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Евразийский Союз Ученых (ЕСУ) #4 (49), 2018
STATE OF T-CELLULAR IMMUNITY DEPENDING ON THE VARIANTS OF COHERENT IMMUNOTHERAPY IN PATIENTS WITH UTERINE CERVICAL
CANCER.
СОСТОЯНИЕ Т-КЛЕТОЧНОГО ИММУНИТЕТА В ЗАВИСИМОСТИ ОТ ВАРИАНТОВ СОПРОВОДИТЕЛЬНОЙ ИММУНОТЕРАПИИ У БОЛЬНЫХ _РАКОМ ШЕЙКИ МАТКИ._
Kamishov Sergey Viktorovich
MD, PhD, senior researcher, chemotherapeutist Republican Specialized Scientific and Practical Medical Center of Oncology and Radiology of the Ministry
of Health of the Republic of Uzbekistan, Tashkent, Uzbekistan
Камышов Сергей Викторович,
к.м.н., старший научный сотрудник отдела химиотерапии Республиканский специализированный научно-практический медицинский центр онкологии и радиологии Министерства здравоохранения Республики Узбекистан, Ташкент
АННОТАЦИЯ
В работе изучались показатели Т-клеточного дисбаланса в процессе сопроводительной иммунотерапии у больных раком шейки матки. Анализ полученных результатов позволил выявить выраженные изменения в клеточном звене иммунитета, которые проявляются подавлением экспрессии CD3+, CD3+CD4+, ИРИ, повышением экспрессии CD3+CD8+ и CD 16+. Как видно, наилучшая ситуация характерна для больных РШМ после ЭИФТ+ПФ, где наблюдается активация Т-клеточного иммунитета. Наблюдается выраженный дисбаланс клеточного иммунитета. Причем, дисбаланс в клеточном звене иммунитета выражается в подавлении ИРИ за счет снижения количества Т-хелперов/индукторов и повышения Т-цитотоксических лимфоцитов. Очевидно, при данной патологии Т - клеточный иммунный ответ существенно слаб и направлен против меньшего числа эпитопов, что позволяет предполагать клональное истощение Т-лимфоцитов. В свою очередь, пониженная иммунореактивность Т - клеточного звена может рассматриваться как результат нарушения представления антигена клеткам иммунной системы, а также нарушение функции самих Т-клеток. Следовательно, выявленная Т-клеточная лимфопения часто характерна на фоне или после ПХТ. Подавление ИРИ свидетельствует о наличии Т-клеточного иммунодефицита, как было сказано выше, в основном за счет снижения количества Т-хелперов/индукторов, которые играют важную роль в реализации иммунного ответа. Повышение Т-цитотоксических лимфоцитов, что свидетельствует о подавлении Т-клеточного иммунитета и о наличии цитотоксического действия на клеточном уровне. Установлена положительная клиническая эффективность применения сочетания иммунотерапии.
Ключевые слова: рак шейки матки, адаптивный клеточный иммунитет, Т- лимфоциты, хелперы/ин-дукторы, цитотоксические лимфоциты
ABSTRACT
We study of T-cell imbalance in the process of accompanying immunotherapy in patients with cervical cancer. Material and methods of investigation. In the study included patients with cervical cancer T2-3N0-1M0 stages (II-III clinical stages). Results. the analysis of the obtained results allowed to reveal pronounced changes in the cell link of immunity, which are manifested by suppression of the expression of CD3 +, CD3 + CD4 +, IRI, increased expression of CD3 + CD8 + and CD16 +. As can be seen, the best situation is typical for patients with cervical cancer after EIPHT + PPh, where activation of T-cell immunity is observed. There is a marked imbalance in cellular immunity. Moreover, the imbalance in the cell link of immunity is expressed in the suppression of IRI by reducing the number of T-helpers / inducers and increasing T-cytotoxic lymphocytes. Obviously, with this pathology T - the cellular immune response is substantially weak and directed against a smaller number of epitopes, which suggests that clonal depletion of T lymphocytes is possible. In turn, the decreased immunoreactivity of the T-cell link can be considered as a result of a disruption in the representation of the antigen to the cells of the immune system, as well as a violation of the function of the T cells themselves. Conclusions. Therefore, T-cell lymphopenia revealed is often characteristic of the background or after PCT. Suppression of IRI indicates the presence of T-cell immunodeficiency, as mentioned above, mainly due to a decrease in the number of T-helpers / inducers, which play an important role in the implementation of the immune response. Increase in T-cytotoxic lymphocytes, which indicates the suppression of T-cell immunity and the presence of cytotoxic action at the cellular level. Positive clinical efficacy of the combination of immunotherapy has been established.
