Tuberculosis Bacteriophage: Development of Obtainment and Application Methods
Tech Area / Field
- MED-DIS/Disease Surveillance/Medicine
3 Approved without Funding
Institute of Tuberculosis and Pulmonology, Georgia, Tbilisi
- Georgian Academy of Sciences / Institute of Bacteriophage, Microbiology and Virology, Georgia, Tbilisi
- Kuzell Institute for Arthritis and Infectious Diseases\California Medical Center Research Institute, USA, CA, San Francisco\nMerlin Technologies, Inc., USA, MA, Boston\nPublic Health Research Institute, New York, USA, USA, New York
Project summaryPurpose of the project: obtaining M. Tuberculosis phage (TB-phage) and its utilization for the development of the alternative methods of improving the efficiency of the measures against Tuberculosis.
In 1990’s the problem of Tuberculosis has regained a substantial momentum. The scale of pandemic proved to be of such extent that in 1993, WHO declared it a worldwide threat (WHO, 1993) and its experts envisage the TB situation in 21st century to outweigh the same in the 1930–1950s, when the incidence has reached the peak (WHO, 1997). The gravity of the present situation is further aggravated by one of the most serious problems modern phthisiology faces – that of the resistance of Mycobacteria to the specific chemotherapy currently in use (Dixie E. et al. 1998, Gill C.O. et al. 1999, Lopez L. et al. 1999). Moreover, preventive, diagnostic, and treatment methods being exercised are progressively antiquating, raising the demand for their exigent modernization. The necessity of the emergence of the advanced technologies to perfect existing methods and means of fight against TB and evolvement of new ones has already long been a pressing issue on the agenda (Ellner J.J. 1998, Iseman M.1998). In this light, research into the possibilities of using bacterophages seems to us to be very promising. To date, application of phage technologies in the fields of treatment, diagnosis, and prophylaxis of various bacterial infections has shown significant success (Zhvania M. Sh. 1974, Anpilov L.I.et al. 1984, Nevsky M. V., 1985, Chanishvili T. G., 1998); however, only a few published sources have touched upon this problem in regard of TB, and the topic of phages in phthisiology has in general been poorly covered as yet. Meanwhile, many currently elucidated properties of the phages imply practicability of both separation of TB-phages and their targeted application. Bacteriophage is a virus with a capacity of infecting a bacterial cell, reproducing inside the host, and causing its lysis – the capacity that conduced to the broad use of phagotherapy in medical practice (Samsygina T. A. et al. 1984, Ponomareva T. R. et al. 1985, Meipariani A. N. et al.1990). The consequences of separation of the TB-phage and its modification into a clinically applicable form are twofold: it could in many senses contribute to the attainment of one of the major goals of modern phthisiatry – that of resistance to antibiotics; and it would widen the prospect of appearance of the optimal preventive tool with no usage restrictions even for resistant forms of disease. Improvement of diagnostic procedures is another indubitable perspective of using TB-phage.
Personnel involved comprises phthisiologists and microbiologists-virologists, all with extensive background in Tuberculosis and phages and with experience in research work in these fields, namely, by areas: Studies of biological and morphological properties of phages and their interaction with host cells – Professor T. G. Chanishvili; Elaboration of methods of developing new prophylactic and medicinal bacteriophage preparations and improvement of existing ones – Professor A. N. Meipariani; TB Chemo prophylaxis – Professor O. G. Batiashvili; Organization of TB services and molecular epidemiology – Professor G. N. Khechinashvili; Surgical treatment of the patients with drug-resistant forms of Tuberculosis and TB in refugees – Professor S. A. Vashakidze; Microbiological aspects of drug-resistance – L. O. Danelishvili, MD, etc.
Development of new TB-phage technologies and viability studies for determination of its applicability will gain new knowledge which will in turn root a new branch in phthisiology, dealing with utilization of the phages for diagnostic, treatment, and prevention purposes. Obtaining medically active forms of the phage will trigger the emergence of qualitatively new TB drug, free of side effects and toxicities for the human organism, whose mode of action will be based on its ability of selective lysis of particular microorganisms.
