Исследование протективных свойств рекомбинантного комплекса белка F наружной мембраны и анатоксина Pseudomonas aeruginosa

Pseudomonas aeruginosa induces the complications after burns, injuries, surgical interventions and appears to be one of the main causative agents of nosocomial infections. This pathogen has the high resistance to the antibacterial preparations, therefore the immunoprophylaxis is considered as one of the major approaches to reduce Pseudomonas infection. Objective: The aim of our investigation is to study the protective properties of the recombinant complex of the outer membrane protein F (OprF) and a non-toxic variant of the exotoxin A (toxoid) against Pseudomonas infection. Methods: The recombinant proteins which contained the additional histidine residues were synthesized into Escherichia coli with isopropyl-βD-thyogalactopyranoside (IPTG). The recombinant proteins were purified by affinity chromatography on Ni-Sepharose. The preparations of recombinant proteins were injected intraperitoneally into the mice. Aluminum hydroxide was used as an adjuvant. For an experimental infection in mice, animals were challenged intraperitoneally by a live virulent culture of P. aeruginosa (PA-103 strain). Results: The best protective effect for the complex containing 25 μg OprF and 50 μg toxoid was identified when we used the double immunization of mice (Index of efficiency of the protective properties in this case was 4.0). Indexes of efficiency of separated recombinant proteins which were injected twice in the same doses were 2.0 for OprF и 2.3 for toxoid. The triple immunization of animals was inefficient for separated recombinant proteins in the same doses. The injection of doses which were lowered twice (12.5 μg for OprF and 25 μg for toxoid) resulted in increased survival of mice immunized by individual proteins (indexes of efficiency: 3 for OprF and и 3,5 for toxoid). However when we administered to the complex of proteins with the same doses Index of efficiency was 2.8. Conclusion: It was shown that the maximum protective effect in a short time is achieved by the combination of double immunization and the mixture of the recombinant proteins OprF and the 25 and 50 μg doses of recombinant toxoid . Pseudomonas aeruginosa вызывает осложнения после ожогов, травм и хирургических вмешательств, являясь одной из основных причин нозокомиальных инфекций. Этот патоген обладает высокой резистентностью к большинству антибактериальных средств, поэтому иммунопрофилактика рассматриваться как один из приоритетных подходов для борьбы с синегнойной инфекцией. Цель исследования: Изучить защитные свойства рекомбинантного комплекса белка F наружной мембраны (OprF) и атоксической формы экзотоксина А (анатоксина) от экспериментальной синегнойной инфекции. Методы: Рекомбинантные белки, содержащие дополнительную шести гистидиновую последовательность, синтезировали в клетках Escherichia coli c помощью изопропил-b-d-тиогалактопиранозида (ИПТГ) и очищали методом аффинной хроматографии с использованием Ni-сефарозы. Препаратами рекомбинантных белков внутрибрюшинно иммунизировали мышей. В качестве адъюванта использовали гидроксид алюминия. Экспериментальное заражение осуществляли внутрибрюшинным введением живой вирулентной культуры P. aeruginosa штамма РА103. Результаты: При двукратной иммунизации мышей комплексом, содержащим 25 мкг OprF и 50 мкг анатоксина, выявлен наилучший протективный эффект (индекс эффективности защитных свойств в этом случае составил 4,0). Индексы эффективности для рекомбинантных белков, вводимых двукратно в тех же дозах по отдельности, соответствовали 2,0 для OprF и 2,3 для анатоксина и не увеличивались после трехкратной иммунизации животных. Уменьшенные в два раза доз (12,5 мкг OprF и 25 мкг анатоксина) при трехкратном введении способствовало увеличению выживаемости мышей иммунизированных отдельными белками (индексы эффективности: 3 для OprF и 3,5 для анатоксина), однако при введении комплекса белков в тех же дозах индекс эффективности не увеличивался и составил 2,8. Заключение: Показано, что максимальный протективный эффект формируется в короткие сроки при использовании комплексной иммунизации двумя рекомбинантными белками OprF и анатоксином в дозах 25 и 50 мкг

Management of adherence-based treatment. Consensus document – Clinical guidelines. English version [упРАВЛЕНИЕ ЛЕЧЕНИЕМ НА ОСНОВЕ пРИВЕРЖЕННОСТИ. КОНСЕНСуСНЫй ДОКуМЕНТ – КЛИНИЧЕСКИЕ РЕКОМЕНДАЦИИ. АНГЛОЯзЫЧНАЯ ВЕРСИЯ]

Consensus Document «Management of adherence-based treatment» developed based on clinical guidelines of Russian Scientific Medical Society of Internal Medicine approved by the XIV National Congress of physicians (Moscow, 20 November 2019). The document is intended primarily to assess adherence to drug therapy, adherence to lifestyle modification, and adherence to medical support for patients who need long-term or permanent treatment. The document’s authors considered the treatment adherence unanimously as the compliance of the patient’s behavior with the recommendations received from the Doctor regarding medication, dieting, and other measures of lifestyle modifications. Insufficient adherence to treatment is a global problem. Assessing adherence as a basis for making medical decisions is an essential element of improving the quality of the healthcare system. Predictions of treatment outcomes cannot be considered effective if individualized levels of adherence are not used to justify project planning and evaluation. In medical practice, quantitative assessment of adherence is preferred, suitable for patients with various diseases, and with the possibility of automated data entry and processing. Therefore, sections on medical interventions based on the assessment of treatment adherence should be reflected in national clinical guidelines. These Joint Recommendations are based on these provisions. © Group of authors, 202

Treatment management based on adherence: Patient recommendation algorithms. Cross-disciplinary guidelines [УПРАВЛЕНИЕ ЛЕЧЕНИЕМ НА ОСНОВЕ ПРИВЕРЖЕННОСТИ: АЛГОРИТМЫ РЕКОМЕНДАЦИЙ ДЛЯ ПАЦИЕНТОВ. МЕЖДИСЦИПЛИНАРНЫЕ РЕКОМЕНДАЦИИ]

The algorithms (sample templates) of recommendations for patients, which are an integral part of adherence-based treatment management technologies, were developed by an interdisciplinary Working Group coordinated by the Treatment Adherence Section of the Russian Scientific Medical Society of Physicians. The Working Group includes opinion leaders and experts in the therapeutic, gastroenterology, cardiology, preventive medicine, gerontology, oncology, hematology, immunology, phthisiopulmonology, infectious diseases, oncohematology, neurology, intensive care, rehabilitation and other specialties, both involved by the section of RSMSIM, and recommended by the Gastroenterological Scientific Society of Russia, National Association of experts in Thrombosis, Clinical Hemostasiology and Hemorheology, Gerontological Society of the Russian Academy of Sciences, Russian Oncohematologists Society, Russian Society of Phthisiatrists, Society of Evidence-based Neurology. The algorithms are based on the materials of «The First Russian Consensus on Quantitative Assessment of adherence to treatment», approved by the XII National Congress of Internal Medicine (2017) and Clinical Guidelines «Treatment Management on the Basis of adherence», approved by the XIV National Congress of Internal Medicine (2019), taking into account the Consenting Document - Interdisciplinary Guidelines of RSMSIM, GSSR, NATH, SEN, GSRAS, ROHS, RSP «Treatment Management on the Basis of Adherence» (2020). The algorithms of treatment strategies are recommended by the Working Group to be used as a basis for inclusion into relevant sections of clinical guidelines and clinical handbooks developed by professional medical communities, as well as for use by practitioners in medical practice. © 2020 Stavropol State Medical University. All rights reserved