Histological changes evoked in mice by Freund's incomplete adjuvant

Abstract

PLATES CLXIII-CLXVI FREUND'S complete adjuvant, a mixture of dead mycobacteria and mineral oils, has been widely used in experimental pathology (Freud, 1947(Freud, , 1951. Although its mode of action is still uncertain, the morphological and immunological responses to this material are well documented (Moore et al., 1963; Schoenberg, Rupp and Moore, 1964; Moore and Schoenberg, 1966; Spector and Lykke, 1966). By contrast, the reactions evoked by incomplete Freund's adjuvant-which does not contain a mycobacterial component-have not received comparable attention.Laufer, Tal and Behar (1959) investigated the morphological changes produced by fractions of Freund's adjuvant injected subcutaneously into mice, guinea-pigs and hamsters, but none of the animals was killed until 5 mth after inoculation and the findings in only six organs-liver, spleen, kidneys, heart, lungs and brainwere presented.During the course of other experiments, we have had occasion to examine tissues from mice given injections of incomplete Freund's adjuvant. The reactions observed at the site of injection and also in the regional and more distant lymph-nodes were sufficiently unusual to warrant a more detailed study. MATERIALS AND METHODSThirty-five male Swiss albino mice were used, aged 4-8 wk and weighing 10-1 5 g. The animals were housed in metal cages and maintained on a pellet diet and water ad libitum.Freund's incomplete adjuvant was obtained from Difco Laboratories, Detroit, Michigan (batch no. 474513). It contained Arlacel A (mannide mono-oleate) 1.5 ml. and Bayol F* (paraffin oil) 8.5 ml.The animals were divided into 2 groups. (I) Twenty-seven mice received an injection of 0.05 ml. incomplete Freund's adjuvant into the left hind foot-pad and were subsequently killed in groups of 3 at the following times after injection: 1, 6, 18, 24, 48 and 72 hr, and 5, 10 and 17 days. (2) Eight mice were used as untreated controls.All mice were killed with ether vapour and the following tissues were removed and h e d in 4 per cent. buffered formaldehyde: hind foot (left); fibromuscular According to White (1967), Bayol F is a mixture of 42.5 per cent. paraffins, 31.4 per cent. monocyclic naphthalenes and 26.1 per cent. polycyclic naphthalenes. J. PATH. BAcT.-VOL. 95 (1968) 41 1

Immunological Adjuvants

“…Complete Freund's adjuvant contains a mineral oil, an emulsifier and whole bacillary bodies. Mineral oil as well as emulsifier cause various histological changes [1,19,45] and the whole bacillary bodies exert diverse biological activities. Therefore, it is not suitable per se for the purpose of analysis of the role and mode of adjuvant action of Mycobacteria tuberculosis though mineral oil enhances adjuvant action and, thus, may be advantageous for a practical use [1,10].…”

Comparative Studies on Effects of Incomplete Oil Adjuvants With Different Physical Properties

“…Freund's complete adjuvant (FCA) is not used in man because of its severe reactogenicity, while Freund's incomplete adj uvant (FIA) has been tested in man for some viral antigens, such as influenza (Henle and Henle, 1945;Medical Research Council, 1964), poliomyelitis (Salk et al, 1951;Cutler, Lesesne and Vaughn, 1962), adeno (Meiklejohn, 1962) and trachoma (Grayston et al, 1964) vaccines, bacterial antigens, such as cholera (Philippines Cholera Committee, 1965;Ogonuki, Hashizume and Abe, 1967) and typhoid (Snyder et al, 1966) vaccines and tetanus toxoid (MacLennan et al, 1965 (Friedewald, 1944;Fischel et al, 1952;Hilleman, Hampil and Sweet, 1963;Carter, Jamison and Vollum, 1968 (Goto et al, 1972(Goto et al, , 1973(Goto et al, , 1974a Adjuvants: Drakeol 6 VR (Pennsylvania Refining Co., Butler, PA, U.S.A.) used was pure light liquid petrolatum. The emulsifier was Arlacel A 71-B (mannide monooleate; Atlas Chemical Industries, Inc., Wilmington, DE, U.S.A.), consisting principally of ester of mannitol and oleic acid.…”