Efficiency of indirect immunofluorescence assay as a confirmatory test for the diagnosis of human retrovirus infection (HIV-1 and HTLV-I/II) in different at risk populations

We compared the indirect immunofluorescence assay (IFA) with Western blot (Wb) as a confirmatory method to detect antibodies anti retrovirus (HIV-1 and HTLV-I/II). Positive and negative HIV-1 and HTLV-I/II serum samples from different risk populations were studied. Sensitivity, specificity, positive, negative predictive and kappa index values were assayed, to assess the IFA efficiency versus Wb. The following cell lines were used as a source of viral antigens: H9 ( HTLV-III b); MT-2 and MT-4 (persistently infected with HTLV-I) and MO-T (persistently infected with HTLV-II). Sensitivity and specificity rates for HIV-1 were 96.80% and 98.60% respectively, while predictive positive and negative values were 99.50% and 92.00% respectively. No differences were found in HIV IFA performance between the various populations studied. As for IFA HTLV system, the sensitivity and specificity values were 97.91% and 100% respectively with positive and negative predictive values of 100% and 97.92%. Moreover, the sensitivity of the IFA for HTLV-I/II proved to be higher when the samples were tested simultaneously against both antigens (HTLV-I-MT-2 and HTLV-II-MO-T). The overall IFA efficiency for HIV-1 and HTLV-I/II-MT-2 antibody detection probed to be very satisfactory with an excellent correlation with Wb (Kappa indexes 0.93 and 0.98 respectively). These results confirmed that the IFA is a sensitive and specific alternative method for the confirmatory diagnosis of HIV-1 and HTLV-I/II infection in populations at different levels of risk to acquire the infection and suggest that IFA could be included in the serologic diagnostic algorithm.

Immunofluorescence assay reactivity patterns of serum samples presenting indeterminate Western blot results for antibodies to HIV-1 and HTLV-I/II in Cordoba, Argentina

Serum samples (n: 110) from blood donors and high risk individuals from Cordoba, Argentina with indeterminate HIV-1 and HTLV-I/II Wb profiles were studied for specific antibodies to HTLV-I/II and HIV-1 by indirect immunofluorescence assay (IFA) and for the presence or absence of HIV-1 and HTLV-I/II specific bands by Wb. This study was carried out in order to characterize their putative reactions with HIV-1 and HTLV-I/II proteins and to resolve the retrovirus infection status of these individuals. Results indicated that blood donors sera displaying indeterminate HIV-1 or HTLV-I/II Wb patterns were not immunoreactive to HTLV-I/II and HIV-1 on IFA. However, a high rate of indeterminate HIV-1 and HTLV-I/II Wb samples from high risk individuals had positive HTLV-I/II and HIV-1 IFA results respectively. Our study supports the growing evidence that HTLV-HIV indeterminate seroreactivity in low risk population is due to a cross reaction against nonviral antigens, and in high risk populations the indeterminate samples show serological cross-recognition between HIV-1 proteins and HTLV-I/II proteins on Wb. These results point out the necessity to investigate the HTLV-I/II reactivity in indeterminate HIV-1 samples and viceversa in order to confirm the diagnosis. Finally, this study shows the potential usefulness of IFA in elucidating the status of HIV-1 and HTLV-I/II infection of individuals with indeterminate Wb profiles, thus enabling resolution of retrovirus infection status.

McCoy cell line as a possible model containing CD4+ receptors for the study of HIV-1 replication

Several studies have recently shown the use of recombinant rabies virus as potential vector-viral vaccine for HIV-1. The sequence homology between gp 120 and rabies virus glycoprotein has been reported. The McCoy cell line has therefore been used to show CD4+ or CD4+ like receptors. Samples of HIV-1 were isolated, when plasma of HIV-1 positive patients was inoculated in the McCoy cell line. The virus infection was then studied during successive virus passages. The proteins released in the extra cellular medium were checked for protein activity, by exposure to SDS Electrophoresis and blotting to nitro-cellulose filter, then reacting with sera of HIV positive and negative patients. Successive passages were performed, and showed viral replication, membrane permeabilization, the syncytium formation, and the cellular lysis (cytopathic effect). Flow cytometry analysis shows clear evidence that CD4+ receptors are present in this cell line, which enhances the likelihood of easy isolation and replication of HIV. The results observed allow the use of this cell line as a possible model for isolating HIV, as well as for carrying out studies of the dynamics of viral infection in several situations, including exposure to drugs in pharmacological studies, and possibly studies and analyses of the immune response in vaccine therapies.

