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Expression of cytokines that regulate immune responses in alpaca (Vicugna pacos) blood leukocytes stimulated by clostridial antigens and retinoic acid

Juan A. More Bayona1*, Alberto G. Manchego1, Hermelinda Rivera2, Nieves Sandoval3, Mercy Ramírez2, Kim Lam Chiok2, Roberto Lazaro1 and Carla Bardález1
  • 1 Laboratorio de Microbiología y Parasitología, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Departamento Académico de Salud Animal, Peru
  • 2 Laboratorio de Microbiología y Parasitología, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Departamento Académico de Salud Animal, Peru
  • 3 Laboratorio de Histología, Embriología y Patología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Departamento Académico de Salud Animal, Peru

Expression levels of cytokines involved in humoral and cellular responses were determined in vitro in blood leukocytes from alpacas, stimulated with Clostridium perfringens (AgCP) antigens and retinoic acid (ATRA) as adjuvant. Different treatments were tested as follows: 10ng AgCP and 0.25mUI ATRA; 1ng AgCP and 2.5mUI ATRA; 100pg AgCP and 25mUI ATRA; 10pg AgCP and 250UI ATRA. Treated and untreated leukocytes were incubated at 37C for 24 hours after which total ARNm (messenger ARN) was extracted. We used real time RT-PCR and relative quantification to assess difference in expression of cytokines in leukocytes in comparison with untreated cell controls. We found increasing expression of IFNγ and Interleukin-2 (IL-2) in all treatments up to 100pg of AgPC and 25UI ATRA (8:1 y 7.5:1, respectively). In case of Tumoral Necrosis Factor alfa (TNF-α), increasing expression was observed until 10pg of agCP and 250mUI of ATRA (10.3:1) were used. As for Interleukin 10 (IL-10), it reached maximum expression when 1ng of AgCP and 2.5mUI of ATRA were used (27.4:1). Taken together, our results suggest clostridial antigens and ATRA confer good stimulation of cellular immune responses in alpaca leukocytes; however, lower concentrations of both, antigen and adjuvant, were able to induce maximum expression of IL-10.

Acknowledgements

We thanks to Consejo Nacional de Ciencia y Tecnología (CONCyTEC – Perú) by financing support.

References

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Keywords: Retinoic acid, Cytokines, Clostridium perfringens, alpaca, Enterotoxemia

Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.

Presentation Type: Abstract

Topic: Translational immunology and immune intervention

Citation: More Bayona JA, Manchego AG, Rivera H, Sandoval N, Ramírez M, Chiok K, Lazaro R and Bardález C (2013). Expression of cytokines that regulate immune responses in alpaca (Vicugna pacos) blood leukocytes stimulated by clostridial antigens and retinoic acid. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00318

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Received: 19 Mar 2013; Published Online: 22 Aug 2013.

* Correspondence: Mr. Juan A More Bayona, Laboratorio de Microbiología y Parasitología, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Departamento Académico de Salud Animal, Lima, Lima, 51, Peru, jmorebayona@yahoo.com