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Es wurden die Ergebnisse einer Untersuchung uber das Vorkommen beta-hamolytischer Streptokokken bei 342 Bonner Schulkindern mitgeteilt. Die Gesamtzahl betrug fur den Herbst-Winter-Zeitraum 1965/1966 20,46% bei 10,52% Gruppe A-Streptokokken. Die weitere Aufschlusselung ergab fur die Gruppe A-Rate einen Schwankungsbereich von 5,78–30%. Daruber hinaus wurden Befunde vorgelegt, welche darauf hinweisen, das die meisten der isolierten Gruppe A-Streptokokkenstamme wahrscheinlich infektios bzw. pathogen waren. Es wurde angeregt, ahnliche Untersuchungen in verschiedenen Teilen Westdeutschlands durchzufuhren, um die Haufigkeit des Vorkommens und die Bedeutung von Gruppe A-Streptokokken bei Schulkindern zu ermitteln und somit einen zuverlassigen Masstab fur Kontrollmasnahmen zu erhalten.

Um eine evtl. Dissoziation von Ag-Ak-Komplexen in vivo (Kaninchen) nachzuweisen, wurden 1. Komplexe injiziert, deren Ak-Teil aus speciesgleichen Tieren (Kaninchen) stammte, 2. Komplexe verwendet, deren Antigenkomponente in ihrer antigenen Wirksamkeit weitgehend bekannt ist (C-Streptokokken) und 3. nur zwei verschiedene Arten von Komplexen (Agglutinat und Pracipitat) gepruft. Bei Injektion des Pracipitates konnten weder gegen den Komplex, noch gegen den Ag-Teil des Komplexes, noch gegen die Antikorperkomponente des Komplexes pracipitierende oder agglutinierende Antikorper nachgewiesen werden. Dagegen gelang es bei Verwendung eines Agglutinates aus inaktivierten C- Streptokokken und dem homologen, artspezifischen Antikorper in den Kaninchen gegen den Ag-Teil des Komplexes wirksame Antikorper festzustellen, wahrend das Serum gegen den Komplex selbst keine Antikorper enthielt. Es scheint somit auch in vivo eine Dissoziation von Ag-Ak-Komplexen unter bestimmten Umstanden vorzukommen.

Die wesentlichen Probleme, welche bei der diagnostischen Anwendung der Imnrunofluorescenz und hier besonders beim Nachweis von den im Gewebe lokalisierten Bakterien auftreten, werden beschrieben.

I. In den Seren von hautverbrannten Kaninchen konnte mit der passiven Hamagglutination nachBoyden der Nachweis humoraler Antikorper gegen einen wasrigen Extrakt aus verbrannter Kaninchenhaut erbracht werden. Diese Antikorper liesen sich mit Bakterien, welche von den verbrannten Hautstellen und aus den Antigen-Extrakten gezuchtet worden waren, nicht absorbieren. Daruberhinaus konnte weder ein Anstieg noch ein Abfall der antibakteriellen Antikorpertiter in den Seren hautverbrannter Kaninchen nach der Verbrennung festgestellt werden. Der negative Ausfall der Absorptionsversuche wird als Hinweis auf die Spezifitat der verbrennungsabhangigen Antikorper angesehen, die nichts mit einer bakteriellen Infektion der Verbrennungswunde zu tun haben. II. Beim Vergleich einer Gruppe von Kaninchen, welche gleichzeitig mit einer experimentellen Hautverbrennung gegen/gb-hamolytische Streptokokken der Gruppe A immunisiert worden war, mit einer Gruppe nicht verbrannter, nur gegen diese Bakterien immunisierter Kaninchen lies sich kein signifikanter Unterschied in der antibakteriellen Antikorperbildung nachweisen. Eine Hemmung des antikorperbildenden Systems infolge Verbrennungen beschrankten Ausmases (15–18% der Korperoberflache) scheint also im Tierversuch nicht vorzuliegen.

Nach intracutaner Injektion von abgetoteten Streptokokken der Gruppe A, zusammen mit inkomplettem Freundschen Adjuvans, wird bei Meerschweinchen ein homologes Hauttransplantat beschleunigt abgestosen. Diese vonRapaport u.Chase gemachte Beobachtung konnte durch die vorliegenden Untersuchungsergebnisse vollauf bestatigt werden. Zusatzlich ergaben unsere Untersuchungen, das gleichzeitig Hautreaktionen nach intradermaler Injektion von unterschiedlich reinen, gruppenspezifischen Kohlenhydratpraparationen und verstarkt gegen einen Extrakt aus Zellmembran von Gruppe A-Streptokokken auftraten, die in ihrer langsamen Auspragung einer Reaktion vom verzogerten Typ, dem histologischen Bild nach einer Mischung von einer Reaktion vom verzogerten und vom Soforttyp (anaphylaktische Reaktion) entsprachen.

