Whole mycobacterial cells, which are used in Freund's complete adjuvant, besides inducing hypersensitivity to tuberculoprotein, also can elicit hyperreactivity to endotoxins, lymphoid hyperplasia, and allergic polyarthritis in rats. The data reported here demonstrate that a potent water-soluble adjuvant obtained from mycobacterial cell walls is also effective in increasing the immune response to viruses, and that it is free of the toxic effects observed with whole mycobacterial cells.
Animal feed additives have for many years constituted the largest and most controversial category of antimicrobial use. The primary concerns addressed have been the generation of antibiotic resistance in animal bacterial and the influence of such resistance on human health. Studies designed to shed light on the controversy generally have yielded information leading to ambiguous conclusions. A 1989 report from the National Academy of Sciences was the latest of a long collection of assessments that have been meant to provide expert guidance to regulatory authorities on this matter. The FDA has been trying for some time to determine whether a regulatory decision on existing products is warranted. After several decades of research directed at this question, most qualified study groups have reported a paucity of appropriate information necessary to judge the reality of a public health risk.
The data reported here demonstrate that a preparation extracted from nonpathogenic mycobacteria such as Mycobacterium smegmatis and hereafter referred to as interphase material protected mice against Ehrlich ascitic carcinoma, L-1210 leukemia, and another syngeneic lymphoid leukemia. Furthermore, mice treated by this preparation were much less susceptible to endotoxins than when stimulated by BCG (bacillus Calmette-Guerin) or M. smegmatis cells. Moreover, guinea pigs treated by interphase material administered in Freund's incomplete adjuvant showed an increased immune response, yet their sensitivity to tuberculin was much weaker than that of controls sensitized with Freund's complete adjuvant.Finally, resistance to Columbia SK virus infection could be demonstrated when interphase material was administered to mice prior to virus challenge.It is now well established that mycobacteria have the ability to stimulate the immunity of the host against tumors (1-2). However, these organisms have also the capacity for rendering mice very susceptible to endotoxins (13) and for inducing delayed hypersensitivity to tuberculin. Our previous studies have shown that delipidated cells as well as purified cell walls of Mycobacterium smegmatis or M. kansasii had greater antitumor activity than killed cells of bacillus Calmette-Guerin (BCG), although they were devoid of several of the noxious effects of whole mycobacterial cells (14).In the following study various biological activities of a preparation extracted from nonpathogenic mycobacteria and hereafter referred to as interphase material (IPM) the press with cooling. The resulting homogenate was centrifuged for 90 min at 5000 X g, yielding a multiphase system consisting of a lower cell debris layer, a lower opaque aqueous layer and an upper hexane layer containing a sedimented gelatinous phase. The aqueous phase was withdrawn from the system by aspiration and discarded. The hexane phase containing the gelatinous sediment was decanted and centrifuged at 10,000 X g for 1 hr. The gelatinous material was separated from the hexane by decantation, suspended in distilled water, and the suspension was freed from residual hexane by heating to about 380 under reduced pressure. The aqueous suspension was freeze-dried to yield 29.0 g of a fluffy, light colored amorphous solid hereafter referred to as M. smegmatis interphase material (IPM). A similar procedure was used for obtaining interphase material from M. phlei and M. kansasii.M. smegmatis IPM is insoluble in neutral aqueous systems and only partly soluble in organic solvents. It is characterized by the following elemental analysis: C, 51-57%; H, 7.5-8.7%; N, 5.8-6.5%; P, 0.5-1.2% and by the following composition: neutral carbohydrates (arabinose, galactose, glucose), 21-23%; amino sugars, 0.8-1.8%; amino acids (as peptides or proteins), 24-28%; free lipids, 40-43%. These data as well as the presence of a,a'-diaminopimelic acid and approximately equimolar amounts of glucosamine and muramic acid indicate that IPM is essentially...
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