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. Author manuscript; available in PMC: 2008 Mar 17.
Published in final edited form as: Cell Immunol. 2007 Jan 25;243(1):19–29. doi: 10.1016/j.cellimm.2006.12.001

Table 2a.

Quantification of CCL5-driven PBMC migration across basal IVBBB and aIVBBB. Parallel transmigration assays across either the basal IVBBB or aIVBBB, with or without 100 ng/mL CCL5 were performed using calcein-AM labeled and unlabeled cells as described in the Materials and Methods section. There was preferential migration of CD14+ monocytes across either IVBBB models, with higher levels seen across the aIVBBB. There was CCR5 up-regulation on monocytes during the transmigration assay (based on increased numbers and percentages of CD14+ CCR5+ cells in the non-migrated and migrated populations without added CCL5, relative to the input), with receptor down-regulation (based on reduced numbers and percentages of CD14+ CCR5+ monocytes) following CCL5-induced migration. CCL5 facilitated an increase in total CD3+ T-cell migration across either IVBBB models that was more efficient for CCR5+ cells (based on the higher percentages of migrating CD3+CCR5+ T-cells relative to the input and cells migrating without added chemokine). Higher numbers of migrating CD3+ T-cells were seen with the aIVBBB, indicating that endothelial activation significantly induced chemokine-driven T-cell migration in vitro. Shown are the means of five separate experiments using different donors. [ ] indicates the mean percentage of input cells that migrated in 3 hours, while ( ) indicates the mean percentages of the parent mononuclear cell population that were CCR5+. Key: aIVBBB: cytokine-activated in vitro blood-brain barrier, IVBBB: in vitro blood-brain barrier, PBMCs: peripheral blood mononuclear cells.

BBB MODEL CONDITION CD14+ CD14+ CCR5+ CD3+ CD3+ CCR5+
INPUT 167,973 12,730 (7.58%) 482,374 103,253 (21.72%)
Basal IVBBB No Chemokine-Non-migrated 67,036 12,531 (18.69%) 476,827 99,582 (20.80%)
Basal IVBBB No Chemokine-Migrated 100,937 [60.1%] 28,661 (28.39%) 5,547 [1.1%] 1,443 (26.01%)
Basal IVBBB + CCL5-Non-migrated 57,762 9,892 (17.13%) 464,544 87,456 (18.83%)
Basal IVBBB + CCL5-Migrated 110,211 [65.6%] 18,055 (16.38%) 17,830 [3.7%] 9,328 (52.33%)
aIVBBB No Chemokine-Non-migrated 61,319 13,344 (21.76%) 475,058 97,866 (20.60%)
aIVBBB No Chemokine-Migrated 106,654 [63.5%] 22,419 (21.02%) 7,316 [1.5%] 2,908 (39.75%)
aIVBBB + CCL5-Non-migrated 48,317 8,504 (17.60%) 450,877 80,916 (17.95%)
aIVBBB + CCL5-Migrated 119,656 [71.2%] 12,730 (10.64%) 31,497 [6.5%] 16,756 (53.20%)