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Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2024.tde-19062024-153449
Document
Author
Full name
Juliana Ferreira Souza Conceição
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2024
Supervisor
Committee
Gomes, Cristina de Oliveira Massoco Salles (President)
Brandão, Paulo Eduardo
Caballero, Otávia Luisa Silva Damas de
Title in Portuguese
Efeitos do Poly-ICLC (Hiltonol®) em células mononucleares caninas: estudo in vitro
Keywords in Portuguese
Avaliação microbicida
Burst oxidativo
Fagocitose
Macrófagos
Modulação imunológica
Abstract in Portuguese
O Poly-ICLC (Hiltonol®) apresenta resultados eficazes como imunomodulador biológico para sinergismo de tratamentos oncológico e infeccioso em diferentes espécies animais. O objetivo deste trabalho foi investigar os efeitos in vitro do Poly-ICLC (Hiltonol®) sobre diferentes células do sistema imune canino. Sendo assim, por meio do tratamento in vitro com este imunomodulador os linfócitos obtidos por meio do isolamento de células mononucleares de sangue periférico foram analisados quanto a proliferação frente ao mitógeno fitohemaglutinina (PHA) e a mensuração de IFN-gama do sobrenadante; macrófagos derivados de monócitos foram analisados quanto a sua capacidade bactericida mensurada por número de UFC (unidades formadoras de colônia) em ensaio de fagocitose com Staphylococcus pseudointermedius; e neutrófilos sanguíneos foram analisados por meio de ensaio de burst oxidativo e fagocitose pela técnica de citometria de fluxo onde foram utilizadas a Staphylococcus aureus e a Echerichia coli. A análise revelou que os linfócitos incubados em diferentes concentrações de Poly-ICLC (Hiltonol®) não tiveram modificações no índice de proliferação e nos níveis de IFN-gama quando comparados com linfócitos cultivados apenas com meio de cultura (controle). Com relação à capacidade bactericida de macrófagos verificou-se menor crescimento de unidades formadoras de colônias em ensaio de fagocitose com diferença significativa ao controle do ensaio (p<0.05) na maior concentração de Poly-ICLC (Hiltonol®) (100 µg/mL; p = 0.003). Além disso, foi verificado em neutrófilos um aumento da capacidade de fagocitose, e também do burst oxidativo de maneira concentração-dependente. Os resultados deste estudo são promissores no sentido de sugerir estudos clínicos para investigar os benefícios do uso clínico de Poly-ICLC (Hiltonol®) em pacientes caninos o que pode contribuir para ampliar a opção terapêutica da classe imunomoduladora, uma vez que os modificadores imunológicos ainda são poucos conhecidos e comercializados na medicina veterinária.
Title in English
Effects of Poly-ICLC (Hiltonol®) on canine mononuclear cells: in vitro study
Keywords in English
Bactericidal evaluation
Burst oxidative
Immune modulation
Macrophages
Phagocytosis
Abstract in English
Poly-ICLC (Hiltonol®) presents effective results as a biological immunomodulator for synergism of oncological and infectious treatments in different animal species. The objective of this work was to investigate the in vitro effects of Poly-ICLC (Hiltonol®) on different cells of the canine immune system. Therefore, through in vitro treatment with this immunomodulator lymphocytes obtained through the isolation of peripheral blood mononuclear cells were analyzed for proliferation against the mitogen phytohemagglutinin (PHA) and the measurement of IFN-gamma in the supernatant; Macrophages derived from monocytes were analyzed for their bactericidal capacity measured by the number of CFU (colony forming units) in a phagocytosis assay with Staphylococcus pseudointermedius; and blood neutrophils were analyzed using an oxidative burst assay and phagocytosis using the flow cytometry technique where Staphylococcus aures or Echerichia coli were used. The analysis revealed that lymphocytes incubated in different concentrations of Poly-ICLC (Hiltonol®) had no changes in the proliferation index and IFN-gamma levels when compared to lymphocytes cultured with culture medium alone (control). Regarding the bactericidal capacity of macrophages, there was less growth of colony-forming units in the phagocytosis assay, with a significant difference compared to the assay control (p<0.05) at the highest concentration of Poly-ICLC (Hiltonol®) (100 µg/mL; p = 0.003). Furthermore, an increase in the phagocytosis capacity and in the oxidative burst was demonstrated in neutrophils in a concentration-dependent manner. The results of this study are promising in suggesting clinical studies to investigate the benefits of the clinical use of Poly-ICLC (Hiltonol®) in canine patients, which may contribute to expanding the therapeutic option of the immunomodulatory class since immunological modifiers are still few known and commercialized in veterinary medicine.
 
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Publishing Date
2024-07-22
 
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