Abstract
Serpins represent one of the most diverse families of serine protease inhibitors. Despite their complexity, they are virtually found in all organisms and play an important role in homeostasis processes such as blood coagulation, inflammation, fibrinolysis, immune responses, chromatin condensation, tumor suppression, and apoptosis. There has recently been particular interest in studying serpin functions in infection and inflammation, especially since more serpins from parasites have been identified and characterized. Among helminths, Trichinella spiralis is one of the few parasites with an extremely strong ability to induce host immune suppression. Previous studies show that serpins are present in Trichinella and are expressed differentially at different parasite stages. More interesting, there is evidence of a recombinant serpin from Trichinella pseudospiralis that alters macrophage polarization in vitro. This finding could be relevant to comprehend the modulation process of the immune response. We expressed Tsp_01570, a putative serpin gene from Trichinella spiralis, in the eukaryotic system Pichia pastoris SMD1168H and evaluated its presence at different parasite stages, finding the serine protease inhibitor in the crude extract of adult worms. The effect of recombinant serpin on THP-1 cells was tested by quantification of IL-12p40, TNF-α, IL-4, and IL-10 cytokines released by ELISA. We also evaluated the expression of the M1 markers, CCR7 and CD86, and the M2 markers, CD163 and CD206, by immunofluorescence staining. This study represents the first insight in elucidating the importance of serpin Tsp_01570 as a potential molecular modulator.
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Acknowledgements
Cortez-de-la-Fuente LJ was a student under a CONACyT National Scholarship for Doctoral Programs (CVU 557895). We also thank Dr. Sergio Lozano Rodríguez for proofreading this manuscript.
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The Mexican National Council for Science and Technology (CONACyT) supported this work through the approval of a Grant application to JPPN (Number 238125) and RHB (Number 255173) from the “Fondo Sectorial de Investigación para la Educación” (SEP-CONACyT).
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José Prisco Palma-Nicolás contributed to the conceptualization, methodology, supervision, resources, project administration and funding acquisition, and writing—review and editing. Luis Jesús Cortez-de-la-Fuente contributed to the investigation, conceptualization, methodology, formal analysis, and writing—original draft. Gerardo García-González contributed to the methodology, investigation, and formal analysis. Romel Hernández-Bello contributed to the resource acquisition, methodology, and formal analysis. Gloria M. González contributed to the supervision, resource acquisition, and writing—review and editing.
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Animal care and protocols were evaluated and approved by the Ethics and Research Committee and the Institutional Committee for the Care and Use of Laboratory Animals, School of Medicine, Universidad Autónoma de Nuevo Leon (protocol number: MB19-00001). All experiments performed in this study followed the official Mexican standard (NOM-062-ZOO-1999), which is in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (NIH) of the United States.
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Cortez-de-la-Fuente, L.J., García-González, G., Hernández-Bello, R. et al. Expression of Trichinella spiralis serpin Tsp_01570 in Pichia pastoris: a first insight of its biomodulatory activity. Parasitol Res 122, 245–255 (2023). https://doi.org/10.1007/s00436-022-07723-0
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DOI: https://doi.org/10.1007/s00436-022-07723-0