[Influence of the coagulase reaction on the leukotactic effect of Staphylococcus aureus in artificial body cavities of rabbits]. / Einfluss der Koagulasereaktion auf die leukotaktische Wirkung von Staphylococcus aureus in "künstlichen Körperhöhlen" von Kaninchen.

It was attempted, to determine the influence of coagulase-induced coagula on the leukotactic effects of Staphylococcus aureus in experimental subcutaneous body-cavities (KK) of rabbits. Purified staphylococcal coagulase mediated a partial coagulation in the KK when fibrinogen had been added. The coagula delayed the leukotaxis into the KK after the injection of (killed) encapsulated and non-encapsulated S. aureus extracted with 6 M guanidiniumchloride (Fig. 1), as well as after the injection of the untreated (viable) staphylococci (Fig. 2). The coagula also delayed phagocytosis, at least during the first hours of the experiments (Fig. 3).

[Production of coagulase from Staphylococcus aureus of different origin (author's transl)]. / Koagulasegewinnung von Staphylococcus aureus-Kulturen unterschiedlicher Herkunft.

Cultivation of Staphylococcus aureus in a fermenter significantly improved production of coagulase (Fig. 1,2,3). Optimal coagulase-activity was reached in the culture supernatant after inoculation with 8 x 10(9) colony-forming S. aureus per ml of brain heart infusion broth and 20 min at 37 degrees C. At this time most of the other enzymes and toxins (particularly protease and staphylokinase) had not been formed or only in small amounts. Characterization of coagulases from respectively 5 S. aureus-cultures from humans, cattle and dogs revealed molecular weights between 56800 and 71000, and isoelectric points between 5.4 and 6.4 (table 1).

[No or little chemotactic activity of staphylococcal substances on macrophages (author's transl)]. / Keine oder geringe chemotaktische Wirkungen von Staphylokokken-Substanzen auf Makrophagen.

Chemotactic effects of staphylococcal substances on macrophages from guinea pigs were studied using modified Boyden-chambers. Filters with an average pore-diameter of 12 micrometer allowed optimal migration of the macrophages. The macrophages were obtained from the peritoneal exsudates 96 h after stimulation with sodium-caseinate (fig. 1) and subsequently concentrated by centrifugation in a ficoll-ronpacon gradient (fig. 2, table 1). Casein had strong chemotactic effects on the macrophages with and without fresh guinea pig serum (fig. 3). Staphylococci, before and after extraction with guanidinium chloride, purified protein A and capsular substances were neither cytotaxic nor cytotaxigenic (in the absence resp. presence of fresh serum). Culture supernatants and staphylococci after incubation with their homologous antiserum proved cytotaxigenic, also lysates of granulocytes from the blood. Lysates of granulocytes from the peritoneal exsudates of guinea pigs were strongly cytotaxigenic for the macrophages.

[Studies on the activation of complement by encapsulated and non-encapsulated staphylococci after their extraction with guanidinium chloride (author's transl)]. / Untersuchungen über die Komplementaktivierung bekapselter und unbekapselter Staphylokokken nach Extraktion mit Guanidinium-chlorid

All demonstrable enzymes and toxins of encapsulated staphylococci (KS) were removed by extraction with guanidinium chloride. The capsules, however, remained apparently intact on the extracted (KS-Gu) staphylococci (fig. 1), as well as clumping factor and protein A. KS and KS-Gu failed to activate complement in the absence of specific antibodies. They showed neither immunadherence (table 1) nor agglutination by an antiserum against C3 (table 2). KS and KS-Gu had no significant chemotactic effects in vitro upon bovine granulocytes (fig. 2).

[Effects of capsules of Staphylococcus aureus on the rosette formation with B-lymphocytes and macrophages (author's transl)]. / Einfluss der Kapseln auf die Rosettenbildung von Staphylococcus aureus bei B-Lymphozyten und Makrophagen

After extraction with guanidium chloride nonencapsulated Staphylococcus aureus (S-Gu) and yeast cells (Hefe-Gu) reduced the hemolytic complement activity of normal human and guinea pig sera much more (fig 1) than encapsulated S. aureus (KS-Gu). Following incubation in the fresh sera S-Gu, Hefe-Gu and guanidinium chloride-extracted S. epidermidis, contrary to KS-Gu, had activated C3 on their surface. Therefore, they formed rosettes with human B-lymphocytes (fig. 2,3,4) and peritoneal macrophages from guinea pigs. The adherence-reactions were less frequent. if the pre-incubation was conducted in the heat-inactivated (30 min, 56 degrees C) sera (table 1,2). After pre-incubation in the absence of serum only the protein A-positive S-Gu formed rosettes with B-lymphocytes.

[Electron microscopic investigations on encapsulated Staphylococci (author's transl)]. / Elektronenmikroskopische Untersuchungen an bekapselten Staphylokokken

Attempts were made to study the surface of encapsulated staphylococci by electron microscopy. After fixation with osmium tetroxide the encapsulated staphylococci showed a "hazy" cell-surface, possibly because of capsular components (Fig. 1). On the other hand, the nonencapsulated staphylococci exposed a "smooth" surface of the cell wall (Fig 1). Similar findings were obtained after the immuneferritintechnique (Fig 2). The staphylococcal capsules could be clearly demonstrated by the "negative contrasting" method with ferritin (Fig 3). The capsules did not significantly enlarge after reaction of the encapsulated staphylococci with their homologous antiserum.

[Possible relationship between bacterial surface structures, immune-adherence, chemotaxis and phagocytosis (author's transl)]. / Einfluss bakterieller Oberflächenstrukturen auf Immunadhärenz, Chemotaxis und Phagozytose

In the present study we attempted to determine the relationship between bacterial surface structures, immune adherence, chemotaxis and phagocytosis. As shown in table 1 encapsulated bacteria (Klebsiella ozanae, K. pneumoniae, Pasteurella multocida, Streptococcus pneumoniae), Mycoplasma pneumoniae and M. fermentans had a much lower immune adherence-activity than non-encapsulated bacteria (Bacillus cereus, Brucella abortus, Erysipelothrix insidiosa, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, S. typhi). Consequently, the former bacteria, in the absece of specific antibodies, activated complement via the C-3-bypass to a smaller extent than the latter. In modified Boyden chambers the encapsulated bacteria and the mycoplasmas were much less cytotaxigenic than the non-encapsulated bacteria (table 3). Corresponding to this was the chemotactic response in vivo. Finally the rates of phagocytosis were considerably lower with the encapsulated bacteria than with the non-encapsulated ones (fig. 1).

[Leukotactic studies with artificial body cavities in rabbits (author's transl)]. / Leukotaktische Versuche in künstlichen Körperhöhlen bei Kaninchen

Artificial body cavities (KK) produced by implanting polyethylene balls in the subcutaneous tissues of rabbits proved to be suitable for the in vivo studies of leukotaxis. Na-caseinate induced a cell content per mm-3 KK-fluid of 7000-8500, "clumping-factor" of Staphylococcus aureus that of 9000 and capsular substances of S. aureus a delayed increase to 5300 cells per mm-3 KK-fluid. Paraffinum liquidum caused a somewhat delayed increase in granulocytes and not in macrophages. In all cases the percentage of granulocytes increased with the cell content and reached values up to 96%.