From 27 till 4 of March 2010 the delegation of TSMU with the head of the rector, honorary worker of Science and Technology of Ukraine, corresponding member of AMS of Ukraine professor Leonid Y. Kovalchuk, visited the Oncology Research Institute of Clemson University

From 27 till 4 of March 2010 the delegation of TSMU with the head of the rector honorary worker of Science and Technology of Ukraine, corresponding member of AMS of Ukraine professor Leonid Y. Kovalchuk; prof. M.Korda, the head of the Medical Biochemistry and Clinical Laboratory Diagnostics department; prof. I.Halaychuk, the head of Oncology, Radiology Diagnostics and Therapy and Radiation Medicine department and ass.prof. N.Khranovska, the head of Experimental oncological laboratory of National Cancer Institute visited the Oncology Research Institute of Clemson University (South Carolina, the USA).

The main aim of Ukrainian delegation’s visit to South Carolina was to acquaint with the achievements of the Oncology Research Institute at the head with Dr. Thomas Wagner, and also to find the ways of cooperating between this institute and TSMU.

Dr. Wagner was educated at Princeton University (A.B. Magna Cum Laude) and Northwestern University (Ph.D.), and has served as Professor of Medicine at Cornell University Medical School, Associate Member of the Sloan-Kettering Cancer Research Institute, founding Chairman of the Department of Molecular and Cellular Biology and founder of the Edison Biotechnology Institute at Ohio University. In 1998 Dr. Wagner moved to South Carolina to establish a new Institute focused upon the application of contemporary molecular (gene therapy) and cellular (cell therapy) biology to medicine. He is presently Director of the Oncology Research Institute of the Greenville Hospital System. Dr. Wagner is also Distinguished Professor of Molecular Medicine at Clemson University. Dr.Wagner led the research team that accomplished the first transfer of a functional gene between species, when in 1979 he produced the first transgenic animal, a mouse with the rabbit hemoglobin gene incorporated into its germline.

The Oncology Research Institute is focused upon the inhibition of tumor angiogenesis, tumor gene-immunotherapy and the treatment of breast cancer by blocking the action of a key breast cancer survival factor. The Oncology Research Institute consists of three laboratories. They are the laboratory of molecular biology, the laboratory of cellular biology and the laboratory of development biology. The staff works on the introduction of innovative approaches to the treatment of cancer.

Human dendritic cell

Clinical research of anticancerous vaccines is held in many oncological centres all over the world. The process of getting the vaccine includes the production of immature autological dendritic cells by the incubation of their precursors with cytokins in different combinations. Then these dendritic cells interact with tumor antigens, incubating by certain factors that stimulate their ripening.

Cultivation of dendritic cells

Dendritic cell is a special type of cell that is a key regulator of the immune system, acting as a professional antigen-presenting cell (APC) capable of activating naпve T cells and stimulating the growth and differentiation of B cells. Dendritic cells are present in small quantities in tissues that are in contact with the external environment, mainly the skin (where there is a specialized dendritic cell type called Langerhans cells) and the inner lining of the nose, lungs, stomach and intestines. They can also be found in an immature state in the blood. Once activated, they migrate to the lymphoid tissues where they interact with T cells and B cells to initiate and shape the adaptive immune response. At certain development stages they grow branched projections, the dendrites, that give the cell its name. However, these do not have any special relation with neurons, which also possess similar appendages. Immature dendritic cells are also called veiled cells, in which case they possess large cytoplasmic 'veils' rather than dendrites.

The important problem is to provide the sufficient gene-immune tumor antigens and to make the conditions for their effective introduction. There is the specific technology of ‘fusion’ of dendritic cells with tumor cells of patient, excreted by biopsy or surgery.

At present in Scientific Institute of Oncology of Clemson University and Hospital System of Greenville immune vaccination is studied efficiently of on the basis of dendritic cells on many types of tumors in an experiment. Vaccination using dendritic/tumor cell hybrids represents a novel, promising cancer immunotherapy. They have developed a technology that can instantly purify the hybrids (dendritomas) from the fusion mixture of DC and tumor cells. Animal studies and a pilot study of stage IV melanoma patients demonstrated that dendritoma vaccination appears to avoid major toxicity and induced some tumor cell specific immunological and clinical responses. In this on going pilot study, seven stage IV renal cell carcinoma patients have been studied. Dendritomas were made from the patient’s autologous DCs and tumor cells and given to the patient by subcutaneous injection. After the initial vaccination, three escalating doses of IL-2 (3, 6, and 9 million units each) were followed within five days. The patients appeared to tolerate this treatment regime well. The most common adverse events were flushing of the skin and rashes. Erythema, chills, joint stiffness of hand and swollen tumor lesions and lethargy were also observed. All these adverse events were grade 1 (total 17). One patient had erythema of the eye (grade 2) which was possibly related to the vaccine. There were no grade 3 or 4 adverse events related to the vaccine. In order to determine whether the vaccine can stimulate tumor cell specific immune response in patient, the percentage of T cells that express interferon- was analyzed using flow cytometry. Seven out of seven patients showed increase of IFN- expressing CD4+ T cells after vaccination(s); while six out of seven patients showed increase of IFN- expressing CD8+ T cells. Clinically, one patient developed partial response and three other patients’ diseases were stabilized after vaccination(s). SD: stable disease; DFD: dead of disease; PD: progressive disease; PR: partial response; N/A: not applicable.

