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Device for Human Lymphocyte Status Monitoring

#B-1040


Device for Human Lymphocyte Status Monitoring in Diseases and in Ecological Effects Based on the Investigation of Lymphocyte Electrophysical Properties

Tech Area / Field

  • MED-DID/Diagnostics & Devices/Medicine
  • ENV-EHS/Environmental Health and Safety/Environment

Status
3 Approved without Funding

Registration date
17.06.2003

Leading Institute
Hematology and Blood Transfusion Research Institute, Belarus, Minsk

Supporting institutes

  • Closed Corporation Design Bureau "Pribor", Belarus, Minsk

Collaborators

  • Johannes Gutenberg-Universitat Mainz / Institut für Physiologische Chemie, Germany, Mainz

Project summary

Project Objective: comparative study of structural, metabolic and electrophysical characteristics of peripheral blood lymphocytes in groups of persons with different morpho-functional peculiarities of lymphoid tissue using complex of methods and development of the device for monitoring of lymphocyte state in disease and upon ecological influences.

Introduction and overview. Functional tests and clinical studies applying for the estimation of the influence of anthropogenic factor at the level of the whole body are complicated for interpreting and standardization of the output. Estimation of the effect of the interaction of the body with external environment by morbidity rate, though the most reliable, is possible only at the late stage of the pathology formation [Svirnovski A.I. Hematological effects of Chernobyl in Belarus: do they really exist ten years later?// Hematology. – 1997. – 2, N 4. – P. 303-312.]. Urgency of the Project is thereby raised from the need of development of standardized scheme for the early estimation of the external environment influence on biochemical and molecular – biological parameters within the framework of a model system. At present, an idea has been actively developing that blood lymphocytes can bear a function of bioindicator at estimation of the organism state due to their unique characteristics [Berridge M.J. Lymphocyte activation in health and disease. // Critical Rev. in Immunol. 1997. 17, N 2. – P.155-178; Burnett K.G. Evaluating intracellular signalling pathways as biomarkers for environmental contaminant exposures // Am.Zoologist. – 1997. – 37, N 6. – P.585-594]. Common tests which are conventionally used for in vitro analysis of lymphocyte reaction to varied stimuli and characterize viability, different apoptotic stages, proliferative state, genotoxic effects and production of the specific products in most cases require considerable expenses being time-, equipment- and reagent- consuming. In this regard, the participants of the Project propose to develop special methods of study which give possibility to characterize lymphocyte state basing on the kinetics of their electrophysical characteristics under short-term cultivation. Due to close interrelation of electroconductivity of cellular suspensions and their рН level with physical and chemical structure of cellular elements and metabolism [Upadhyay P., Bhaskar S. Real time monitoring of lymphocyte proliferation by an impedance method. // J Immunol Methods. - 2000. - 244, N 1-2. - P.133-137] it is likely that registration of kinetics of rapid metabolic response of cells based on the given parameters will characterize activation of cells, potential ability to performance of their specific function or intensity of apoptotic processes upon physical, chemical and biological influences.

The impact of the Project on the progress in the field: the above-stated enables to believe that on the basis of dynamic conductometry (impedancimetry) and рН - measurements it is possible to develop a system for early detection of lymphoid cell response to activating, proliferative and apoptotic stimuli. Differences in dynamic changes of extracellular рН and electric resistance of peripheral blood lymphocytes during their response to a complex of stimuli will allow to differentiate between normal state and various autoimmune and lymphoproliferative diseases, to estimate inpidual sensitivity to certain stimulus and to reveal groups at risk for the given pathology among the healthy population and the persons continuously exposed to adverse ecological factors due to their residence (inhabitants of radioactive-polluted territories) or occupation. At the same time, specificity of screening investigations on lymphocytic cultures which apply рН - and conductometric methods will result in the development of highly productive measuring device (a know-how of participants of the Project).

