O Estado de São Paulo, situado na região Sudeste do Brasil, apresenta esporadicamente casos autóctones de malária que se caracterizam pela presença de quadro clínico benigno com parasitemias baixas e sintomatologia branda, identificados como malária vivax. Pouco se sabe a respeito da sintomatologia e resposta imune desenvolvidas pelo ser humano para as variantes Plasmodium vivax VK247 e Plasmodium vivax-like humano. Estas variantes são transmitidas pelo mosquito Anopheles (Kerteszia) cruzii, uma das espécies mais abundantes no Sudeste brasileiro. O objetivo deste trabalho foi verificar a infecção em anofelinos desta região, através do teste imunoenzimático ELISA com utilização de anticorpos monoclonais específicos dirigidos contra as regiões repetitivas da proteína circunsporozoíta de P. vivax clássico, P. brasilianum/P. malariae e P. vivax VK247. Coletas entomológicas foram realizadas no período de 1991 a 1993 em São Vicente e Juquitiba, municípios localizados em área remanescente da Mata Atlântica do Estado de São Paulo. A Mata Atlântica é rica em plantas da família Bromeliaceae, criadouros de formas imaturas de anofelinos do subgênero Kerteszia. De um total de 1117 espécimes de An. (Ker.) cruzii capturados no Município de São Vicente, 0,179% foram positivos para P. vivax clássico. Em Juquitiba, dentre 1161 An. (Ker.) cruzii pesquisados, 0,086% foram positivos para o P. vivax VK247, o que demonstra a presença da variante na região. Embora o índice de infecção encontrado seja baixo, a alta densidade destes mosquitos e sua voracidade (picam durante as 24 h do dia) poderiam compensar a baixa porcentagem de espécimes infectados.

Sporadic cases of autochthonous malaria have been recorded in São Paulo State, located in the Southeast region of Brazil. These cases are characterized by their benign course, low parasitemia, and mild symptomatology and have been identified as vivax malaria. Little is known about the symptoms and immune response elicited in humans by the variants Plasmodium vivax VK247 and P. vivax-like human malaria parasites. These variants are transmitted by Anopheles (Kerteszia) cruzii, one of the most common species of mosquitoes in the Southeast of Brazil. The objective of the study described in this paper was to investigate infection in anophelines using ELISA immunoenzymatic assay with specific monoclonal antibodies directed against the repetitive regions of the circumsporozoite protein in classic P. vivax, P. brasilianum/P. malariae, and P. vivax VK247. Between 1991 and 1993, mosquitoes were collected in São Vicente and Juquitiba, municipalites located in a remnant of the Brazilian Atlantic forest in São Paulo State, an ecosystem rich in plants of the Bromeliaceae family. These plants function as nurseries for immature forms of anophelines of the subgenus Kerteszia. Of 1117 An. (Ker.) cruzii captured in São Vicente, 0.179% were positive for classic P. vivax. In Juquitiba, of 1161 An. (Ker.) cruzii, 0.086% were positive for P. vivax VK247, confirming the presence of this variant in the region. Although the infection rate is low, the high density of these mosquitoes and their voracity (they exhibit 24-h biting activity) could compensate for the low percentage of infected specimens.

During 1992-1994, 33 malaria cases were reported in two regions in Brazil where few sporadic atypical cases occur, most of them in home owners, who are weekenders, while home caretakers live there permanently. Indirect Fluorescent Antibody Test (IFAT), with Plasmodium vivax, and Enzime Linked Immunosorbent Assay (ELISA) with repeat peptides of the circumsporozoite (CS) proteins of the 3 known P. vivax variants and P. malarie/P. brasilianum, were performed on 277 sera, obtained within a 5 to 10 km range of malaria cases. Very rarely did any of these donors recall typical malaria episodes. Blood smears of all but 5 were negative. One of the 5 malaria cases included in our serology was of a home owner, 1 of a permanent resident, 3 from Superintendência de Controle de Endemias employees who went there to capture mosquitoes. In Region 1 the prevalence of IFAT positive sera was 73% and 28% among caretakers, 18% and 9.6% among home owners. In Region 2 (3 localities) no distinction was possible between caretakers and home owners, IFAT positivity being 38%, 28% and 7%. The relative percentage of positive anti-CS repeats ELISA, differed for each of the peptides among localities. Dwellings are in the vicinity of woods, where monkeys are frequently seen. The origin of these malaria cases, geographical differences and high seropositivity is discussed

Calomys callosus a wild rodent, is a natural host of Trypanosoma cruzi. Twelve C. callosus were infected with 10(5) trypomastigotes of the F strain (a myotropic strain) of T. cruzi. Parasitemia decreased on the 21 st day becoming negative around the 40th day of infection. All animals survived but had positive parasitological tests, until the end of the experiment. The infected animals developed severe inflammation in the myocardium and skeletal muscle. This process was pronounced from the 26 th to the 30th day and gradually subsided from the 50 th day becoming absent or residual on the 64 th day after infection. Collagen was identified by the picro Sirius red method. Fibrogenesis developed early, but regression of fibrosis occurred between the 50th and 64th day. Ultrastructural study disclosed a predominance of macrophages and fibroblasts in the inflammatory infiltrates, with small numbers of lymphocytes. Macrophages had active phagocytosis and showed points of contact with altered muscle cells. Different degrees of matrix expansion were present, with granular and fibrilar deposits and collagen bundles. These alterations subsided by the 64th days. Macrophages seem to be the main immune effector cell in the C. callosus model of infection with T. cruzi. The mechanisms involved in the rapid fibrogenesis and its regression deserve further investigation.