Another direction which is widely developed in Scientific Institute of Oncology is research of breast cancer. In particular by an interesting project, which Ukrainian delegation familiarized with, and which, according to dr. Vagner, has a good prospect, there is a study of influencing of hormone of protactinium in a complex with chemotherapy, on proliferation of breast cancer. It is known that prolaktina stimulates growth of breast tissues and, accordingly, stimulates proliferation of tumor cells. However, at the same time, it promotes the sensitiveness of maligned cells, especially immature, undifferentiated, or poorly differentiated, to the chemotherapy, in particular, to doxorubicin. That is why, the use of doxorubicin together with a lactate hormone results in the increase of chemotherapy, considerably anymore oppression of growth of tumor, than application of chemotherapy, especially sharply the amount of youths, undifferentiated carcinoma cells diminishes at such treatment.

Human Macrophage

It is known that basic function of macrophage this fagocitation of extraneous matters which got into organism, and also cells which lost viability or mutations were added, in particular tumor cells. It appears that if to “acquaint” macrophage with the cage of tumor which perished after the mechanism of necrosis, such macrophage acquires protective, properties of antitumor’s. Through the products of certain citochine it activates and mobilizes immune antitumor defense. If to “acquaint” macrophage with the cage of tumor which perished after the mechanism of apoptosis, such macrophage, opposite, stimulates proliferation of cells, activates angiogenesis and, as a result, has a antitumor action.

Different effect of apopic and nectotic tumor cells on macrophage control of tumor growth

Human monocytes, isolated from whole blood, were seeded onto tissue culture grade polystyrene (PS) and three polycarbonate-based polyurethanes (PCNUs) (synthesized with either 1,6-hexane diisocyanate (HDI) or 4,4′-methylene bis-phenyl diisocyanate (MDI), poly(1,6-hexyl 1,2-ethyl carbonate) diol (PCN) and 1,4-butanediol (BD) in different stoichiometric ratios (HDI:PCN:BD 4:3:1 or 3:2:1 and MDI:PCN:BD 3:2:1) (referred to as HDI431, HDI321 and MDI321, respectively). Following their differentiation to monocyte-derived macrophages (MDMs) the cells were trypsinized and reseeded onto each of the PCNUs synthesized with either 14C-HDI or 14C-BD and degradation was measured by radiolabel release (RR). When the differentiation surface was MDI321, there was more RR from 14C-HDI431 than from any other surface (p<0.0001) whereas the amount of esterase (identified by immunoblotting) as well as the esterase activity was the greatest in MDM differentiated on PS, reseeded on 14C-HDI431 (p<0.0001). The effect of potential degradation products (methylene dianiline (MDA) and BD) from the PCNUs was carried out to determine possible links between products and substrate-induced activation of MDM. MDA was found to inhibit RR 60% from MDM seeded on 14C-MDI321B (p<0.0001), ∼20% from 14C-HDI431 (p=0.002) and no effect from 14C-HDI321B. MDA inhibited esterase activity 30% from MDM only on 14C-MDI321B (p=0.003), but no effect on esterase activity was observed for the other two polymers. BD had no inhibitory effect on RR from any PCNU, but did inhibit esterase activity in MDM on 14C-HDI431 (p=0.025). This study indicates that the degradation of a specific material is a multi-factorial process, dictated by its susceptibility to hydrolysis, the effect of specific products generated during this course of action, and perhaps not as well appreciated, the material’s inherent ability to influence enzyme synthesis and release.

