International Conference on Biomedical and Cancer Research
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Accepted Abstracts

Extracellular traps and mitochondrial dynamic in stimulated autologous leukocyte cultures from healthy human blood samples

Rinero R1 , Reyna MV1 , Rodríguez FM1 , Carabajal Miotti CL2 , Ruiz de Frattari S2 , Vargas AH2 , Gonzalez Silva N2 , Novak ITC*1
1 Institute of Cell Biology, National University of Cordoba (UNC), Argentina.
2 Institute of Hematology and Hemotherapy, National University of Cordoba (UNC), Argentina

Citation: Rinero R , Reyna MV , Rodríguez FM, Gonzalez Silva N , Novak ITC etal (2019) SciTech Biomrf-Cancer Sciences 2019. Tokyo: Japan

Received: June 18, 2019         Accepted: June 26, 2019         Published: June 26, 2019

Abstract

Extracellular traps (ETs) are structures of chromatin and intracellular proteins which are extruded in leukocytes under inflammatory conditions. The presence of beta-tubulin and HLA-DR have not been reported in the traps. Not all ETs are created equal, this depends on the source of stimulation. On the other hand, phenotypes adopted by immune cells in their responses are influenced by "mitochondrial dynamic" (MD). MD is the set of characteristics of shape, position and size of mitochondria. These organelles are currently considered as regulating functions of innate and adaptive immunity. Mitocondrial damage was reported in dendritic cells and macrophages exposed to NETs. Objectives: to generate ETs in cultures of leukocytes challenged with LPS or fMLP and to carry out the beta-tubulin, HLA-DR labeling, on the other hand, to observe the morphological characteristics of the mitochondria in lymphocytes in the LPS, fMLP or OVA assays. Methods: Autologous cultures from healthy human blood samples (n = 10) with ethical consent (HNC, FCM), anticoagulated with heparin were stimulated with 25 ng/ml LPS or 0,25 ng/mL fMLP or 100 ug/mL OVA, 30 minutes. Immunofluorescence technique with anti-beta tubulin and anti HLA-DR antibodies, DNA staining with DAPI. Paired blood samples provided the controls. Cells of the cultures were studied with transmission electron microscopy. Results: beta-tubulin and HLA-DR molecules were localized in the ETs. Alterations of mitochondrial morphology of lymphocytes were observed in the samples with LPS with increase in size and complexity of the cristaes with electrolucid images (t-test for paired samples, p <0.0001). Differences of mitocondrial areas were observed in OVA assay (p<0.005) and in fMLP assay were not observed differences. Conclusions: the expression of beta-tubulin and HLA-DR contributes to the better understanding of the composition of the ETs generated by different stimulators and may have significance as a therapeutic target. HLA-DR release in ETs may influence enviroment contributing to antigen presentation class II pathway. The ETs would affect the DM of the surrounding cells, influencing cellular function. Key words Extracellular traps, mitochondrial dynamics, human leukocytes, lymphocytes, beta tubulin, HLA-DR.