Results of a sample-to-cutoff ratio using Abbott Architect rHTLV-I/II assay allow to predict detection of HTLV-1 and HTLV-2 proviral DNA by real-time PCR / Resultados da razão sinal da amostra sobre o cutoff da reação no ensaio Abbott Architect rHTLV-I/II permite prever a detecção de DNA proviral por PCR em tempo real

The present study aims to correlate the sample-to-cutoff ratios (S/CO) distributions of reactive results for HTLV-1/2 antibodies with the detection of proviral DNA in a population of blood donor candidates. It was carried out a retrospective data search of 632 HTLV-1/2 reactive samples, submitted to confirmatory testing from January 2015 to December 2019. Serological screening was performed by chemiluminescent microparticle immunoassay Architect rHTLV-I/II, whereas confirmatory testing was performed by in-house real-time polymerase chain reaction method. 496 out of 632 samples (78%) had undetectable HTLV-1/2 proviral DNA and 136 (22%) had detectable proviral DNA. HTLV infection was not confirmed in any individual for whom the S/CO ratio value was <4, and proviral DNA detection rates gradually escalated as S/CO ratio values increased. The sensitivity and predictive positive value found for the Architect rHTLV-I/II was 100% and 22%, respectively. The receiver operating characteristic (ROC) curve analysis showed that the optimal S/CO ratio value for predicting the presence of HTLV-1/2 was 18.11. High S/CO ratios were more associated with the detection of proviral DNA. The S/CO ratio value <4 suggests excluding true HTLV infection and the risk of blood transmission (AU).

O estudo tem como objetivo correlacionar às distribuições das razões sample-to-cutoff (S/CO) de resultados reagentes para anticorpos HTLV-1/2 com a detecção de DNA proviral em uma população de candidatos à doação de sangue. Realizou-se uma busca retrospectiva de dados de 632 amostras reagentes para HTLV-1/2 submetidas à testagem confirmatória entre janeiro de 2015 a dezembro de 2019. A triagem sorológica foi realizada pelo imunoensaio quimioluminescente de micropartículas Architect rHTLV-I/II, enquanto o teste confirmatório foi realizado pelo método de PCR em tempo real in-house. 496 de 632 amostras (78%) apresentaram DNA proviral indetectável e 136 (22%) apresentaram DNA proviral detectável. A infecção por HTLV não foi confirmada em nenhum indivíduo com valor de S/CO <4 e as taxas de detecção de DNA proviral escalonaram gradualmente à medida que as razões S/CO aumentaram. A sensibilidade e valor preditivo positivo encontrados para o Architect rHTLV-I/II foram 100% e 22%, respectivamente. Utilizando análise de curva ROC, o valor de razão S/CO ideal para predizer a presença de DNA proviral foi de 18,11. Razões S/CO elevadas foram mais associadas à detecção de DNA proviral. Em suma, o valor de S/CO <4 sugere a exclusão de infecção por HTLV e o risco de transmissão pelo sangue (AU).

Seroprevalence of HTLV-1 and 2 in organ donors and kidney transplant recipients. Colombia 2010-2017

Abstract Since 2004, in Colombia, the detection of human T-cell lymphotropic virus type 1 and 2 (HTLV-1-2) has been recommended for organ donors and recipients. The prevention of HTLV-1 and 2 infection in recipients is important due to its relationship with lymphoproliferative and inflammatory diseases and opportunistic infections. The objective of this study was to describe the seroprevalence of HTLV-1 and 2 among organ donors and kidney transplant recipients between 2010 and 2017 in Colombia. Methods: this was a descriptive study which included 1979 organ donors and 3,311 kidney transplant recipients from the donation and transplant network from 2010 to 2017. The seroprevalence of HTLV-1 and 2 was calculated, and serological and demographic variables were described. Results: out of 1979 donors, detection of antibodies against HTLV-1 was performed in 92% (1820), with a seroprevalence of 0.2%; 50% of the cases were from the Pacific region (an endemic zone in Colombia). Ninety percent of the donors were examined for HTLV-2, with a seroprevalence of 0.2%. Of the 3311 kidney recipients between 2010 and 2017, only 44% were evaluated for HTLV-1 and 43% for HTLV-2. The seroprevalence for both viruses was 0.3%. Two of the HTLV-1 cases were positive for HLADRB1*01. Conclusions: the seroprevalence found in organ donors and kidney transplant recipients was similar to that previously reported in Colombia. Screening of all potential donors must be adhered to prevent transmission of this virus. (Acta Med Colomb 2021; 46. DOI:https://doi.org/10.36104/amc.2021.2001).

