БЕЗОПАСНОСТЬ ИСПОЛЬЗОВАНИЯ РЕТРОВИРУСНЫХ ВЕКТОРОВ В ГЕННОЙ ТЕРАПИИ

Retroviral vectors are widely used in gene therapy and found to be an effective tool for the delivery of genetic constructs into cells. A unique feature of these vectors is the ability to incorporate therapeutic genes into a chromosome that ensures its passage to all progeny cells and enables to cure the diseases requiring genetic correction of dividing cells such as hematopoietic cells or skin cells. Retroviral vectors have been successfully used in gene therapy clinical trials for the treatment of 2 forms of severe combined immunodeficiencies and some other hereditary blood disorders. However, the integration of the vector into the chromosome was accompanied by genotoxicity and caused development of hematologic malignancies in several patients. Later it was shown that genotoxicity is not a general feature of retroviral vectors but it depends on many factors. In the present article we discuss safety issues concerning the use of different retroviral vectors in gene therapy. The description of modern vectors which designed to avoid the genotoxicity and other possible side effects are given. Ретровирусные векторы широко используют в генотерапевтических исследованиях, они признаны эффективным инструментом для доставки генетических конструкций в клетки. Уникальной особенностью данных векторов является способность эффективно встраивать терапевтические гены в хромосому, что обеспечивает передачу генов дочерним клеткам и позволяет лечить заболевания, требующие генетической коррекции активно делящихся клеток, таких как гемопоэтические клетки или клетки кожи. Ретровирусные векторы были успешно использованы в клинических испытаниях препаратов, направленных на лечение 2 форм тяжелых комбинированных иммунодефицитов и некоторых других врожденных заболеваний крови. Однако в нескольких случаях интеграция вектора в хромосому сопровождалась генотоксичностью и привела к развитию онкогематологического заболевания. В дальнейшем было показано, что генотоксичность не является неотъемлемой чертой всех ретровирусных векторов, а зависит от многих факторов. В настоящей статье рассматриваются вопросы безопасности применения различных ретровирусных векторов в генной терапии. Приведено описание современных векторов, обладающих свойствами, позволяющими избежать генотоксичности и других возможных побочных эффектов.

ПРИРОДНЫЙ РЕЗЕРВУАР ФИЛОВИРУСОВ И ТИПЫ СВЯЗАННЫХ С НИМИ ЭПИДЕМИЧЕСКИХ ВСПЫШЕК НА ТЕРРИТОРИИ АФРИКИ

Family Filoviridae includes a set of etiological agents of human hemorrhagic fevers distributed in Africa: Zaire ebolavirus (ZEBOV), Sudan ebolavirus (SUDV), Bundibugyo ebolavirus (BDBV), Taï Forest ebolavirus (TAFV), Marburg marburgvirus (MMARV). Historiography and recent taxonomical structure of Filoviridae family are considered in the review. The discussed data of laboratory and ecological-virological field researches demonstrate the presence of a natural reservoir of filoviruses among fruit-bats (Chiroptera, Megachiroptera) which carry filovirus infection without clinical signs but allocate viruses with urine, saliva, excrements, and sperm, as well as contain viruses in blood and internals. The potential hosts of filoviruses are various mammal species including the higher primacies (Anthropoidea) and the humans (Homo sapiens sapiens). A brief comparison of anatomic and morphologic features of fruit bats and bats (Chiroptera, Microchiroptera) belonging to another suborder of chiropterans is presented. The description of the basic characteristics of the four types of epidemic outbreaks linked with Filoviridae-associated fevers — speleological (from Ancient Greek σπήλαιον — cave), forest, rural, and urban are given; their possible transformation directions are considered as well.Семейство Filoviridae содержит ряд возбудителей геморрагических лихорадок человека, распространенных на территории Африки: эболавирус Заир (Zaire ebolavirus, ZEBOV), эболавирус Судан (Sudan ebolavirus, SUDV), эболавирус Бундибугё (Bundibugyo ebolavirus, BDBV), (Taï Forest ebolavirus, TAFV), марбургвирус Марбург (Marburg marburgvirus, MMARV). В обзоре представлены историография и рецентная таксономическая структура семейства Filoviridae; выполнен этимологический анализ устаревших и современных названий представителей этого семейства; обсуждаются данные лабораторных и полевых эколого-вирусологических исследований, свидетельствующих о том, что резервуаром филовирусов является подотряд Крыланов (Chiroptera, Megachiroptera), которые переносят филовирусную инфекцию инаппарантно, но выделяют вирусы с мочой, слюной, фекалиями и спермой, а также содержат вирусы в крови и внутренних органах. Потенциальными хозяевами филовирусов является широкий спектр видов млекопитающих, включая высших приматов (Anthropoidea) и человека (Homo sapiens sapiens). В работе приводится краткое сравнение анатомических и морфологических особенностей крыланов и летучих мышей (Chiroptera, Microchiroptera), входящих в другой подотряд отряда Рукокрылых. Дано описание основных характеристик четырех типов эпидемических вспышек филовирусных лихорадок — спелеологического (от др.-греч. σπήλαιον — пещера), лесного, деревенского и городского, а также возможные направления трансформации в процессе их развития и масштабирования

