Journal Article•
Studies on eosinophil leucocyte migration. II. Factors specifically chemotactic for eosinophils and neutrophils generated from guinea-pig serum by antigen-antibody complexes.
TL;DR: The properties associated with guinea-pig eosinophil chemotactic activity were similar to previously published data for a fragment cleaved from the 5th component of complement.
Abstract: Chemotactic activity for eosinophils and neutrophils has been studied using guinea-pig serum activated by preformed antigen–antibody complexes.
Rabbit complexes or complexes prepared either with guinea-pig IgG2 or IgG1 were equally capable of generating a heat stable activity from guinea-pig serum that was chemotactic for guinea-pig eosinophils and for neutrophils of both the guinea-pig and the rabbit. This was distinguished from a relatively heat-labile chemotactic activity present in untreated guinea-pig serum.
The heat-stable complex-mediated chemotactic activity was thought to be complement dependent since chemotaxis for eosinophils or neutrophils could not be generated from heated serum, ammonia treated serum or from serum treated with complexes in the presence of 0·01 M EDTA.
Guinea-pig sera, activated either with rabbit, guinea-pig IgG1 or IgG2 complexes, was fractionated by sucrose-density gradient ultracentrifugation and by Sephadex chromatography. In all experiments two peaks of cell specific chemotactic activity could be separated. The peak of activity for guinea-pig neutrophils was approximately 4·5S and in the fractionation range of proteins having a molecular weight of between 65,000 and 85,000. The peak of guinea-pig eosinophil chemotactic activity was 1·5S–2S and in the molecular weight range of between 15,000 and 20,000. Those fractions which were chemotactic for guinea-pig neutrophils did not attract rabbit neutrophils. Rabbit neutrophils migrated towards those fractions of guinea-pig serum chemotactic for guinea-pig eosinophils; therefore, the properties associated with guinea-pig eosinophil chemotactic activity were similar to previously published data for a fragment cleaved from the 5th component of complement.
Citations
More filters
TL;DR: This chapter discusses the structural and functional characterization of anaphylatoxins, spasmogenic substances released during complement activation that elicit a variety of cellular responses, which implies that they play a significant role in inflammation and acute allergic reactions.
Abstract: Publisher Summary This chapter discusses the structural and functional characterization of anaphylatoxins, spasmogenic substances released during complement activation. These low molecular weight peptide fragments of C3 and C5 elicit a variety of cellular responses, which implies that they play a significant role in inflammation and acute allergic reactions. Their stimulation of multiple cell types at pico- and femtomolar concentrations suggests a hormone-like action via specific cell surface receptors. By using the information available defining the amino acid sequences of these molecules it is possible to synthesize oligopeptides with the properties of anaphylatoxins and these compounds serve as effective tools for analyzing further the mechanism(s) by which these potent substances function. They are functionally defined by their actions on the vasculature, smooth muscle, mast cells, and certain types of peripheral blood cells. The most important contribution of anaphylatoxin research to biology is the realization that complement reaction products act as hormone-like messengers that profoundly affect the function of certain cells. A variety of cell types is activated by the complement reaction products C3a and C5a, whether they are acting as spasmogens, chemotaxins, or cause release reactions.
448 citations
TL;DR: The results indicate that in addition to the classic inhibitory DP1 receptor, eosinophils possess a second, novel DP2 receptor that is associated with PGD2-induced cell activation and may play a role in eos inophil recruitment in asthma.
316 citations
TL;DR: The observed refractoriness of eos inophils from eosinophilic subjects to both directional migratory and priming effects of IL-5 in vitro, may reflect a deactivation process resulting from prior exposure in vivo.
247 citations
TL;DR: The molecules in question, namely C3a, C4a, and C5a, are bioactive fragments that are generated in vivo as a result of complement activation and have recently been identified as immunomodulators capable of regulating aspects of the humoral and cellular immune response.
