Mariana Cavalheiro Magri

The best algorithm to confirm the diagnosis of HTLV-1 and HTLV-2 in at-risk individuals from São Paulo, Brazil

The ability to confirm the diagnosis of human T-lymphotropic virus types 1 and 2 (HTLV-1 and HTLV-2) in at-risk individuals in São Paulo, Brazil by Western blotting (WB), conventional polymerase chain reaction (tax and pol PCR) and real-time PCR (pol) is compared. Seventy-three blood samples that were reactive in HTLV-1/2 serological screening enzyme immunoassays (EIAs) were evaluated. HTLV-1/2 was confirmed in 53 blood samples: 48 were positive by WB, 41 were positive by PCR and 42 scored positive by real-time PCR assays (37 of 48 WB-positive samples plus five WB-indeterminate samples that were further confirmed by sequencing). Although WB was able to detect more cases of HTLV-1/2 infection, the real-time PCR assay was able to discriminate between these two viruses and confirm an individual HTLV-1/HTLV-2 diagnosis in two HTLV WB-untyped samples and five WB-indeterminate samples. Because of the large number of WB-indeterminate samples and the cost of the WB assay in Brazil, it is proposed an algorithm that employs two EIAs for screening and then real-time PCR to confirm the infection, followed by testing any PCR-negative samples with the WB assay. This strategy reduces costs and improves the accuracy of the diagnosis of HTLV-1/2.

Prevalence of Human T-Cell Lymphotropic Virus (HTLV-1/2) in Individuals from Public Health Centers in Mozambique

The prevalence of human T-cell lymphotropic viruses types 1 and 2 (HTLV-1/2) in Mozambique is not known. The present study examined blood samples from 208, 226, and 318 individuals from Northern, Central, and Southern Mozambique, respectively, of all socioeconomic and demographic strata attending public health centers in Mozambique for HTLV-1/2-specific antibodies. Serum samples were assessed for HIV- and HTLV-1/2-specific antibodies by using enzyme immunoassays, and infections with HTLV-1 and -2 were confirmed by using Western blot. An overall HTLV-1/2 prevalence of 2.3% (2.9% in female and 1.1% in male subjects) was observed, and the prevalence of infection increased with age. Regional variation in the prevalence of HIV and HTLV-1/2 was observed; 32.2%, 65.5%, and 44% of individuals tested HIV positive in Northern, Central, and Southern Mozambique, respectively, and 2.4%, 3.9%, and 0.9% tested HTLV-1/2 positive in the same regions. HTLV-1 infection was confirmed in these individuals. No association between HTLV-1 infection and sociodemographic variables or HIV status was detected, although the low number of HTLV-1-positive cases did not allow robust statistical analyses. The results obtained suggest different risk factors and epidemiologic correlates of HIV and HTLV-1 transmission in Mozambique. Furthermore, our results suggested that North and Central Mozambique should be considered endemic regions for HTLV-1 infection. As no cases of HTLV-2 were detected, HTLV-2 appears to have not been introduced into Mozambique.

Latent human herpesvirus - 8 (HHV-8) infection in female commercial sex workers from Imbituba, Santa Catarina, Brazil

Human herpesvirus 8 (HHV-8) infection was identified in 6 out of 90 (6.7%) female commercial sex workers from Imbituba, Santa Catarina, and was associated to age. Frequencies of 5.6% of anti-latent and 3.3% of anti-lytic antibodies were detected. Considering non-endemic areas from Brazil, the anti-latent antibodies frequency seems elevated and requires further investigation on referent female population.

Long terminal repeat sequence analysis of HTLV-2 molecular variants identified in Southern Brazil.

In Brazil, human T-lymphotropic virus type 2 (HTLV-2) is endemic in Amerindians and epidemic in intravenous drug users (IDUs). The long terminal repeat (LTR) is the most divergent genomic region of HTLV-2, therefore useful to characterize subtypes. Nucleotide sequence and restriction fragment length polymorphism (RFLP) analysis of LTR genomic segments of fourteen HTLV-2 strains isolated from HIV-infected patients of Londrina, Southern Brazil, were carried out. Molecular analysis disclosed that all HTLV-2 strains belonged to 2a subtype, and RFLP detected the presence of the a4, a5, and a6 subgroups according to Switzers nomenclature. RFLP correlated with nucleotide sequence, and phylogenetic analysis clustered HTLV-2 sequences of IDUs into subgroups a5 and a6. HTLV-2 sequences from individuals of sexual risk factor clustered into the a4 subgroup. These results extend the knowledge of the genetic diversity of HTLV-2 circulating in Brazil and provide insights into HTLV-2 transmission and virus movement in this geographic area.

