Venom components of the scorpion Centruroides limpidus modulate cytokine expression by T helper lymphocytes: Identification of ion channel-related toxins by mass spectrometry

• Published in: International immunopharmacology 2020-07, Vol.84, p.106505, Article 106505
• Main Authors: Cota-Arce, Julián M., Zazueta-Favela, Daniela, Díaz-Castillo, Fernando, Jiménez, Samanta, Bernáldez-Sarabia, Johanna, Caram-Salas, Nadia L., Dan, Kee W.L., Escobedo, Galileo, Licea-Navarro, Alexei F., Possani, Lourival D., De León-Nava, Marco A.
• Format: Article
• Language: English
• Subjects: Animals; CD4 antigen; Cell proliferation; Cell Proliferation - drug effects; Cells, Cultured; Centruroides limpidus; Chromatography; Chromatography, Liquid; Cytokines; Cytokines - immunology; Helper cells; High performance liquid chromatography; Immunomodulation; Immunomodulators; Interleukin 10; Interleukin 4; Ion channels; Liquid chromatography; Lymphocytes; Lymphocytes T; Male; Mass spectrometry; Mass spectroscopy; Membrane proteins; Mice, Inbred BALB C; Potassium; Potassium channels; Potassium channels (voltage-gated); Scientific imaging; Scorpion venom; Scorpion Venoms - chemistry; Scorpion Venoms - toxicity; Scorpions; Sodium channels (voltage-gated); Spectroscopy; T helper cell; T-Lymphocytes, Helper-Inducer - drug effects; T-Lymphocytes, Helper-Inducer - immunology; Tandem Mass Spectrometry; Toxins; Venom; γ-Interferon
• ISSN: 1567-5769; 1878-1705
• DOI: 10.1016/j.intimp.2020.106505

•Toxins of the scorpion Centruoides limpidus increase T helper cytokine production.•Venom fractions of the scorpion C. limpidus has no effect on T cell proliferation.•NaScTx and KScTx toxins are involved in the effector function of T lymphocytes.•NaScTx and KScTx could be used as immunomodulatory molecule in the near future. The study of the effector mechanisms of T helper cells has revealed different phenotypic characteristics that can be manipulated for designing new therapeutic schemes in different pathological scenarios. Ion channels are significant targets in T lymphocyte modulation since they are closely related to their effector activity. Remarkably, some toxins produced by scorpions specifically affect the function of these membrane proteins. For that reason, these toxins are important candidates in the search for new immunomodulators. Here, the effect of two venom fractions of the scorpion Centruroides limpidus was assessed on T lymphocyte proliferation and cytokine production. The venom fractions ClF8 and ClF9 were separated by reversed-phase high-performance liquid chromatography (RP-HPLC) and cultured at 25 and 35 µg/ml with murine T lymphocytes. The results indicate that the fraction ClF8 increased both production and secretion levels of IFN-γ, IL-4, IL-17A and IL-10 by CD4+ T cells at 24 h. In contrast, fraction ClF9 only promoted the secretion of IL-17A and IL-10 at its highest concentration (35 µg/ml). Both fractions did not show any effect on T cell proliferation. Subsequent analyses by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed seventeen toxins in the fraction ClF8 and five toxins in the fraction ClF9, most of them with voltage-gated sodium (NaScTx) and potassium (KScTx) channels as molecular targets. These toxins might probably interact with ion channels involved in T lymphocyte activity. Our findings suggest that the difference in composition between the two fractions could be related to the observed effects, and the components identified could be isolated to search for possible immunomodulatory molecules.