Purified Protoplasmic Peptides of Mycobacteria: In Vivo and In Vitro Comparison of the Species Specificity of Purified Protoplasmic Peptides and Purified Protein Derivatives of Mycobacterial Culture Filtrates

Abstract

The specificity of purified protein derivatives (PPD) prepared from the culture filtrates of Mycobacterium tuberculosis (PPD), M. kansasii (PPD-Y), M. intracellulare (PPD-B), and M. scrofulacewn (PPD-G) were compared to comparable protoplasmic extracts (PPP) of the same organisms by gel diffusion and delayed hypersensitivity reactions in sensitized guinea pigs. PPD and, to a lesser degree, PPD-Y demonstrated specificities sufficient to enable identification of homologously sensitized guinea pigs in the above group of four mycobacteria. PPD-B and PPD-G did not always elicit the largest reaction in homologously sensitized animals. The PPP sensitins from M. tuberculosis and M. kansasii produced as good skin reactions at 24 and at 48 hr as did their PPD counterparts. The PPP from M. scrofulaceum and M. intracellulare were more specific and more reactive than corresponding PPD, regardless of the time of comparison. Although based on different immunological mechanisms, the specificity of these two groups of sensitins, as demonstrated by delayed hypersensitivity, correlated well with serological comparisons in the gel diffusion test. The low degree of specificity of PPD-B and PPD-G in contrast to that of corresponding PPP was reflected in the precipitin bands in agar gel. (P8), M. scrofulaceum (T-72-5), and M. intracellulare

Mycobacterial antigens: a review of their isolation, chemistry, and immunological properties.

“…The observation by Stottmeier et al that the 4,000to 5,000-molecular-weight peptide that they isolated possessed significant species specificity (190) is noteworthy because the amino acid composition, although not the sequence, of this peptide was determined. It seems likely that the first identification of the structure of a my-…”

“…It probably contained 3 to 4 residues of glucose, 12 residues each of aspartic and glutamic acid, and smaller amounts of six other amino acids (189). It proved to be active in the eliciting of delayed skin test reactions in sensitized animals and to have greater specificity in this regard than the corresponding PPD preparation from the same species of mycobacteria (190).…”

“…Moreover, this work clearly demonstrated that delayed skin tests can be evoked with mycobacterial products which, like haptens, are inadequate to serve as immunogens or complete antigens in other systems. Stottmeier et al prepared protoplasmic peptides from several species of mycobacteria by disruption of cells in a cell press, ultracentrifugation, and gel ifitration (24,189,190). Their product had a molecular weight of 4,000 to 5,000.…”

Comparison of antigens in sonic and pressure cell extracts of Mycobacterium tuberculosis

“…Typical delayed hypersensitive skin reactions have been produced with bacillary extracts in sensitized animals (8,13,15). Protoplasmic preparations have been found to be as specific and sensitive as PPD for detection of delayed hypersensitivity (4, 12,14), and purified protoplasmic components have been claimed to be even more specific skin test antigens than PPD (24). Bacillary extracts also have been used serologically to differentiate and group mycobacterial species (9-11).…”

Purified Protoplasmic Peptides of Mycobacteria: Chemical Composition of a Tuberculin-Active Glycopeptide

“…In an earlier publication, we described the biological activity of tuberculin-active skin test antigens isolated from the protoplasm of mycobacteria (13). For this study we fractionated protoplasmic peptides of Mycobacterium tuberculosis strain H37Rv (PPP-TB) and subsequently investigated the chemical composition of the tuberculin-active fraction.…”