Between the Ukrainian side and guidance of Scientific Institute of Oncology negotiations about a possible collaboration were conducted. Sides agreed about organization of joint activity in scientific and practical collaboration in industry of experimental and clinical oncology, in particular in relation to carrying out researches, ant carcinogenic vaccines related to creation on the basis of dendritic cells, and by their clinical application. Also both Ukrainian and American sides consented to take part in scientific researches of commons in other directions of oncology. As a result of the conducted negotiations agreement about collaboration was signed, from the American side director of Scientific Institute of Oncology dr, Wagner signed the agreement and from Ukrainian side- rector of Ternopil state medical university, corresponded member of AMS of Ukraine, prof. Kovalchuk L. Ya. From the Ukrainian side a project will be carried out under an aegis National Institute of Cancer (director – prof. I.B. Shepotin). It is necessary to mark, that in National Institute of Cancer of Ukraine workers of researches already for a few years successfully are engaged in creation of autologic tumor vaccines on the basis of dendritic cells and already attained in this direction enough considerable successes both in an experimental and in clinical plan. Except National Institute of Cancer and Ternopil medical university, Ciber Clinic of Spizhenko (akad. Ю.П.Спіженко) and medical center of “Medivit” of private medical college of Ternopil will participatate in scientific researches of common scientific researches.

The fusion of dendritic cells with tumor cells to form hybrid cells – dendritomas

One of Dr.Wagner achievements is using of magnetic nanoparticles (dendritic cells, tumor cells and dendritomas are marked by them) and using of special magnetic columns to obtain the pure fraction of hybrid cells. The pure fraction is the vaccine, which is injected the patient, stimulating the immune system to identify malignant cells in the organism. Then the immune system attacks tumor cells and kills them.

Microscopic image of tumor cell (A), dendritic cells (B), mix of cells after fusion, consisting of dendritomas, tumor and dendritic cells (C), pure fraction by dendritomas ,obtained after sorting on the magnetic column (D).

The general scheme of tumor vaccine therapy

Director of the Scientific Oncological Institute Dr. Vagner presents his achievements in creation of anticancer vaccines on the basis of dendritic cells.

Professor of Biology Department of the University of Klemson Dr. Ven Chen tells his Ukrainian collegues about his scientific project concerning the perspectives of prolactin usage in breast cancer treatment.

At the molecular biology laboratory of the Scientific Oncological Institute.

Rector of Ternopil State Medical University professor Kovalchuk L. Ya. made a decision on the creation of groups which would work on three scientific projects in coordination with American scientists. These are: “Treatment of cancer with the help of antitumoral autologous vaccines on the basis of hybrid cells by dendrite”, “The rise of effectiveness of cytostatic influence of chemical substances at experimental breast carcinoma” and “Double role of macrophages in the immune protection from the tumours and regeneration of tissues”. Those who are interested in these projects, young scientists, students can address themselves to the scientific department for consultations.

At the last day of their visit to South Carolina Ukrainian guests acquainted themselves with the work of laboratories of the Hospital System of Grinville and visited one of the hospitals of this system – Grig Memorial Hospital that was built only two years ago.

Laboratory complex that serves all the hospitals of the Hospital System of Grinville, and other hospitals of the region that do not belong to this system consists of virusologic, bacteriologic, general clinicalal, biochemical, haematological, toxicological laboratories. Here thousands of samples of biological material are analysed, tens of thousands of various analyses are made. All the work is made by robots and automatical analysators. All the information about the analyses is recorded and analysed on computer then it is printed or in electronic variant is given to the doctor in charge.

Computer-aided lines of biochemical laboratory

Biochemical analisator

High-perfomance liquid chromatograph

Visit to the Grig Memorial Hospital was very interesting. This is one of the hospitals that is a part of the Hospital System of Grinville. As far as the hospital was built only two years ago everything in it from the design to internal filling meets world standarts.

Grig Memorial Hospital has very beautiful architecture

Interior of Grig Memorial Hospital

President of Grig Memorial Hospital John Mansure tells rector of Ternopil State Medical University professor Kovalchuk L. Ya.about his staff

At the obstetrics department. A woman gives birth in a separate room which is equiped with all the necessary facilities.

Here in the ward there is a place for a baby…

… and a sofa for the father that can stay here with his wife and baby

Each room has a TV, DVD, fridge, bathroom. After the birth of each baby there sounds pleasant music at the hospital

Surgical wings are equiped with modern apparatus

Nursing post (one for two ward)

Center for senior people also belongs to the Grig Memorial Hospital. Twelve houses where senior people live are situated not far from the hospital. In this center stay senior people who need regular care, i. e. who suffer from chronic diseases or have some mental disorders. Here they get qualified and professional help; nurces, physical therapist and other specialists work with them. Volunteers help the center too.

Center for senior people

Every house contains 12 rooms

In the center of each room there is a kitchen, dining-room, and hall where old people gether together, have rest, watch TV.

Rector of Ternopil State Medical University professor Kovalchuk L. Ya. and president of the Center for senior people