Structure-function Studies of Human Cytokines and their Mutant Analogs
Tech Area / Field
- PHY-PFA/Particles, Fields and Accelerator Physics/Physics
- BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
- MED-DIS/Disease Surveillance/Medicine
3 Approved without Funding
Institute of Immunological Engineering, Russia, Moscow reg., Lyubuchany
- Scientific Research Institute of Epidemiology, Microbiology and Infectious Diseases, Kazakstan, Almaty\nState Research Center of Virology and Biotechnology VECTOR / Scientific Research, Design and Technology Institute of Biologically Active Substances, Russia, Novosibirsk reg., Koltsovo\nInstitute of Bioorganic Chemistry, Russia, Moscow
- University of Turku / Finnish-Russian Joint Biotechnology Laboratory, Finland, Turku
Project summaryThe goal of the project
The goal of the project is studying the structural and functional properties of recombinant gamma-interferon, tumor necrosis factor alpha and their mutant analogs for selection of new gene-engineering preparations with a definite biological activity. The obtaining of the information on molecular mechanisms of cytokine functioning.
Main tasks to reach the above objective
-Structure studies using mainly 1H-NMR spectroscopy as well as CD and microcalorimetry of recombinant human IFN-g, TNF-a and their analogs under physiological conditions and upon changing the external factors (temperature, pH of solution, action of denaturants).
- Assay of biological activity of mutant analogs of human gamma-interferon and TNF-a;
- Design of new hybrid proteins on the basis of human IFN-g and study of their structural and biological properties;
- Carrying out the preclinical trials of TNF-a as a medicinal preparation for treatment and prophylaxis of brucellosis;
- Comparative assay of structure-function relations of mutant analogs of recombinant human IFN-g and TNF-a.
- new data on molecular mechanisms of cytokine functioning;
- as a result of the structure-function studies of mutant analogs of gamma-interferon and TNF-a and their comparative assay, the grounds will be provided for designing gene-engineering preparations with a definite biological activity;
- the design genes of new hybrid proteins on the basis of human IFN-g and study of their structural and biological properties;
- preclinical trials of TNF-a as a medicinal preparation against brucellosis will allow one to determine its possible application for treatment and prophylaxis of this disease.
Technical Approach and Methodology
It is proposed to study the structural characteristics of cytokines using mainly NMR spectroscopy and other physical methods such as CD and microcalorimetry.
- Experiments are proposed for assaying the following types of biological activity of the cytokines studied: antiviral, antiproliferative, cytolytic and immunomodulating activities.
- Radioreceptor analysis will be applied to assay the receptor-binding activity of cytokines.
A number of experiments, with the aid of which the effect of TNF-a on formation and course of infectious brucellosis will be studied, are planned to be carried out using a model of acute brucellosis in laboratory animals. Besides, efficiency of the treatment will be evaluated by objective characteristics of the state of the animals (weight, integument etc.) and using bacteriological procedures. It is also planned to use serologic methods, Burne alergotest, opsonophagocytic reaction with brucella and determination of the functional activity of the phagocytic system.
International cooperation under the project submitted to the ISTC
It is planned to exchange the information received under the project, to prepare joint publications, participation in symposium and conference works, to make visits to laboratories of collaborators aiming at carrying out the works under the project.