Prevalence of infection by human T Cell lymphotropic viruses (HTLV-1/2) in adult population in Vitoria-ES

ABSTRACT Introduction: Brazil has a high number of HTLV-1/2 infections which are unequally distributed in the country. Most prevalence studies have focused on specific populations, such as blood donors and pregnant women. Some areas, for example the state of Bahia, have robust information about HTLV-1/2 infection, however there is no information available about this infection in the general population of Vitoria, Espírito Santo, Brazil. Objective: To determine the prevalence of HTLV-1/2 infection in adults from the municipality of Vitoria, ES. Methods: A cross sectional study was performed from September 2010 to December 2011, in individuals of both sexes, aged 18 or older living in Vitoria-ES. Venous blood samples were collected and tested for anti-HTLV-1/2 antibodies by chemiluminescent immunoassay (CMIA). Individuals with CMIA reactive results were submitted to a new blood collection for retesting by CMIA, followed by PCR to confirm infection and discriminate the viral type. Results: From 1502 tested samples, eight were reactive in CMIA and all were confirmed by PCR. Therefore, the prevalence of HTLV-1/2 was 0.53% (8/1502, 95% CI: 0.2-1.0%). The infection rate was 0.7% in men (5/711, 95% CI: 0.17-1.51%), and 0.38% in women (3/791, 95% CI: 0 -0.81%). Conclusions: The prevalence of HTLV-1/2 infection was 0.53% (8/1502; 95% CI: 0.2 -0.9%). Confirmatory test using real-time PCR (qPCR) identified seven individuals positive for HTLV-1 and one for HTLV-2. Considering the risk of infected individuals to develop high morbidity and mortality diseases, it would be important to implement public health policies aimed at stopping transmission of these viruses in this municipality. 2021 Sociedade Brasileira de Infectologia. Published by Elsevier España, S.L.U. This is an open access article under the CC BY-NC-ND license

HTLV-2 infection in Manaus, Brazil: first description of HTLV-2c subtype in an urban area of the Western Amazon region

Abstract INTRODUCTION: We investigated the prevalence of human T-cell lymphotropic virus types 1 and 2 (HTLV-1/2) infection in patients with hematological diseases from the western Amazon region of Brazil. METHODS: Samples from 306 patients were submitted for the molecular diagnosis of HTLV-1/2 infection by real time PCR (qPCR), with amplification, sequencing, and phylogenetic analysis of the long terminal repeat (LTR) region. RESULTS: A 29-year-old male carrier of sickle cell anemia with a history of multiple blood transfusions was diagnosed with the HTLV-2c subtype. CONCLUSIONS: This study describes the first known occurrence of HTLV-2c in the urban area of Brazil's western Amazon region.

Provirus Mutations of Human T-Lymphotropic Virus 1 and 2 (HTLV-1 and HTLV-2) in HIV-1-Coinfected Individuals

Provirus mutations of human T-lymphotropic virus 1 (HTLV-1), mostly the lack of the 5= long terminal repeat (LTR) genomic region, have been described and associated with severe adult T cell leukemia/lymphoma (ATLL), non-sense point mutations with low proviral load, and Western blotting indeterminate results. Until now, no information concerning provirus mutations of HTLV-2 and its consequences, as well as those of HTLV-1/2 in HIV-coinfected individuals, had been described. Therefore, we searched for these mutations in provirus samples of 44 HIV/HTLV-1- and 25 HIV/HTLV-2-coinfected individuals. Using protocols well established for amplification and sequencing of segments of the LTR, env, and tax regions, we searched for defective type 1 particles that retain LTRs and lack internal sequences and type 2 particles that lack the 5=LTR region. In addition, using as references the prototypes ATK (HTLV-1) and Mo (HTLV-2), we searched for point mutations in the LTR and synonyms and nonsynonymous mutations and non-sense mutations in env and tax regions. Defective HTLV-1 and HTLV-2 provirus type 1 or 2 was detected in 31.8% of HIV/HTLV-1- and 32.0% of HIV/HTLV-2-coinfected individuals. Synonymous and nonsynonymous mutations were identified mostly in HTLV-2 and associated with lower levels of specific antibodies. No non-sense mutations that resulted in premature termination of Env and Tax proteins were detected. On the contrary, mutation in the stop codon of Tax2a produced a long protein characteristic of the HTLV-2c subtype. The clinical significance of these mutations in coinfected individuals remains to be defined, but they confirmed the lower sensitivity of serological and molecular diagnostic tests in HIV/HTLV-1/2 coinfections. IMPORTANCE HTLV-1 and HTLV-2 are endemic to Brazil, and they have different effects in HIV/AIDS disease progression. HIV/HTLV-1 has been described as accelerating the progression to AIDS and death, while HIV/HTLV-2 slows the progression to AIDS. Provirus mutations of HTLV-1 were implicated in severe leukemia development and in problems in the diagnosis of HTLV-1; in contrast, provirus mutations of HTLV-2 had not been confirmed and associated with problems in HTLV-2 diagnosis or disease outcome. Nevertheless, data obtained here allowed us to recognize and understand the false-negative results in serologic and molecular tests applied for HTLV-1 and HTLV-2 diagnosis. Defective proviruses, as well as synonymous and nonsynonymous mutations, were associated with the diagnosis deficiencies. Additionally, since HIV-1 and HTLV-1 infect the same cells (CD4 positive), the production of HIV-1 pseudotypes with HTLV-1 envelope glycoprotein during HIV/HTLV-1 coinfection cannot be excluded. Defective provirus of HTLV-2 and Tax2c is speculated to influence progression to AIDS. (AU)

