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. 2024 Jul 22;33(3):e002624.
doi: 10.1590/S1984-29612024039. eCollection 2024.

Allergenic response induced by Pterobothrium crassicolle (Cestoda: Trypanorhyncha) extracts in murine model

Danuza Pinheiro Bastos Garcia de Mattos  1 , Maurício Afonso Verícimo  2 , Sérgio Carmona de São Clemente  3 , Michelle Cristie Gonçalves da Fonseca  4 , Marcelo Knoff  4
Affiliations

Allergenic response induced by Pterobothrium crassicolle (Cestoda: Trypanorhyncha) extracts in murine model

Danuza Pinheiro Bastos Garcia de Mattos et al. Rev Bras Parasitol Vet. .

Abstract
in English, Portuguese

The aim of this study was to determine the allergenic activity of components present in crude extracts of Pterobothrium crassicolle plerocerci (CPE) and blastocysts (CBE) obtained from Micropogonias furnieri in a murine model. Two groups of seven animals each received 50 µg of CPE or CBE on days 1, 35 and 120. Serum samples were tested by ELISA and Immunoblotting. Specific IgG and IgE levels were detected by ELISA, showing specific humoral responses for the primary immunization for both immunoglobulins and continuously growing titers for IgE. Positive Passive Cutaneous Anaphylaxis tests in rats sensitized with anti-CBE sera and tested by CBE, showed biologically, the allergenic activity of the extracts. The CPE and CBE showed some different recognition regions but both experimental groups recognized all regions of the extracts when tested for cross reactions, showing that CPE and CBE could share antigenic recognition sites.

O objetivo deste estudo foi determinar a atividade alergênica de componentes presentes em extratos crus de plerocercos (CPE) e de blastocistos de Pterobothrium crassicolle (CBE), obtidos de Micropogonias furnieri, em modelo murino. Dois grupos de sete animais receberam cada um 50 µg de CPE ou CBE nos dias 1, 35 e 120. As amostras de soro foram testadas por ELISA e Imunoblot. Os níveis específicos de IgG e IgE foram detectados por ELISA, mostrando respostas humorais específicas para a imunização primária para ambas as imunoglobulinas e títulos crescentes de IgE. Testes positivos de Anafilaxia Cutânea Passiva em ratos sensibilizados com soros anti-CBE e testados por CBE, demonstraram biologicamente, a atividade alergênica dos extratos. O CPE e o CBE evidenciaram algumas regiões de reconhecimento diferentes, mas ambos os grupos experimentais reconheceram todas as regiões dos extratos, quando testados para reações cruzadas, mostrando que o CPE e o CBE poderiam compartilhar locais de reconhecimento antigênico.

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Conflict of interest statement

Conflict of interest: The authors declare they have no conflict of interest.

Figures

Figure 1
Figure 1. Dynamics of the specific IgG (A) and IgE (B) serum levels. Two groups, each with 7 mice, received intraperitoneally 50 µg of crude extract of Pterobothrium crassicolle plerocerci - CPE (square) or blastocysts - CBE (triangle) associated with 2 mg Al(OH)3, on days 0, 35 and 120 (arrow). A control group (circle) with 5 animals, received saline solution with 2 mg Al(OH)3 on the same days using the same pathway. The values indicate the means of the sums of the optical densities (OD). +/- standard error of the mean of each group. As from day 14, the IgG and IgE levels of both experimental groups were p<0.001 (exception *p<0.01) when compared to the control. a p<0.05, b p<0.01, c p<0.001 between groups.
Figure 2
Figure 2. The passive cutaneous anaphylaxis assay (PCA). PCA reaction using Lou-H rat as the receptor of anti-CBE sera from BALB/c mice. Positive PCA reactions for mice sera after 127 (a) and 135 (b) days induced by the CBE. Increased blood influx (arrows); Sera without reaction (asterisks); Crude extract of Pterobothrium crassicolle blastocysts (CBE). Bar = 10 mm.
Figure 3
Figure 3. Recognition of immunogenic proteins by immunoblotting. (A) SDS-PAGE (12%) staining by Coomassie brilliant blue showing: the molecular weight (kDa) (a.1), protein profile of CPE (a.2) and CBE (a.3). (B) Immunoblot showing IgG recognition of the immunogenic proteins of CPE (b.1, b.2) and CBE (b.3) in pooled sera from all sensitized mice, seven days after the second immunization (42nd experimental day). Crude extract of Pterobothrium crassicolle plerocerci (CPE) and blastocysts (CBE).
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