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The Chemokine CCL4 (MIP-1β) Evokes Antinociceptive Effects in Mice: a Role for CD4+ Lymphocytes and Met-Enkephalin

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Abstract

In the present study, we characterize the antinociceptive effects produced by the chemokine CCL4 in mice. The intraplantar administration of very low doses of CCL4 (0.1–3 pg) produced bilateral antinociception assessed by the unilateral hot-plate test (UHP) without evoking chemotactic responses at the injection site. Moreover, the subcutaneous administration of CCL4 (3–100 pg/kg) also yielded bilateral antinociception in the UHP and the paw pressure test and reduced the number of spinal neurons that express Fos protein in response to noxious stimulation. The implication of peripheral CCR5 but not CCR1 in CCL4-evoked antinociception was deduced from the inhibition produced by systemic but not intrathecal, administration of the CCR5 antagonist DAPTA, and the inefficacy of the CCR1 antagonist J113863. Besides, the inhibition observed after subcutaneous but not intrathecal administration of naloxone demonstrated the involvement of peripheral opioids and the efficacy of naltrindole but not cyprodime or nor-binaltorphimine supported the participation of δ-opioid receptors. In accordance, plasma levels of met-enkephalin, but not β-endorphin, were augmented in response to CCL4. Likewise, CCL4-evoked antinociception was blocked by the administration of an anti-met-enk antibody. Leukocyte depletion experiments performed with cyclophosphamide, anti-Ly6G, or anti-CD3 antibodies indicated that the antinociceptive effect evoked by CCL4 depends on circulating T lymphocytes. Double immunofluorescence experiments showed a four times more frequent expression of met-enk in CD4+ than in CD8+ T lymphocytes. CCL4-induced antinociception almost disappeared upon CD4+, but not CD8+, lymphocyte depletion with selective antibodies, thus supporting that the release of met-enk from CD4+ lymphocytes underlies the opioid antinociceptive response evoked by CCL4.

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Acknowledgements

Grants were provided by the Ministerio de Economía, Industria y Competitividad, and FEDER (European Union) (SAF2017-86799-R). IUOPA is supported by Obra Social Fundación Cajastur-Liberbank (Asturias, Spain). M.G.D. is granted by a FPU predoctoral fellowship (FPU13-04585, MECD, Spain). A.R.F. is recipient of a Ramón y Cajal Fellowship.

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Authors and Affiliations

  1. Laboratorio de Farmacología, Facultad de Medicina, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Universidad de Oviedo, C/ Julián Clavería 6, 33006, Oviedo, Asturias, Spain

    Mario García-Domínguez, Ana Lastra, Agustín Hidalgo, Luis Menéndez & Ana Baamonde

  2. Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Universidad de Oviedo, 33006, Oviedo, Asturias, Spain

    Alicia R. Folgueras

  3. Área de Biología Celular, Departamento de Morfología y Biología Celular, INEUROPA (Instituto de Neurociencias del Principado de Asturias), Universidad de Oviedo, C/ Julián Clavería 6, 33006, Oviedo, Asturias, Spain

    Rafael Cernuda-Cernuda

  4. Unidad de Histopatología Molecular en Modelos Animales de Cáncer, IUOPA, Universidad de Oviedo, C/ Julián Clavería 6, 33006, Oviedo, Asturias, Spain

    María Teresa Fernández-García

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  1. Mario García-Domínguez
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García-Domínguez, M., Lastra, A., Folgueras, A.R. et al. The Chemokine CCL4 (MIP-1β) Evokes Antinociceptive Effects in Mice: a Role for CD4+ Lymphocytes and Met-Enkephalin. Mol Neurobiol 56, 1578–1595 (2019). https://doi.org/10.1007/s12035-018-1176-8

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