Abstract: Biologic characterization and structural analysis of the complement-derived factors known as anaphylatoxins have advanced markedly over the past decade. For example, structure analyses have progressed to a point that detailed features of these The number designation used throughout the text for amino acid residues is that of the human C3a sequence unless otherwise stated factors have been elucidated at the molecular level. The molecules in question, namely C3a, C4a, and C5a, are bioactive fragments that are generated in vivo as a result of complement activation. The physiologic role of these anaphylatoxins in host defense mechanisms is becoming more and more apparent as their many and varied biologic activities are better defined and understood. These humoral mediators induce numerous inflammatory responses and have recently been identified as immunomodulators capable of regulating aspects of the humoral and cellular immune response [87, 88]. In a very real sense the anaphylatoxins are local hormones, assuming that one accepts the definition for a local hormone of “a substance with hormone-like properties, produced from blood or other body fluids precisely when and where it is needed, that is rapidly destroyed.” The anaphylatoxins fulfill these criteria by acting at the cellular level via surface receptors to stimulate responses, are generated in blood either after tissue injury or during immunologic damage, and are under the strict control of a serum enzyme (e. g., carboxypeptidase N) [4, 68].
239 citations
TL;DR: ECF-A represents a hitherto undescribed agent which selectively attracts eosinophil leukocytes and is distinguishable from a previously described complement-dependent eOSinophilotactic factor ( ECF-C).
Abstract: The capacity of actively or passively sensitized guinea pig lung to react with antigen to release a factor specifically chemotactic for eosinophil leukocytes (ECF-A) has been demonstrated. The release of ECF-A was also accompanied by the elaboration of both histamine and SRS-A and the appearance of all these mediators exhibited a similar response in terms of the time course of passve sensitization, the effect of antigen dose, the time course of release, divalent cation dependence and enhancement by the presence of succinate or maleate. Decomplementation by the administration of purified cobra venom factor had no effect on the antigen-induced release of ECF-A from actively or passively sensitized lung fragments.
When fragments of guinea pig lung were passively sensitized with fractions of guinea pig 7S IgG, only the IgG1-containing fractions prepared tissue for the antigen-induced release of ECF-A. Histamine, SRS-A, bradykinin, serotonin, and the prostaglandins PGE1, PGE2, and PGF2α were not eosinophilotactic per se; neither was ECF-A detected following the incubation of these agents with sensitized lung in the absence of antigen.
Both eosinophilotactic activity and SRS-A survived extraction in 80% ethanol and evaporation to dryness. SRS-A, however, withstood boiling in alkaline solution for 20 min, whereas ECF-A activity was abolished by this procedure. SRS-A and ECF-A could also be separated by gel filtration. ECF-A activity was completely recovered following its passage through a column of Sephadex G-25 and had an estimated molecular weight of between 500 and 1000. On the basis of size and a formation mechanism independent of the complement system, ECF-A is distinguishable from a previously described complement-dependent eosinophilotactic factor (ECF-C). Thus, ECF-A represents a hitherto undescribed agent which selectively attracts eosinophil leukocytes.
232 citations
Cites background or methods from "Studies on eosinophil leucocyte mig..."
...Material selectively attracting neutrophils, also of complement origin (1) but having a larger molecular size, is termed NCF-C....
[...]
...The antiserum was fractionated by diethylaminoethyl (DEAE)-cellulose chromatography (1)....
[...]
...release an eosinophilotact ic agent after the addi t ion of specific an t igen and whether such an agent is the same or different f rom tha t produced from the in te rac t ion of se rum with preformed a n t i g e n - a n t i b o d y complexes (1) or zymosan3 For this purpose, an in v i t ro model invo lv ing the release of chemical media tors f rom guinea pig lung was used, which has been employed b y numerous workers as a model of immedia te - type hypersens i t iv i ty in the guinea pig (4-7)....
[...]
...Using this technique, it was shown that guinea pig IgG1 or IgG2, as preformed antigen-antibody complexes, could both generate from whole serum a factor which was specifically chemotactic for eosinophils (ECF-C) 1 and was apparently identical to C5a (1)....
[...]
...Guinea pig eosinophils were obtained by peritoneal lavage from animals which had received multiple injections of horse serum (1)....
[...]