Chemical structure and immunomodulating activities of peptidoglycan from Actinobacillus actinomycetemcomitans

The chemical structure and immunomodulating activities of the cell wall peptidoglycans isolated from Actinobacillus actinomycetemcomitans were investigated. Peptidoglycans were isolated from A. actinomycetemcomitans strains Y4 and ATCC 29522 by boiling in 4% sodium dodecyl sulfate and by digestion with pronase, trypsin and alpha-amylase. Analysis of amino acids and amino sugars of the peptidoglycans revealed that glucosamine, muramic acid, D-glutamic acid, D-alanine, and meso-2, 6-diaminopimelic acid (A2pm) were the principal components. Serine and glycine were not found. Dinitrophenylation method revealed that about half of A2pm residue had a free aminogroup, and analysis by hydrazinolysis showed that a small part of alanine and A2pm located at the C-terminal. The above results indicate that one of the amino groups of A2pm residue at one strand of the stem peptide subunit crosslinked to the carboxyl group of alanine of the neighboring strand. It was thus revealed that the peptidoglycans of A. actinomycetemcomitans belonged to the Al gamma type of the classification by Schleifer and Kandler. Peptidoglycans isolated from A. actinomycetemcmitans strain Y4 and ATCC 29522 were found to be definitely adjuvant-active in induction of delayed type hypersensitivity against ovalbumin when administered to guinea pigs as water-in oil emulsion and stimulation of increase serum antibody levels was found in both peptidoglycans. Regarding mitogenicity on splenocytes of BALB/c and BALB/c nu/nu mice, peptidoglycans from two strains of A. actinomycetemcomitans were markedly enhanced the uptake [3H] thymidine in dose of 10 micrograms/10(5) cells, however thymocytes were not reactive. Stimulation effects on peritoneal macrophages from a guinea pig to incorporation of 14C-glucosamin were not exhibited on addition of 100 micrograms of both peptidoglycans. These findings indicate that peptidoglycan of A. actinomycetemcomitans might eventually be responsible for destruction of periodontal tissue by host mediated activities.