Analysis of the specificity of natural human antibody reactive to Actinomyces.

The specificity of a frequently-occurring precipitin response to soluble antigens from cell-walls and culture filtrates of A. viscosus ATCC 19246 was examined. After precipitation with isopropanol (50-75% v/v), antigen fractions of different charge and molecular weight were isolated by ion exchange and gel filtration. When heated in mineral acid or alkali above 0.15 M, each of the purified antigens lost precipitating activity, but now inhibited the precipitin reaction between serum and exogenous unheated antigen. The inhibitor was isolated over Biogel P30 and characterized as a peptide fragment (mol. wt about 2 kd) containing approximately 50 moles of ornithine and 6-12 moles, respectively, of aspartate, serine, threonine, glutamate, glycine, alanine and histidine per 100 moles amino acids. The inhibitor was totally destroyed by heating for 1.0 hr in 2.0 M HCl. Variability in the number of fragments and differences in the non-antigenic portions probably accounted for the complexity of the antigens. Ornithine, putrescine, N-acetyl putrescine and various sugars had little or no effect on the precipitin reaction with intact antigen at high concentrations (200 mM), whereas the fragment inhibited completely at 0.4 mM. This indicates that neither ornithine nor its side-chain amides are exclusively recognized by antibody. However, ornithine may be part of a larger sequence and/or important in forming the configuration recognized by the human antibodies.