Medicinal forms of TB-phage (autophage, polyvalent phage), being exceedingly prospective from the point of view of resolution of drug resistance problem, will cause important consequences.
One of these is using phages as a highly effective prevention method in high-risk groups of population, such as children, contacts of patients, inmates and personnel of penitentiaries, patients of specialized medical institutions (hospitals, hospices), etc. It is to be underscored that the phagoprophylaxis can be implemented in cases where currently used BCG vaccination is contraindicated, i.e., in severe cases and in tuberculin- and HIV-positive groups, as well as among the patients with drug-resistant forms of disease. Phagodiagnostics may also become an alternative modality of bacteria detection in TB patients, offering certain advantages as compared to the traditional methods of smear microscopy and bacteriology based on selective media in terms of sensitivity and promptness of result availability.
Should the project be successfully implemented, high-scale industrial technologies may ensue, with excellent commercial potential.
This project fully meets the ISTC objectives: it enables the scientists involved in Defense industry in CIS (in this case, in Georgia) to redirect their research field, expedites their integration into the global scientific community, contributes its share to fundamental research, and bears a significant medical and humanitarian importance as well as a good commercial potential which will strengthen the new Democracies through facilitation of the introduction of market economy.
Research will include search for and separation of TB-phage (use of lung resection materials, sputum, and environmental specimens – soil, sewage, etc. - is planned), study of its morphological, serological, and biological properties in vitro and in vivo, determination of possibility if its use for phagodiagnostics of Tuberculosis, development of an experimental preparation, as well as integrated assessment of its prospective use for the elaboration of alternative clinical methods of diagnosis, treatment, and prevention of Tuberculosis.
Scale of collaboration with foreign collaborators: information exchange during the implementation of the project, joint studies of specimens (Control mycobacteriophage specimen).
On the first stage of research, Polymerase chain reaction (PCR) shall be used to achieve high sensitivity, specificity, and rapidity of detection of M. Tuberculosis in clinical specimens of pathological material.
In order to obtain test cultures for TB-phage detection and to assemble the bank of M. Tuberculosis cultures, bacterial cultural method will be used, which will also facilitate the solution of drug-sensitivity and bacterial identification issues. Drug resistance will be determined with absolute concentrations method, and rapid and precise identification of Mycobacterial species will be performed using Gen-Probe (ACCUPROBE) method.
To separate TB-phages from pathology and environmental specimens, Redmond and Ward method will be employed. Upon separation, morphology, serological properties, lythic range, and specific affinity for various standard (H37Rv, Academia, Bovis 8, M. avium P, M. smegmatis 607) and wild (sensitive and resistant) cultures of Mycobacterium will be studied, and to clarify the feasibility of future use of TB-phage for diagnostic purposes, phagotyping and reaction of phage titer increment will be used.
On the penultimate stage, in vivo studies in Guinea pigs will be performed, and the efficiency of treatment will be evaluated based on histomorphological and bacteriological examination of the viscera and survival time of infected animals. Comparative analysis of these data in respective groups will comprise the ground for determination of the degree of antibacterial activity of TB-phages. Upon completion of this study, creation of an experimental preparation is planned. Apelman and Gracia method will be used to evaluate its activity.
The research described above will provide data to be further analyzed to apprize the prospects of using TB-phages for the development of alternative methods of diagnosis, treatment, and prevention of Tuberculosis.
The International Science and Technology Center (ISTC) is an intergovernmental organization connecting scientists from Kazakhstan, Armenia, Tajikistan, Kyrgyzstan, and Georgia with their peers and research organizations in the EU, Japan, Republic of Korea, Norway and the United States.
ISTC facilitates international science projects and assists the global scientific and business community to source and engage with CIS and Georgian institutes that develop or possess an excellence of scientific know-how.