INFECTION AND IMMUNITY, Mar. 1977, p. 997-998 Copyright © 1977 American Society for Microbiology
Vol. 15, No. 3 Printed in U.S.A.
Effectiveness of Parenteral and Oral Typhoid Vaccination in Mice Challenged with a Salmonella typhi-Salmonella typhimurium Hybrid B. B. DIENA,* A. RYAN, R. WALLACE, E. M. JOHNSON, L. S. BARON, AND F. E. ASHTON Laboratory Center for Disease Control, Health Protection Branch, Health and Welfare Canada, Ottawa, Canada,* and Department of Bacterial Immunology, Walter Reed Army Institute of Research,
Washington, D.C. 20012
Received for publication 13 September 1976
Live Salmonella typhi administered intraperitoneally, acetone-killed S. typhi administered intraperitoneally, and live S. typhi given orally, with their effectiveness decreasing in that order, protected Swiss white mice against death from challenge with a virulent Salmonella typhimurium hybrid expressing S. typhi antigens.
We reported previously (3-5) the develop- of 0.5 ml (500 jig/dose), with a 1-week interval ment of an assay system for differentiating the between the first and second injection. The protective activities of various typhoid vac- same doses of vaccines AK, live, and Vi were cines. It uses Swiss Webster white mice as the administered orally to other groups of mice. test animals and mouse-virulent Salmonella Animals vaccinated with AK and Vi vaccines typhimurium hybrids that express Salmonella either i.p. or orally were challenged i.p. 2 typhi antigens as the challenge strains. In ear- weeks after the last vaccination, as described lier studies with this system (4, 5), we showed previously (4), with 2,500 organisms (0.5 ml) of that it can demonstrate differences among var- S. typhimurium hybrid H42. This hybrid exious kinds of typhoid vaccines with respect to presses the S. typhi antigens 9, 12, Vi, and d their ability to confer protection against death and has a mean lethal dose of less than 50 of the animals. Our studies also provided evi- organisms. Animals injected with live vaccines dence that the Salmonella somatic antigens are either i.p. or orally were challenged after 5 important in conferring this protection, weeks in the same manner. The longer interval whereas the Vi antigen was seen to play no between live immunization and challenge has been recommended by other investigators (8, significant role (3). In these earlier investigations, the vaccines 10). we examined were, in all cases, nonliving, and From the results of these experiments, as the immunizing doses were administered intra- presented in Table 1, it is apparent that the peritoneally (i.p.). In the present study, we best protection against death from the hybrid have used our assay system to investigate the challenge organism was afforded by the live protective capabilities of living as well as ace- and AK vaccines when administered i.p. The tone-treated (AK) S. typhi vaccines adminis- live vaccine was slightly better than the AK tered both by the oral route and i.p. We have vaccine in this comparison (P < 0.05), a result also examined a purified S. typhi Vi antigen that is in accordance with the findings of a preparation administered both i.p. and orally. number of investigators using various other The AK vaccine was prepared from S. typhi experimental systems (1, 2, 6-9). Several workas described previously (4). The purified Vi an- ers have pointed out that the superiority of live tigen, prepared from S. typhi strain no. 59, was vaccines is particularly well demonstrated generously supplied by S. Marcus of the Uni- when they are administered orally (2, 6, 7), and versity of Utah, Utah Medical Center, Salt the present results would appear to bear this Lake City. Swiss Webster white mice (50 per out; oral administration of the live cells afgroup), HPB strain, random bred, 16 to 18 g, forded significant protection against death, were inoculated i.p. with 0.5 ml of the AK whereas oral administration of the AK vaccine vaccine (equivalent to 5 x 108 organisms) or did not. It might be worth noting, however, with 0.5 ml of a live vaccine (107 organisms) that the degree of protection afforded by the prepared from a 16-h culture ofS. typhi TY2. Vi live cells administered orally did not, in this antigen vaccine was administered in two doses system, match that afforded by the AK vaccine 997
998
NOTES
TABLE 1. Effectiveness ofAK, live, and Vi antigen vaccines in mice challenged with S. typhimurium hybrid H42 Interval between last vac- No. of Vaccinated Vaccinating Route cination surviwith dose and vors/no. chalinjected lenge (weeks) 2 i.p. 5 x 10" cells AK 30/50a 2 AK 5 x 10" cells Oral 7/50 5 TY2 live 107 cells i.p. 40/50a 5 TY2 live 107 cells Oral 21/50b 2 Vi 2 x 500,ug i.p. 9/50 2 Vi 2 x 500 Ag Oral 3/50 Control 8/50 a Significantly better (P < 0.005) than the controls. bSignificantly better (P < 0.01) than the controls.
administered i.p. Whether or not a similar situation would be observed in immunizing against human typhoid fever with live, attenuated, orally administered vaccines, as opposed to killed vaccines parenterally administered, remains to be determined. In a previous study (5), we tested purified Vi antigen prepared from Citrobacter freundii for its protective capability (administered i.p.) in this system and found it afforded no protection against death from the hybrid challenge organism. As might have been expected, the presently tested Vi antigen, prepared this time from S. typhi, also failed to protect the animals, whether administered i.p. or orally.