Anticorpos anti-RESA/Pf155 foram testados em soros de indivíduos de três localidades (Laranjal do Jari, Vila Padaria e Vila Paraíso) localizadas no Estado do Amapá, Brasil, durante as estações de chuvas "longas" e "curtas". Todos os esfregaços apresentaram-se negativos para malária. A maioria dos soros coletados foram positivos na reação de Imunofluorescência Indireta (IFI) para P. falciparum, não apresentando variação sazonal. Uma alta porcentagem destes soros (62% a 100%) foram positivos para RESA na reação de Imunofluorescência Indireta Modificada (IFIM), com aumento na média geométrica dos títulos significante (p < 0.05) durante a estação das chuvas "curtas", época de maior transmissão da doença. O teste de ELISA com os três peptídeos repetitivos do RESA: (EENV)3(4x3), (EENVEHDA)2 (8x2) e (DDEHVEEPTVA)2(11x2), não revelou variações sazonais estatisticamente significantes, embora um pequeno aumento na positividade tenha sido observado, na época de chuvas "curtas", em V. Padaria (15,3 para 38,8%) com o peptídeo 8x2 e em V. Paraíso com os peptídeos 4x3 e 8x2, decrescendo com o peptídeo 11x2. Os títulos de IFIM pareceram se correlacionar principalmente com o peptídeo 4x3 e este mostrou-se imunodominante nas três localidades.

Anti-RESA/Pf155 antibodies were assayed in sera of individuals from three localities (Laranjal do Jari, Vila Padaria and Vila Paraíso) in the State of Amapá, Brazil, during the long-rains and short-rains seasons. All of these had negative blood smears for malaria. Most of the sera collected were positive in Indirect Fluorescent Antibody (IFA) with P. falciparum parasites, with no seasonal variation. A high percentage of these sera (62% to 100%) was RESA positive by Modified Indirect Fluorescent Antibody (MIFA), with a significant (p < 0.05) increase of geometric mean titers during the short-rains season, when the transmission of the disease is highest. ELISA with three repetitive RESA peptides (EENV)3 (4x3), (EENVEHDA)2 (8x2) and (DDEHVEEPTVA)2(11x2) did not reveal statistically significant seasonal variations, although a small enhancement of positivity was observed in V. Padaria (15.3 to 38.8%) in the short-rains season with the 8x2 peptides, and with 4x3 and 8x2 peptides in V. Paraíso, with a decrease in 11x2. MIFA titers appeared to be correlated mainly to the peptide 4x3 and it was the immunodominant in the three localities.

Macrófagos obtidos do peritoneo de camundongos após estímulo, com peptona, foram cultivados em lamínulas, infectados com tripomastigotas das cepas F e Y de T. cruzi, obtidos de cultivo de tecidos ou do sangue de camundongos infectados. Os parasitas, obtidos de cultivo de tecidos, tanto da cepa Y como os da cepa F, são interiorizados por macrófagos em proporção muito mais elevada do que os sanguícolas. Parasitas de cultivo de tecidos incubados com soro de camundongos normais, ou soro híperimune específico em diluição sub-aglutinante, comportam-se essencialmente como parasitas não opsonizados. Foram observadas diferenças a nível ultraestrutural na fase inicial de interação entre macrófagos e tripomastigotas das duas origens. Após 30 minutos, tripomastigotas de cultivo de tecidos localizam-se em agrupamentos na área de contato com os macrófagos. Enquanto os tripomastigotas sanguícolas estão na maioria das vezes no interior de vacúolos fagocíticos largos, após 3 horas de interação os tripomastigotas de cultura situam-se em um único vacúolo estreito. Tanto as formas de cultivo de tecidos quanto os tripomastigotas sanguícolas da cepa Y multiplicam-se em macrófagos; os tripomastigotas sanguícolas da cepa F são destruídos no interior da célula hospedeira, enquanto os tripomastigotas de cultivo de tecidos desta cepa são capazes de multiplicar-se.

Mouse peritoneal elicited macrophages cultured on coverslips were infected with Trypanosoma cruzi trypomastigotes from both the F strain and Y strain obtained either from tissue culture or from the bloodstream of infected mice. Both the Y strain and F parasites obtained from tissue culture were interiorized by macrophages at a much higher rate than bloodstream trypomastigotes. Tissue culture parasites incubated with normal mouse serum, mouse plasma obtained at the 7th day after infection, or specific hyperimmune serum at subagglutinating concentration, behaved essentially as non-opsonized parasites. Ultrastructural differences were seen at the early interaction phase between macrophages and trypimastigotes from both sources. After 30 minutes, tissue culture trypomastigotes were located in clusters at the area of contact with macrophages. While bloodstream trypomastigotes, at 3 hours post-infection were most frequently enclosed in a loose phagocytic vacuole, tissue culture trypomastigotes were enclosed in single tight vacuoles. Both tissue culture and bloodstream trypomastigotes of the Y strain multiplied within macrophages; F strain bloodstream trypomastigotes did not develop within the host cells, while tissue culture trypomastigotes multiplied.