Elsevier

Vaccine

Volume 23, Issue 28, 25 May 2005, Pages 3687-3696
Vaccine

Multicomponent Lyme vaccine: Three is not a crowd

https://doi.org/10.1016/j.vaccine.2005.02.006Get rights and content

Abstract

Lyme disease is caused by the spirochete Borrelia burgdorferi and it is the most common vector-borne disease in the United States. Disseminated spirochetes can persist in various tissues and can result in a variety of different disease manifestations. Vaccination trials testing various lipoprotein candidates have yielded mixed results despite the generation of robust antibody titers. Data presented in this report demonstrate that a combination vaccine composed of DbpA, BBK32 and OspC is more effective than single or double component formulations and that the ratio of each component dramatically impacts vaccine efficacy when tested in protection experiments against Borrelia following needle inoculation.

Introduction

Lyme disease is caused by the spirochete Borrelia burgdorferi sensu lato; a family of organisms that includes B. burgdorferi sensu stricto, B. garinii, and B. afzelii. The spirochetes are transmitted to humans during the blood meal of infected ticks [1], [2]. During a blood meal, an infected tick will deposit the bacteria in the dermis of the mammalian host, and in a process that can take several weeks, the bacteria subsequently disseminate throughout the body of an infected individual. The severity of the disease can vary substantially following dissemination. Symptoms can be multisystemic or localized to specific tissues, and Lyme disease is often difficult to diagnose [1], [3], [4]. Late stages of Lyme disease can include neurological, ocular, cutaneous, and cardiac disease in addition to arthritis [1]. Early antibiotic therapy is often an effective treatment of Lyme disease, however, up to 10% of affected individuals do not respond to antibiotic therapy and are classified as treatment resistant patients [3], [5]. In the absence of antibiotic intervention, disseminated B. burgdorferi can persist in an individual for months if not years even in the face of an apparently strong immune response against the spirochete.

Despite a plethora of highly antigenic and apparently immunologically accessible Borrelia surface proteins [6], [7], [8], [9], [10], [11], [12], [13], [14], [15], [16], [17], [18], the only vaccine formulation tested and approved for human use was derived from the outer surface protein A (OspA) [19]. This formulation (LYMErix), however, has been removed from the market (Spring, 2002) primarily because of low sales resulting from claims alleging that LYMErix alone induced arthritis in vaccinated patients uninfected with the Lyme spirochete [20], [21], [22], [23].

Although OspA-based vaccines have been the focus of many immunization trials [11], [15], [24], [25], [26], [27], various other lipoproteins have also been tested as vaccine candidates against B. burgdorferi infection with varying results [10], [28], [29], [30], [31], [32], [33], [34], [35], [36], [37], [38], [39], [40]. Table 1 describes various active or passive vaccination trials using either the decorin binding protein (DbpA) [9], [41], the fibronectin-binding protein (BBK32) [42], [43], or the outer surface protein (OspC) [44] (DBO) that yielded mixed results with respect to vaccine efficacy. The DBO components were chosen for several reasons: DbpA and BBK32, are Borrelia adhesins or MSCRAMMs (Microbial Surface Components Recognizing Adhesive Matrix Molecules) [9], [41], [42], [43], [45] and OspC appears to be involved in spirochete migration from the tick midgut into the salivary glands during a blood meal [30], [46], [47]. Because MSCRAMMs like DbpA and BBK32 mediate bacterial adherence to the host's extracellular matrix (ECM) components (decorin and fibronectin, respectively), MSCRAMM-based vaccines have become attractive targets for vaccine design because anti-MSCRAMM antibodies can serve as conventional opsonins as well as inhibitors of bacterial adherence to host ECM components [48]. In addition, regions of MSCRAMMs critical for ligand binding must remain conserved if protein function is to be maintained; therefore, protective antibodies generated against these regions should be more likely to protect against infections by heterologous Borrelia strains. All three of the selected proteins are highly antigenic and are significantly up-regulated during the transition between the tick vector and the mammalian host [49] and combination vaccines composed of DbpA/OspA or OspC/OspA have been tested, demonstrating increased vaccine efficacy compared to single component immunizations [31], [50].

Humoral immunity has been long believed to be protective against B. burgdorferi infections [18], [26], [40], [51], [52], and numerous antigens can elicit high antibody titers without conferring protection, however, the reasons for this are unknown [10], [29], [30], [32], [40]. One explanation is the prozone effect [53], a phenomenon that describes high antibody titers but no protection or enhanced infection. Prozone effects have been observed primarily in passive vaccine studies for other pathogens such as Cryptococcus neoformans, Brucella abortus, Pseudomonas aeruginosa, Escherichia coli and Varicella-Zoster [54], [55], [56], [57], [58] and may be a reason for the lack of protection observed in the presence of anti-Borrelia antibodies directed to specific antigens (Table 1). Data presented in this report suggests that multicomponent formulations composed of recombinant forms of DbpA, BBK32, and OspC administered at defined concentrations are more effective than single or double component formulations of any combination.

Section snippets

Mice

Specific pathogen-free (MTV−) BALB/c mice were obtained from an in-house BALB/c breeding colony. The animals were maintained in facilities approved by the American Association for Accreditation of Laboratory Animal Care in accordance with current regulations and standards of the United States Department of Agriculture, Department of Health and Human Services, and National Institutes of Health. All animal procedures were approved by the Institutional Animal Care and Use Committee. Female mice

Single, double, and triple component vaccinations

The data presented in Fig. 1a–c represents pooled data (428 BALB/c mice total) collected from each vaccination trial involving single, double or triple vaccine component experiments, respectively. Since the infection only and complete Freund's adjuvant/incomplete Freund's adjuvant (CFA/IFA) groups were the only groups that remained constant throughout the course of the study, these groups have a significantly higher pooled number of mice (72 and 96 mice total for CFA/IFA and infection only

Discussion

One hypothesis that can be put forth to explain the differences in vaccine efficacy observed for the different DBO immunization groups relates to the concentrations of antigen-specific antibodies and their ability to confer protection following each different DBO vaccination. Although a minimum amount of specific serum IgG is necessary for protection against certain pathogens [74], there is also evidence that antibody-protective efficacy can decline in conditions of antibody excess. This

Acknowledgments

This work was supported by the Department of Health and Human Services, grants from the Center for Disease Control (U50/CCU618387-02) and the Fondren Foundation to E. Brown.

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    Present address: Department of Microbiology and Immunology, University of California, San Francisco, 521 Parnassus Ave, San Francisco, CA 94143-0654, USA.

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