INFECT. IMMUN. LITERATURE CITED 1. Blanden, R. V., G. B. Mackaness, and F. M. Collins. 1966. Mechanisms of acquired resistance in mouse typhoid. J. Exp. Med. 124:585-600. 2. Collins, F. M., and P. B. Carter. 1972. Comparative immunogenicity of heat-killed and living oral Salmonella vaccines. Infect. Immun. 6:451-458. 3. Diena, B. B., L. S. Baron, E. M. Johnson, R. Wallace, and F. E. Ashton. 1974. Role of typhoid antigens in protection and pathogenicity for mice. Infect. Immun. 9:1102-1104. 4. Diena, B. B., E. M. Johnson, L. S. Baron, R. Wallace, and L. Greenberg. 1973. Assay of typhoid vaccines with Salmonella typhosa-Salmonella typhimurium hybrids. Infect. Immun. 7:5-8. 5. Diena, B. B., A. Ryan, R. Wallace, F. E. Ashton, E. M. Johnson, and L. S. Baron. 1975. Ineffectiveness of Vi and chemically treated endotoxins as typhoid vaccines in mice challenged with a Salmonella typhosaSalmonella typhimurium hybrid. Infect. Immun. 12:1470-1471. 6. Germanier, R. 1972. Immunity in experimental salmonellosis. III. Comparative immunization with viable and heat-inactivated cells of Salmonella typhimurium. Infect. Immun. 5:792-797. 7. Germanier, R., and E. Furer. 1975. Isolation and characterization of Gal E mutant Ty 21a of Salmonella typhi: a candidate for a live, oral typhoid vaccine. J. Infect. Dis. 131:553-558. 8. Kawakami, M., H. Nakata, and S. Mitsuhashi. 1969. Experimental Salmonellosis: immunizing effect of live vaccine prepared from various mutants of Salmonella having different cell wall polysaccharides. Jpn. J. Microbiol. 13:315-324. 9. Kiefer, W., P. Gransow, G. Schmidt, and 0. Westphal. 1976. Salmonellosis in mice: immunization experiments with Salmonella-Escherichia coli hybrids. Infect. Immun. 13:1517-1518. 10. Mitsuhashi, S., M. Kawakami, Y. Yamaguchi, and M. Nagai. 1958. Studies on the experimental typhoid: 1. A comparative study of living and killed vaccines against the infection of mice with S. enteritidis. Jpn. J. Exp. Med. 28:249-258.
Vol. 15, No. 3 Printed in U.S.A.
Effectiveness of Parenteral and Oral Typhoid Vaccination in Mice Challenged with a Salmonella typhi-Salmonella typhimurium Hybrid B. B. DIENA,* A. RYAN, R. WALLACE, E. M. JOHNSON, L. S. BARON, AND F. E. ASHTON Laboratory Center for Disease Control, Health Protection Branch, Health and Welfare Canada, Ottawa, Canada,* and Department of Bacterial Immunology, Walter Reed Army Institute of Research,
Washington, D.C. 20012
Received for publication 13 September 1976
Live Salmonella typhi administered intraperitoneally, acetone-killed S. typhi administered intraperitoneally, and live S. typhi given orally, with their effectiveness decreasing in that order, protected Swiss white mice against death from challenge with a virulent Salmonella typhimurium hybrid expressing S. typhi antigens.
We reported previously (3-5) the develop- of 0.5 ml (500 jig/dose), with a 1-week interval ment of an assay system for differentiating the between the first and second injection. The protective activities of various typhoid vac- same doses of vaccines AK, live, and Vi were cines. It uses Swiss Webster white mice as the administered orally to other groups of mice. test animals and mouse-virulent Salmonella Animals vaccinated with AK and Vi vaccines typhimurium hybrids that express Salmonella either i.p. or orally were challenged i.p. 2 typhi antigens as the challenge strains. In ear- weeks after the last vaccination, as described lier studies with this system (4, 5), we showed previously (4), with 2,500 organisms (0.5 ml) of that it can demonstrate differences among var- S. typhimurium hybrid H42. This hybrid exious kinds of typhoid vaccines with respect to presses the S. typhi antigens 9, 12, Vi, and d their ability to confer protection against death and has a mean lethal dose of less than 50 of the animals. Our studies also provided evi- organisms. Animals injected with live vaccines dence that the Salmonella somatic antigens are either i.p. or orally were challenged after 5 important in conferring this protection, weeks in the same manner. The longer interval whereas the Vi antigen was seen to play no between live immunization and challenge has been recommended by other investigators (8, significant role (3). In these earlier investigations, the vaccines 10). we examined were, in all cases, nonliving, and From the results of these experiments, as the immunizing doses were administered intra- presented in Table 1, it is apparent that the peritoneally (i.p.). In the present study, we best protection against death from the hybrid have used our assay system to investigate the challenge organism was afforded by the live protective capabilities of living as well as