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Infection and Immunity, July 2002, p. 3324-3329, Vol. 70, No. 7
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.7.3324-3329.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Protection of Mice against Brucellosis by Intranasal Immunization with Brucella melitensis Lipopolysaccharide as a Noncovalent Complex with Neisseria meningitidis Group B Outer Membrane Protein

Apurba K. Bhattacharjee,^1* Lillian Van De Verg,^1, Mina J. Izadjoo,² Liang Yuan,^1, Ted L. Hadfield,³ Wendell D. Zollinger,¹ and David L. Hoover¹

Department of Bacterial Diseases, Walter Reed Army Institute of Research, Washington, D.C. 20307-5100,¹ American Registry of Pathology,² Department of Infectious and Parasitic Diseases, Armed Forces Institute of Pathology, Washington, D.C. 20306-6000³

Received 10 January 2002/ Returned for modification 22 March 2002/ Accepted 27 March 2002

Intranasal immunization of mice with purified Brucella melitensis lipopolysaccharide (LPS) as a noncovalent complex with Neisseria meningitidis group B outer membrane protein (GBOMP) elicited a high-titer anti-LPS systemic antibody response and a significant mucosal antibody response. The anti-LPS immunoglobulin G (IgG) antibody was predominantly of the IgG1 subtype, although there was some response of the IgG2a, IgG2b, and IgG3 subtypes. The antibody titer remained high for 16 weeks postimmunization. Immunized mice and sham-immunized control mice were challenged intranasally with 10⁴ CFU of virulent B. melitensis strain 16 M 4 weeks after the second dose of vaccine. The numbers of bacteria in lungs, livers, and spleens at 3 days, 9 days, and 8 weeks postchallenge were determined. Bacteria were found in lungs of all mice on day 3, but there was no disseminated infection of liver or spleen. By day 9, 40% of the mice had infected spleens and livers. At 8 weeks postchallenge, spleens of 25 of 62 immunized mice were infected, compared to 61 of 62 control mice (P < 0.0001). The livers of 12 of 43 immunized mice were infected, compared to 22 of 36 control mice (P = 0.005). In contrast, the lungs of 26 of 46 immunized mice were still infected, compared to 27 of 44 control mice. The numbers of bacterial CFU in lungs of immunized and control animals were identical. These studies show that intranasal immunization with B. melitensis LPS-GBOMP subunit vaccine significantly protects mice against intranasal challenge with virulent B. melitensis. Vaccination reduces bacterial dissemination to spleen and liver but has no effect on the course of lung infection.

Editor: R. N. Moore

Present address: Joint Vaccine Acquisition Program, USAMRMC, Fort Detrick, MD 21702-5041.

Present address: New England Medical Center, Tufts University School of Medicine, Boston, MA 02111.

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