Immunity to asexual blood stage malaria and vaccine approaches

The development of a malaria vaccine seems to be a definite possibility despite the fact that even individuals with a life time of endemic exposure do not develop sterile immunity. An effective malaria vaccine would be invaluable in preventing malaria-associated deaths in endemic areas, especially amongst children less than 5 years of age and pregnant women. This review discusses our current understanding of immunity against the asexual blood stage of malaria - the stage that is responsible for the symptoms of the disease - and approaches to the design of an asexual blood stage vaccine.

Secretory pathway of trypanosomatid parasites

The Trypanosomatidae comprise a large group of parasitic protozoa, some of which cause important diseases in humans. These include Tryanosoma brucei (the causative agent of African sleeping sickness and nagana in cattle), Trypanosoma cruzi (the causative agent of Chagas' disease in Central and South America), and Leishmania spp. (the causative agent of visceral and [muco]cutaneous leishmaniasis throughout the tropics and subtropics). The cell surfaces of these parasites are covered in complex protein- or carbohydrate-rich coats that are required for parasite survival and infectivity in their respective insect vectors and mammalian hosts. These molecules are assembled in the secretory pathway. Recent advances in the genetic manipulation of these parasites as well as progress with the parasite genome projects has greatly advanced our understanding of processes that underlie secretory transport in trypanosomatids. This article provides an overview of the organization of the trypanosomatid secretory pathway and connections that exist with endocytic organelles and multiple lytic and storage vacuoles. A number of the molecular components that are required for vesicular transport have been identified, as have some of the sorting signals that direct proteins to the cell surface or organelles it? the endosome-vacuole system. Finally, the subcellular organization of the major glycosylation pathways in these parasites is reviewed. Studies on these highly divergent eukaryotes provide important insights into the molecular processes underlying secretory transport that arose very early in eukaryotic evolution. They also reveal unusual or novel aspects of secretory), transport and protein glycosylation that may be exploited in developing new antiparasite drugs.

Prospects for a vaccine against malaria [Commentary]

No Abstract

Anomalies in the developing neural and visceral head skeleton of the Australian lungfish, Neoceratodus forsteri

Several anomalies occur in the developing neural and visceral head skeleton of young specimens of Neoceratodus forsteri that have been reared under laboratory conditions. These include anomalies of the basicranium and its derivatives, aberrations of the anterior mandible and hyoid apparatus, and abnormalities in the articulation of the jaws and the elements that produce them. Apart from the occasional absence of the basihyal, and failure of the quadrate processes to form, the anomalies are not deficiencies. Most involve malformations of parts of the neurocranium and visceral skeleton, inappropriate articulations or fusions between elements, disunity in structures that are normally fused and the appearance of supernumerary elements. The incidence of chondral anomalies, generally higher than aberrations that occur in the dermal skeleton in juvenile lungfish, ranges from 1-10% in laboratory reared individuals that have not been subjected to experimental interference. The anomalies differ from those found in many amphibian populations, in the field and in the laboratory, because they involve the cranium, and not the limbs, and the lungfish have not been exposed to the factors that cause anomalies in the amphibians. It is unlikely that the existence of those anomalies, if it is reflected in the wild population, places a selective pressure on the lungfish, because, in a normal season, less than 1% of the total number of eggs produced survive to be recruited into the adult population.

Kunjin virus replicon vectors for human immunodeficiency virus vaccine development

We have previously demonstrated the ability of the vaccine vectors based on replicon RNA of the Australian flavivirus Kunjin (KUN) to induce protective antiviral and anticancer CD8(+) T-cell responses using murine polyepitope as a model immunogen (I. Anraku, T. J. Harvey, R. Linedale, J. Gardner, D. Harrich, A. Suhrbier, and A. A. Khromykh, J. Virol. 76:3791-3799, 2002). Here we showed that immunization of BALB/c mice with KUN replicons encoding HIV-1 Gag antigen resulted in induction of both Gag-specific antibody and protective Gag-specific CD8(+) T-cell responses. Two immunizations with KUNgag replicons in the form of virus-like particles (VLPs) induced anti-Gag antibodies with titers of greater than or equal to1:10,000. Immunization with KUNgag replicons delivered as plasmid DNA, naked RNA, or VLPs induced potent Gag-specific CD8(+) T-cell responses, with one immunization of KUNgag VLPs inducing 4.5-fold-more CD8(+) T cells than the number induced after immunization with recombinant vaccinia virus carrying the gag gene (rVVgag). Two immunizations with KUNgag VLPs also provided significant protection against challenge with rVVgag. Importantly, KUN replicon VLP vaccinations induced long-lasting immune responses with CD8(+) T cells able to secrete gamma interferon and to mediate protection 6 to 10 months after immunization. These results illustrate the potential value of the KUN replicon vectors for human immunodeficiency virus vaccine design.

