Caracterização imunológica da formulação vacinal baseada em VLP (Virus Like Particle) da Proteína Circumsporozoíta de Plasmodium vivax / Immunological characterization of the VLP (Virus Like Particle) vaccine formulation fused to Circumsporozoite Protein of Plasmodium vivax

O Plasmodium vivax é a espécie mais comum de parasita causador da malária humana encontrada fora da África, com maior endemicidade na Ásia, América Central e do Sul e Oceania. Embora o Plasmodium falciparum cause a maioria do número de mortes, o P. vivax pode levar à malária grave e resultar em morbimortalidade significativa. O desenvolvimento de uma vacina protetora será um passo importante para a eliminação da malária. Recentemente, uma formulação contendo as três variantes alélicas da proteína circumsporozoíta de P. vivax (PvCSP - All epitopes) induziu proteção parcial em camundongos após desafio com esporozoíto híbrido Plasmodium berghei (Pb), no qual as repetições centrais do PbCSP foram substituídas por repetições PvCSP-VK210 (esporozoítos Pb/Pv). No presente estudo, a proteína quimérica PvCSP contendo as variantes alélicas (VK210, VK247 e P. vivax-like) fusionadas com a proteína de nucleocapsídeo do vírus da caxumba (formando partículas semelhantes a nucleocapsídeos ou do inglês, NLP - Núcleo Like Particles) na ausência (NLP-CSPR) ou na presença do domínio C-terminal (CT) conservado da PvCSP (NLP-CSPCT). Para a realização do estudo selecionamos os adjuvantes Poly (I:C), um RNA sintético de dupla fita, agonista do receptor Toll do tipo 3 (TLR3) ou o adjuvante Montanide ISA 720, uma emulação óleo em agua. Para obter uma forte resposta imune, a levedura Pichia pastoris foi usada para expressar as proteínas recombinantes na forma de NLPs. Camundongos foram imunizados com cada uma das proteínas recombinantes em combinação com os adjuvantes citados. Embora ambas as NLPs tenham sido capazes de gerar uma forte resposta imune, com altos níveis de títulos e longevidade, apenas a formulação contendo a proteína NLP-CSPCT na presença do adjuvante Poly (I:C) foi selecionada para ser explorada em experimentos futuros. Esta proteína em combinação com o adjuvante Poly (I:C) induziu alta frequência de células secretoras de anticorpos específicas para o antígeno homólogo nos dias 5 e 30, no baço e na medula óssea, respectivamente. Altos títulos de IgG contra as 3 variantes de PvCSP foram detectados nos soros. Posteriormente camundongos imunizados com NLP-CSPCT foram desafiados com esporozoítos Pb/Pv e a parasitemia no 5º dia demonstrou proteção estéril em 30% dos camundongos desafiados. Portanto, a formulação vacinal gerada neste estudo tem potencial para ser explorada no desenvolvimento de uma vacina universal contra a malária causada por P. vivax