Infección por virus HTLV-1/2 confirmada por serología y detección de provirus en pacientes ecuatorianos de paraparesis espástica tropical / HTLV-1/2 virus infection confirmed by serology and provirus DNA detection in tropical spastic paraparesis Ecuadorian patients

Introduccion: La infección con el virus linfotrópico de células T humano (HTLV) de tipo 1 y 2 es endémica en América del Sur. Existen muy pocos reportes clínicos de este retrovirus en pacientes de Ecuador y nunca confirmaron la presencia del virus con el aislamiento o la detección de su ADN. Esta infección se encuentra desatendida por las autoridades de salud pública, sin que existan estudios epidemiológicos de prevalencia a nivel del país. Objetivos: Este estudio tiene como objetivo la detección de infección por HTLV -1/2 en individuos sintomáticos de paraparesis espástica tropical (TSP) utilizando por primera vez en Ecuador diagnóstico serológico y la detección del provirus por biología molecular. Diseño del estudio. Se tomaron muestras de nueve pacientes con un diagnóstico de mielopatía y sospechoso de TSP, que fueron analizadas para la detección del virus HTLV-1/2 usando tres metodologías: ELISA comercial, ensayo de inmunofluorescencia indirecta (IF) y PCR anidada. Resultados: Cinco de los 9 (55.5%) pacientes fueron positivos tanto para la prueba de ELISA como para IF y PCR anidada. Conclusión: La alta prevalencia de infección por HTLV-1/2 entre individuos sintomáticos de mielopatía muestra la endemicidad de este retrovirus en Ecuador, la asociación de HTLV-1/2 con TSP y la necesidad de implementar estrategias de control y prevención para evitar la diseminación de esta enfermedad infecciosa desatendida.

Background: Infection with the Human T-cell lymphotropic virus (HTLV) type 1 and 2 is endemic in South America. There are only a few clinical reports of this retrovirus in patients in Ecuador, without employing virus-isolation methods or viral DNA detection. The infection is usually neglected in health care settings and by public health authorities and no country wide prevalence data are available. Objectives: This study aims to detect the HTLV-1/2 infection in symptomatic individuals for tropical spastic paraparesis (TSP) using for the first time in Ecuador both immunological and molecular biology methodologies. Study Design: Blood samples of 9 symptomatic individuals with myelopathy were analyzed for the detection of the HTLV-1/2 virus with three methods: ELISA, an indirect immunofluorescence (IF) assay and a nested-PCR. Results: Five out of 9 (55.5%) myelopathy patients were positive for ELISA, IF and nested- HTLV-1/2 PCR. Conclusion. The high prevalence of HTLV-1/2 infection among myelopathy patients shows the endemicity of this retrovirus in Ecuador, the association of HTLV-1/2 infection with TSP and the need of the implementation of control and prevention interventions to overcome the further dissemination of this neglected infectious disease.

Hepatitis C viral load in HCV-monoinfected and HCV/HIV-1-, HCV/HTLV-1/-2-, and HCV/HIV/HTLV-1/-2-co-infected patients from São Paulo, Brazil

ABSTRACT Co-infections of hepatitis C virus (HCV) and either human immunodeficiency virus type 1 (HIV-1), human T-cell lymphotropic virus type 1 (HTLV-1) or type 2 (HTLV-2) have been described as having an impact on HCV viremia and subsequent disease progression. HCV load in serum samples from 622 patients (343 males, 279 females; median age 50.8 years) from São Paulo/southeast Brazil was analyzed using the Abbott Real Time HCV assay (Abbott Molecular Inc., IL, USA). Samples were obtained from HCV-monoinfected (n = 548), HCV/HIV-1- (n = 41), HCV/HTLV-1- (n = 16), HCV/HTLV-2- (n = 8), HCV/HIV/HTLV-1- (n = 4), and HCV/HIV/HTLV-2-co-infected (n = 5) patients, and results were compared among the groups and according to sex. The median HCV load in HCV-monoinfected patients was 5.23 log10 IU/mL and 0.31 log10 higher in men than in women. Increases in viral load of 0.51 log10, 0.54 log10, and 1.43 log10 IU/mL were detected in HCV/HIV-1-, HCV/HTLV-1- and HCV/HIV/HTLV-1-co-infected individuals, respectively, compared with HCV-monoinfected counterparts. In contrast, compared to HCV/HIV co-infected patients, HCV/HTLV-2-co-infected patients had an HCV load of 5.0 log10 IU/mL, whereas HCV/HIV/HTLV-2-co-infected patients had a median load 0.37 log10 IU/mL lower. Significant differences in HCV loads were detected, with males and HCV/HIV-1- and HCV/HIV/HTLV-1-co-infected patients presenting the highest values. Conversely, females and HCV/HTLV-2-co-infected patients exhibited lower HCV loads. Overall, HCV viremia is increased in HIV and/or HTLV-1-co-infection and decreased in HTLV-2 co-infection.