DNA vaccine coding for the full-length infectious Kunjin virus RNA protects mice against the New York strain of West Nile virus

A plasmid DNA directing transcription of the infectious full-length RNA genome of Kunjin (KUN) virus in vivo from a mammalian expression promoter was used to vaccinate mice intramuscularly. The KUN viral cDNA encoded in the plasmid contained the mutation in the NS1 protein (Pro-250 to Leu) previously shown to attenuate KUN virus in weanling mice. KUN virus was isolated from the blood of immunized mice 3-4 days after DNA inoculation, demonstrating that infectious RNA was being transcribed in vivo; however, no symptoms of virus-induced disease were observed. By 19 days postimmunization, neutralizing antibody was detected in the serum of immunized animals. On challenge with lethal doses of the virulent New York strain of West Nile (WN) or wild-type KUN virus intracerebrally or intraperitoneally, mice immunized with as little as 0.1-1 mug of KUN plasmid DNA were solidly protected against disease. This finding correlated with neutralization data in vitro showing that serum from KUN DNA-immunized mice neutralized KUN and WN,viruses with similar efficiencies. The results demonstrate that delivery of an attenuated but replicating KUN virus via a plasmid DNA vector may provide an effective vaccination strategy against virulent strains of WN virus.

Immunogenicity of recombinant HBsAg/HCV particles in mice pre-immunised with hepatitis B virus-specific vaccine

Due to their spatial structure virus-like particles (VLPs) generally induce effective immune responses. VLPs derived from the small envelope protein (HBsAg-S) of hepatitis B virus (HBV) comprise the HBV vaccine. Modified HBsAs-S VLPs, carrying the immunodominant hypervariable region (HVR1) of the hepatitis C virus (HCV) envelope protein E2 within the exposed 'a'-determinant region (HBsAg/HVR1-VLPs), elicited HVR1-specific antibodies in mice. A high percentage of the human population is positive for anti-HBsAg antibodies (anti-HBs), either through vaccination or natural infection. We, therefore, determined if pre-existing anti-HBs could influence immunisation with modified VLPs. Mice were immunised with a commercial HBV vaccine, monitored to ensure an anti-HBs response, then immunised with HBsAg/HVR1-VLPs. The resulting anti-HVR1 antibody titre was similar in mice with or without pre-existing anti-HBs. This suggests that HBsAg/HVR1-VLPs induce a primary immune response to HVR1 in anti-HBs positive mice and, hence, they may be used successfully in individuals already immunised with the HBV vaccine. (C) 2003 Elsevier Science Ltd. All rights reserved.

Therapeutic LMP1 polyepitope vaccine for EBV-assolciated Hodgkin disease and nasopharyngeal carcinoma

Development of an epitope-based vaccination strategy designed to enhance Epstein-Barr virus (EBV)-specific CD8(+) cytotoxic T lymphocytes (CTLs) is increasingly being considered as a preferred approach for the treatment of EBV-associated relapsed Hodgkin disease (HD) and nasopharyngeal carcinoma (NPC). EBV-encoded latent membrane proteins, LMP1 and LMP2, are the only target antigens available for therapeutic augmentation of CTL responses in patients with HD and NPC. Here, we describe preclinical studies using a recombinant poxvirus vaccine that encodes a polyepitope protein comprising 6 HLA A2-restricted epitopes derived from LMP1. Human cells infected with this recombinant polyepitope construct were efficiently recognized by LM1-specific CTL lines from HLAA2 healthy individuals. Furthermore, immunization of HLrA A2/K-b mice with this polyepitope vaccine consistently generated strong LMP1 -specific CTL responses to 5 of the. 6 epitopes, which were readily detected by both ex vivo and in vitro assays. More important, this polyepitope vaccine successfully reversed the outgrowth of LMP1-expressing tumors in HLA A2/Kb mice. These studies provide an important platform for the development of an LMP-based polyepitope vaccine as an immunotherapeutic tool for the treatment of EBV-associated HD and NPC. (C) 2003 by The American Society of Hematology.

Investigação epidemiológica de um caso de leishmaniose visceral autóctone da Grande São Paulo, Brasil

Foi descrita uma investigação epidemiológica realizada na zona urbana de Diadema, município da Grande São Paulo (Brasil), com a finalidade de elucidar a fonte de infecção e o mecanismo de transmissão de um caso de leishmaniose visceral autóctone da área. Foram realizados inquéritos sorológicos através da reação de imunofluorescência indireta na população humana (591 soros) e na população canina (55 soros), e levada a efeito pesquisa entomológica no local da residência do doente e em uma área de reserva florestal situada a 500m desta residência.

Leishmaniose visceral adquirida no Estado de São Paulo (Brasil)

Foi descrita a notificação de um caso de leishmaniose visceral em uma criança com dois anos de idade, tendo sempre residido em Capão Redondo, localidade situada na Grande São Paulo (Brasil). Apesar de transmissão congênita ou por transfusão de sangue ter sido cogitada, é provável que a infecção tenha decorrido de mecanismo habitual, consubstanciando ocorrência autóctone relativa do Estado de São Paulo.

