Impact of Epstein-Barr virus co-infection on natural acquired Plasmodium vivax antibody response.

BACKGROUND: The simultaneous infection of Plasmodium falciparum and Epstein-Barr virus (EBV) could promote the development of the aggressive endemic Burkitt's Lymphoma (eBL) in children living in P. falciparum holoendemic areas. While it is well-established that eBL is not related to other human malaria parasites, the impact of EBV infection on the generation of human malaria immunity remains largely unexplored. Considering that this highly prevalent herpesvirus establishes a lifelong persistent infection on B-cells with possible influence on malaria immunity, we hypothesized that EBV co-infection could have impact on the naturally acquired antibody responses to P. vivax, the most widespread human malaria parasite. METHODOLOGY/PRINCIPAL FINDINGS: The study design involved three cross-sectional surveys at six-month intervals (baseline, 6 and 12 months) among long-term P. vivax exposed individuals living in the Amazon rainforest. The approach focused on a group of malaria-exposed individuals whose EBV-DNA (amplification of balf-5 gene) was persistently detected in the peripheral blood (PersVDNA, n = 27), and an age-matched malaria-exposed group whose EBV-DNA could never be detected during the follow-up (NegVDNA, n = 29). During the follow-up period, the serological detection of EBV antibodies to lytic/ latent viral antigens showed that IgG antibodies to viral capsid antigen (VCA-p18) were significantly different between groups (PersVDNA > NegVDNA). A panel of blood-stage P. vivax antigens covering a wide range of immunogenicity confirmed that in general PersVDNA group showed low levels of antibodies as compared with NegVDNA. Interestingly, more significant differences were observed to a novel DBPII immunogen, named DEKnull-2, which has been associated with long-term neutralizing antibody response. Differences between groups were less pronounced with blood-stage antigens (such as MSP1-19) whose levels can fluctuate according to malaria transmission. CONCLUSIONS/SIGNIFICANCE: In a proof-of-concept study we provide evidence that a persistent detection of EBV-DNA in peripheral blood of adults in a P. vivax semi-immune population may impact the long-term immune response to major malaria vaccine candidates.

Rotavirus G serotypes and p[],G genotypes identified in cases of reinfection among children participating in a trial with rhesus-human reassortant tetravalent vaccine (RRV-TV) in Belém, Brazil.

Group A rotaviruses are the most important agents of severe diarrhea in children and infants worldwide. The aim of present study was to identify rotavirus G serotypes and P[],G genotypes in cases of reinfection among children who participated in a vaccine trial with the tetravalent rhesus-human reassortant rotavirus vaccine (RRV-TV 4 x 10(4) pfu/dose) in Belém, Brazil. From July 1990 to June 1992, 540 children received, at their first, third and fifth months of life, oral doses of either vaccine or placebo. A total of 90 rotavirus diarrheal episodes among children who completed the three-dose vaccination schedule were recorded. We studied 11 reinfection rotavirus cases among five children (three female and two male). Fecal specimens were tested by using a enzyme immunoassay (IDEIA Rotavirus), followed by EIA with monoclonal antibodies to determine infecting serotypes Gl, G2, G3 and G4 and subgroups I and II. The viral dsRNA was extracted and electrophoresed through polyacrylamide gel and then subjected to reverse-transcription-polymerase chain reaction and nested-PCR for the determination of Gl, G2, G3, G4, G5 and G9 and P[4], P[6], P[8] and P[9] rotavirus genotypes. A total of 11 cases of reinfection (12 per cent) occurred among five children, three from the placebo group and two from the vaccine group. In four of the cases of reinfection G serotypes and P[],G genotypes were as follows: for the first and second infections, respectively: (1) G2/P[4],G2 and Gl/P[4],G1; (2) G2/P[4],G2 and G2/P[6],G5; (3) G2/P[4],G2 and G1/P[8],G1; and (4) G2/P[8],G1 and G1/P[8],G1. A fifth child had three successive infections caused by serotypes/genotypes G1/P[8],G1, in the first and second infections, and G2/P[4],G2 in the third infection. The common genotypes and unusual genotypes were detected in 8 (73 per cent) and 3 (27 per cent) of the isolates, respectively. With regards to the clinical severity, in two children a score indicated moderate/severe disease in both first and second infections. One child had three successive infections; the first episode was moderate/severe, the second very severe and the third was not available. In contrast, in two other children, the first episode was very severe, and the second episode was moderate/severe in one child and data was not available for the other child. The results obtained in the present investigation underscore the need to broaden our knowledge of the immunity in rotavirus reinfection. This should be useful regarding future rotavirus vaccination strategies in Brazil.

