Complement and Fc receptors on cord blood and adult neutrophils.

The percentage of neutrophils forming erythrocyte-antibody-complement complex (EAC) and with erythrocyte-antibody complex (EAG) was determined for 30 normal newborns and 10 normal adults. The cord values ranged from 84-100% EAC binding cells with a mean of 95%. The adult values ranged from 89-100% EAG binding cells with a mean of 96% rosetted neutrophils. The mean percentage of EAG binding cells in the cord samples was 81%, whereas in the seven adult samples the mean was 87%.

Characteristics of cells present in peritoneal fluids of mice injected intraperitoneally with Bordetella pertussis.

Peritoneal fluids obtained from mice after the intraperitoneal administration of Bordetella pertussis vaccine, heated vaccine, an extract of the organisms, killed Escherichia coli, or thioglycolate medium were examined in terms of total cells and percentage that adhered to glass cover slips during 2-h incubation period. All these substances were found to increase the number of leukocytes in peritoneal fluid within 1 to 2 days after the injection. This increase appeared to be due to an influx of macrophages and polymorphonuclear leukocytes with relative proportions at a given time dependent upon the material involved in the induction of the response. The initial increases after pertussis vaccine seemed to be due mainly to an influx of monomuclear cells, whereas with E. coli neutrophils constituted the major portion of the cell population. The percentage of peritoneal cells that attached to glass was also found to be markedly reduced in preparations obtained from mice after the injection of B. pertussis or E. coli. There appeared to be differences in persistence of this phenomenon, with preparations containing the histamine-sensitizing factor being the most active in affecting adherence properties. Thus these data would suggest that the action of B. pertussis on macrophages (or precursors) and neutrophils is not expressed in terms of suppression of emigration properties, as has been reported by others for lymphocytes, but is manifested in the alteration of glass-adherence characteristics. Within experimental limitations, it is believed that macrophages are possibly more involved in terms of altered function than are the polymorphonuclear cells.

Influence of Bordetella pertussis on Adenosine Diphosphate-Induced Aggregation of Mouse Platelets.

Several lines of evidence support the concept that Bordetella pertussis-induced histamine hypersensitivity in mice represents a malfunction in beta-adrenergic receptors of the autonomic nervous system. In this study mouse platelets were used as an in vitro test for such receptor dysfunction. It was found, to our knowledge for the first time, that platelets of normal mice were subject to the aggregative action of adenosine diphosphate (ADP) with the response pattern resembling that reported for cells of certain other species. These "normal" platelets were not aggregated by epinephrine, norepinephrine, or isoproterenol (isuprel). However, pretreatment with these catecholamines potentiated the platelet response to ADP. Potentiation by norepinephrine was suppressed in the presence of dibenzyline (an alpha-receptor antagonist); blockade of beta-type receptors with propranolol reduced isuprel enhancement. An extract of B. pertussis influenced aggregation initiated by a combination of ADP and isuprel in a way quite similar to propranolol pretreatment. These observations suggested the possible role of adrenergic receptors in at least the enchancement phenomena and, moreover, pointed to those types of experiments which might distinguish a receptor disturbance. Platelets derived from pertussis-vaccinated mice were more responsive to ADP than were those from normal animals. This alteration appeared to be a property of the cells and not of the suspending medium. ADP-initiated aggregation was also enhanced in the presence of epinephrine and norepinephrine. However, pretreatment with isuprel did not have the same effect as noted for normal donor platelets. These results point to a disturbance at the beta-adrenergic receptor level and lend support to the concept of beta-blockade in pertussis-treated mice.

Bordetella pertussis-induced alteration of the normal hyperglycemic response of mice to 3',5'-adenosine phosphate.

Blockade of the adrenergic receptors or the injection of Bordetella pertussis vaccine prevented the normal hyperglycemic response of CFW mice to exogenous cyclic 3',5'-adenosine phosphate (3',5'-AMP). The cyclic nucleotide was also ineffective in offsetting the histamine hypersensitivity of the beta-adrenergically blocked and the pertussis-sensitized groups. These observations suggest that the autonomic malfunction occurs at a point(s) subsequent to release of 3',5'-AMP. Neither the hyperglycemic effect of epinephrine or 3',5'-AMP, or both, nor the histamine sensitivity of pertussis-pretreated animals was influenced by the subcutaneous administration of theophylline.

Protein-polyelectrolyte interactions. The concanavalin A precipitin reaction with polyelectrolytes and polysaccharide derivatives.

1. Concanavalin A formed precipitates with polyelectrolytes such as fucoidan, RNA, heparin and bacterial lipopolysaccharides. 2. Precipitate formation also occurred between ficoll and concanavalin A. 3. Precipitate formation between concanavalin A and dextran or soluble starch was induced by the incorporation of phosphate groups into the unreactive glucans. 4. Introduction of polar groups, such as acetate, formate and phosphate, into glycogen resulted in enhanced precipitation with concanavalin A, whereas the opposite effect was noted on incorporation of hydrophobic (methyl) centres. 5. Neutral sugars and salt partially inhibited complex-formation between polyelectrolytes and concanavalin A. 6. Concanavalin A-glycogen complexes could be dissociated with 5% (w/v) trichloroacetic acid or 44% phenol-water. 7. Concanavalin A lost its glycogen-complexing ability after phenol treatment. 8. Evidence is presented for the existence of common binding sites on concanavalin A for both neutral polysaccharides and polyelectrolytes. 9. Hydrogen bonding appeared to play a major role in neutral polysaccharide-concanavalin A precipitate formation, whereas both hydrogen bonding and electrostatic forces were implicated in polyelectrolyte-concanavalin A complex-formation.

In vivo and in vitro manifestations of adrenergic blockade in Bordetella pertussis-vaccinated mice.

The in vivo metabolic events which follow the administration of epinephrine, norepinephrine, or isoproterenol were examined in normal, Bordetella pertussis-vaccinated, and alpha and beta adrenergically blocked mice. The normal hyperglycemic response to epinephrine was suppressed in all experimental groups. The pertussis-sensitized and beta-blocked animals produced similar split patterns of altered response not duplicated by the alpha-blocking compounds. Those catecholamines that normally increase free fatty acids and lactic acid in the circulation failed to do so in the pertussis-sensitized and beta-blocked animals; the inhibition of free fatty acid mobilization was also demonstrated with adipose tissue incubated in vitro. An extract of the pertussis organism added to incubation media prevented the catecholamine-induced free fatty acid response. The epinephrine-stabilizing effect of bovine serum albumin (Cohn-fraction V) was observed. The results of these studies further emphasize a correlation between pertussis-sensitized and beta-adrenergically blocked mice.