Cell Therapy of Cancer and Autoimmune Diseases
Design of the Novel Approach for Immunotherapy of Cancer and Autoimmune Diseases by the Use of the Allospecific Cytotoxic Lymphocytes
Tech Area / Field
- BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
3 Approved without Funding
Cancer Research Center, Russia, Moscow
- Institute of Immunological Engineering, Russia, Moscow reg., Lyubuchany
- Moffit Cancer Center & Research Institute, USA, FL, Tampa\nSaskatoon Cancer Center, Canada, SK, Saskatoon\nUS Department of Health & Human Services / National Institute of Health / Laboratory of Immunology, USA, MD, Bethesda\nUS Department of Health & Human Services / National Institute of Health / National Cancer Institute / Frederick Cancer Research and Development Center, USA, MD, Frederick\nUniversity of Turku / Finnish-Russian Joint Biotechnology Laboratory, Finland, Turku
Project summaryDisregulation of immune response is often the key factor in development of many infectious diseases also as malignant tumors, autoimmune disorders, etc. Cytotoxic T lymphocytes (CTL) are the basic T-cell subpopulation protecting the organism against tumor growth by recognizing and deleting mutant or malignantly transformed cells. Tumor-specific CTL are widely used in clinical practice as the main effectors in immunotherapy of tumor diseases. However the results of recent studies indicate that neoplastic cells can often escape or be resistant to specific immune response. Moreover, autoimmune reactions to host antigens, leading to generalized involvement of various organs, are often observed during generation of specific immune response to malignant cells. Hence, search for new approaches to cellular immunotherapy of tumor diseases is an important task.
Our studies of allospecific CTL (allo-CTL) lines demonstrated new regulatory functions of these cells. It was found that allo-CTL (TCR+CD8+CD25+FasL+), exhibiting strict specific effect in testing of their killer activity of fibroblastic cell lines, demonstrated the capacity to nonspecifically inhibit the proliferation of activated lymphocytes of different haplotypes, including syngeneic. The presence of allo-CTL led to virtually complete suppression of the immune response induced in vitro in mitogen-dependent blast-transformation and mixed lymphocyte culture (MLC) reactions. On the other hand, analyzed allo-CTL practically did not modulate the intensity of proliferation in syngeneic MLC and did not modify the spontaneous proliferation of intact cells. We have determined that the regulatory effect of allo-CTL can manifest only on condition of their physical contact with responding cells, which were activated. Allo-CTL, having no specific antitumor direction, intensely suppressed the growth of a wide spectrum of transformed tumor cells of different haplotypes and different origin such as: myelomas X-63, NS-1, thymomas EL-4, RDM-4, BW5147 ect.( ratio effector:target = 0,25:1-1:1).
The data obtained allows to suggest allo-CTL, along with innate immunity cells- natural killers, macrophages, etc., to be an additional line of defense against tumor transformation in the organism. This defense can manifest at the initial stage of the process, when targets are recognized by the signs of activation and high proliferative activity.
The results of our study of the mechanisms of regulatory effect of allo-CTL showed that suppression of proliferation of both normal activated and malignant cells can be due to induction of apoptosis process in target cells. It was shown that the presence of FasL molecule on the surface of allo-CTL is not the decisive factor of their apoptotic effect. However the molecular mechanisms of the described suppressive effect remain unclear up to the present time. In order to determine the mechanism of target cell killing by allo-СTL, the induction of apoptosis signal pathways initiated by granzyms, TNF-R, TRAIL, IFN-γ-R also as mitochondrial types of apoptosis (expression of bcl-2 family proteins) will be studied. Further determination of the regulatory mechanisms of allo-CTL effect on activated and intensely proliferating cells can contribute to development of new strategies for the treatment of diseases caused by disregulation of cell activation and proliferation, such as cancer and autoimmune diseases.