Key words: cervical cancer, adaptive cellular immunity, T-lymphocytes, helpers / inducers, cytotoxic lymphocytes.
Topicality. It is well known that immunosuppression of cellular factors of adaptive immunity, which play an important role in the formation and progression of malignant processes, is often the main causes of ineffectiveness of chemotherapy and long-term results of treatment of cervical cancer (cervical cancer) patients [4, -P. 1064;
12,-P. 178; 13, -P.203]. Until now, studies are underway to improve the results of traditional therapy in oncology and to introduce immunological methods of treatment. The results of such studies suggest that the evaluation of the immune profile of the tumor can make a difference in the conditions of personalized medicine. It is known that the therapeutic approaches of antitumor immunotherapy are based on the stimulation of antitumor immunity as a result of action on the nonspecific or adaptive effector link of the immune system. It is believed that the immune system recognizes the tumor process, forming specific antibodies and a pool of specific cytotoxic immunocompetent cells, which is an important condition for activation and implementation of antitumor immunity [1, -P.120;4, -P. 1064; 6,7, -P.925;
13, -P.203]. However, up to now, data on the effectiveness of immunotherapy as an accompanying immunotherapy for cervical cancer have not been adequately described or described in the literature. It should be noted that dysfunctions of the cellular part of the immune system, in particular, a violation of the effector function of T-lymphocytes and the balance of "pro-tumor", "antitumor" and "regulatory" mediators, were revealed in cancer patients [5,11,15]. At the same time, the severity and mechanisms of the development of immunodepression inherent in any oncological disease are different at different stages of tumor progression [8, -P.18538; 9,-P. 18538]. In this connection, the studies conducted in the field of studying various variants of immunotherapy in cervical cancer are an important and topical direction used in medicine, and leaves much hope [9,-P. 18538]. Approaches to its implementation, the described methods, the timing of implementation, the possibility of combining with other methods of conservative and surgical treatment remain insufficiently studied and developed. As shown in the literature, mechanisms of immune imbalance that affect the effectiveness of therapy and predict the course of the disease are not sufficiently disclosed. When evaluating the results of immunotherapy, one should take into account their influence on the key cellular mechanisms of antitumor immunity. It is important to note that great achievements in the field of molecular genetic studies stimulated a broad study of the possibilities of immunotherapeutic methods for the treatment of cancer patients. In the literature it is shown that the use of immunotherapy is aimed at the induction of both innate and adaptive immunity of the organism for the realization of antitumor activity.
The aim of the study was to study the T-cell imbalance in the process of accompanying immunotherapy in patients with cervical cancer.
Material and methods of the research. In the study included patients with cervical cancer T2-3N0-1M0 stages (II-III clinical stages), who were in the departments of oncogynecology and chemotherapy of RSSPMCOR MH RUz. The cervical cancer patients were randomized to the following groups: the 1 group - 38 practically healthy individuals; the 2 group - 38 patients with cervical cancer before treatment; the 3 group - 44 patients with cervical cancer who received immunotherapy in the form of extracorporeal immunopharmacotherapy (EIPHT); the 4 group - 42 patients with cervical cancer who received immunotherapy in the form of extracorporeal immunopharmacotherapy and plasmapheresis (EIPHT + PPh); the 5 group (control) - 54 patients with cervical cancer without immunotherapy.