Confirming high prevalence of human herpesvirus 8 infection in chronic kidney disease patients in São Paulo, Brazil

Sir, Human herpesvirus 8 (HHV-8) is frequently associated with Kaposis sarcoma. It can be transmitted through organ transplantation or reactivated by immunosuppressive therapy. Chronic kidney disease (CKD) patients are at risk of this infection [1]. The present study aimed to determine the seroprevalence of HHV-8 in CKD patients in Sao Paulo, Brazil. The study was approved by the research ethics committees at participating institutions. Blood samples collected from 805 CKD patients attended Hospital do Rim e Hipertensao and/or Santa Casa de Misericordia de Sao Paulo (Sao Paulo, Brazil) were tested for latent and lytic HHV-8-specific antibodies using indirect immunofluorescence assays at Instituto Adolfo Lutz in Sao Paulo, Brazil, as previously described [2]. The chi-square test and/or Fishers exact test were performed for comparing categorical variables and HHV-8 serum status, using SPSS for Windows. Of the 805 CKD patients, 61.4% were males, 61.5% white, 35.5% black/pardum and 3.0% yellow. The mean age was 58 years (18–91). Two hundred ninety-five patients were on haemodialysis (HD), 54 on peritoneal dialysis (PD) and 456 not yet on renal replacement therapy (RRT). One hundred forty-five (18.0%) CKD patients were found HHV-8-seropositive, of whom 56 (18.9%) were on HD, 8 (14.8%) on PD and the remaining 81 (17.7%) were not on any RRT. Examination of these different groups revealed no statistical significant differences (P = 0.963). Further statistical analyses were conducted without this sub-grouping by RRT received. Table ​Table11 discloses the comparison between HHV-8-seronegative and HHV-8-seropositive groups. Patients HHV-8-seropositive had a higher prevalence of previous transplant as well as higher prior exposure to sexually transmitted diseases. Of note, 57.0% of CKD patients who had syphilis also had HHV-8-seropositivity (P = 0.021). Other variables showed lack of association with HHV-8 serological results. Table 1 Comparison between HHV-8-seronegative and HHV-8-seropositive groups High HHV-8 seroprevalence was observed in CKD patients in Sao Paulo, Brazil [3]. Interestingly, in the present study, similar proportions of HHV-8-seropositivity were observed in pre-dialysis and dialysis patients suggesting that dialysis proceedings were not related to HHV-8 transmission/acquisition. Another result was the strong association between HHV-8-seropositivity and previous transplant presenting another route of viral transmission as previously reported [4]. The association between HHV-8-seropositivity and syphilis could suggest that the syphilis lesions facilitate the entrance of the virus during sexual intercourse. Finally, several studies conducted worldwide have attempted to find the best immunosuppressive therapy for use with HHV-8-seropositive transplant recipients [5]. It is not yet defined, but it is certain that these patients need an appropriated attendance to avoid iatrogenic KS and organ rejection, giving them perhaps a better quality of life after transplant. Therefore, due to the high HHV-8 seroprevalence found in the present study and the seriousness of the HHV-8-associated diseases, the authors suggest that screening for HHV-8 must be performed in CKD patients, even those in pre-dialysis.

Prevalence of HTLV-1/2 and HIV-1/2 in individuals attending Public Health Center in Mozambique. One HTLV-1 symptomatic case report

An overall HTLV-1/2 prevalence of 2.3% was observed, and increased with age. Regional variation in the prevalence of HIV and HTLV-1/2 was observed; 32.2%, 65.5% and 44% of individuals tested HIV-positive in Northern, Central and Southern MZ, respectively, and 2.4%, 3.9% and 0.9% tested HTLV-1-positive in the same regions. No association between HTLV-1 infection and sociodemographic variables or HIV status was detected. One woman aged 54 years, born in Northern MZ, HIV-negative, who complaints of rheumatic pain, presented high HTLV-1 proviral load, and phylogenetic analysis of env and LTR sequences clustered HTLV-1-isolate into Cosmopolitan subtype, transcontinental subgroup A (GenBank Accession numbers HM770441 and JF271853). Discussion and conclusions These data suggest different risk factors and epidemiologic correlates of HIV and HTLV-1 transmission in MZ. HTLV-1 is endemic in this country, and arthritis was associated with HTLV-1 infection in one patient.