Human T-cell lymphotropic virus types I and II infections in a cohort of patients with neurological disorders in Belém, Pará, Brazil

Serum- and/or- cerebrospinal fluid (CSF) samples obtained from 190 patients suffering from chronic, progressive neurological disease were screened for the presence of human T-cell lymphotropic viruses type I (HTLV-I) and type II (HTLV-II) antibodies over a six-year period (1996 to 2001) in Belém, Pará, Brazil. Patients were of both sexes (male subjects, 52%) with ages ranging from 2 to 79 years (mean, 35.9). Overall, 15 (7.9%) subjects - of whom 12 (80%) were female adults - reacted HTLV-I/II-seropositive when screened by enzyme-linked immunosorbent assay (ELISA). Serum samples from 14 of these patients were also analyzed using a recombinant Western blot (WB) assay that yielded HTLV-I-, HTLV-II-, and HTLV-I/II- reactivities for 10 (71.4%), 3 (21.4%) and 1 (7.2%) of them, respectively. The yearly rates of HTLV-I/II antibodies ranged from 2.6% (2001) to 21.7% (2000), with progressively increasing seropositivities from 1998 to 2000. Altogether, walking difficulty (n = 5 subjects), spasticity (n = 4) and leg weakness (n = 3) accounted for 80% of symptoms recorded among the 15 patients whose sera had antibodies to HTLV-I/II as detected by ELISA. These findings provide evidence that both HTLV-I and HTLV-II play a role in the development of chronic myelopathy in Belém, Pará, Northern Brazil.

Clinical and epidemiological aspects of HTLV-II infection in São Paulo, Brazil: presence of Tropical Spastic Paraparesis/HTLV-Associated Myelopathy (TSP/HAM) simile diagnosis in HIV-1-co-infected subjects

In this study, the epidemiological and clinical features observed in solely HTLV-II-infected individuals were compared to those in patients co-infected with HIV-1. A total of 380 subjects attended at the HTLV Out-Patient Clinic in the Institute of Infectious Diseases "Emilio Ribas" (IIER), São Paulo, Brazil, were evaluated every 3-6 months for the last seven years by infectious disease specialists and neurologists. Using a testing algorithm that employs the enzyme immuno assay, Western Blot and polymerase chain reaction, it was found that 201 (53%) were HTLV-I positive and 50 (13%) were infected with HTLV-II. Thirty-seven (74%) of the HTLV-II reactors were co-infected with HIV-1. Of the 13 (26%) solely HTLV-II-infected subjects, urinary tract infection was diagnosed in three (23%), one case of skin vasculitis (8%) and two cases of lumbar pain and erectile dysfunction (15%), but none myelopathy case was observed. Among 37 co-infected with HIV-1, four cases (10%) presented with tropical spastic paraparesis/HTLV-associated myelopathy (TSP/HAM) simile. Two patients showed paraparesis as the initial symptom, two cases first presented with vesical and erectile disturbances, peripheral neuropathies were observed in other five patients (13%), and seven (19%) patients showed some neurological signal or symptoms, most of them with lumbar pain (five cases). The results obtained suggest that neurological manifestations may be more frequent in HTLV-II/HIV-1-infected subjects than those infected with HTLV-II only.

