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Lyme disease vaccine

a technology for lyme disease and vaccine, applied in the field of lyme disease vaccine and diagnostic, can solve the problems of ineffective use of full-length ospa, high socio-economic cost of lyme disease, and inability to provide an efficacious vaccine for use in the prevention and/or treatment of lyme disease, etc., and achieve the effect of broad protection against infection

Inactive Publication Date: 2011-10-27
VIRGINIA COMMONWEALTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0011]In order to address prior art problems with Lyme disease vaccines, the OspA protein from Borrelia burgdorferi has been epitope mapped by assessing the reactivity of sera generated during murine infection with recombinant OspA subfragments. The epitope map demonstrated several linear epitope-containing regions of OspA. While conformational epitopes of OspA have been mapped and described, linear epitopes have not previously been reported for use in a vaccine. The use of one or more of these small, defined epitope-containing sequences in a polypeptide vaccine formulation allows the avoidance of specific regions of OspA that have been implicated in vaccine-mediated adverse events when full-length OspA is used. The inclusion of linear epitopes from a plurality of phyletic groups of Borrelia results in a vaccine that provides broad protection against infection over large geographical areas.

Problems solved by technology

In addition, Lyme disease has significant socio-economic costs, manifested by reductions in outdoor recreational and social activities due to concerns about tick exposure.
However, a major drawback to the use of full-length OspA was the potential (whether real or perceived) for adverse events secondary to vaccination, such as the development of arthritis caused by immunological cross-reactivity with human proteins (e.g. LFA-1).
Despite the above-referenced technologies, to date the prior art has failed to provide an efficacious vaccine for use in the prevention and / or treatment of Lyme disease.

Method used

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Examples

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example 1

Identification of Linear Immunodominant Epitopes of Borrelia OspA Protein

Materials and Methods

[0073]Recombinant protein production

[0074]Fragments of the outer surface protein A (OspA) gene were amplified by PCR using high fidelity DNA polymerase (Phusion HF, New England Biolabs), and the amplicons purified by agarose gel electrophoresis and gel extraction. Overhangs were generated by treatment with T4 polymerase to allow ligase-independent annealing to the pET-32 Ek / LIC vector. The annealed vector was transformed into Escherichia coli, and the transformants were selected for ampicillin resistance, and were screened for the presence of the OspA gene fragment by PCR. Plasmids were then extracted from the transformants and the sequence confirmed by DNA sequencing (MWG Biotech). The plasmids were then used to transform E. coli BL21 (DE3) cells for protein production.

[0075]To generate recombinant proteins, BL21(DE3) cells were grown at 37° C. to an OD600 of 0.5, then 1 mM IPTG was added ...

example 2

[0083]When administered to test mammals, this chimeric polypeptide construct comprising at least one 221-240 epitope containing region, and usually two or more 221-240 epitope containing regions from of different phyletic types, is found to elicit a robust immune response, and to provide protection from the development of Lyme disease.

[0084]While the invention has been described in terms of its preferred embodiments, those skilled in the art will recognize that the invention can be practiced with modification within the spirit and scope of the appended claims. Accordingly, the present invention should not be limited to the embodiments as described above, but should further include all modifications and equivalents thereof within the spirit and scope of the description provided herein.

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Abstract

Antigenic polypeptides comprising linear immunodominant epitopes of Borrelia outer surface protein A (OspA) or Borrelia outer surface protein C (OspC) are useful as vaccines against Lyme disease, and as diagnostics for detecting Borrelia infections. The OspA and OspC antigenic polypeptides typically comprise a plurality of peptides representing epitope containing regions from multiple distinct phyletic groups. The antigenic polypeptides may also include epitopes from both Borrelia OspA and Borrelia OspC.

Description

BACKGROUND OF THE INVENTION[0001]1. Field of the Invention[0002]The invention generally relates to a vaccine and diagnostic for Lyme disease. In particular, the invention provides a Lyme disease vaccine and diagnostic that includes linear Borrelia outer surface protein A (OspA) epitopes and / or Borrelia outer surface protein C (OspC) epitopes, usually from multiple distinct phyletic groups.[0003]2. Background of the Invention[0004]Lyme disease is the most common arthropod-borne disease in North America and Europe, where in some areas up to 3% of the population is infected annually. Lyme disease is caused by the spirochetes Borrelia burgdorferi, B. garinii and B. afzelii. Transmission to mammals occurs through the bite of infected Ixodes ticks. Infection results in a multi-systemic inflammatory disease with early stage symptoms that may include erythema migrans, low-grade fever, arthralgia, myalgia, and headache. Late stage clinical manifestations can be severe and may include in part...

Claims

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Application Information

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IPC IPC(8): A61K39/02C07K14/195A61P37/04C07K16/12A61P31/04C07K7/08G01N33/566
CPCA61K39/0225C07K14/20C07K16/1207G01N2333/20G01N2469/20G01N33/56911A61P31/04A61P37/04Y02A50/30A61K39/00C07K14/001C07K2319/00C07K2319/21
Inventor EARNHART, CHRISTOPHER G.MARCONI, RICHARD T.
Owner VIRGINIA COMMONWEALTH UNIV
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