Immunotherapy was performed in the hospital, when patients were admitted to chemotherapy. In total, patients received 2 EIPHT sessions at the beginning of admission to hospital and before discharge from the hospital. Polyoxidonium (azoxime bromide) was used as an immunostimulant preparation. Polyoxidonium has a complex action: immunomodulating, detoxifying, antioxidant, moderate anti-inflammatory. Manufacturer - NPO PETROVAKS PHARM, LTD (Russia). Immunological studies included the study of cellular and humoral parameters of the immune system in patients with ovarian cancer. Determination of cellular immunity (CD3 +, CD3 + CD4 +, CD3 + CD8 +, CD16 +, CD20 +), and identification of activation markers of lymphocytes (CD38 + and CD95 +) was performed by flow cytometry on Accuri C6 (USA) using MCAT. During the statistical analysis of the data presented in the work, the results of the research were entered into databases prepared in Microsoft Excel XP. Numerical (continuous) values were presented as mean arithmetic mean values and mean error (M ± m). A comparison of the quantitative traits was carried out with the help of the Student's test, for continuous variables - the paired Student test. As a boundary comparative criterion for the statistical significance of reliability, p <0.05 was assumed.
The results and discussion. It is known that to the main factors of immunity, which have been widely studied to this day, is T-cell immunity [6]. The study of the state of T-cell immunity in patients with cervical cancer is an important factor in determining the depth of immunodeficiency, predicting the disease and, most importantly, identifying the most radical ways of therapy, including immunotropic therapy. Despite a significant deepening in the last decade of ideas in the etiology, immunopathogenesis, progression and progression of malignant processes, many questions concerning the mechanisms of development of the pathological process and its course, assessing the effectiveness of treatment remain open. Based on the foregoing, we performed an evaluation of T-cell immunity in female cervical cancer, depending on the use of different approaches to immunotherapy in the scheme of accompanying therapy under stationary conditions. It is known that phenotypic markers of T-lymphocytes include such markers as CD3 +, CD3 +
CD4 +, CD3 + CD8 +. In the literature, the information that the activation and regulation of the effectiveness of the immune response is largely determined by the specific antigen of T lymphocytes is widely reported. Responsible for this function are antigen-recognizing receptors - ARR. It is known that the degree of surface expression of CD3 + receptors on the T-lymphocyte membrane reflects its transmissive function and allows the total number of T-lymphocytes to be identified [3.-P.942]. Thus, the analysis of the expression of CD3 + T-lymphocytes in patients with cervical cancer, depending on the type of immunotherapy, showed that the presence of significant suppression of CD3 + expression on T-lymphocytes is observed in all groups of patients with cervical cancer, compared with the control group, and there are significant differences between groups of patients p <0.05). It was shown that the lowest value of CD3 + expression was observed in the group of cervical cancer patients after the application of PCTs without immunotherapy. Moreover, reliable suppression of CD3 + expression in the group of patients after PCT without immunotherapy is observed in comparison with the values of patients with cervical cancer in groups where EIPHT and EIPHT + PPh were used. Consequently, in the group of patients after PCT without immunotherapy, a decrease in CD3 + expression was noted, which is most likely a toxic and depressive effect of PCT on factors of cellular immunity. Obviously, a decrease in the total pool of T-lymphocytes (CD3 +) was noted by suppressing the expression of CD3 + CD4 + T-helper / inducers. The study of expression of CD3 + CD4 + on T-lymphocytes, which are the main regulatory cells of immunity, showed that the lowest value of T-helpers / inducers was observed in groups of patients with cervical cancer without immunotherapy and before the initiation of therapy (p <0.05). It has been shown in the literature that CD4 + T-cell response to tumor proteins is an important cellular mechanism for protecting the macroorganism, since CD4 + T-helpers stimulate the production of antibodies by B lymphocytes and activate CD8 + T lymphocytes specific for tumor cells [4,-P. 1064; 8, -P.18538;9, -P.18538]. The analysis showed that in the group of patients with cervical cancer without immunotherapy, the expression of CD3 + CD4 + was 22.6 ± 1.2%, while in the groups of patients after EIPHT - 26.9 ± 1.4%, after EIPHT + PPh - 28.2 ± 0,9%, and in the group of patients before the start of therapy -20,7 ± 1,1%, in the group of practically healthy persons - 36,8 ± 1,2%. It is known that cytotoxic CD3 + CD8 + T-lymphocytes play an important role in the pathogenesis of cancer [2,-P. 227; 3, -P.942, 7,-P. 925; 10,-P.91]. It is proved that the function of these cells is the recognition of antigens on the cell surface in a complex with MHC molecules of class 1. Since they are present on almost all nuclear cells of the body, any cell carrying MHC class 1 molecules in combination with an antigenic peptide can activate a clone of cytotoxic T lymphocytes. The biological role of this activation is the removal of mutant cells. Analysis of the expression of CD3 + CD8 + on T-lymphocytes revealed a significant increase in all groups of patients with cervical cancer compared with the value of a group of
practically healthy individuals. It should be noted that the maximum increase in CD3 + CD8 + was detected in the group of patients before and after PCT therapy without immunotherapy (p <0.05). When analyzing the expression of CD3 + CD8 + on T-lymphocytes between the investigated groups of cervical cancer patients, it was seen that before the treatment the expression of CD3 + CD8 + was significantly increased and amounted to 35.7 ± 2.2%, in the group of patients after PCT without immunotherapy was on average 32,6 ± 1,2%, and in the groups of patients after the application of EIPHT and EIPHT + PPh a significant decrease in the number of cytotoxic T-lymphocytes and an approximation to the values of the regulatory group is observed, which indicates a decrease in immunosuppression against the background of the use of various immunotherapy variants using polyoxidonium Which is a drug detoxification and has pronounced immunotropic properties.
