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. 2019 Nov 22;10:1188. doi: 10.3389/fgene.2019.01188

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Copyright © 2019 Tsai, Lin, Tsai, Lin, Ho, Lu, Weng, Lin, Lin and Li

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Using in vitro leukocyte transendothelial migration (LTEM) assay to investigate whether serum promotes leukocyte to penetrate BBB endothelial layer. We developed the specific LTEM model to study autoimmune encephalitis (AE) by making the endothelial layer with the cells of BBB endothelial cell line (hCMEC). (A) We first determined the specified FSC-A and SSC-A values which would be applied to gate the cells with LTEM assay. (B) When treated with HC serum, 239 of the treated Toledo cells were detected. (C) When the Toledo cells were treated with AE serum, we collected 358 ones. (D) AE serum significantly promoted Toledo cells to penetrate the BBB endothelial layer by almost 1.5 times than HC serum did (N = 4). (E) When fresh B cells were applied to replace Toledo cell, consistent and significant result was observed (N = 4). (F) No significant difference was observed when neutrophil-like cells were applied (N = 4). Data was presented as mean ± S.D. * denoted p-value < 0.05.