Intravenous injections of glucan simultaneously with Formalin-killed erythrocytic stages of Plasmodium berghei elicited a greater degree of resistance in mice against subsequent infection with viable parasites than injections of killed erythrocytic stages alone. In two experiments with P. berghei strain NK 65, 100% of mice immunized with the glucan-dead parasite preparation survived challenge, whereas only 28.6% of mice receiving dead parasites alone survived. In the third experiment, using P. berghei strain NYU-2, the same proportion of mice survived after immunization with glucan and dead parasites as with dead parasites alone (i.e., 10 of 11 in each group), but mice immunized with the glucan-dead parasite preparation experienced parasitemias of significantly less intensity and shorter duration than mice which received only dead parasites before infection. Inoculation of glucan alone or with normal erythrocytes conferred no protection against challenge.Glucan is a polysaccharide constituent of the inner cell wall of bakers' yeast, Saccharomyces cerevisiae, consisting of a chain of glucopyranose units united by a ,81,3-glucosidic linkage. Stimulation with glucan elicits nonspecific resistance in experimental animals against several infectious agents, including Candida albicans (15), RES J. Reticuloendothel. Soc. 20:66A, 1976), Mycobacterium leprae (3), Staphylococcus aureus (7), murine viral hepatitis (18), and Leishmania donovani (2).Recent studies have shown that glucan exerts an adjuvant effect in experimental immunization trials in visceral leishmaniasis (T. W. Holbrook, J. A. Cook, and B. W. Parker, Am. J. Trop. Med. Hyg., in press) and Venezuelan equine encephalitis (14). In this report, we describe a strong adjuvant effect of glucan injected simultaneously with Formalin-killed erythrocytic stages of the rodent malaria parasite Plasmodium berghei.
MATERLALS AND METHODSParasite and experimental animals. P. berghei strain NK 65 was used in the initial two experiments in the study. The strain was obtained from Richard Roth, Rutgers University, and has been maintained here by serial passage in mice for several years. For a third experiment, P. berghei strain NYU-2 was recently acquired as a frozen stabilate from the American Type Culture Collection (no. 30090). The parasite strain is presently maintained by serial intraperitoneal passage of infected blood in mice. Female ICR mice were obtained from the Department of Laboratory Animal Medicine at this institution. Animals were 8 weeks old at the start of experiments and were housed in air-conditioned quarters; they were allowed food and water ad libitum.Immunization method. Parasitized erythrocytes (RBC) used for immunization were obtained by retroorbital bleeding of mice exhibiting at least 50% parasitemia. RBC were washed three times in Earle balanced salt solution and killed by suspension in 0.1% Formalin for 30 min at room temperature and kept at 4°C overnight. Cells were washed three times and resuspended in Earle balanced salt solution at a concentration ...