Hepatitis C viral load in HCV-monoinfected and HCV/HIV-1-, HCV/HTLV-1/-2-, and HCV/HIV/HTLV-1/-2-co-infected patients from São Paulo, Brazil

Co-infections of hepatitis C virus (HCV) and either human immunodeficiency virus type1 (HIV-1), human T-cell lymphotropic virus type 1 (HTLV-1) or type 2 (HTLV-2) have beendescribed as having an impact on HCV viremia and subsequent disease progression. HCVload in serum samples from 622 patients (343 males, 279 females; median age 50.8 years)from São Paulo/southeast Brazil was analyzed using the Abbott Real Time HCV assay(Abbott Molecular Inc., IL, USA). Samples were obtained from HCV-monoinfected (n = 548),HCV/HIV-1- (n = 41), HCV/HTLV-1- (n = 16), HCV/HTLV-2- (n = 8), HCV/HIV/HTLV-1- (n = 4), andHCV/HIV/HTLV-2-co-infected (n = 5) patients, and results were compared among the groupsand according to sex. The median HCV load in HCV-monoinfected patients was 5.23 log10IU/mL and 0.31 log10higher in men than in women. Increases in viral load of 0.51 log10, 0.54log10, and 1.43 log10IU/mL were detected in HCV/HIV-1-, HCV/HTLV-1- and HCV/HIV/HTLV-1-co-infected individuals, respectively, compared with HCV-monoinfected counterparts. Incontrast, compared to HCV/HIV co-infected patients, HCV/HTLV-2-co-infected patients hadan HCV load of 5.0 log10IU/mL, whereas HCV/HIV/HTLV-2-co-infected patients had a medianload 0.37 log10IU/mL lower. Significant differences in HCV loads were detected, with malesand HCV/HIV-1- and HCV/HIV/HTLV-1-co-infected patients presenting the highest values.Conversely, females and HCV/HTLV-2-co-infected patients exhibited lower HCV loads. Over-all, HCV viremia is increased in HIV and/or HTLV-1-co-infection and decreased in HTLV-2co-infection. (AU)

Prevalência de infecção pelos vírus linfotrópicos de células T humanas dos tipos 1 e 2 (HTLV-1 e HTLV-2) e vírus da imunodificiência humana do tipo 1 (HIV-1) em população infectada pelos vírus da hepatite B (HBV) e hepatite C (HCV)

Os vírus linfotrópicos de células T humanas dos tipos 1 (HTLV-1) e 2 (HTLV2),assim como o vírus da imunodeficiência humana (HIV) e os vírus dashepatites B (HBV) e C (HCV) compartilham vias de transmissão, portantocoinfecções por estes vírus podem acontecer e alterar o curso das doençasa eles relacionadas. O presente estudo avaliou a prevalência de infecção porHTLV-1/2 em população com hepatite B e C, infectada ou não pelo HIV, e oimpacto das coinfecções na viremia HBV e HCV. O estudo foi realizado em1.244 amostras de plasma/soro enviadas ao Instituto Adolfo Lutz de SãoPaulo para determinação de carga viral (CV) de HBV e HCV: 622 depacientes com HBV (G1, 327 homens e 295 mulheres, média de idade 45,8anos) e 622 de pacientes com HCV (G2, 343 homens e 279 mulheres, médiade idade de 50,8 anos). A triagem de HTLV-1/2 foi realizada por ensaioimunoenzimático (EIA HTLV-I/II, Gold ELISA, REM) e confirmadas porWestern Blot (HTLV BLOT 2.4, MP Biomedicals) e imunoensaio de linha(INNO-LIA HTLV-I/II, Fujirebio). A pesquisa de infecção por HIV foi realizadapor teste imunocromatográfico (kit Rapid Check HIV 1 e 2, NDI, UniversidadeFederal do Espírito Santo, Brasil) seguido do EIA (GS HIV-1/HIV-2 Plus OEIA, Bio-Rad). A infecção por HTLV-1 foi confirmada em 25 amostras (cincono G1 e 20 no G2)...