РАЙОНИРОВАНИЕ АФРИКАНСКОЙ ПРИРОДНО-ОЧАГОВОЙ ПРОВИНЦИИ В ОТНОШЕНИИ ФИЛОВИРУСНЫХ ЛИХОРАДОК

The review presents the following division of the African natural foci province into districts: I. Upper Guinea natural focus region includes the following individual natural foci: I.1. Kazamans; I.2. North Guinea; I.3. Volta; I.4. Adamawa; I.5. Sao Tome. II. Central Africa: II.1. Southern Guinea; II.2. Katanga; II.3. Congo; II.4. Azande; II.5. Ruwenzori. III. South-Eastern Africa: III.1. Mafungabusi; III.2. Drakensberg; III.3. Comoros; III.4. Madagascar; III.5. Seychelles; III.6. Mascarenes. For each natural focus the article describes the characteristic landscapes and species list of fruit-bats (Chiroptera, Megachiroptera) which are natural reservoir for viruses belonging to Filoviridae family, the types of epidemic outbreaks and the intensity of filovirus circulation. Possible explanations of narrowness of filoviruses areal in comparison with fruit-bats areal in Africa are discussed.В обзоре предложено районирование африканской природно-очаговой провинции, в составе которой выделены: I. Верхнегвинейский природно-очаговый район, включающий следующие природные очаги: I.1. Казамaнс; I.2. Северо-Гвинейский; I.3. Вόльта; I.4. Адамaва; I.5. Сан-Томe. II. Центральноафриканский: II.1. Южно-Гвинейский; II.2. Катaнга; II.3. Кόнго; II.4. Азaнде; II.5. Рувензόри. III. Юго-Восточноафриканский: III.1. Мафунгабус́ и; III.2. Дракόновы горы; III.3. Комόры; III.4. Мадагаскaр; III.5. Сейшeлы; III.6. Маскарeны.Для каждого природного очага описаны характерные ландшафты, видовой состав крыланов (Chiroptera, Megachiroptera), представляющих собой природный резервуар для вирусов семейства Filoviridae, а также типы эпидемических вспышек и интенсивность циркуляции филовирусов. Обсуждаются возможные объяснения узости ареала филовирусов по сравнению с ареалом крыланов в Африке

Current approaches and prospects for the development of laboratory diagnosis of measles

Measles virus causes an acute infectious disease with high contagiousness. It is possible to limit the spread of measles virus only with a sufficiently wide coverage of the population by vaccination. Despite the success of measles elimination programs, many countries have seen an increase in the incidence of measles in recent years, making early diagnosis increasingly important. The importance of laboratory diagnosis is related to the difficulties of clinical differential diagnosis of measles in the early stages of the disease. This review is devoted to an analysis of existing methods for diagnosing measles. It demonstrates the limitations of the most commonly used method, the enzyme immunoassay, and the need to develop and implement alternative diagnostic methods. Particular attention in the review is paid to molecular diagnostic methods, the sensitivity of which is reviewed for different types of biological sampled at different stages of the disease. Characteristics of the measles virus that are of key importance in the development of PCR tests are described. Studies evaluating the significance of introducing PCR in the routine diagnosis of measles are presented. The main advantages of molecular methods are the possibility of early detection of the virus and the possibility of simultaneous detection of several pathogens, which allows differential diagnosis of diseases with a similar clinical presentation. The development and implementation of rapid and accurate approaches based on molecular diagnostic methods into the health care system is an urgent need in the implementation of global and local programs for the elimination of measles

The reproducibility of qualitative measurement of RNA HIV-1, RNA hcv and RNA hbv using amplisens reagents' kits

