scbmtБИОМЕДИЦИНАJournal Biomed2074-59822713-0428Scientific center of biomedical technologies of Federal Medical and Biological Agency10.33647/2074-5982-20-2-32-44scbmt-1582Research ArticleТРАНСГЕНОЗ И ГУМАНИЗИРОВАННЫЕ ЖИВОТНЫЕTRANGENOSIS AND HUMANIZED ANIMALSДоказательство наличия целевых белков — β2m hom и HLA у гуманизированных трансгенных мышей линий HLA-A*02:01, HLA-B*07:02 и HLA-C*07:02Evidence for the Presence of β2m hom Target Proteins and HLA in Humanized Transgenic Mice of HLA-A*02:01, HLA-B*07:02, and HLA-C*07:02 LinesКаркищенкоВ. Н.KarkischenkoV. N.

Каркищенко Владислав Николаевич, д.м.н., проф.

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Vladislav N. Karkischenko,  Dr. Sci. (Med.), Prof.

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

cbmt@yandex.ruБерзинаА. Г.BerzinaA. G.

Берзина Ася Григорьевна, к.б.н.,

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Asya G. Berzina, Cand.  Sci.  (Biol.)

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

berzina07@mail.ruПетроваН. В.PetrovaN. V.

Петрова Наталья Владимировна

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Nataliya V.  Petrova

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

m-sklad@yandex.ruПомыткинИ. А.PomytkinI. A.

Помыткин Игорь Анатольевич, к.х.н.

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Igor A. Pomytkin, Cand. Sci. (Chem.)

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

ipomytkin@mail.ruГлотоваЕ. С.GlotovaE. S.

Глотова Елена Сергеевна

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Elena S. Glotova

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

savelaine@gmail.comПетровД. В.PetrovD. V.

Петров Дмитрий Валерьевич

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Dmitry V. Petrov

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

1941-65@mail.ruТабояковаЛ. А.TaboyakovaL. A.

Табоякова Лидия Александровна

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Lidiya A. Taboyakova

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

lida-vet@mail.ruБолотскихЛ. А.BolotskihL. A.

Болотских Любовь Александровна,  к.с.-х.н.

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Lubov’  A.  Bolotskih

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

lyuba.bolotskikh@mail.ruЛарюшинаН. А.LaryushinaN. А.

Ларюшина Надежда Андреевна

143442, Московская обл., Красногорский р-н, п. Светлые горы, 1

Nadezhda A. Laryushina

143442, Moscow Region, Krasnogorsk District, Svetlye Gory V illage, 1

kichi09@mail.ruФГБУН «Научный центр биомедицинских технологий ФМБА России»РоссияScientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of RussiaRussian Federation2024230720242023244Copyright © Каркищенко В.Н., Берзина А.Г., Петрова Н.В., Помыткин И.А., Глотова Е.С., Петров Д.В., Табоякова Л.А., Болотских Л.А., Ларюшина Н.А., 20242024Каркищенко В.Н., Берзина А.Г., Петрова Н.В., Помыткин И.А., Глотова Е.С., Петров Д.В., Табоякова Л.А., Болотских Л.А., Ларюшина Н.А.Karkischenko V.N., Berzina A.G., Petrova N.V., Pomytkin I.A., Glotova E.S., Petrov D.V., Taboyakova L.A., Bolotskih L.A., Laryushina N.А.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.scbmt.ru/jour/article/view/1582

Человеческий лейкоцитарный антиген играет первостепенное значение в формировании иммунного ответа и патогенезе заболеваний различной этиологии, в т.ч. при развитии негативных побочных эффектов на фармакологические препараты. Современные стандарты фармакобезопасности требуют совершенствования существующих тест-систем для проведения качественных доклинических исследований. В НЦБМТ ФМБА России был разработан и создан ряд гуманизированных трансгенных линий мышей, несущих гибридные молекулы HLA I класса на поверхности клеток, которые соответствуют аллельным вариантам человека HLA-A*02:01, HLA-B*07:02 и HLA-C*07:02. В данной статье представлены экспериментальные данные по количественному определению белка β2-микроглобулина человека и результаты определения HLA «сэндвич»-методом ИФА у мышей, несущих разные аллели генов HLA I класса. Полученные данные подтверждают наличие целевых функциональных белков (трансгенность) у гуманизированных трансгенных мышей, что согласуется с данными, полученными нами при определении первичной последовательности трансгена методом секвенирования по Сэнгеру. Кроме того, в работе рассматривается научно-практическая значимость таких биомоделей и область их применения.

Human leukocyte antigen plays a primary role in the formation of immune response and pathogenesis of   diseases of various etiologies, including the development of negative side effects induced by pharmacological agents. Modern pharmacosafety standards require improvement of existing test systems to conduct high-quality preclinical studies. A number of humanized transgenic mouse lines with hybrid HLA I class molecules on the cell surface, which correspond to the human allelic variants HLA-A*02:01, HLA-B*07:02, and  HLA-C*07:02, were developed at the Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of Russia. In this article, we present experimental data on quantitative determination of β2-microglobulin protein and HLA by the “sandwich” ELISA method in mice with different alleles of   HLA I class genes. The results obtained confirm the presence of target functional proteins (transgenicity) in humanized transgenic mice, which is consistent with our previous data obtained when determining the primary sequence of the transgene using Sanger sequencing. We also discuss the scientific and practical significance of such biomodels, as well as the scope of their application.

иммунопероксидазный конъюгатИФА«сэндвич»-методгуманизированные трансгенные мышиβ2-микроглобулинHLA A-B-C-immunoperoxidase conjugateELISAsandwich methodhumanized transgenic miceβ2-microglobulinHLA A-B-C-государственное задание по теме «Получение новой линии гуманизированных трансгенных мышей, несущих ген HLA-A*pX человека и нокаут комплекса H-2K мыши» (шифр: «Транснокаут-2024») ФГБУН НЦБМТ ФМБА России.the research topic “Generation of a new line of humanized transgenic mice carrying the human HLA-A*pX gene and knockout of the mouse H-2K complex” (code: “Transknockout-2024”) of state assignment of Scientific Center of Biomedical Technologies of FMBA of Russia.ReferencesКаркищенко В.Н., Болотских Л.А., Капанадзе Г.Д., Каркищенко Н.Н., Колоскова Е.М., Максименко С.В., Матвеенко Е.Л., Петрова Н.В., Рябых В.П., Ревякин А.О., Станкова Н.В., Семёнов Х.Х. Создание линий трансгенных животных-моделей с генами человека NAT1 и NAT2. 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The authors declare that there are no conflicts of interest present.