Effects of preparative procedures and of cell activation on flow of white cells through micropore filters

Abstract

Using newly developed filtration techniques to analyse the flow resistance of white blood cells (WBC), the effects of preparative procedures and of various treatments (including activation) have been tested. Flow rates of WBC were measured using both 5 and 8 micron pore filters. It was found that EDTA was the anticoagulant of choice over heparin and sodium citrate, and that calcium-containing media should be avoided because they promoted cell-cell aggregation. Exposure to density separation media did not significantly alter filtration rates, but storage of prepared WBC for even 1 h caused deterioration in flow. Storage of whole blood before preparation of the WBC had a much lesser effect. Storage appeared to be linked to cell activation, and use of the activating agents phorbol ester (PE, 4.5 x 10(6) M/l) and f-methyl-leucyl-phenylalanine (FMPL, 10(-7) and 10(-9) M/l) also increased WBC resistance to pore transit. The effect of PE was greater, causing rapid pore blockage, but was non-specific, while the lesser effect of FMLP was restricted to granulocytes and was concentration dependent. Cell volume was increased by these agents, but measurement of the filterability of hypotonically swollen granulocytes showed that volume changes alone only partly explained the activation effects. These results suggest that activation in vivo would have a significant rheological effect, detectable by filtration methods.