Unpredicted HTLV-1 infection in female sex worker from Imbituba, Santa Catarina, Brazil

Recently, some of us presented data concerning HIV frequency in female sex workers (FSW) from the port city of Imbituba, Santa Catarina (SC), Southern Brazil. Out of 90 FSW, six (6.7%) were found to be HIV-positive, and significant association were found between HIV infection and the number of clients attended per day (p = 0.008), and sexual relations without frequent use of condoms (p = 0.015). Now, in order to add some information concerning to other human retroviruses (human T-lymphotropic virus type 1 HTLV-1, and human T-lymphotropic virus type 2 HTLV-2) which share the same routes of virus transmission/acquisitions (i.e., mother-to-child transmission, sexual contact, blood transfusion, contaminated needles sharing among injecting drug users IDU), we conducted the present study. Using enzyme immunoassays (EIA) screening tests ((VironostikaTM HTLV-I/II, BioMeriéux, Bostel, The Netherlands, and MurexTM HTLV-I+II, Abbott, Dartford Kent England, UK), and Western Blotting (WB) confirmatory test (HTLV BLOT 2.4TM, Abbott Murex, Singapore System, Abbott, Delkenhein, Germany), we were able to detect one case of HTLV-1 infection. The socio-demographic as well the comportamental characteristics of the HTLV-1-infected FSW are as follow: female of 25 years of age, white color, birth place in Imbituba, four-year old son who was breastfed, incomplete elementary school, no steady partner, one month in prostitution, one client per day, vaginal and oral sexual practices, no sex during menses, no use of condoms, use of alcohol, sex with IDU, and no blood transfusion. All these data were obtained after she signed the informed consent, and based on the interview and the answers of a questionnaire. Subsequently, when she returned to receive the HIV and HTLV serological results, another blood sample was collected and confirmed HTLV-1 infection. At this time, she was attended by physicians and no HTLV-1-associated disease was detected. On the other hand, she was pregnant, and she was advised not to breastfeed her offspring. Since then, she has been visited by the non-governmental organization called Industry of Solidarity (ISO) working group, and medical care has been conducted in an outpatient site in Tubarão, SC, Brazil. As previously described, HTLV-1 is associated with several diseases such as adult T-cell leukemia/lymphoma (ATL), HTLV-1-associated myelopathy (HAM) also known as tropical spastic paraparesis (TSP), infective dermatitis in children, and uveitis. Differently to those occurring in elderly patients from Japan where HTLV-1 has been endemic, in Bahia, Brazil HTLV-1 has been detected in younger people, including some cases diagnosed in adolescents who presented severe eczema during childhood. In Brazil, the great number of HTLV-1 and HTLV-2 seropositive individuals besides the severity of HTLV-1-associated diseases, prompted the government and physicians to elaborate in 2004, a guideline for counseling persons infected with HTLV-1 and HTLV-2. Of note, Santa Catarina presents the lowest prevalence rate of HTLV-1/2 in Brazil, estimated in 0.4/1,000 inhabitants. We do not know the means of HTLV-1 acquisition by this FSW from Imbituba, SC, but since she denied the use of intravenous illicit drugs and previous blood transfusion, and because she does not belong to HTLV-1 endemic populations from Brazil or elsewhere, we could speculate that the sexual practices (vaginal and oral sex without use of condom) was the major via of virus acquisition. In accordance to this, during the interview she mentioned the presence of ulcers in vagina, which could propitiate the virus entrance during a sexual contact with an HTLV-1infected man. Several reports pointed unprotected sexual contact mostly with IDU, and histories of sexually transmitted diseases as risk factors for acquiring such retroviruses; these seem to be the causes of infection in the index case. Interestingly, this FSW resulted HIV-1-noninfected; in spite of this, she was counseled regarding HIV-1, HTLV-1 and HTLV-2 infections, especially concerning sexual and vertical transmission.