The evaluation of reproducibility of three reagents' kits AmpliSens for quantitative measurement of RNA HIV-1, RNA HCV and RNA HBV using retrospective analysis of protocols of output control of reagents' kits produced by the central research institute of epidemiology of Rospotrebnadzor in 2014-2016. For every reagents' kits more than 100 produced series are analyzed. The total number of tested samples for every reagents' kits was at least 590. For every sample a difference between passport and experimental value of decimal algorithm of concentration of RNA/DNA of virus (dLg) was calculated. The obtained values of dLg were used to determine median and standard deviation (SD). The obtained values of SD were accepted as a reproducibility. The calculations were implemented using Microsoft Excel 2010. The average reproducibility according samples with various concentration of RNA HIV-1 made up to 0,218 Lg, RNA HCV - 0,191 Lg and I'M HBV- 0,174 Lg. The least reproducibility of measurement was observed for concentrations corresponding to lower limit of linear range of qualitative measurement and it made up to for RNA HIV1 0,275 Lg. RNA HCV - 0,206 Lg and RNA HBV- 0,211 Lg. The obtained reproducibility of the given reagents' kits AmpliSens is comparable with reproducibility of reagents' kits of foreign manufacturers. There is no necessity to enhance achieved reproducibility because, taking into account actual recommendations on treatment of patients with HIV and viral hepatitis B and C, according to them a therapy is considered effective when viral load decreases up to 10-100 times and even up to non-detectable levels

The reproducibility of qualitative measurement of RNA HIV-1, RNA hcv and RNA hbv using amplisens reagents' kits

The evaluation of reproducibility of three reagents' kits AmpliSens for quantitative measurement of RNA HIV-1, RNA HCV and RNA HBV using retrospective analysis of protocols of output control of reagents' kits produced by the central research institute of epidemiology of Rospotrebnadzor in 2014-2016. For every reagents' kits more than 100 produced series are analyzed. The total number of tested samples for every reagents' kits was at least 590. For every sample a difference between passport and experimental value of decimal algorithm of concentration of RNA/DNA of virus (dLg) was calculated. The obtained values of dLg were used to determine median and standard deviation (SD). The obtained values of SD were accepted as a reproducibility. The calculations were implemented using Microsoft Excel 2010. The average reproducibility according samples with various concentration of RNA HIV-1 made up to 0,218 Lg, RNA HCV - 0,191 Lg and I'M HBV- 0,174 Lg. The least reproducibility of measurement was observed for concentrations corresponding to lower limit of linear range of qualitative measurement and it made up to for RNA HIV1 0,275 Lg. RNA HCV - 0,206 Lg and RNA HBV- 0,211 Lg. The obtained reproducibility of the given reagents' kits AmpliSens is comparable with reproducibility of reagents' kits of foreign manufacturers. There is no necessity to enhance achieved reproducibility because, taking into account actual recommendations on treatment of patients with HIV and viral hepatitis B and C, according to them a therapy is considered effective when viral load decreases up to 10-100 times and even up to non-detectable levels

Decline of antibodies to hepatitis c virus during antiviral therapy

The goal of this study was to investigate interrelationship between changes in anti-hepatitis C virus antibody and response to antiviral therapy. The comparative quantitative analysis of antibodies to individual structural and nonstructural viral proteins was done during two years in three patient groups: initial responders, non-responders and a reference group (without therapy). All patients in the treated groups exhibited decrease in the analyzed antibodies to HCV proteins, but with different patterns. The first statistically significant difference in the decrease of virus-specific antibody between initial responders and non-responders was observed within the first three months of therapy beginning. Some treated patients demonstrated decrease in antibody levels to HCV proteins after the end of therapy

The decline in antibodies to hepatitis C virus during antiviral therapy

The goal of the study was to analyze B-cell response to hepatitis C virus during antiviral therapy among responders and non-responders. The content of antibodies to individual structural and non-structural HCV proteins was investigated during two years in three groups of patients: initial responders, non-responders and a reference group (without therapy). Treated patients in all groups exhibited the decrease in antibodies to analyzed HCV proteins, but with different patterns. The first statistically significant differences in the decline of the virus-specific antibodies between initial responders and non-responders were observed within the first three months after the beginning of therapy. Some treated patients demonstrated the decrease in antibody levels to HCV proteins after the end of therapy. © Pleiades Publishing, Ltd. 2009