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Enhancement of Chemotactic Responsiveness in Vitro by C1 INH and Heparin

The Department of Anatomy and Department of Microbiology and Public Health, Michigan State University, East Lansing, Mich.

Abstract

Employing a modification of the Boyden technique for studying neutrophil chemotaxis in vitro, we investigated the effect of C1 inhibitor (C1 INH) and heparin on leukotaxis of human neutrophils. Leukocytes of normal adults were obtained from heparin anticoagulated plasma, and were washed in Hanks' solution (HBSS). The compartments of the chemotaxis chambers were separated by Millipore filters of 3 µ pore size. When C1 INH (7 units/ml) was added to the cell compartment of chemotaxis chambers containing 1 x 10⁶ neutrophils suspended in HBSS and 10% heparinized human plasma, there was a 3-fold enhancement (p < 0.001) in leukotactic response to lipopolysaccharide (LPS)-activated human plasma as compared to chambers without C1 INH. Subsequent experiments revealed the following results: 1) The enhancement of chemotaxis occurred also in response to zymosan and antigen-antibody-activated human plasma or serum; 2) no enhancement or increase in random motility occurred in the absence of a chemotactic stimulus; 3) enhancement could be completely prevented by prior exposure of the medium to a monospecific anti-C1 INH immunoadsorbent; 4) maximum enhancement occurred with 7 units of C1 INH while 4 units produced no effect; 5) the effect of C1 INH was reversible; 6) increasing levels of chemotactic stimulant increased the magnitude of enhancement.