Nutritional status in relation to the efficacy of the rhesus-human reassortant, tetravalent rotavirus vaccine (RRV-TV) in infants from Belém, pará state, Brazil.

The rhesus-human reassortant, tetravalent rotavirus vaccine (RRV-TV) was licensed for routine use in the United States of America but it was recently withdrawn from the market because of its possible association with intussusception as an adverse event. The protective efficacy of 3 doses of RRV-TV, in its lower-titer (4 x 10(4) pfu/dose) formulation, was evaluated according to the nutritional status of infants who participated in a phase III trial in Belém, Northern Brazil. A moderate protection conferred by RRV-TV was related to weight-for-age Z-scores (WAZ) greater than -1 only, with rates of 38% (p = 0.04) and 40% (p = 0.04) for all- and- pure rotavirus diarrhoeal cases, respectively. In addition, there was a trend for greater efficacy (43%, p = 0.05) among infants reaching an height-for-age Z-score (HAZ) of > -1. Taking WAZ, HAZ and weight-for-height Z-score (WHZ) indices 0.05) if both placebo and vaccine groups are compared. There was no significant difference if rates of mixed and pure rotavirus diarrhoeal cases are compared in relation to HAZ, WAZ and weight-for-height Z-score (WHZ) indices. Although a low number of malnourished infants could be identified in the present study, our data show some evidence that malnutrition may interfere with the efficacy of rotavirus vaccines in developing countries.

Association between human parvovirus B19 and arthropathy in Belém, Pará, North Brazil.

A total of 220 patients with arthropathy were selected in Belém, Pará between January 1994 and December 2000, and screened for the presence of human parvovirus B19 IgM and IgG antibodies by enzyme-linked immunosorbent assay (ELISA). A subgroup (n = 132) of patients with high levels of antibodies (either IgM+/IgG+ or IgM-/IgG+) were examined for the presence of DNA by polymerase chain reaction/nested PCR. Recent/active infection (detection of IgM and/or IgG-specific antibodies and presence of viral DNA) was identified in 47.7% of the 132 individuals with arthropathy. In our study, women were significantly more affected (59.7%) than men (35.4%) (P = 0.0006). The age group of 11-20 years (84.6%), among female patients, and 21-30 years (42.1%), among male, were those with the highest incidence rates. The analysis of the temporal distribution of B19-associated arthropaties showed a cyclic pattern, with peak incidence rates occuring at 3-5 year intervals. Significant diference (P = 0.01) was observed when comparing both the highest (39.0%) and the lowest (11.0%) seropositivity rates for the years of 1995 and 2000, respectively. The interfalangial joints of hands and feet were mostly affected, with 50.0% and 48.0% of cases among both women and men, respectively. In a smaller proportion, other joints such as those of knee, ankle, pulse and shoulder were affected. As for the duration, symptoms lasted 1 to 5 days in 54.0% of the individuals, whereas in 46.0% of them the disease lasted 6-10 days, if considered the subgroup (n = 63) of patients with recent/active infection by parvovirus B19. In our study, joint clinical manifestations occurred symmetrically. Our results indicate that B19 may be an important agent of arthropathies in our region, and this underscores the need for specific laboratory diagnosis when treating patients suffering from acute arthropathy, mainly pregnant women.

Nutritional status in relation to the efficacy of the rhesus-human reassortant, tetravalent rotavirus vaccine (rrv-tv) in infants from Belém, Para state, Brazil

The rhesus-human reassortant, tetravalent rotavirus vaccine (RRV-TV) was licensed for routine use in the United States of America but it was recently withdrawn from the market because of its possible association with intussusception as an adverse event. The protective efficacy of 3 doses of RRV-TV, in its lower-titer (4 x 10(4) pfu/dose) formulation, was evaluated according to the nutritional status of infants who participated in a phase III trial in Belém, Northern Brazil. A moderate protection conferred by RRV-TV was related to weight-for-age Z-scores (WAZ) greater than -1 only, with rates of 38 percent (p = 0.04) and 40 percent (p = 0.04) for all- and- pure rotavirus diarrhoeal cases, respectively. In addition, there was a trend for greater efficacy (43 percent, p = 0.05) among infants reaching an height-for-age Z-score (HAZ) of > -1. Taking WAZ, HAZ and weight-for-height Z-score (WHZ) indices <= -1 together, there was no significant protection (p > 0.05) if both placebo and vaccine groups are compared. There was no significant difference if rates of mixed and pure rotavirus diarrhoeal cases are compared in relation to HAZ, WAZ and weight-for-height Z-score (WHZ) indices. Although a low number of malnourished infants could be identified in the present study, our data show some evidence that malnutrition may interfere with the efficacy of rotavirus vaccines in developing countries