The human T-cell lymphotropic virus types 1 and 2 (HTLV-1 and HTLV-2) aswell as the human immunodeficiency virus (HIV) and the hepatitis B virus(HBV) and hepatitis C virus (HCV) share routes of virus transmission; thusco-infections with such viruses can occur and alter the course of subsequentdiseases. The present study aimed at evaluating the prevalence of HTLV-1/-2 in blood samples of individuals with hepatitis B and C, infected or not byHIV, and the impact of co-infections on the HBV and HCV viremia. The studywas conducted with 1,244 plasma/serum samples sent to Instituto AdolfoLutz of São Paulo for measuring HCV and HBV viral load (VL): 622 fromHBV-infected patients (G1, 327 male and 295 female, median age 45.8years), and 622 from HCV-infected patients (G2, 343 male and 279 female,median age 50.8 years). HTLV-1/-2 antibodies were screened by enzymeimmunoassay (EIA, HTLV-I/II, Gold ELISA, REM), and confirmed by Westernblot (HTLV BLOT 2.4, MP Biomedicals), and line immunoassay (INNO-LIAHTLV-I/II, Fujirebio). The HIV infection was detected byimmunochromatographic assay (Rapid Check HIV 1 e 2, NDI, UniversidadeFederal do Espírito Santo, Brasil) and by EIA (GS HIV-1/HIV-2 Plus O EIA,Bio-Rad). HTLV-1 was confirmed in 25 samples (5 in G1 and 20 in G2)...

Desafios no diagnóstico da infecção pelos Vírus Linfotrópicos de Células T Humanas do tipo 1 e tipo 2 (HTLV-1 e HTLV-2) em pacientes infectados com o HIV-1 / Challenges in the diagnosis of Human T-Cell Lymphotropic Virus type 1 and type 2 (HTLV-1 and HTLV-2) infection in patients infected with HIV-1

Desde a década de 90 o Instituto Adolfo Lutz de São Paulo (IAL) tem realizado o diagnóstico da infecção por Vírus Linfotrópicos de Células T Humanas dos tipos 1 e 2 (HTLV-1 e HTLV-2) e, desde então, têm sido reportadas as dificuldades principalmente no diagnóstico de HTLV-2, em especial em pacientes infectados pelo HIV-1. O presente trabalho teve como objetivo avaliar várias técnicas de diagnóstico disponíveis no momento atual (kits comerciais e testes in house) e estabelecer o melhor algoritmo para ser empregado no diagnóstico de pacientes infectados pelo HIV-1. A população analisada foi composta por dois grupos provenientes de Serviços de Assistência Especializados em HIV/AIDS de São Paulo: um pioneiro [Grupo 1 (G1), n=1.608] e outro com histórico mais recente [Grupo 2 (G2), n=1.383]. Ambos os grupos foram formados, na maioria, por indivíduos do sexo masculino... (AU).

Since the 90 decade, the Instituto Adolfo Lutz (IAL) has performed the diagnosis of Human T-cell Lymphotropic Virus type 1 and type 2 (HTLV-1 and HTLV-2), and thenceforth the difficulties in diagnosing HTLV-2 have been reported, mostly in HIV-infected patients. The present study aimed at evaluating the several diagnostic techniques currently available (commercial kits and in-house assays), and to establish the best algorithm to be employed for diagnosing HTLV-1/-2 in patients infected with HIV-1. The study population was composed by two patient groups attended at HIV/AIDS specialized services care in São Paulo: the pioneer one [Group 1 (G1), n=1,608], and the other with the most recent historical health setting [Group 2 (G2), n=1,383. The majority of the both groups were composed by male patients...(AU).

Origin and prevllence of human t-lymphotropic vrus type 1 (htlv-1) and type 2 (htlv-2) among indigenous populations in the americas / Origem e prevalência do vírus linfotrópico de células T humanas em populações indígenas das Américas

Human T-lymphotropic virus type 1 (HTLV-1) is found in indigenous peoples of the Pacific Islands and the Americas, whereas type 2 (HTLV-2) is widely distributed among the indigenous peoples of the Americas, where it appears to be more prevalent than HTLV-1, and in some tribes of Central Africa. HTLV-2 is considered ancestral in the Americas and is transmitted to the general population and injection drug users from the indigenous population. In the Americas, HTLV-1 has more than one origin, being brought by immigrants in the Paleolithic period through the Bering Strait, through slave trade during the colonial period, and through Japanese immigration from the early 20th century, whereas HTLV-2 was only brought by immigrants through the Bering Strait. The endemicity of HTLV-2 among the indigenous people of Brazil makes the Brazilian Amazon the largest endemic area in the world for its occurrence. A review of HTLV-1 in all Brazilian tribes supports the African origin of HTLV-1 in Brazil. The risk of hyperendemicity in these epidemiologically closed populations and transmission to other populations reinforces the importance of public health interventions for HTLV control, including the recognition of the infection among reportable diseases and events.