Plasmodium vivax is the most common species of human malaria parasite found outside Africa, with high endemicity in Asia, Central and South America, and Oceania. Although Plasmodium falciparum causes the majority of deaths, P. vivax can lead to severe malaria and result in significant morbidity and mortality. The development of a protective vaccine will be a major step toward malaria elimination. Recently, a formulation containing the three allelic variants of the P. vivax circumsporozoite protein (PvCSP--All epitopes) showed partial protection in mice after a challenge with the hybrid Plasmodium berghei (Pb) sporozoite, in which the PbCSP central repeats were replaced by the VK210 PvCSP repeats (Pb/Pv sporozoite). In the present study, the chimeric PvCSP allelic variants (VK210, VK247, and P. vivax-like) were fused with the mumps virus nucleocapsid protein (assembling into nucleo like particles - NLP) in the absence (NLP-CSPR) or presence of the conserved C-terminal (CT) domain of PvCSP (NLP-CSPCT). To carry out the study, we selected the adjuvants Poly (I:C), a synthetic double-stranded RNA, Toll-like receptor 3 (TLR3) agonist or Montanide ISA 720 adjuvant, an oil-water emulation. To elicit stronger immune response, Pichia pastoris yeast was used to produce the NLPs. Mice were immunized with each recombinant protein in combination with above. Although both NLPs were able to generate stronger immune response, with high antibodies titer levels and longevity, formulation containing NLP-CSPCT in the presence of Poly (I:C) was selected to be explored in future experiments. NLP-CSPCT with Poly (I:C) adjuvant presented a high frequency of antigen-specific antibody-secreting cells (ASCs) on days 5 and 30, respectively, in the spleen and bone marrow. Moreover, high IgG titers against all PvCSP variants were detected in the sera. Later, immunized mice with NLP-CSPCT were challenged with Pb/Pv sporozoites. Sterile protection was observed in 30% of the challenged mice. Therefore, this vaccine formulation use has the potential to be a good candidate for the development of a universal vaccine against P. vivax malaria.

Increased platelet distribution width and reduced IL-2 and IL-12 are associated with thrombocytopenia in Plasmodium vivax malaria

BACKGROUND Thrombocytopenia in malaria involves platelet destruction and consumption; however, the cellular response underlying this phenomenon has still not been elucidated. OBJECTIVE To find associations between platelet indices and unbalanced Th1/Th2/Th17 cytokines as a response to thrombocytopenia in Plasmodium vivax infected (Pv-MAL) patients. METHODS Platelet counts and quantification of Th1/Th2/Th17 cytokine levels were compared in 77 patients with uncomplicated P. vivax malaria and 37 healthy donors from the same area (endemic control group - ENCG). FINDINGS Thrombocytopenia was the main manifestation in 55 patients, but was not associated with parasitaemia. The Pv-MAL patients showed increases in the mean platelet volume (MPV), which may be consistent with larger or megaplatelets. Contrary to the findings regarding the endemic control group, MPV and platelet distribution width (PDW) did not show an inverse correlation, due the increase in the heterogeneity of platelet width. In addition, the Pv-MAL patients presented increased IL-1β and reduced IL-12p70 and IL-2 serum concentrations. Furthermore, the reduction of these cytokines was associated with PDW values. MAIN CONCLUSIONS Our data demonstrate that an increase in MPV and the association between reductions of IL-2 and IL-12 and PDW values may be an immune response to thrombocytopenia in uncomplicated P. vivax malaria.

On cytoadhesion of Plasmodium vivax: raison d'être?

It is generally accepted that Plasmodium vivax, the most widely distributed human malaria parasite, causes mild disease and that this species does not sequester in the deep capillaries of internal organs. Recent evidence, however, has demonstrated that there is severe disease, sometimes resulting in death, exclusively associated with P. vivax and that P. vivax-infected reticulocytes are able to cytoadhere in vitro to different endothelial cells and placental cryosections. Here, we review the scarce and preliminary data on cytoadherence in P. vivax, reinforcing the importance of this phenomenon in this species and highlighting the avenues that it opens for our understanding of the pathology of this neglected human malaria parasite.

Infecciones maláricas en individuos asintomáticos en la población indígena Jivi, Amazonas, Venezuela / Asymptomatic malaria infection in the indigenous Jivi population, Amazonas state, Venezuela

Se evaluó la presencia de infecciones maláricas en individuos asintomáticos en la población Jivi de Puente Parhueña. El estudio fue de tipo prospectivo en tres momentos. El diagnóstico parasitológico se realizó mediante el examen convencional de gota gruesa y extendido (GGE) y la reacción en cadena de la polimerasa (PCR). El diagnóstico por microscopia indicó 2% (5/261) de láminas positivas en Abril, 1% (3/274) en Septiembre y 4% (5/135) en Diciembre. La PCR para Plasmodium spp., fue 46% (26/57) en abril, 49% (28/57) en Septiembre y 35% (20/57) en Diciembre. En los tres momentos predominó la presencia de P. vivax. La prueba de ELISA demostró 72% (41/57) seroreactivos en Abril, 53% (30/57) en Septiembre y 60% (34/57) en Diciembre. En Puente Parhueña habitan individuos con infecciones maláricas asintomáticos, con persistencia de anticuerpos antimalaricos, que probablemente representan un reservorio de gametocitos dentro de la comunidad.