Induction of phagocytic activity and nitric-oxide production in natural populations of Trypanosoma Cruzi I and II from the state of Paraná, Brazil

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

TOLL-LIKE RECEPTORS (TLR) 2 AND 4 EXPRESSION OF KERATINOCYTES FROM PATIENTS WITH LOCALIZED AND DISSEMINATED DERMATOPHYTOSIS

There are few studies on the role of innate immune response in dermatophytosis. An investigation was conducted to define the involvement of Toll-Like Receptors (TLRs) 2 and 4 in localized (LD) and disseminated (DD) dermatophytosis due to T. rubrum. Fifteen newly diagnosed patients, eight patients with LD and seven with DD, defined by involvement of at least three body segments were used in this study. Controls comprised twenty skin samples from healthy individuals undergoing plastic surgery. TLR2 and TLR4 were quantified in skin lesions by immunohistochemistry. A reduced expression of TLR4 in the lower and upper epidermis of both LD and DD patients was found compared to controls; TLR2 expression was preserved in the upper and lower epidermis of all three groups. As TLR4 signaling induces the production of inflammatory cytokines and neutrophils recruitment, its reduced expression likely contributed to the lack of resolution of the infection and the consequent chronic nature of the dermatophytosis. As TLR2 expression acts to limit the inflammatory process and preserves the epidermal structure, its preserved expression may also contribute to the persistent infection and limited inflammation that are characteristic of dermatophytic infections.

Imunização de camundongos com «vacina» viva avirulenta de Trypanosoma cruzi: II Ensaio da avaliação do tempo de eficiência da vacina

Camundongos "vacinados" com vacina viva avirulenta de T. cruzi se mostraram protegidos contra infecção virulenta, até 20 semanas após a imunização. Essa proteção foi inferior à observada avós 4 semanas de imunização e menor ainda do que a apresentada por animais infectados 4 semanas após a "vacinação" e reinfeetados 15 semanas depois. Neste último caso, a primeira infecção parece ter atuado como uma dose de reforço. O Autor assinala a dificuldade em determinar o tempo de validade da vacina, em virtude da resistência natural apresentada com a idade, mesmo pelos animais mais sensíveis.

Levantamentos seccionais sobre a esquistossomose no Nordeste do Brasil: II. Estado do Rio Grande do Norte

Os autores apresentam os resultados de levantamentos seccionais sóbre a prevalência e a mortndade da esquistossomose mansônica no Estado do Rio Grande do Norte. Os levantamentos foram, feitos em 44 localidades, urbanas e rurais. A prevalência da esquistossomose e de outras helmintoses intestinais foi muito variável. As formas hépato-esplênicas da esquistossomose variaram de 0 a 4%.

Avaliação sorológica da vacina oral, tipo Sabin, contra a poliomielite em região semi - rural: II - Aspectos quantitativos dos anticorpos formados e anticorpos para outros enterovirus

Os autores apresentam dados quantitativos sôbre os anticorpos presentes em vacinados com vacina oral trivalente, contra a poliomielite (Tabela 1). Apos a 3.ª dose de vacina verificou-se um aumento do título geométrico médio da população em relação aos títulos obtidos com duas doses de vacina. Infecções naturais devem ter contribuído para formação de anticorpos para poliomielite na população, ao lado da vacina. Anticorpos para enterovírus não-pólio (Coxsackie B e alguns tipos de vírus ECHO) são apresentados na Tabela II e referem-se a amostras de sôro colhidas quando da 1.ª dose de vacina. Os autores chamam a atenção para a incidência de enterovírus na região, embora poucos sejam os dados ainda disponíveis.

Diagnóstico laboratorial de varíola: II. resultados do segundo ano de atividades, de maio de 1969 a maio de 1970

Os autores relatam os resultados obtidos no diagnóstico laboratorial de varíola, durante o segundo ano de funcionamento de uma unidade montada no Instituto Presidente Castello Branco, da Fundação Instituto Oswaldo Crus, no Rio de Janeiro. O exame de 105 espécimens de crostas e de 76 de líquido vesicular /pustular, forneceu 39 e 34 amostras de vírus da varíola, respectivamente (Tabela 1). A demora em chegar ao laboratório influencia significativamente a taxa de isolamento de vírus (Tabela 2). Foi encontrada estreita relação entre os diagnósticos clínico e laboratorial (Tabela 3), quando possível compará-los. A inoculação em ovos embrionados após 1 a 2 horas do abaixamento da mebrana cório-alantóica, foi considerada como adequada às condições em que são realizados os exames. O laboratório continua a receber regularmente mais especimens para diagnóstico.

