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The Chemotactic Activity of Normal and Complement‐Activated Serum as Measured by the Leading‐Front Method Using a Boyden Chamber
Author(s) -
HÅKANSSON L.,
VENGE P.
Publication year - 1984
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1984.tb00900.x
Subject(s) - chemotaxis , sephadex , size exclusion chromatography , chemistry , complement system , filtration (mathematics) , chromatography , biochemistry , immunology , antibody , biology , receptor , enzyme , statistics , mathematics
The chemotactic activity in serum, defined as the attractant effect of serum on the migration of neutrophil granulocytes (PMN) has been investigated for the purpose of characterizing the major chemotactic factors in serum as measured by the leading‐from technique, using a modified Boyden chamber. The chemotactic activity was measured in fresh and heated normal and activated serum and in serum fractions thereof separated by gel filtration. By gel filtration on Sephacryl S‐200 a partly heat‐labile C3‐C5‐associated chemotactic factor with molecular weight between 70,000 and 150,000 was isolated from fresh normal serum. The heat‐labile chemotactic activity was destroyed by pronounced complement activation. Gel filtration of complement‐activated serum on a Sephacryl S‐200 column showed the existence of one C5‐associated chemotactic factor with ˜70,000 molecular weight and one unidentified factor with ˜ 150.000 molecular weight, whereas no low molecular weight chemotactic activity was demonstrated. On the other hand, get filtration of activated serum on a Sephadex G‐75 column demonstrated one C5‐associaied chemotactic factor of ˜ 70,000 molecular weight and one 10,000–50,000 molecular weight factor active only in the presence of 2% normal serum. This investigation suggests that the chemotactic activity in fresh normal serum is mediated by a partly heat‐labile C3–C5‐associated complex. In activated serum three chemotactic factors were demonstrated, one unidentified factor with 150,000 mol wt and two CS‐dependcnt factors with 70,000 and 10,000–50,000 mol wt, the latter probably corresponding to C5a desarg . Accordingly, this study also suggests that C5a is not the only chemotactic factor generated in serum.