Investigação epidemiológica de, um novo caso de leishmaniose visceral ocorrido na Grande São Paulo, Brasil

Descreve-se uma investigação epidemiológica realizada em zona urbana do município de São Paulo, Brasil, para esclarecer um caso de leishmaniose visceral ocorrido em criança de 2 anos de idade, nascida e sempre residente no local. Afastou-se a possibilidade de transmissão por via transfusional e por vetor biológico, tendo como base os dados levantados da anamnese do doente, os resultados de inquéritos realizados na área em população humana, utilizando testes de imunofluorescência indireta, hemaglutinação passiva e intradermoreação de Montenegro, em população canina com o teste de imunofluorescência indireta, além de pesquisa entomológica em mata residual.

Ecological aspects of American cutaneous leishmaniasis: 4. Observations on the endophilic behavior of the sandfly and the vectorial role of Psychodopygus intermedius in the Ribeira Valley region of the S.Paulo State, Brazil

The invasive tendency of Psychodopygus intermedius in the home environment, observed initially by Forattini et al. (1976), has now been confirmed by the demonstration of its high endophilic ability and by the use of human residences for shelter. Populations such as Lutzomyia migonei and Pintomyia fischeri were also present in that environment, though their low densities registered during this investigation could be an indication of their poor ability to overcome the barriers raised by the artificial environment. An objective epidemiological analysis based on the variables here given showed that human infection takes place in the extraforest environment, and the principal vectorial function falls, without doubt, on P. intermedius.

Epidemia de leishmaniose visceral no Estado do Piauí, Brasil, 1980-1986

Analisou-se a epidemia de calazar ocorrida no Estado do Piauí, no período de 1980-1986. Os dados foram coletados pela SUCAM-Piauí, órgão do Ministério da Saúde para o controle de endemias, pela busca ativa na rede de assistência à saúde do Estado. A epidemia iniciou-se em municípios do centro e do norte, em 1980. No interior, ao contrário do período endêmico, quando predominou em áreas de maior altitude e clima semi-árido, a epidemia grassou nos vales de rios e em região mais úmida, de clima tropical. A capital do Piauí, Teresina, foi atingida em 1981, com pico epidêmico em 1984 e tendo sido responsável por mais de 60% dos 1.509 casos de todo o Estado. Foram feitas tentativas de controle pelo uso intensivo de inseticidas e eliminação de cães. Nas outras regiões do Piauí, borrifadas intensivamente para o controle da doença de Chagas e da malária, a epidemia foi pouco importante e cedeu espontaneamente. Nem a casuística e nem a população flebotomínica de Teresina apresentaram variações sazonais significativas, mas estiveram moderadamente correlacionadas entre si. Houve maior prevalência em menores de cinco anos, principalmente nos anos de maior incidência, e menor em maiores de 40 anos. A distribuição geográfica do processo epidêmico e a concomitância de seu início com seca prolongada acompanhada de emigração de pessoas e animais domésticos procedentes de regiões endêmicas para aquelas epidêmicas, sugerem que estes movimentos migratórios desencadearam a epidemia. O fato de o processo epidêmico ter cedido espontaneamente em áreas onde não se tentou o seu controle indica que não se pode atribuir apenas às medidas de controle o fim da epidemia. A partir da análise dos coeficientes de incidência específicos por faixa etária, é discutida a possiblilidade da progressiva redução na proporção de suscetíveis, determinada tanto por um grande número de infecções assintomáticas como pela ocorrência de imunidade duradoura, ter contribuído para a extinção da epidemia.

Physiological age in Lutzomyia youngi (Diptera: Psychodidae) populations from an endemic area for cutaneous leishmaniasis, Venezuela

Batches of sylvatic females of Lutzomyia youngi (Phlebotominae) captured in a Shannon trap on twelve occasions over one year in a locality where subcutaneous leishmaniasis is endemic, near the city of Trujillo, Venezuela, were used to study: 1) the percentages of parous females according to previously established criteria and 2) the average number of eggs laid spontaneously by isolated females during 7 days after feeding on hamsters. The data on the batches of females captured on nights previous to the rainy period (prepluvial) were compared with those on females captured after the rains (postpluvial) . Significant differences were detected by variation analysis for two variables and different number of N, as also were consistent groupings by Duncan's Test for pre-and postpluvial lots of females. The females captured on nights prior to the rainy periods (January-March and August-September) presented higher rates of nulliparity (86-72%) and contained or laid a greater number of eggs (71-67) than those captured after the rains (March-June and November-December) which presented lower rates of nulliparity (60-24%) and a smaller number of eggs (50-30). The rainfall peaks occurred in April and September-October, respectively. It is considered that these differences can be used by epidemiological studies as a means of estimating the physiological age of female populations of L. youngy.