Health survey on the Ilha da Conceição, state of Rio de Janeiro, II: selected health characteristics in children

As part of the health survey on the Ilha da Conceição, a 25 percent random sample of households was identified and a health guestionnaire completed at 236 households. These households contained 536 children, of whom 239 were under age six. Prenatal care had been obtained by 70 percent of the mothers during the pregnancies of the youngest children and 83 percent of these children had been born in hospital. The use of available health facilities was reported more frequently for the younger children in comparison to the older children. Over 90 percent of the children had been vaccinated against one or more diseases but only 50 to 60 percent of the children had complete vaccination against pertussis, diphtheria and tetanus. Almost two-thirds of the stool specimens from the children revealed evidence of parasites and were most commonly found in children two to three years of age. Low hemoglobin values were found commonly under age three and hemoglobin leveis ábove 12 grams were not commonly found until age six. Compared with a North American standard for height and weight, proportionately more children on the Ilha da Conceição were found below the 25th and 3rd percentiles. These findings suggest that an improved health status for the children on the Ilha da Conceição would result from a household health record maintained at the island clinic including current information on vaccination status of all children, and a health education program focused on expectant mothers and the well baby clinic program.

Microepidemia de histoplasmose na zona rural de Brasília - DF - 1967: II - Estudos Epidemiológico e Parasitológico da Fonte de Infecção

Neste trabalho são relatadas pesquisas parasitológicas e epiâemiológicas realizadas numa provável fonte de infecção de Histoplasmose da área rural do D. F. - Brasília, onde 14 pessoas contraíram a doença. Os estudos clínico, imunológico e radiológico foram anteriormente descritos. Os autores conseguiram isolar o H. capsulatum do solo da caverna e das vísceras e do sangue de morcegos (Phyllostomus hastatus hastatus, Pallas 1767) que nela habitavam. Resultaram negativas as tentativas de isolamento do fungo de animais sentinelas (cobaios), assim como não se obteve neles a viragem dos testes intradérmicos com histoplasmina. Em impressões de vísceras dos morcegos, constataram-se formas semelhantes as do T. gondii que posteriormente foram isoladas, em camundongos jovens, por inoculação de vísceras maceradas e sangue. Foram encontrados 2 ectoparasitos nos morcegos: Boophilus microplus e um díptero da família Streblidae. O ácaro albergava tripomastigotos do tipo cruzi, não sendo porém conseguido seu isolamento. No tubo digestivo dos quirópteros, foram retirados nematódeos (Histiostrongylus octaeantus) e cestódeos do gênero Mathevotaenia. Foram capturados, em torno da entrada da caverna, 2 exemplares de Cercomys cunicularis apereoide não sendo examinados sob o ponto de vista parasitológico. Testes intradérmicos realizados em 826 habitantes da área resultaram positivos em 184 (22,27%). A gruta, fonte da infecção, está localizada em uma formação calcárea, pertencente à série Bambuí, acreditando-se, pelos aspectos tectônicos, ser da idade siluriana. No Brasil, o isolamento de H. capsulatum de solo, guanos de morcegos e vísceras de roedores já tinham sido realizados; contudo, esta foi a primeira vez que se conseguiu isolá-lo do solo de uma caverna, fonte de infecção, e das vísceras e sangue de morcegos. Os resultados obtidos com os testes intradérmicos com histoplasmina demonstraram a prevalência de histoplasmose na área rural do DF e, conseqüentemente, a possibilidade de ocorrerem outra microepidemia ou aparecimento de casos isolados de histoplasmose-doença.

Contribuição ao estudo da história natural de enteroparasitas em uma comunidade fechada: II. Mecanismos de transmissão de enteroparasitas em uma comunidade fechada

Para estabelecer os mecanismos de transmissão de enteroparasitas entre a população de um orfanato, os autores pesquisaram a presença de ovos de helmintos e cistos de protozoários nas mãos, unhas e roupas das internas; nos objetos e peças de banheiros coletivos e poeira de dormitórios. Realizaram também exame bacteriológico da água que abastece a instituição. Os resultados são apresentados e discutidos.