Expressão heteróloga da EMA-2 (equi merozoite antigen) de Theileria equi em Pichia pastoris com potencial utilização em imunobiológicos

Vianna, Ana Muñoz; Gonçales, Relber Aguiar; Lara, Ana de Souza Stori de Paula; Pinto, Luciano da Silva; Nizoli, Leandro Quintana

doi:10.1590/0103-8478cr20131003

Cited by 5 publications

(10 citation statements)

References 21 publications

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“…The method chosen to perform the serological evaluation of animals in this study was indirect ELISA. The protocol used had been already tested in a previous study and had showed 83.3% specificity and 90.9% sensitivity (Vianna et al 2014). Several variations of ELISA with different antigens have been successfully used in the diagnosis of and in studies with T. equi (Kumar et al 2013, El-Sayed et al 2015, Kumar et al 2015.…”

Section: Discussionmentioning

confidence: 99%

“…The use of the protein rEMA-2 in the vaccine formulation and as an antigen for the ELISA test is justifiable by the fact that EMA-2 is expressed in several stages of the protozoan life cycle. In addition, it is one of the first antigens to be recognized by the immune system through its expression on the membrane and cytoplasm of erythrocytes (Vianna et al 2014).…”

Section: Discussionmentioning

confidence: 99%

“…Vaccination schedule and sampling. The EMA-2 was obtained as per the protocol described by Vianna et al (2014). The vaccine was prepared by adding 200μg of the recombinant protein in 10% aluminum hydroxide adjuvant.…”

Section: Methodsmentioning

confidence: 99%

“…Laboratorial analysis. Samples (serum, amniotic fluid, and colostrum) were analyzed by enzyme linked immunonosorbent assay (ELISA), as described previously by Vianna et al (2014). The serum of an experimentally infected horse was used as positive control, while the serum of a newborn foal delivered by a mare negative for T. equi infection was used as negative control.…”

Section: Methodsmentioning

confidence: 99%

“…These antigens are expressed at different stages of the protozoan cycle. EMA-1 and EMA-2 are expressed at several stages; however, EMA-2 is one of the first antigens to be recognized by the immune system (Kumar et al 2013), which makes it potentially useful for the development of prophylactic tools such as vaccines (Kumar et al 2013, Vianna et al 2014.…”

Section: Introductionmentioning

confidence: 99%

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Dynamics of humoral immune response in pregnant mares and foals vaccinated with Theileria equi recombinant EMA-2

et al. 2018

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Theileria equi is an infectious hemoprotozoan agent of equine piroplasmosis, a disease that has severe economic and sanitary impact internationally. In addition to its common clinical features, piroplasmosis can cause gestational losses and neonatal damage, which makes neonates susceptible to this disease. The aim of this study was to evaluate the dynamics of humoral immune response to recombinant EMA-2 of T. equi in pregnant mares and foals, as well as the transfer of vaccine antibodies through the colostrum ingested by sucking foals. For vaccine production, the EMA-2 expression gene was cloned and expressed in the yeast species, Pichia pastoris. Thirty-six horses were used, of which 18 were pregnant mares and 18 were foals. The mares were divided into control and vaccinated groups, and the vaccinated group received three doses of rEMA-2 every 21 days starting at 300 days of gestation. Foals from vaccinated and control groups were evaluated until the sixth month of life. The production of antibodies by foals on the rEMA-2 vaccination schedule was also evaluated from the second month of life. Foals in the vaccinated group had received three doses of the vaccine every 21 days. The method used to evaluate serum and colostrum samples was indirect ELISA, and plates were sensitized with the rEMA-2 protein. At the end of the vaccination schedule, vaccinated mares showed a 2.3-fold increase in antibody levels when compared to baseline values. The colostrum of vaccinated mares presented antibody levels of 1.0432±0.33. Foals delivered by vaccinated mares presented levels of antibodies greater than those of foals delivered by control mares after their first time sucking (at about twelve hours after birth). Foals vaccinated in the second month of life showed an 8.3-fold increase in antibody levels when compared to baseline values. The vaccination schedule with rEMA-2 was able to stimulate humoral immunity in pregnant mares. Vaccine immunoglobins were concentrated in the colostrum of vaccinated mares and foals delivered by these mares showed an increase in serum levels of vaccine antibodies after the first-time sucking.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%