Immunoregulatory index (IRI), which is the ratio of CD3 + CD4 + / CD3 + CD8 + values, is of significant importance in secondary immunodeficiency states. It is known that, in healthy IRI, an average of 1.52 ± 0.02. Obviously, suppression of CD3 + CD4 + expression against the background of increased expression of CD3 + CD8 + leads to a decrease in IRI. According to our data, the least decrease in IRI is observed in the group of patients before and after treatment without the use of immunotherapy. It was noted that in the group of patients with cervical cancer who underwent EIPHT after PCT too, there was a decreased IRI in comparison with the data of patients receiving EIPHT + PPh. Thus, the lowest value of IRI in the group of patients after PCT without immunotherapy was 0.68 ± 0.04, and the highest value was noted in the group of patients after EIPHT + PPh and was 1.38 ± 0.05 (p <0.05 ). Consequently, pronounced immunosuppression was characteristic of patients with cervical cancer in the groups of patients prior to treatment and after PCT without immunotherapy. Obviously, a decrease in IRI is an important criterion for the depth of the T-cell immunodeficiency state, especially when assessing the effectiveness of treatment for cervical cancer. Further expression of CD16 + on killer cells was studied, which is the third population of lymphocytes providing maintenance of genetic homeostasis, which phenotypically and functionally differ significantly from T and B lymphocytes. Killer lymphocytes are classified as the main effectors of natural or innate immunity, which are capable of lysing target cells or carrying out antibody-dependent cellular cytotoxicity. It is their inherent performance of the functions of the first line of defense before the emergence of immune T-lymphocytes and specific antibodies. We have studied killer cells with the phenotypes CD16 +. A significant increase in CD16 + expression was revealed in all groups of patients with cervical cancer. It was shown that the greatest expression of CD16 + was observed in the group of patients with cervical cancer before and after PCT without immunotherapy, which was significantly increased in comparison with other groups of patients (p <0.05). Thus, in the group of patients before treatment, expression of CD16 + was
Евразийский Союз Ученых (ЕСУ) # 4 (49), 2018
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26.1 ± 1.2%, in the group of patients after PCT without immunotherapy - 22.8 ± 1.19%, in the group of patients after EIPHT - 21.2 ± 0.98% , in the group after EIPHT + PPh - 18.1 ± 1.1%, in the group of practically healthy persons - 16.8 ± 1.2%. Consequently, the greatest expression of CD16 + was observed in the groups of patients with cervical cancer before and after PCT without immunotherapy. As you can see, immunotherapy has a beneficial effect on the immune system, reducing its tension. Thus, the analysis of the results obtained revealed pronounced changes in the cell link of immunity, which are manifested by suppression of the expression of CD3 +, CD3 + CD4 +, IRI, increased expression of CD3 + CD8 + and CD16 +. As can be seen, the best situation is typical for patients with cervical cancer after EIPHT + PPh, where activation of T-cell immunity is observed. There is a marked imbalance in cellular immunity. Moreover, the imbalance in the cell link of immunity is expressed in the suppression of IRI by reducing the number of T-helpers / inducers and increasing T-cytotoxic lymphocytes. Obviously, with this pathology T - the cellular immune response is substantially weak and directed against a smaller number of epitopes, which suggests that clonal depletion of T lymphocytes is possible. In turn, the decreased immunoreactivity of the T-cell link can be considered as a result of a violation of the antigen presentation to immune system cells, as well as a violation of the function of the T-cells themselves. Thus, the detected T-cell lymphopenia is often characteristic of the background or after PCT. Suppression of IRI indicates the presence of T-cell immunodeficiency, as mentioned above, mainly due to a decrease in the number of T-helpers / inducers, which play an important role in the implementation of the immune response. Increase in T-cytotoxic lymphocytes, which indicates the suppression of T-cell immunity and the presence of cytotoxic action at the cellular level. Positive clinical efficacy of the combination of immunotherapy has been established.