ace- and AK vaccines when administered i.p. The tone-treated (AK) S. typhi vaccines adminis- live vaccine was slightly better than the AK tered both by the oral route and i.p. We have vaccine in this comparison (P < 0.05), a result also examined a purified S. typhi Vi antigen that is in accordance with the findings of a preparation administered both i.p. and orally. number of investigators using various other The AK vaccine was prepared from S. typhi experimental systems (1, 2, 6-9). Several workas described previously (4). The purified Vi an- ers have pointed out that the superiority of live tigen, prepared from S. typhi strain no. 59, was vaccines is particularly well demonstrated generously supplied by S. Marcus of the Uni- when they are administered orally (2, 6, 7), and versity of Utah, Utah Medical Center, Salt the present results would appear to bear this Lake City. Swiss Webster white mice (50 per out; oral administration of the live cells afgroup), HPB strain, random bred, 16 to 18 g, forded significant protection against death, were inoculated i.p. with 0.5 ml of the AK whereas oral administration of the AK vaccine vaccine (equivalent to 5 x 108 organisms) or did not. It might be worth noting, however, with 0.5 ml of a live vaccine (107 organisms) that the degree of protection afforded by the prepared from a 16-h culture ofS. typhi TY2. Vi live cells administered orally did not, in this antigen vaccine was administered in two doses system, match that afforded by the AK vaccine 997
998
NOTES
TABLE 1. Effectiveness ofAK, live, and Vi antigen vaccines in mice challenged with S. typhimurium hybrid H42 Interval between last vac- No. of Vaccinated Vaccinating Route cination surviwith dose and vors/no. chalinjected lenge (weeks) 2 i.p. 5 x 10" cells AK 30/50a 2 AK 5 x 10" cells Oral 7/50 5 TY2 live 107 cells i.p. 40/50a 5 TY2 live 107 cells Oral 21/50b 2 Vi 2 x 500,ug i.p. 9/50 2 Vi 2 x 500 Ag Oral 3/50 Control 8/50 a Significantly better (P < 0.005) than the controls. bSignificantly better (P < 0.01) than the controls.
administered i.p. Whether or not a similar situation would be observed in immunizing against human typhoid fever with live, attenuated, orally administered vaccines, as opposed to killed vaccines parenterally administered, remains to be determined. In a previous study (5), we tested purified Vi antigen prepared from Citrobacter freundii for its protective capability (administered i.p.) in this system and found it afforded no protection against death from the hybrid challenge organism. As might have been expected, the presently tested Vi antigen, prepared this time from S. typhi, also failed to protect the animals, whether administered i.p. or orally.
INFECT. IMMUN. LITERATURE CITED 1. Blanden, R. V., G. B. Mackaness, and F. M. Collins. 1966. Mechanisms of acquired resistance in mouse typhoid. J. Exp. Med. 124:585-600. 2. Collins, F. M., and P. B. Carter. 1972. Comparative immunogenicity of heat-killed and living oral Salmonella vaccines. Infect. Immun. 6:451-458. 3. Diena, B. B., L. S. Baron, E. M. Johnson, R. Wallace, and F. E. Ashton. 1974. Role of typhoid antigens in protection and pathogenicity for mice. Infect. Immun. 9:1102-1104. 4. Diena, B. B., E. M. Johnson, L. S. Baron, R. Wallace, and L. Greenberg. 1973. Assay of typhoid vaccines with Salmonella typhosa-Salmonella typhimurium hybrids. Infect. Immun. 7:5-8. 5. Diena, B. B., A. Ryan, R. Wallace, F. E. Ashton, E. M. Johnson, and L. S. Baron. 1975. Ineffectiveness of Vi and chemically treated endotoxins as typhoid vaccines in mice challenged with a Salmonella typhosaSalmonella typhimurium hybrid. Infect. Immun. 12:1470-1471. 6. Germanier, R. 1972. Immunity in experimental salmonellosis. III. Comparative immunization with viable and heat-inactivated cells of Salmonella typhimurium. Infect. Immun. 5:792-797. 7. Germanier, R., and E. Furer. 1975. Isolation and characterization of Gal E mutant Ty 21a of Salmonella typhi: a candidate for a live, oral typhoid vaccine. J. Infect. Dis. 131:553-558. 8. Kawakami, M., H. Nakata, and S. Mitsuhashi. 1969. Experimental Salmonellosis: immunizing effect of live vaccine prepared from various mutants of Salmonella having different cell wall polysaccharides. Jpn. J. Microbiol. 13:315-324. 9. Kiefer, W., P. Gransow, G. Schmidt, and 0. Westphal. 1976. Salmonellosis in mice: immunization experiments with Salmonella-Escherichia coli hybrids. Infect. Immun. 13:1517-1518. 10. Mitsuhashi, S., M. Kawakami, Y. Yamaguchi, and M. Nagai. 1958. Studies on the experimental typhoid: 1. A comparative study of living and killed vaccines against the infection of mice with S. enteritidis. Jpn. J. Exp. Med. 28:249-258.