Association between human parvovirus B19 and arthropathy in Belém, Pará, North Brazil

A total of 220 patients with arthropathy were selected in Belém, Pará between January 1994 and December 2000, and screened for the presence of human parvovirus B19 IgM and IgG antibodies by enzyme-linked immunosorbent assay (ELISA). A subgroup (n = 132) of patients with high levels of antibodies (either IgM+/IgG+ or IgM-/IgG+) were examined for the presence of DNA by polymerase chain reaction/nested PCR. Recent/active infection (detection of IgM and/or IgG-specific antibodies and presence of viral DNA) was identified in 47.7 percent of the 132 individuals with arthropathy. In our study, women were significantly more affected (59.7 percent) than men (35.4 percent) (P = 0.0006). The age group of 11-20 years (84.6 percent), among female patients, and 21-30 years (42.1 percent), among male, were those with the highest incidence rates. The analysis of the temporal distribution of B19-associated arthropaties showed a cyclic pattern, with peak incidence rates occuring at 3-5 year intervals. Significant diference (P = 0.01) was observed when comparing both the highest (39.0 percent) and the lowest (11.0 percent) seropositivity rates for the years of 1995 and 2000, respectively. The interfalangial joints of hands and feet were mostly affected, with 50.0 percent and 48.0 percent of cases among both women and men, respectively. In a smaller proportion, other joints such as those of knee, ankle, pulse and shoulder were affected. As for the duration, symptoms lasted 1 to 5 days in 54.0 percent of the individuals, whereas in 46.0 percent of them the disease lasted 6-10 days, if considered the subgroup (n = 63) of patients with recent/active infection by parvovirus B19. In our study, joint clinical manifestations occurred symmetrically. Our results indicate that B19 may be an important agent of arthropathies in our region, and this underscores the need for specific laboratory diagnosis when treating patients suffering from acute arthropathy, mainly pregnant women

Outbreaks of human-herpes virus 6 (HHV-6) infection in day-care centers in Belém, Pará, Brazil

A total of 730 children aged less than 7 years, attending 8 day-care centers (DCCs) in Belém, Brazil were followed-up from January to December 1997 to investigate the occurrence of human-herpes virus 6 (HHV-6) infection in these institutional settings. Between October and December 1997 there have been outbreaks of a febrile- and -exanthematous disease, affecting at least 15-20 percent of children in each of the DCCs. Both serum- and- plasma samples were obtained from 401 (55 percent) of the 730 participating children for the detection of HHV-6 antibodies by enzyme-linked immunosorbent assay (ELISA), and viral DNA amplification through the nested-PCR. Recent HHV-6 infection was diagnosed in 63.8 percent (256/401) of them, as defined by the presence of both IgM and IgG-specific antibodies (IgM+/IgG+); of these, 114 (44.5 percent) were symptomatic and 142 (55.5 percent) had no symptoms (p = 0.03). A subgroup of 123 (30.7 percent) children were found to be IgM-/IgG+, whereas the remaining 22 (5.5 percent) children had neither IgM nor IgG HHV-6- antibodies (IgM-/IgG-). Of the 118 children reacting strongly IgM-positive ( > or = 30 PANBIO units), 26 (22.0 percent) were found to harbour the HHV-6 DNA, as demonstrated by nested-PCR. Taken the ELISA-IgM- and- nested PCR-positive results together, HHV-6 infection was shown to have occurred in 5 of the 8 DCCs under follow-up. Serological evidence of recent infections by Epstein-Barr virus (EBV) and parvovirus B19 were identified in 2.0 percent (8/401) and 1.5 percent (6/401) of the children, respectively. Our data provide strong evidence that HHV-6 is a common cause of outbreaks of febrile/exanthematous diseases among children attending DCCs in the Belém area

Imunidade para varicela entre habitantes de Belém, PA, Brasil / Chickenpox immunity among Belém (PA) Brazil inhabitants

Objetivo: Conhecer o estado imunitário para a varicela em indivíduos de diversas faixas etárias da Cidade de Belém, no Estado do Pará, norte do Brasil, para a tomada de decisäo quanto à indicaçäo de vacinaçäo. Método: Os soros de 525 indivíduos residentes na Cidade de Belém, separados aleatoriamente da soroteca de dois laboratórios privados de análise clínicas e do Instituto Evandro Chagas, foram testados pelo método imunoenzimático (ELISA). Resultados: Dos 525 soros testados 22,2 por cento, 5l,7 por cento , 72,9 por cento e 92,5 por cento foram positivos nas faixas etárias de 0-4, 5-14, 15-24 e > e igual 25 anos, respectivamente. Conclusöes: Näo obstante a grande concentraçäo da varicela nos primeiros dez anos de vida, cerca da metade dos adolescentes e adultos jovens persistem sem imunidade. Este achado sugere que há indicaçäo da vacinaçäo contra a varicela para as crianças, mas também, e principalmente, para os adolescentese adultos jovens, sem história consistente de doença correlata, uma vez que nesses grupos de faixas etárias maiores, a infecçäo pode evoluir com doença mais grave, com risco de complicaçöes e óbito.

