Interest in the bacterial endotoxins arises, not only from their specific antigenicity and from the phenomenon of tolerance, but, equally, from the remarkable ability of these pyrogenic and highly toxic complexes to stimulate, at microgram doses, substantial refractoriness to a variety of ordinarily lethal insults.Endotoxin treatment has been shown to protect against the consequences of infection with Gram-negative, Gram-positive, or acid-fast bacteria (1-4), and to promote resistance to the lethality of hemorrhagic or traumatic shock (5-7) or wholebody x-irradiation (8, 9). Despite their wide spectrum of activities (10, 11), no unequivocal evidence for experimental dissociation of the in ~ivo non-specific properties of endotoxins by chemical modification of the molecule has been reported.The presence of lipid, carbohydrate, and, usually, protein moieties within the endotoxin molecule has encouraged hydrolytic approaches designed to recover one or another of these components. Hydrolysis with acid or base leads to destruction of all in ~ivo activities, but, where active material has been recovered following hydrolysis, the whole range of properties, toxic and protective, has been present. This is perhaps best illustrated by the current difference of opinion on the role of the lipid component (12, 13). To avoid hydrolytic or oxidative alterations, studies on the chemical modification of endotoxin were undertaken using reactions resulting in substitution at functional groups known to be present in the molecule. This and the following paper (14) describe the acylation of endotoxin, with recovery of a fraction exhibiting in the gross reduced pyrogenicity and lethality but retaining the ability to stimulate so-called non-specific host resistance to infection. A preliminary report of our findings has been published (15). While our manuscripts were in preparation, Noll and Braude (16) reported the detoxification of endotoxin by reductive cleavage of ester bonds yielding a preparation of high immunogenic potency, which, unlike our preparation, confers tolerance to the parent endotoxin.
Materials and MethodsChemicaL--Several endotoxin preparations have been used as starting materials for a variety of substitution reactions. This and the following report (14) describe results obtained with wcetylafion of a Boivin-type endotoxin derived from Salmonella typhosa 0-901 (Difco 929 on