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Blood, Vol. 95 No. 9 (May 1), 2000:
pp. 2954-2959
Importance of L-selectin-dependent
leukocyte-leukocyte interactions in human whole blood
Debra J. Mitchell,
Pauline Li,
Paul H. Reinhardt, and
Paul Kubes
From the Immunology Research Group, University of Calgary,
Calgary, Alberta, Canada.
The objective of this study was to investigate whether leukocytes
could be recruited by rolling leukocytes in a human whole blood model
system. In all experiments, either neutrophils, whole blood, or diluted
blood was perfused over immobilized E-selectin. With isolated
neutrophils (2 × 105/mL), the free-flowing neutrophils
were captured by attached neutrophils to form secondary interactions
that resulted in lines of rolling leukocytes. These secondary tethers
accounted for 50% to 60% of all interactions and were eliminated by
an L-selectin antibody, which also eliminated the lines of rolling
leukocytes. Perfusion of whole blood or diluted blood revealed no lines
of rolling leukocytes. The addition of red blood cells to isolated
neutrophils either in a 1000:1 or a 10:1 ratio also inhibited lines of
rolling leukocytes. Leukocytes were fluorescently labeled with
rhodamine-6G so that leukocyte-leukocyte interactions could be studied
in whole blood. A small number of secondary tethers (less than 20%)
occurred and could be reduced by more than 80% with an L-selectin
antibody. However, the overall impact on leukocyte recruitment was
negligible. Similar experiments were performed using murine whole blood
or isolated murine leukocytes. In the absence of red blood cells, murine leukocytes also formed lines of rolling leukocytes on
E-selectin, and secondary tethers accounted for 50% of total
interactions. However, when murine blood (diluted 1:5 with buffer) was
perfused over E-selectin, secondary tethers accounted for only 13% of
total interactions. These interactions were completely absent when
blood was used from L-selectin-deficient mice. These data demonstrate for the first time that the importance of L-selectin-dependent leukocyte-leukocyte interactions is greatly reduced in whole blood and
does not enhance overall recruitment of leukocytes in this physiologic milieu.
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