Emergence - Emergence

Pre-clinical development of a live vaccine against plague – PLAGVAC

Development of therapeutic monoclonal antibodies against plague using memory B cells from survivors

Plague is a potential bioterrorist weapon, and monoclonal antibodies could be an efficient way to fight antibiotics-resistant strains of Yersinia pestis. A comprehensive plague survivor T and B cell epitope maping will be performed, which will allow to derive cell lines from healed patients' PBMCs, producing monoclonal antibodies with neutralizing capacity against the bacillus. They will then be produced at the pilot scale level to perform validation studies in mouse models of plague.

Production of monoclonal antibodies to fight plague, a potential bioterrorist weapon and disease for which antibiotics-resistant strains of Yersinia pestis have been observed in nature.

It is feared that antibiotics might rapidly be insufficient against plague and no vaccine is available at present. Plague is a potential bioterrosist weapon and has been classified as 'Tier 1' by US CDC. Monoclonal antibodies are more and more used to treat different medical conditions: oncology, autoimmune diseases, and microbial infections. As recently confirmed for COVID-19, they constitute an efficient prophylaxis or treatment opportunity that could be used for Plague notably in case of antibiotiotics- multi-resistant strains. The present project aims to identify, in patients cured of plague, the antigenic targets whose recognition by the immune system is correlated with resistance to infection. We will do a fine characterization of the sites (epitopes) recognized by B and T lymphocytes, and their link with protection will be estimated by a bacteria neutralization test. Protective monoclonal antibodies (mAbs) against plague will be produced by EBV transformation of B cells from cured patients carrying protective antibodies, selection of the specific clones of the identified protective epitopes, then selection of the active agents in the neutralization test. Cloning of the VH and VL fragments will allow large-scale production and testing in animal models of plague.

Institut Pasteur Madagascar (IPM) has a unique acces to plague patients. We will use the library of samples (serum, PBMCs) from Plague patients with various clinical profiles to perform a comprehensive B cell and T cell epitope mapping. Healed patients with highly protective Abs will be identified. We will then use their blood cells to produce immortalized cell lines, and select the monoclonal antibodies with best neutralizing activity, evaluated by bacteria neutralization tests in vitro. The best antibodies will be produced in pilot scale and tested for their ability to protect against infection in animal models of bubonic and pneumonic plague.

The new and strong partnership between IP Paris and Madagascar in a 'Pasteur International Plague Unit' allows to develop this ambitious project. We will identify protective monoclonal antibodies against plague, validate their use in animal models and have them ready for large scale production and potentialy therapeutic applications. These antibodies might represent a new tool against antibio-resistance in Yersinia pestis and industrial partnerships will be searched to patent, produce and commercialize them.

Antibodies against plague would not only serve to fight plague as a bioterrorist weapon, but also to fight the public health problem represented by Plague in countries where it is endemic. These countries include USA, China, India, Brazil, etc. and our technology could be valuable internationally. The development of the methods used in this project will also open the possibility to develop antibodies against other diseases in places of the world where the International Pasteur Institute Network is present.

We expect to produce 3 validated monoclonal antibodies againt Y. pestis with a therapeutic potential.
Cell lines will be immortalized to save them and they will be patented. This collaborative project will be also the source of several publications and communications for public dissemination of the results.

Yersinia pestis, the causative agent of plague, killed millions of humans during three pandemics and is among the deadliest bacteriological agents affecting man. Plague has recently been included in the list of re-emerging diseases and Y. pestis is classified as a potential biological weapon for terrorist use. Because antibiotic resistant strains of Y. pestis have been observed or could be engineered for evil use, vaccination against plague might become the only means to fight against the disease. No safe and effective plague vaccine is currently available. Most efforts made in the recent years were focused on subunit formulations combining the capsular F1 antigen and the V antigen (LcrV). Such vaccines however require the use of an adjuvant such as the controversial Aluminium hydroxyde, and repeated injections to protect, and this protection can be easily circumvented by genetical engineering Y. pestis for bioterrorist use. We followed a vaccine strategy based on oral vaccination with a live, attenuated strain of Yersinia pseudotuberculosis, as this species is genetically almost identical to Y. pestis, but of much lower pathogenicity and higher genomic stability. We have recently constructed an attenuated Y. pseudotuberculosis strain, designated V674TnF1, by deletion of genes coding for three essential virulence factors from the IP32953 strain which genome is known. It was also induced to stably produce the Y. pestis F1 pseudocapsule to increase its immunogenicity. Upon a single oral vaccination, this strain confers 100% protection against a lethal pneumonic plague challenge with a dose of up to 3000 x LD50 of the fully virulent Y. pestis CO92 strain. V674TnF1 also provides 100% protection against bubonic plague, even when an infectious dose of 104 x LD50 is administered. To our knowledge, the protection against bubonic and pneumonic plague conferred by a single dose of V674TnF1 is probably one of the most efficient -if not the most efficient- ever reported. A patent application procedure is currently ongoing.
The present project consist in achieving the steps preliminary to clinical trials in humans. These are 1) the production of a clinical-grade form of the vaccine strain, by reconstructing the vaccine strain without antibiotics –resistance cassettes, as previously used to construct the V674Tn-F1 strain.
2) the verification that the new strain, which will be named V.Yptb-F1, retained the desired vaccinial properties. This implies that it has a strong attenuation of virulence, the ability to persist in vivo in the gut, and the ability to induce a humoral and cellular immune response resulting in 100% protection of animals against bubonic and pneumonic plague.
3) the setup of assays able to predict protective immunity by the measurement of immunological parameters named “correlates of immunity“, whic can be Yersinia pestis-specific antibodies or cell populations.
4) the evaluation of the human immune response in humanized mice, induced by vaccination, to confer protection against plague, before starting clinical trials in humans, by vaccinating and evaluating humanized HIS mice.
5) to protect the intellectual property of the vaccine and the development of scenarios to exploit the potential commercial value of the vaccine, by collaborations with the industry.

Project coordinator

Monsieur Christian DEMEURE (Institut Pasteur - Unité de Recherche Yersinia) – cdemeure@pasteur.fr

The author of this summary is the project coordinator, who is responsible for the content of this summary. The ANR declines any responsibility as for its contents.

Partner

UII (IP) Institut Pasteur - Unité d'Immunité Innée
DARRI (IP) Institut Pasteur - Direction des Applications de la Recherche et des Relations Industrielle
URY (IP) Institut Pasteur - Unité de Recherche Yersinia

Help of the ANR 240,232 euros
Beginning and duration of the scientific project: December 2012 - 24 Months

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