Es wird uber einen 58jahrigen Porzellandreher mit einer Lungensilikose (II) berichtet, bei dem zusatzlich noch eine fibrosierende Pankarditis und eine vernarbende Thyreoiditis vorhanden waren. Quarzkristalle liesen sich in den Narben der Schilddruse und des Herzens nicht feststellen. Im Leichenserum waren Antikorper gegen menschliches Schilddruseneiweis nachweisbar. Ein moglicher immunpathologischer Zusammenhang zwischen der Lungensilikose und dem fibrosierenden Prozes in Herz und Schilddruse konnte jedoch nicht bewiesen werden.

Staphylococcus epidermidis peptidoglycans solubilized by sonication or lysozyme digestion, and synthetic peptidoglycan analogs such as HSA-carboxymethyl-Gly-L-Ala-L-Ala-D-Ala-D-Ala (HSA-pentapeptide) or L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala (pentapeptide) have been labeled with 125I and tested for their applicability in the radioactive antigen binding assay. Use of radioiodinated Staph. epidermidis peptidoglycans was found to be considerably impeded by the presence of at least 2 different antigenic sites on such molecules, the pentapeptide and the glycan determinant. Application of labeled HSA-pentapeptide was limited by the necessity to use PEG for precipitation of Ag-Ab-complexes and by short linear potions of binding curves. However, the synthetic pentapeptide hapten, radioiodinated by the active ester method of BOLTON and HUNTER, proved to be a most useful regent for the selective measurement of pentapeptide antibody. Inhibition studies indicated that the immunological specificity of the labeled hapten was retained. Pentapeptide binding curves were linear from 15-500 g/ml of antibody. Generally, there was good agreement between pentapeptide antibody concentrations measured by radioimmunoassay and quantitative precipitation.

The purification and characterization of the streptolytic exo-enzyme from the Maxted-McCarty strain of Streptomyces albus is described. This enzyme was shown to be an endo-N-acetylmuramidase with a molecular weight of 10 to 12,000 and optimal activity at pH 8 and 45 degrees C. The enzyme is lytic for streptococci of various groups, Micrococcus lysodeikticus, Staphylococcus aureus, as well as Escherichia coli. It closely resembles the F1 endo-N-acetylmuramidase described by Ghuysen et al. (1966) except for small differences in the products of lysis of streptococcal cell walls and the resistance of Escherichia coli to lysis by the F1 enzyme. Lysates of group A and A variant streptococcal cell walls prepared with purified Streptomyces albus muramidase contained serologically active M protein and C carbohydrate-peptidoglycan complexes. The chemical and immunological characteristics of these enzymmatic products of streptococcal cell walls are reported and their utility as immunologic reagents is described.

A brief survey on the ultrastructure, the chemical composition and the immunological properties of bacterial cell wall peptidoglycan (mucopeptide) is presented. This paper deals with the various recently discovered biological activities of peptidoglycan. These could be divided into three different groups, namely: 1. Endotoxin-like properties: pyrogenicity, induction of the local Shwartzman reaction, increase in non-specific resistance to bacterial infection, release of 5-hydroxytryptamine from rabbit platelets, complement activation,gelation of amebocyte lysate. 2. Inflammatory reactions of skin and internal organs, aggressin activity (virulence factor), inhibition of phagocytosis of bacteria by granulocytes and macrophages, inhibition of growth of cell cultures, cytotoxity to granulocytes and macrophages. 3. Potentiation of humoral and cellular immune responses (adjuvant), enhancement of tumor defense in experimental animals. The potential mechanisms of action of peptidoglycan are discussed and attention is focused on the implications of the various peptidoglycan activities for medicine.

A study was performed to find out whether or not RNA and DNA isolated from Group A streptococcal cytoplasm does possess adjuvant activity. The findings obtained indicate that the adjuvancy of streptococcal RNA is very similar to that of poly I:C. The adjuvant activity of poly I:C is characterized by its capacity to increase the number of pre-existing hemolysin-producing spleen cells and by its ability to increase the development of 19S and 7S producers early in the 12-day-period of primary immune response. The adjuvancy of streptococcal DNA was considerably less pronounced than that of RNA. Neither poly I:C nor streptococcal RNA and DNA were found to be capable of increasing the process of priming for the secondary immune response.