References
1. Antoni P.A. CD4+CD25+ T regulatory cells, immunotherapy of cancer, and interleukin-2// J. Immu-nother.- 2005. -Vol.28.-P.120-128.
2. Chen L, Flies DB. Molecular mechanisms of T cell co-stimulation and co-inhibition. Nat Rev Immunol. 2013;13:227-42.
3. Curiel TJ, Coukos G, Zou L, Alvarez X, Cheng P, Mottram P, et al. Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nature medicine. 2004;10:942-9.
4. Gubser, P. M. et al. Rapid effector function of memory CD8+ T cells requires an immediate-early glycolytic switch. Nat. Immunol. 14, 1064-1072 (2013).
5. Hamanishi J, Mandai M, Iwasaki M, Oka-zaki T, Tanaka Y, Yamaguchi K, et al. Programmed cell death 1 ligand 1 and tumor-infiltrating CD8+ T lymphocytes are prognostic factors of human ovarian cancer. Proceedings of the National Academy of Sciences of the United States of America. 2007;104:3360-5.
6. Hassan M., Watari H., AbuAlmaaty A., Ohba Y., Sakuragi N. Apoptosis and molecular targeting therapy in cancer. Biomed Res Int 2014; 2014: 150845, http://dx.doi. org/10.1155/2014/150845.
7. Kandalaft L.E., Powell D.J. Jr., Singh N., Coukos G. Immunotherapy for ovarian cancer: what's next? J ClinOncol 2011;29:925-33.
8. Sato E., Olson S.H., Ahn J. et al. Intraepithe-lial CD8+ tumor-infiltrating lymphocytes and a high CD8+/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer. ProcNatlAcadSci USA 2005;102:18538-43.
9. Sato E, Olson SH, Ahn J, Bundy B, Nishi-kawa H, Qian F, et al. Intraepithelial CD8+ tumor-infiltrating lymphocytes and a high CD8+/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer. Proceedings of the National Academy of Sciences of the United States of America. 2005;102:18538-43.
10. Sierro, S., Romero, P. &Speiser, D. E. The CD4-like molecule LAG-3, biology and therapeutic applications. Expert Opin. Ther. Targets 15, 91-101 (2011).
THE EVALUATION OF PROVECTORAL CYTOKINS OF THE IMMUNE SYSTEM DEPENDING ON THE VARIANTS OF COHERENT IMMUNOTHERAPY IN PATIENTS WITH UTERINE CERVICAL CANCER. ОЦЕНКА ПРОВОСПАЛИТЕЛЬНЫХ ЦИТОКИНОВ ИММУННОЙ СИСТЕМЫ
В ЗАВИСИМОСТИ ОТ ВАРИАНТОВ СОПРОВОДИТЕЛЬНОЙ _ИММУНОТЕРАПИИ У БОЛЬНЫХ РАКОМ ШЕЙКИ МАТКИ._
Kamishov Sergey Viktorovich
MD, PhD, senior researcher, chemotherapeutist Republican Specialized Scientific and Practical Medical Center of Oncology and Radiology of the Ministry
of Health of the Republic of Uzbekistan, Tashkent.
Камышов Сергей Викторович, к.м.н., старший научный сотрудник отдела химиотерапии Республиканский специализированный научно-практический медицинский центр онкологии и радиологии Министерства здравоохранения Республики Узбекистан, Ташкент