O vírus linfotrópico de células T humanas do tipo 1 (HTLV-1) é encontrado em populações indígenas de países do Pacífico e Américas enquanto o tipo 2 (HTLV-2) é amplamente distribuído entre as populações indígenas das Américas, nas quais aparenta ser mais prevalente que o HTLV-1, e em algumas tribos da África Central, sendo considerado ancestral nas Américas e transmitido à população geral e de usuários de drogas injetáveis a partir da população indígena. No continente americano o HTLV-1 teria mais de uma origem, sendo trazido na era paleolítica pelos imigrantes através do estreito de Bering, através do tráfico de escravos no período colonial e com a imigração japonesa a partir do início do século XX, enquanto para o HTLV-2 teria sido trazido pelos imigrantes através do estreito de Bering. A endemicidade do HTLV-2 entre os indígenas do Brasil tornam a região amazônica brasileira a maior área endêmica do mundo para sua ocorrência e a revisão da infecção pelo HTLV-1 em todas as tribos brasileiras apoiam a origem africana do HTLV-1 no Brasil. O risco de hiperendemicidade nestas populações epidemiologicamente fechadas e de transmissão a outras populações reforçam a importância de medidas no âmbito da saúde pública para seu controle, incluindo o reconhecimento da infecção entre os agravos e eventos de notificação compulsória.

Vírus linfotrópicos de células T humanas dos tipos 1 e 2 (HTLV-1 e HTLV-2): estudo de segmentos do genoma proviral obtidos de pacientes com HIV/Aids de São Paulo e de Londrina e região / Human T-lymphotropic virus type 1 and 2 (HTLV-1 and HTLV-2): study on proviral genome segments obtained from patients with HIV/Aids from Sao Paulo and Londrina and vicinities

O Brasil é considerado o país com o maior número absoluto de indivíduos infectados pelos vírus linfotrópicos de células T humanas dos tipos 1 e 2 (HTLV-1 e HTLV2), perto de 2,5 milhões; além disso, é também considerado epidêmico para o HIV e, portanto, casos de coinfecção HIV/HTLV são frequentes no país. O presente trabalho efetuou o seqüenciamento das regiões LTR, env e tax do genoma proviral do HTLV-1 e do HTLV-2 isolados das amostras de sangue de pacientes coinfectados pelo HIV-1 de Londrina e região (n=34) e de São Paulo (n=20), para realizar a caracterização molecular e determinar subtipos virais. Foram utilizadas na análise das sequências as ferramentas Sequencher 4.7, BLAST, Genotyping-NCBI, Subtyping-REGA, BioEdit 7.0.5.3, ClustalW, GenBank, PAUP 4.0.b10, Modeltest 3.7, TreeView 1.6.6 e MEGA4. As diversas análises confirmaram como subtipos prevalentes o HTLV-1a, subgrupo Transcontinental A, e o HTLV-2a (variante -2c). Foram detectadas assinaturas moleculares nos isolados do Brasil. Detectou-se o genótipo brasileiro taxA para o HTLV-1 e para o HTLV-2 a Tax longa, a qual é característica da variante HTLV-2c. Houve também a confirmação da troca de aminoácido S1909P no env dos HTLV-2. Especulou-se sobre duas entradas do HTLV-1 no Brasil e sobre a disseminação do HTLV-2c em grupos distintos quanto ao comportamento de risco e região geográfica. O estabelecimento de métodos laboratoriais otimizados para isolados brasileiros de HTLV-1 e HTLV-2 possibilitou melhor compreensão da diversidade genômica e da origem e disseminação dos HTLVs em populações coinfectadas pelo HIV no Brasil

Brazil is considered the country with the major absolute number of individuals infected with human T-lymphotropic virus types 1 and 2 (HTLV-1 and HTLV-2), close to 2,500,000; moreover, it is also considered epidemic for HIV/Aids =and therefore HIV/HTLV coinfection is frequent in the country. This study aimed at sequencing the LTR, env and tax regions of the proviral genome of HTLV-1 and HTLV-2 isolated from blood samples obtained from patients coinfected with HIV-1 from Londrina and vicinities (n=34) and São Paulo (n=20), in order to perform the molecular characterization and viral subtyping. For sequences analysis, several bioinformatics tools were employed: Sequencher 4.7, BLAST, Genotyping-NCBI, Subtyping-REGA, BioEdit 7.0.5.3, ClustalW, GenBank, PAUP 4.0.b10, Modeltest 3.7, TreeView 1.6.6 and MEGA4. The results confirmed as prevalent the HTLV-1a subtype, the Transcontinental subgroup A, and the HTLV-2a (variant-2c). Molecular signatures characteristic of Brazilian isolates were detected: taxA Brazilian genotype in HTLV-1, and the long Tax which is characteristic of the HTLV-2c in HTLV-2. Also, it was confirmed the S1909P amino acid change in the env region of HTLV-2c. It was speculated on two entrances of HTLV-1 in Brazil, and on the spread of HTLV-2c in distinct groups related to risk factors and geographic region. The establishment and optimization of laboratory methods performed in this study allowed to get a better understanding on HTLVs genomic diversity, and to give insights on the origin and spread of HTLVs in populations coinfected with HIV in Brazil