This review deals with those biological activities of peptidoglycan that are not directly analogous to the properties of gram-negative bacterial endotoxin. The report is divided into 3 major parts: 1. A survey of peptidoglycan activities such as the induction of inflammatory skin reactions, lesion-enhancing activity (virulence factor), inhibition of phagocytosis of bacteria, inhibition of cell migration, cytotoxicity to mammalian cells, potentiation of the humoral and cellular immune response (adjuvant activity) and enhancement of tumor defense in experimental animals. 2. A presentation of factors which may influence these biological activities of peptidoglycan. 3. A brief discussion of the potential mechanisms of action of peptidoglycan.

The gelation of a lysate prepared from the amebocytes of Limulus polyphemus, the horseshoe crab, has been used to detect endotoxin-like material in clinical studies and in "in vitro" experiments. The investigation of blood samples from 54 hospitalized patients suspected of having endotoxemia, revealed a positive limulus test in 14. Infections due to gram-positive organisms were not associated with positive assays. These results were in agreement with the observation that living gram-positive microorganisms - in contrast to gram-negative bacteria - did not initiate lysate gelation when studied "in vitro". Only very high concentrations of peptidoglycan, isolated from the cell walls of various gram-positive bacteria, induced the reaction. Therefore, these findings support the view, that the limulus test is relatively specific for the detection of gram-negative bacterial endotoxin and endotoxemia.

A major portion of the humoral immune response to peptidoglycans is directed against the non-cross-linked pentapeptide side chains of these ubiquitous bacterial antigens. At present, no specific and sensitive assay for pentapeptide antibody determination is available. Therefore, a radioimmunoassay has been developed which employs the synthetic pentapeptide hapten L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala, labeled by the active ester method of Bolton and Hunter to high specific activities (6.74 to 18.18 muCi/mug) with 125I, and used as a reagent for measuring pentapeptide antibody. A-variant streptococcal antisera, known to contain pentapeptide antibodies as shown by quantitative precipitation, would bind more than 95% of the radiolabeled hapten in contrast to 2 to 3% by preimmune rabbit sera. Specificity of the binding reaction was demonstrated by inhibition experiments imploying various synthetic oligopeptides related or unrelated to the pentapeptide in the radioimmunoassay. Binding curves established with serial dilutions of peptidoglycan antiserum were linear from 15 to 500 mug/ml of antibody permitting pentapeptide antibody measurement within this range. Comparative data on pentapeptide antibody determinations by quantitative precipitation and radioimmunoassay are given and the time course of the production of this antibody in 14 rabbits hyperimmunized with A-variant streptococcal vaccine is reported.

Previous studies have shown that peptidoglycans from group A and B streptococci inhibit the migration of peritoneal exudate cells from non-sensitized rats and guinea pigs. In the present studies peptidoglycans from S. aureus and S. epidermidis were found to inhibit the migration of cells to the same extent as group A streptococcal peptidoglycan. In contrast, HSA-pentapeptide, an immunologically active synthetic analog of the peptide moiety of peptidoglycan which is free of the intrinsic toxicity of naturally occurring peptidoglycans, did not induce migration inhibition of peritoneal exudate cells from non-sensitized guinea pigs. Sensitization of animals with 200 mug HSA-pentapeptide emulsified in incomplete Freund's adjuvant significantly reduced the inhibitory effect of streptococcal and staphylococcal peptidoglycan; HSA-pentapeptide again showed no activity. However, when HSA-pentapeptide was tested against cells from animals sensitized with 200 mug HSA-pentapeptide incorporated in complete Freund's adjuvant, a strong inhibition of migration was evident. Skin tests performed in these animals, in contrast to the dermonecrotic reaction elicited by streptococcal or staphylococcal peptidoglycan, revealed a characteristic delay hypersensitivity to HSA-pentapeptide.

A recently described latex agglutination test was used to determine peptidoglycan antibody titers in sera from healthy human subjects and in sera from patients with a history of streptococcal infections or rheumatoid arthritis. Using latex particles coated with group A streptococcal peptidoglycan 32.8% of the sera from 961 healthy donors reacted positively with titers ranging from 1 : 5 to 1 : 320. Peptidoglycan antibodies were more frequently present in the younger population (45.1% in the 20-29 years old) and considerably decreased in advanced age (15.7% in the 70 years or older). Sera from 82 patients with elevated ASO-titers showed detectable peptidoglycan antibody levels in 40.2%; a statistically significant correlation between ASO and peptidoglycan antibody titers could not be substantiated. Sera from 25 patients with rheumatoid arthritis gave a high incidence (56-92%) of positive results. However there was evidence that this may be due to the action of rheumatoid factor present in such sera.