Project Title: „Usage of conservative borrelia protein as vaccine target”

Funding: European Regional Development Fund (ERDF), Measure 1.1.1.1 “Support for applied research”

Project No.: 1.1.1.1/20/A/048

Period: 1 March 2021 – 30 November 2023

Project costs: 540 540.54 EUR

Principle Investigator: Dr. biol. Kaspras Tārs

Cooperation partner: AS “Biotehniskais centrs”

Project summary:

The aim of the project is to produce chromosome encoded conservative Borrelia protein and test its efficacy as vaccine candidate against Lyme disease in mice model.

To achieve our aim, we propose to use borrelial protein BBH0689 as antigen. BBH0689 will be produced as recombinant proteins in E. coli. To enhance the immune response, we plan to conjugate the obtained protein to virus-like particles (VLPs) of ssRNA phages.

Lyme disease is the most prevalent tick-born disease in Europe and North America, with about 400,000 registered cases annually. Disease may have different complications – pain in joints, neurological disorders, symptoms like multiple sclerosis and arthritis. Although the disease can be cured with antibiotics, symptoms may persist for years even after antibiotic treatment. Currently, no vaccine against Lyme disease is available in the market. In 1998, Smith Kline Beecham Biologicals (now part of GlaxoSmithKline) developed LYMErix anti-Lyme disease vaccine, but it was removed from the market due to complaints about side-effects and multiple lawsuit cases.

Information published 01.03.2021.

Progress of the project:

1 March 2021 – 31 May 2021

1. Produced and purified bacteriophage AP205 and Qβ virus-like particles (VLPs)
2. Produced and purified Borrelia burgdorferi surface protein BB0689

Information published 31.05.2021.

Progress of the project:

1 June 2021 – 31 August 2021

1. Produced bacteriophage AP205 virus-like particle genetic fusion conjugate with Borrelia burgdorferi surface protein BB0689
2. Produced Borrelia burgdorferi surface protein BB0689 chemical conjugate with bacteriophage Qβ virus-like particles

Information published 31.08.2021.

Progress of the project:

1 September 2021 – 30 November 2021

1. Started production of conserved Borrelia burgdorferi surface proteins BB028, BB0158, BB0171, BB0213, BB0352, BB0823 and BB058
2. Started search for Borrelia burgdorferi surface protein BB0689 interacion partners in mice bood serum and tissue homogenates

Information published 30.11.2021.

Progress of the project:

1 December 2021 – 28 February 2022

1. Produced conserved Borrelia burgdorferi surface proteins BB028 and BB0158
2. Produced conserved Borrelia burgdorferi surface proteins BB028 and BB0158 as genetic fusions with bacteriophage AP205 VLPs
3. Continued interaction partner search of Borrelia burgdorferi protein BB0689 in mice serum and organ homogenates using electrophoresis, chromatography and mass spectrometry techniques
4. Started immunization of mice with Borrelia burgdorferi BB0689 protein, BB089 protein in genetic fusion with bacteriophage AP205 VLPs and BB0689 protein chemical conjugate with bacteriophage Qβ VLPs

Information published 28.02.2022.

Progress of the project:

1 March 2022 – 31 May 2022

1. Produced conserved Borrelia burgdorferi surface proteins BB0171 and BB0213
2. Produced conserved Borrelia burgdorferi surface proteins BB0171 and BB0213 as genetic fusions with bacteriophage AP205 VLPs
3. With aid of mass spectrometry apolipoprotein A-1 identified as potential interaction partner of Borrelia burgdoreri surface protein BB0689
4. Finished immunization of mice with Borrelia burgdorferi BB0689 protein, BB089 protein in genetic fusion with bacteriophage AP205 VLPs and BB0689 protein chemical conjugate with bacteriophage Qβ VLPs

Information published 31.05.2022.

Progress of the project:

1 June 2022 – 31 August 2022

At this period the following has been accomplished in the project:

1. Produced conserved Borrelia burgdorferi surface proteins BB0352 and BB0823
2. Produced conserved Borrelia burgdorferi surface proteins BB0352 and BB0823 as genetic fusions with bacteriophage AP205 VLPs
3. With aid of mass spectrometry apolipoprotein A-1 identified as potential interaction partner of Borrelia burgdoreri surface protein BB0689
4. Finished infection and evaluation of disease progress after vaccination of mice with Borrelia burgdorferi BB0689 protein, BB089 protein in genetic fusion with bacteriophage AP205 VLPs and BB0689 protein chemical conjugate with bacteriophage Qβ VLPs

Information published 31.08.2022.

Progress of the project:

1 September 2022 – 30 November 2022

From 01.09.2022 to 30.11.2022 the following has been accomplished in the project:

1. Optimized conditions for bacteriophage Qβ VLP upscale fermentation
2. Produced Borrelia burgdorferi bacterial culture for downstream bactericidity experiments
3. Started mice immunization with proteins BB028, BB0158 and their chemical conjugates with bacteriophage Qβ VLP

Information published 30.11.2022.

Progress of the project:

From 01.12.2022 to 28.02.2023 the following has been accomplished in the project:

1. Three-dmensional structure of protein BB0158 established
2. Finished BALB/c mice immunuzation with proteins BB028, BB0158 and their chemical conjugates with bacteriophage Qβ VLP
3. Started bactericidity experiments with sera from immunized mice

Information published 28.02.2023.

Progress of the project:

From 01.03.2023 to 31.05.2023 the following has been accomplished in the project:

1. Written part of manuscript about three-dmensional structure of BB0158 protein
2. In order to try to establish interaction partners of BB0689 and BB0158 proteins, obtained crystals after treatment of proteins with various physiological fluids
3. Finished bactericidity experiments with sera from immunized mice

Information published 31.05.2023.

Progress of the project:

From 01.06.2023 to 31.08.2023 the following has been accomplished in the project:

1. Written manuscript on the three-dimensional structure of BB0158 protein
2. A Latvian patent was written and submitted on the potential use of Borrelia surface protein in the construction of a Lyme disease vaccine
3. A manuscript has been written on the use of several Borrelia surface proteins in the construction of a Lyme disease vaccine.

Information published 01.09.2023.

Progress of the project:

1 September 2023 – 30 November 2023

From 01.09.2023 to 30.11.2023. the following has been accomplished in the project:

1. Manuscript on three-dimensional structure of BB0158 protein submitted to journal, revisions made, manuscript accepted for publication
2. Written and submitted for publication a manuscript on mathematical modeling of Qβ VLP mass production
3. Manuscript on the use of multiple Borrelia surface proteins in the construction of a Lyme disease vaccine has been submitted to the journal, revisions have been made, and the manuscript has been accepted for publication

Information published 30.11.2023.