or at risk for it. The study population included 70 homosexual men (11 of whom were prostitutes), 58 bisexual men (19 of whom were prostitutes), 101 female prostitutes from three socioeconomic groups, 13 wives of men with hemophilia who were seropositive for HIV-1 antibodies, and 47 blood donors with positive Venereal Disease Research Laboratory tests for syphilis, all from Rio de Janeiro; 86 female prostitutes from two rural towns in Minas Gerais; 133 patients with AIDS from S?o Paulo; and 196 men with bleeding disorders who were seropositive for HIV-1 antibodies on enzyme-linked immunosorbent assay, from S?o Paulo and Rio de Janeiro. The prevalence of HIV-1 infection was highest in the homosexual male prostitutes (45 percent), the wives of patients with hemophilia (38 percent), the bisexual men (28 percent), the homosexual men who were not prostitutes (19 percent), and the female prostitutes from the lower class (9 percent). Combined HIV-1 and HIV-2 infection was found in 3 percent of the patients with AIDS and in 1 percent of the homosexual men. The prevalence of HTLV-I infection ranged from 1 percent in rural female prostitutes to 13 percent in HIV-1-positive men with bleeding disorders in Rio de Janeiro. Combined HIV-1 and HTLV-I infection occurred in 1 to 11 percent of some male subgroups. We conclude that in Brazil HIV-1 infection is already well established among homosexuals, bisexuals, and lower-class female prostitutes, with bisexual men probably acting as a bridge between the heterosexual and homosexual communities, that HTLV-I infection is prevalent in groups at risk for AIDS, and that HIV-2 infection has already been introduced into the country.     

Primary lymphoma of the central nervous system and HTLV-I infection

Only a few cases of AIDS-related primary lymphomas of the central nervous system (CNS) show a T-cell phenotype. We have recently studied two intravenous drug users with HIV infection who had primary CNS T-cell lymphomas. In both cases, the enzyme immunoassay (EIA) for HTLV gave a positive result. In the first case, study by western-blot (WB) and specific PCR confirmed the human T-cell lymphotropic virus type I (HTLV-I) infection and serological study by EIA for HTLV of his mother was negative. In the second case, analysis of ante-mortem serum samples by two different WBs showed an indeterminate pattern suggestive of HTLV-I infection, but adequate samples for PCR were not available. We speculate about the possibility that the horizontal transmission of HTLV-I infection could have facilitated the devepolment of a primary CNS T-cell lymphoma in these HIV patients, although they cannot be strictly considered as ATLL cases.