Investigations aimed at development of cell therapy strategies as a method of cancer control traditionally paid greater attention to autologous tumor-specific CTL. However it was shown that the use of these cells can be limited because their cross reaction against the antigens expressed on normal tissues and cause autoimmune disorders.
Data project suggests using syngeneic allospecific CTL, carrying receptors which recognize the major histocompatibility complex of allogenic haplotype molecules, as a means of non-specific protection from intensively proliferating autoreactive and malignant cells. The main advantage of the suggested method is that allo-CTL generated in vitro and injected in vivo suppress the growth of tumor cells without inducing autoimmune disorders in the patient due to the absence of alloantigen in the host. Data approach reduces the risk of side effects of such immunotherapy and can be used for cancer treatment and correction of autoimmune diseases.
The goal of the project is search for approaches to correction of pathological states caused by infringement of cell activation and proliferation control. Basing on the obtained data on molecular and cellular mechanisms of allo-CTL regulatory effects evaluation of the possibility of using allospecific CTL for therapy of cancer and autoimmune diseases.
Scientific level of the project executors. Laboratory of Regulation Mechanisms in Immunity, Carcinogenesis Institute has necessary base and methods for comprehensive evaluation of immunological parameters, experimental models of tumor growth, and is experienced in manipulations with long living T-cell lines. Department of Molecular and Cellular Biology, Institute of Immunology, carries out structural and functional investigations of proteins also as investigations of apoptosis pathways induced by different factors in cancer and normal cells. Due to their experience and findings, the scientists of Institute of Immunology will effectively cooperate with their colleagues at Laboratory of Regulation Mechanisms in Immunity in solving the tasks of data project.
Results of theoretical importance:
· Description of a new phenomenon of regulatory functions of allospecific cytotoxic T lymphocytes.
· Detection of the mechanisms of regulation and interactions of the immune system components with estimation of CTL role in the total system of regulatory effects.
Results of applied and commercial importance:
· Development of a new strategy for immunotherapy of cancer, based on non-specific allo-CTL-dependent regulation of malignant cell proliferation.
· Development of new approaches to the treatment of autoimmune diseases associated with pathologically high activity and proliferation of immunocompetent cells.
· Development of new drugs for the treatment of diseases caused by infringement of cell activation and proliferation control and aimed at stimulation or neutralization of regulatory cell-to-cell interactions in cancer and autoimmune diseases.
Meeting ISTC Goals and Objectives.
Realization of this project will promote the decision of an important social problem: the scientists and engineers of the Institute of Engineering Immunology of Joint Stock Company “Biopreparat”, who previously worked within the framework of state orders in the sphere of biological defense, will be able to carry out fundamental and applied research aimed at solution of international research and technological problems of biology and medicine.
Scope of Activities.
The main tasks of the project are:
· Detailed analysis of cellular and humoral mechanisms determining the realization of regulatory function of allo-CTL.
· Detection of molecular mechanisms of allo-CTL-induced suppression in cancer and normal target cells.
· Analysis of sensitivity of malignant cells of different origin and of radio- and chemotherapy-resistant cells in order to detect the targets for allo-CTL suppression.
· Study of allo-CTL effect on normal immune response in experimental models in vivo. Detection of allo-CTL effect on activated CD4+25+ regulatory T cells.
· Development of experimental models in vivo for evaluating the possibility of using allospecific CTL for the treatment of cancer and autoimmune diseases.
Role of foreign collaborators.
The following types of collaboration are intended: information exchange and collaborative research within the framework of the project.
Technological approach and methodology.
Isolation of cell populations and analysis of the markers will be carried out using knockout mice and magnetic beads covered with antibodies to analyzed structures. The evaluations will be carried out by flow cytometry. Synthesis and production of humoral factors will be evaluated by PCR and ELISA. Apoptosis mechanisms will be studied using annexin V and propidium iodide, appropriate kits also as mice and cell lines transgenic and knockout of analyzing protein genes. Autoimmunity models will be studied on mice genetically predisposition to these diseases.
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