The study was carried out to determine the present malaria infection in the asymptomatic Jivi people of Puente Parhueña. The study was prospective over three periods of time. The parasitological diagnoses were from thick and thin blood smears (GGE) and polymerase chain reaction (PCR). The antibody search was performed by ELISA. Microscopy of the slides detected the following positive results: 2% (2/261) April, 1% (3/274) September and 4% (5/135) December. Detection of Plasmodium by PCR was 46% (26/57) in April, 49% (28/57) in September y 35% (20/57) in December. Plasmodium vivax infected individuals predominated during these 3 times. Positives for ELISA were 72% (41/57) in April, 53% (30/57) September and 60% (34/57) December. The study demonstrated that people living in Puente Parhueña presented asymptomatic malaria infection with malaria antibodies persistence which likely represents a gametocyte potential reservoir for infection among the population.

Plasmodium vivax malaria presen with spontaneous intrasplenic Haemorrhage

A 6-year-old child with pallor and abdominal pain associated with fever. proved to have spontaneous intrasplenic haemorrhage, as shown by abdominal ultrasonography and computerised tomogram [CT], as the initial manifestation of acute Plasmodium vivax malaria infection She responded well to supportive care and antimalarial therapy and did not progress to splenic rupture. This case is presented to increase the awareness of physicians to this rare life-threatening condition that patients afflicted with malaria might present with

Malaria por Plasmodium vivax: aspectos clínicos y hematológicos / Malaria due to Plasmodium vivax. Clinical and hematological aspects

Se reportan los hallazgos clínicos, de laboratorio e histopatológicos, presentes en 20 pacientes que ingresaron al Hospital Hipólito Unanue en el año 1984, con el diagnóstico de malaria. Se revisa la fisiopatología con énfasis en los casos de malaria por Plasmodium vivax. El agente etiológico más frecuente, en esta serie, fue el Plasmodium vivax ( 19 pacientes ). Fiebre y escalofrío se presentó en el 100 por ciento, hepatomegalia en el 90 por ciento, esplenomegalia en el 70 por ciento, ictericia en el 30 por ciento, esta última fue muy severa en un caso asociada a hepatitis crónica activa. La anemia que se observó en el 85 por ciento, fue de origen multifactorial; la trombocitopenia estuvo presente en el 83.3 por ciento de casos, rara vez se acompañó de sangrado. Otros hallazgos fueron: linfopenia ( 70 por ciento ), leucopenia leve ( 45 por ciento ), neutropenia leve ( 35 por ciento ). La histopatología en médula ósea mostró: hiperplasia de las tres series y cambios reactivos inespecíficos. En hígado encontramos: depósito de pigmento malárico en el 42.8 por ciento de casos. Dada las actuales condiciones sanitarias y epidemiológicas del país, es impostergable reforzar la lucha contra esta enfermedad

Pathological rupture of malarial spleen.

Two cases of spontaneous rupture of malarial spleen are reported here. One of them was a male who was on chloroquine for an acute attack of malaria. While on therapy, he complained of pain in left hypochondrium followed by palpitations. The other patient was a female who was admitted for continuous dull aching pain and fever. In both the patients, exploratory laparotomy revealed an enlarged spleen with tear. Splenectomy was performed. Histopathological examination revealed dilated congested sinusoid with follicular atrophy, and RBCs with malarial parasites. The post-operative course was smooth in both patients.