Competence of the Project team: experts of two institutions will take part in performance of the Project: Hematology and Blood Transfusion Research Institute (HBTRI) and Close Corporation Design Bureau “Pribor” (CCDB "Pribor”). The medical-biologic group of participants of the Project (HBTRI) which consists of 13 persons (1 – Prof., M.D., Ph.D., Dr. Sci (med), 3 – Ph.D., 5 – physicians and biologists, 4 – supporting personnel) has an operational experience in the area connected to performance of Project tasks 1, 3, 5, 6 (the analysis of lymphocyte properties), that is reflected in the published works (see 12.2). The engineering - design group (CCDB "Pribor”) which consists of 7 person (4 – Cand. Tech. Sci., of them 1 – Prof. and 2 – Associate Prof., 1 – expert without a degree, 2 – supporting personnel) participates in performance of tasks 2-6 and has the overall experience in weapon expertise areas of 138 years in the fields of design, construction and performance of missiles (techniques for guidance and control). By employees of the group 482 scientific works are published and 108 copyright certificates are received.

Expected results. Categories of Technology Development of Research: applied research and technology development.


New data on connection between the basic processes in lymphocytes in normal and pathological conditions (task 1).
New methodical approach to the analysis of lymphocyte properties (task 2).
Informative characteristics for the estimation of lymphocyte state and organism state as a whole (task 3).
Automated device for the characterization of lymphocyte properties (task 4).
Risk estimation of potential danger of environmental contamination by various factors, Chernobyl-related ones included, and estimation of inpidual sensitivity to the action of damage factors and of occupational safety (at admission to work) for persons working in unfavorable conditions (task 5).
Test for in vitro determination of sensitivity of patients with autoimmune, immunodeficient and lymphoproliferative diseases to potential drugs and selection of inpidual treatment (task 6).

Global result – to hallmark the early lymphocyte response to various factors and device development for this criterion estimation in order to provide not only for lymphocyte properties evaluation but also for the assessment of body-environment interaction and treatment option in lymphoid tissue pathology.

Scientific significance of the Project: new data on lymphocyte properties that allow to establish quantitative regularities connecting early lymphocyte responses (among them production of reactive oxygen species, change of extracellular рН and electroconductivity of cellular suspension) to an exogenous stimulus and manifestations of genotoxic effects, induction of apoptotic program and proliferative state.

Applied significance: definition of those special parameters which can be used for estimation of early lymphocyte responses to any influences and implementation of economically and technically accessible device for registration of the most informative and reproducible response and for choice of the optimal methods to effect autoimmune, immunodeficient and lymphoproliferative diseases.

Scope of Project results: medicine, namely hematology, therapy, immunology, occupational pathology, hygiene, as well as ecology.

Commercial significance: development of the effective device, cost of which appears acceptable even in the countries with transitive economy. It is prospected to certificate the device, to carry out wide clinical and laboratory research of the device and methods, to carry out special search for investments for manufacturing of the device on the basis of Close Corporation Design Bureau “Pribor”. It is supposed to keep the intellectual property under the Project, would-be users: research and patient care institutions, clinical, diagnostic and hygienic laboratories.

Meeting ISTC goals and objectives: re-orientation of experts, who were in the past connected with weapon expertise areas, to civilian activities by involving them in the development of a device that is designed for blood cell analysis; integration of the Project participants in the international scientific community by their collaboration with foreign scientists and publication of the results of their work in international journals; support of the applied research and development of technologies for the peaceful purposes in the field of body-environment interaction by the creation of a special device for lymphocytes and organism status analysis; promotion of transition to market economy that is provided for by search of buyers of technology and of the device with subsequent self-financing of the Project participants.

Project duration and total person-months of Project efforts: duration of the Project is 36 months. The amount of people-months is 344 (HBTRI – 173.5; CCDB “Pribor” – 170.5). Solution of 6 interrelated tasks.

Participation of foreign collaborators: participation in preparing of detailed work plan, discussions and checking of the results of work, presentation of some scientific data..

Technical approach and methodology: abundant experience of weapon and medical biology specialists in their fields, usage of lymphocytes as test-cells for the evaluation of body-environment interaction and a set of known and original methods for the estimation of lymphocyte properties and hardware for device design based on the estimation of cell suspension electroconductivity variable with time and conditions (know-how): 5 channels, run time 1-3 hours, temperature span 35-38 °C, temperature accuracy 0.2 °C, sample volume up to 1 ml.


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