CD45RO and mitochondrial dynamics in human autologous leukocyte cultures stimulated in vitro.
The phenotypes adopted by immune cells in their responses are influenced by the "mitochondrial dynamics" (MD): set of characteristics of shape, position and size of mitochondria. These organelles are currently considered to have regulatory functions upon innate and adaptive immunity....
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| Autores principales: | , , , , , , , |
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| Formato: | Artículo revista |
| Lenguaje: | Español |
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Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2019
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| Materias: | |
| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/25863 |
| Aporte de: |
| Sumario: | The phenotypes adopted by immune cells in their responses are influenced by the "mitochondrial dynamics" (MD): set of characteristics of shape, position and size of mitochondria. These organelles are currently considered to have regulatory functions upon innate and adaptive immunity. The T-lymphocytes intervene in the adaptive response and upon activation they express the CD45RO marker molecule. The endotoxemia produced by endotoxins such as lipopolysaccharide (LPS) in the blood circulation lead to inflammation in multiple organs. T-lymphocytes respond innately to LPS with signaling via the TLR4 receptor, it does not affect proliferation or cytokine secretion but increases adherence and inhibits their migration. The aims of the present work were to observe the morphological characteristics of mitochondria in lymphocytes in cultures of leukocytes exposed to LPS, formylated peptides (fMLP) or ovalbumin (OVA) and mark CD45RO in tests with LPS or OVA.
Autologous leukocyte cultures from healthy heparin anticoagulated human blood samples (n = 10) with ethical consent (HNC, FCM), were stimulated with 25 ng / ml of LPS or 0.25 ng / ml of fMLP or 100 ug / ml of OVA, for 30 minutes. Immunofluorescence technique with anti-CD45RO, DNA staining with DAPI. Paired blood samples provided the controls. Cultured cells were studied with transmission electron microscopy.
Alterations of the mitochondrial morphology of the lymphocytes were observed in the samples with LPS with an increase in the size of the mitochondria and the complexity of the cristae, with electrolucid images (t test for paired samples, p <0.0001). Significant differences were observed in the increased mitochondrial areas in the OVA test (p <0.005), but not in the fMLP assay. Increased positivity was observed for CD45RO in trials with LPS (p <0.005) or OVA (p <0.0001) with respect to paired samples without stimulation.
The increase of the mitochondrial area has been related to early activation of T-cells in the OVA assay, coinciding with CD45RO positivity. The various activators influence the MD and the phenotypes of the immune cells and such MD is important both in the metabolic and immune functions in said cells.
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