Characterization of rotavirus P genotypes circulating among paediatric inpatients in northern Brazil

De novembro de 1992 a agosto de 1993, 28 amostras fecais positivas para rotavirus, obtidas de pacientes pediatricos hospitalizados em Belem, Brasil, com idades inferiores a 4 anos, foram testadas por RT-PCR visando a determinacao dos genotipos P. Com excecao de 7 criancas nao diarreicas, todos os pacientes apresentavam diarreia a admissao ou a desenvolviam enquanto internados no hospital. Cepas de rotavirus com especifidades antigenicas P correspondentes aos genotipos P1B[4] e P1A[8]...

Rotaviruses as a cause of nosocomial, infantile diarrhoea in northern Brazil: pilot study

Faecal samples were obtained from 190 children, aged 0 to 5 years, admitted to a public hospital in Belém, Pará, Brazil. These patients were placed in a pediatric ward with 40 beds distributed in six rooms. Case were classified into three groups: (a) nosocomial: children who developed gastroenteritis 72 hr or later after admission; (b) community-acquired: patients admitted either with diarrhoea or who had diarrhoea within 72 hr following admission; (c) non-diarrhoeic: those children who had no diarrhoea three days before and three days after collection of formed faecal sample. Specimens were routinely processed for the presence of rotaviruses, bacteria and parasites. Rotaviruses were detected through enzyme-linked immunosorbent assay (ELISA) and subsequently serotyped/electrophoretyped. Rotaviruses were the most prevalent enteropathogens among nosocomial cases, accounting for 39 porcento (9/23) of diarrhoeal episodes; on the other hand, rotaviruses ocurred in 8.3 por cento (11/133) and 9 por cento (3/34) of community-acquired and non-diarrhoeic categories, respectively. Mixed infections involving rotavirus and Giardia intestinalis and rotavirus plus G. intestinalis and Entamoeba histolytica were detected in frequencies of 8.6 and 4.3 por cento, respectively, in the nosocomial group. The absence of bacterial pathogens in this category, and the unusual low prevalence of these agents in the other two groups may reflect the early and routine administration of antibiotics following admission to this hospital. Rotavirus serotype 2 prevailed over the other types, accounting for 77.8 por cento of isolates from nosocomial diarrhoeal episodes. In addition, at least five different genomic profiles could be observed, of which one displayed an unusual five-segment first RNA cluster. Dehydration was recorded in all cases of hospital-acquired, rotavirus-associated diarrhoea, whereas in only 57 por cento of nosocomial cases of other aetiology. It was also noted that nosocomial, rotavirus-associated diarrhoeal episodes ocurr earlier (7 days), following admission, if compared with those hospital-acquired cases of other aetiology (14 days).

Epidemiology of rotavirus gastroenteritis in the Amazon region and the need for a specific vaccine

In the Amazon region, rotaviruses account for at least 30 per cent of all episodes of acute gastroenteritis among hospitalized children and are associated with nearly 1O per cent of cases of infantile acute diarrhea at community level. All four rotavirus serotypes are shown to infect children in our region, serotype l being predominant (about 50 per cent). Sequential infections in the same child, caused by different serotypes, are commonly noted. No clear seasonal variation on the occurrence of rotavirus diarrhea has been recorded, as cases are readily detected throughout the year. Rotavirus diarrhea cases have been found to be, in general, more severe than those of other aetiology. On the other hand, it has been noted that early (children less than 4 months of age) rotavirus infections are more likely to be asymptomatic (p = 0.021). Occurrence of rotavirus infections among Amazonian Indian populations seems to be very common. An explosive outbreak of rotavirus diarrhea affected possibly 88 per cent of both children and adults of the Tiryió population, Northern Pará State. In addition, rotavirus antibody was detected in 54.7 per cent of 1,299 sera collected from Amerinds belonging to 13 relatively isolated communities in the Amazon region. In the light of the above mentioned findings it was suggested that our region would be suitable for a field trial with a rotavirus-candidate vaccine. A study is therefore underway aiming to compare safety, immunogenicity and efficacy of a rhesus-human reassortant rotavirus (RRV-tetravalent) vaccine and placebo in 500 healthy infants living in the peripheral area of Belém.