Diagnóstico de infecção por vírus linfotrópicos de células T humanas dos tipos 1 (HTLV-1) e -2 (HTLV-2) em população de risco: passado, presente e futuro / Diagnosis of human T-cell lymphotropic virus type 1 (HTLV-1) and type 2(HTLV-2) infection in at-risk populations: past, present and future

O Brasil é o país com o maior número de pessoas infectadas pelos vírus linfotrópicos de células T humanas dos tipos 1e -2 (HTLV-1 e HTLV–2) com mais de 2,5 milhões de indivíduos infectados. Em 1993, a realização de testes sorológicos específicos tornou-se obrigatória em Bancos de Sangue. O HTLV-1 causa leucemia/linfoma de células T do adulto e mielopatia associada ao HTLV-1/paraparesia espástica tropical além de outras doenças, enquanto o HTLV-2 pode causar alguns quadros neurológicos e alterar a evolução de HIV/Aids. Os testes sorológicos que identificam anticorpos específicos disponíveis no mercado têm falhado no diagnóstico, principalmente de infecção por HTLV-2. Vários algoritmos de testes de triagem e confirmatórios têm sido propostos, mas nenhum deles se mostrou 100% eficiente com casuística de alto risco. Muitos soros resultam em padrão indeterminado no Western blot, e os isolados virais utilizados na composição dos kits podem ser a causa desses resultados. As técnicas de biologia molecular têm sido descritas como testes confirmatórios, mas não têm sido empregadas na rotina. Desde 1991, a Seção de Imunologia do Instituto Adolfo Lutz tem estudado a infecção por HTLV-1/2, contribuindo para o diagnóstico sorológico e molecular, e tem como desafio implantar um teste laboratorial capaz de detectar infecção causada por cepas brasileiras de HTLV-2.

Confirming the presence of HTLV-1 infection and the absence of HTLV-2 in blood donors from Arequipa, Peru / Confirmação da presença de infecção por HTLV-1 e ausência de infecção por HTLV-2 em doadores de sangue de Arequipa, Peru

Epidemiological studies conducted in Peru disclosed HTLV-1 to be prevalent in different ethnic groups, and found HTLV-2 in some Amazonian Indians and in men who have sex with men. No data concerning HTLV-1/2 infection in blood donors from Arequipa, a highlands region in southern Peru, is available. We searched for the presence of HTLV-1 and HTLV-2 antibodies in 2,732 serum samples obtained from blood donors from this geographic area. HTLV-1/2-specific antibodies were detected using an enzyme-linked immunosorbent assay (ELISA) and were confirmed by Western blot (WB). Reactive sera had their blood bags discarded from donation, and the demographic characteristics of the donors were analyzed. Thirty-five sera (1.2 percent) were HTLV seroreactive by ELISA, and 25 were confirmed HTLV-1-positive by WB. One serum disclosed HTLV-positivity, and the remaining nine serum samples showed indeterminate results by WB; three of which had an HTLV-1 indeterminate Gag profile. The median age of HTLV-positive individuals was 34.6 years; 27 were male and eight were female. All individuals were from southern Peru: 27 from Arequipa, five from Puno, and three from Cuzco. HTLV co-positivity with hepatitis B (five sera) and syphilis (one serum) were detected. Previous transfusion and tattooing were observed in two and one individuals, respectively. No serum was positive for HTLV/HIV co-infection. This study confirmed, for the first time, HTLV-1 infection and the absence of HTLV-2 infection in blood donors from Arequipa, Peru and suggests vertical transmission as the major route of HTLV-1 transmission and acquisition in this geographic region.

Estudos epidemiológicos conduzidos no Peru apontam a infecção por HTLV-1 como prevalente em diferentes grupos étnicos e por HTLV-2 restrita a alguns índios da região Amazônica e a homens que fazem sexo com homens. Não existem dados sobre a infecção por HTLV-1/2 em doadores de sangue de Arequipa, região montanhosa do sul do Peru. Portanto, o presente estudo pesquisou anticorpos anti-HTLV-1 e HTLV-2 em 2.732 doadores de sangue desta região geográfica. Foram utilizados na triagem sorológica os testes imunoenzimáticos (ELISA) e para confirmação dos resultados o Western Blot (WB). Soros reagentes no ELISA tiveram suas bolsas de sangue descartadas. Os resultados obtidos foram analisados de acordo com características demográficas dos indivíduos. Trinta e cinco soros (1,2 por cento) resultaram HTLV-1/2 reagentes no ELISA, 25 confirmaram infecção por HTLV-1 no WB. Um soro resultou HTLV positivo e os nove soros restantes resultaram em padrão indeterminado no WB: três com perfil HTLV-1 Gag indeterminado. A média de idade dos indivíduos HTLV positivos foi de 34,6 anos; 27 do gênero masculino e oito do gênero feminino. Todos eram da região sul do país: 27 de Arequipa, cinco de Puno e três de Cuzco. Foi detectada co-positividade HTLV com hepatite B (cinco soros) e sífilis (um soro). Nenhum soro resultou positivo para a co-infecção HIV/HTLV. Havia dois indivíduos com transfusão prévia e um com tatuagem. Este trabalho confirma pela primeira vez infecção por HTLV-1 e ausência de infecção por HTLV-2 em doadores de sangue de Arequipa, sul do Peru e sugere que a transmissão vertical seja a principal via de transmissão/aquisição de HTLV-1 nesta região geográfica.

Difficulties in the diagnosis of HTLV-2 infection in HIV/AIDS patients from Brazil: comparative performances of serologic and molecular assays, and detection of HTLV-2b subtype

The current diagnosis of human T-lymphotropic virus type-2 (HTLV-2) infection is based on the search of specific antibodies; nevertheless, several studies conducted in Brazil pointed deficiencies of the commercially available kits in detecting HTLV-2, mostly in HIV/AIDS patients. This study searched for the presence of HTLV-1 and -2 in 758 HIV/AIDS patients from Londrina, Paraná, Brazil. Serum samples were screened for HTLV-1/2 antibodies using two EIA kits (Vironostika and Murex), and confirmed by WB (HTLV Blot 2.4, Genelabs). The results obtained by EIA disclosed 49 (6.5 percent) reactive sera: 43 positive by both EIA kits, and six with discordant results. WB confirmed HTLV-1 infection in seven samples (0.9 percent) and HTLV-2 in 21 sera (2.8 percent). Negative and indeterminate results were detected in four (0.5 percent) and 16 (2.1 percent) sera, respectively. Blood from 47 out of 49 HTLV seroreactive patients were collected and analyzed for the presence of env, LTR and tax genomic segments of HTLVs by PCR. PCR confirmed six cases of HTLV-1 and 37 cases of HTLV-2 infection (14 out of 16 that were found to be WB indeterminate). Restriction analysis of the env PCR products of HTLV-2 disclosed 36 isolates of HTLV-2a/c subtype, and one of HTLV-2b subtype. These results emphasize the need of improving serologic tests for detecting truly HTLV-2 infected patients from Brazil, and confirm the presence of HTLV-2b subtype in the South of this country.

O diagnóstico de infecção por HTLV-2 se baseia na pesquisa de anticorpos específicos, entretanto, vários estudos conduzidos no Brasil têm apontado falhas nos kits sorológicos disponíveis no mercado em detectar HTLV-2, principalmente nos pacientes com HIV/aids. Este trabalho avaliou a presença de infecção por HTLV-1 e -2 em 758 pacientes HIV/aids de Londrina, Paraná, Brasil. Amostras de soro foram analisadas quanto à presença de anticorpos anti-HTLV-1/2 por dois kits de EIA (Vironostika e Murex) e confirmados por WB (HTLV Blot 2.4, Genelabs). Os resultados obtidos pelos testes sorológicos mostraram 49 (6,5 por cento) soros reagentes: 43 positivos para ambos os kits e seis com resultados discordantes. O WB confirmou infecção por HTLV-1 em sete soros (0,9 por cento) e HTLV-2 em 21 soros (2,8 por cento). Resultados negativos e indeterminados foram detectados, respectivamente, em quatro (0,5 por cento) e 16 (2,1 por cento) soros. Amostras de sangue de 47 dos 49 pacientes com sorologia reagente foram avaliadas quanto à presença de segmentos do genoma dos HTLVs (env, LTR e tax), usando a técnica de PCR. As PCRs confirmaram seis casos de infecção por HTLV-1 e 37 casos por HTLV-2 (14 dos 16 cuja sorologia resultou WB indeterminada). A subtipagem de HTLV-2 por análise de restrição enzimática de produtos da PCR env mostrou 36 isolados de subtipo HTLV-2a/c e um HTLV-2b. Esses resultados reforçam a necessidade de melhorar o diagnóstico de infecção por HTLV-2 no Brasil e confirmam a presença do subtipo HTLV-2b na região sul do país.