title: ÐÑÐ¾ÑÐ¸Ð²Ð¾Ð²Ð¸ÑÑÑÐ½Ð°Ñ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð¼Ð ÐÐ, ÐºÐ¾Ð´Ð¸ÑÑÑÑÐ¸Ñ Fab-ÑÑÐ°Ð³Ð¼ÐµÐ½ÑÑ Ð°Ð½ÑÐ¸ÑÐµÐ» Ðº Ð½ÑÐºÐ»ÐµÐ¾Ð¿ÑÐ¾ÑÐµÐ¸Ð½Ñ Ð²Ð¸ÑÑÑÐ° Ð³ÑÐ¸Ð¿Ð¿Ð° Ð, in vitro
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Antiviral activity of mRNAs encoding Fab-fragments of antibodies to nucleoprotein of influenza B virus, in vitro
Creators: name: ÐÐµÐ¼Ð°ÐµÐ², ÐÐ»ÐµÐºÑÐµÐ¹
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2024
Subjects: Ð²Ð¸ÑÑÑ Ð³ÑÐ¸Ð¿Ð¿Ð°
Ð½ÑÐºÐ»ÐµÐ¾Ð¿ÑÐ¾ÑÐµÐ¸Ð½
Ð¼Ð ÐÐ
Ð¼Ð ÐÐ ÑÐµÑÐ°Ð¿Ð¸Ñ
Ð»Ð¸Ð¿Ð¾ÑÐ¾Ð¼Ñ
ÑÑÐ°Ð³Ð¼ÐµÐ½ÑÑ Ð°Ð½ÑÐ¸ÑÐµÐ»
influenza virus
nucleoprotein
mRNA
mRNA therapeutics
liposomes
antibody fragments

description: ÐÐ¾Ð¸ÑÐº Ð¿ÑÐ¾ÑÐ¸Ð²Ð¾Ð²Ð¸ÑÑÑÐ½ÑÑ Ð¿ÑÐµÐ¿Ð°ÑÐ°ÑÐ¾Ð², Ð°ÐºÑÐ¸Ð²Ð½ÑÑ Ð² Ð¾ÑÐ½Ð¾ÑÐµÐ½Ð¸Ð¸ ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÑ ÑÑÐ°Ð¼Ð¼Ð¾Ð² Ð²Ð¸ÑÑÑÐ° Ð³ÑÐ¸Ð¿Ð¿Ð°, ÑÐ²Ð»ÑÐµÑÑÑ ÑÑÐµÐ·Ð²ÑÑÐ°Ð¹Ð½Ð¾ Ð°ÐºÑÑÐ°Ð»ÑÐ½Ð¾Ð¹ Ð·Ð°Ð´Ð°ÑÐµÐ¹. Ð ÑÐ¾ Ð²ÑÐµÐ¼Ñ ÐºÐ°Ðº Ð¼Ð ÐÐ-Ð²Ð°ÐºÑÐ¸Ð½Ñ, Ð¾Ð´Ð¾Ð±ÑÐµÐ½Ð½ÑÐµ Ðº ÑÐºÑÑÑÐµÐ½Ð½Ð¾Ð¼Ñ Ð¿ÑÐ¸Ð¼ÐµÐ½ÐµÐ½Ð¸Ñ Ð² Ð¿ÐµÑÐ¸Ð¾Ð´ COVID-19, Ð¿ÑÐ¾Ð´ÐµÐ¼Ð¾Ð½ÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð»Ð¸ Ð±ÐµÐ·Ð¾Ð¿Ð°ÑÐ½Ð¾ÑÑÑ Ð¸ Ð²ÑÑÐ¾ÐºÑÑ ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ, ÑÐµÑÐ°Ð¿ÐµÐ²ÑÐ¸ÑÐµÑÐºÐ¸Ð¹ Ð¿Ð¾Ð´ÑÐ¾Ð´, Ð¾ÑÐ½Ð¾Ð²Ð°Ð½Ð½ÑÐ¹ Ð½Ð° ÑÐµÑÐ½Ð¾Ð»Ð¾Ð³Ð¸Ð¸ Ð¼Ð ÐÐ, Ð¿ÑÐµÐ´ÑÑÐ°Ð²Ð»ÑÐµÑ ÑÐ¾Ð±Ð¾Ð¹ Ð¼Ð½Ð¾Ð³Ð¾Ð¾Ð±ÐµÑÐ°ÑÑÑÑ Ð°Ð»ÑÑÐµÑÐ½Ð°ÑÐ¸Ð²Ð½ÑÑ ÑÑÑÐ°ÑÐµÐ³Ð¸Ñ. ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° in vitro Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð»ÐµÑÐµÐ±Ð½Ð¾-Ð¿ÑÐ¾ÑÐ¸Ð»Ð°ÐºÑÐ¸ÑÐµÑÐºÐ¾Ð³Ð¾ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»Ð° Ð¼Ð ÐÐ, ÐºÐ¾Ð´Ð¸ÑÑÑÑÐ¸Ñ Fab-ÑÑÐ°Ð³Ð¼ÐµÐ½ÑÑ (2/3-Fab) Ð½ÐµÐ½ÐµÐ¹ÑÑÐ°Ð»Ð¸Ð·ÑÑÑÐ¸Ñ Ð°Ð½ÑÐ¸ÑÐµÐ» Ðº Ð²Ð½ÑÑÑÐµÐ½Ð½ÐµÐ¼Ñ ÐºÐ¾Ð½ÑÐµÑÐ²Ð°ÑÐ¸Ð²Ð½Ð¾Ð¼Ñ Ð±ÐµÐ»ÐºÑ Ð²Ð¸ÑÑÑÐ° Ð³ÑÐ¸Ð¿Ð¿Ð° B â Ð½ÑÐºÐ»ÐµÐ¾Ð¿ÑÐ¾ÑÐµÐ¸Ð½Ñ. Ð ÑÐ¾Ð´Ðµ Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ Ð¾ÑÐ½Ð¾Ð²Ð½ÑÑ ÑÐ²Ð¾Ð¹ÑÑÐ² Ð²Ð¸ÑÑÑÐ° Ð³ÑÐ¸Ð¿Ð¿Ð° B/Austria/1359417/2021 Ð±ÑÐ» Ð¿Ð¾Ð»ÑÑÐµÐ½ ÐµÐ³Ð¾ ÑÐ¸ÑÑ Ð² ÐµÐ´Ð¸Ð½Ð¸ÑÐ°Ñ 50%-Ð½Ð¾Ð³Ð¾ ÑÐ¸ÑÐ¾Ð¿Ð°ÑÐ¸ÑÐµÑÐºÐ¾Ð³Ð¾ Ð´ÐµÐ¹ÑÑÐ²Ð¸Ñ Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ ÑÐµÐ°ÐºÑÐ¸Ð¸ Ð³ÐµÐ¼Ð°Ð³Ð³Ð»ÑÑÐ¸Ð½Ð°ÑÐ¸Ð¸ (Ð ÐÐ) Ð¸ Ð¸Ð¼Ð¼ÑÐ½Ð¾ÑÐµÑÐ¼ÐµÐ½ÑÐ½Ð¾Ð³Ð¾ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° (ÐÐ¤Ð). Ð¤Ð»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð°Ñ Ð¼Ð¸ÐºÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ñ Ð¸ Ð»ÑÑÐ¸ÑÐµÑÐ°Ð·Ð½ÑÐ¹ ÑÐµÑÑ Ð¿Ð¾Ð´ÑÐ²ÐµÑÐ´Ð¸Ð»Ð¸ ÑÐ¿Ð¾ÑÐ¾Ð±Ð½Ð¾ÑÑÑ Ð»Ð¸Ð¿Ð¾ÑÐ¾Ð¼ 2X3-DOPE ÑÑÐ¿ÐµÑÐ½Ð¾ ÑÑÐ°Ð½ÑÑÐ¸ÑÐ¸ÑÐ¾Ð²Ð°ÑÑ ÐºÐ»ÐµÑÐºÐ¸ MDCK Ð¸ Ð´Ð¾ÑÑÐ°Ð²Ð»ÑÑÑ Ð² Ð½Ð¸Ñ Ð¼Ð¾Ð´ÐµÐ»ÑÐ½ÑÐµ Ð¼Ð ÐÐ Ð·ÐµÐ»ÐµÐ½Ð¾Ð³Ð¾ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ° eGFP Ð¸ Ð¼Ð ÐÐ Ð»ÑÑÐ¸ÑÐµÑÐ°Ð·Ñ ÑÐ²ÐµÑÐ»ÑÑÐºÐ° (FLuc) ÑÐ¾Ð¾ÑÐ²ÐµÑÑÑÐ²ÐµÐ½Ð½Ð¾. Ð£ÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½ ÑÐ°ÐºÑ Ð²Ð½ÑÑÑÐ¸ÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¹ Ð½Ð°ÑÐ°Ð±Ð¾ÑÐºÐ¸ Ð°Ð½ÑÐ¸ÑÐµÐ» 2/3-Fab Ð¸ Ð¾Ð¿Ð¸ÑÐ°Ð½Ð° Ð´Ð¸Ð½Ð°Ð¼Ð¸ÐºÐ° Ð¸Ñ Ð½Ð°ÐºÐ¾Ð¿Ð»ÐµÐ½Ð¸Ñ. ÐÑÐ°ÑÐ°ÐºÑÐµÑÐ¸Ð·Ð¾Ð²Ð°Ð½Ð¾ Ð²Ð»Ð¸ÑÐ½Ð¸Ðµ ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑÐ¾Ð² Ð»Ð¸Ð¿Ð¾ÑÐ¾Ð¼ 2X3-DOPE Ð¸ Ð¼Ð ÐÐ (FLuc) Ð½Ð° Ð·Ð°ÑÐ°Ð¶ÐµÐ½Ð¸Ðµ ÐºÐ»ÐµÑÐ¾Ðº MDCK Ð²Ð¸ÑÑÑÐ¾Ð¼ Ð³ÑÐ¸Ð¿Ð¿Ð° B/Austria/1359417/2021. ÐÐ°Ðº ÑÐ»ÐµÐ´ÑÑÐ²Ð¸Ðµ, Ð¾Ð¿ÑÐ¸Ð¼Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°Ð½Ñ ÑÑÐ»Ð¾Ð²Ð¸Ñ Ð´Ð¾ÑÑÐ°Ð²ÐºÐ¸ Ð¼Ð ÐÐ-2/3-Fab, Ð¿ÑÐ¸ ÐºÐ¾ÑÐ¾ÑÑÑ ÑÑÐ°Ð½ÑÐ¿Ð¾ÑÑ Ð»Ð¸Ð¿Ð¾Ð¿Ð»ÐµÐºÑÐ½ÑÑ ÑÑÑÑÐºÑÑÑ Ð½ÐµÐ¹ÑÑÐ°Ð»ÐµÐ½ Ð² Ð¾ÑÐ½Ð¾ÑÐµÐ½Ð¸Ð¸ Ð¸Ð½ÑÐ¸ÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ, ÑÐ¾ ÐµÑÑÑ Ð½Ðµ Ð½Ð¾ÑÐ¸Ñ Ð½Ð¸ Ð¿ÑÐ¾Ð²Ð¸ÑÑÑÐ½ÑÐ¹, Ð½Ð¸ Ð¿ÑÐ¾ÑÐ¸Ð²Ð¾Ð²Ð¸ÑÑÑÐ½ÑÐ¹ ÑÐ°ÑÐ°ÐºÑÐµÑ. ÐÐ° Ð¾ÑÐ½Ð¾Ð²Ð°Ð½Ð¸Ð¸ Ð¿Ð¾Ð´Ð¾Ð±ÑÐ°Ð½Ð½ÑÑ Ð²ÑÐµÐ¼ÐµÐ½Ð½ÑÑ Ð¿Ð°ÑÐ°Ð¼ÐµÑÑÐ¾Ð² Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½ ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½Ñ Ð¿Ð¾ Ð¾Ð¿ÑÐµÐ´ÐµÐ»ÐµÐ½Ð¸Ñ Ð¿ÑÐ¾ÑÐ¸Ð²Ð¾Ð²Ð¸ÑÑÑÐ½Ð¾Ð³Ð¾ ÑÑÑÐµÐºÑÐ° Ð¾Ñ Ð¿ÑÐ¸Ð¼ÐµÐ½ÐµÐ½Ð¸Ñ ÑÐµÑÐ°Ð¿ÐµÐ²ÑÐ¸ÑÐµÑÐºÐ¸Ñ Ð¼Ð ÐÐ-2/3-Fab, Ð¿Ð¾Ð´ÑÐ²ÐµÑÐ´Ð¸Ð²ÑÐ¸Ð¹ ÑÑÑÐµÑÑÐ²ÐµÐ½Ð½Ð¾Ðµ Ð¿Ð¾Ð´Ð°Ð²Ð»ÐµÐ½Ð¸Ðµ Ð²Ð¸ÑÑÑÐ½Ð¾Ð¹ ÑÐµÐ¿Ð»Ð¸ÐºÐ°ÑÐ¸Ð¸. Ð¼Ð ÐÐ-2/3-Fab Ð·Ð½Ð°ÑÐ¸ÑÐµÐ»ÑÐ½Ð¾ ÑÐ½Ð¸Ð¶Ð°ÑÑ ÑÐ¸ÑÑ Ð¸ Ð¿ÑÐ¸Ð²Ð¾Ð´ÑÑ Ðº Ð¿Ð¾ÐºÐ°Ð·Ð°ÑÐµÐ»ÑÐ¼, ÑÐ¾Ð¿Ð¾ÑÑÐ°Ð²Ð¸Ð¼ÑÐ¼ Ñ Ð´ÐµÐ¹ÑÑÐ²Ð¸ÐµÐ¼ Ð·Ð°Ð½Ð°Ð¼Ð¸Ð²Ð¸ÑÐ°. ÐÐ¾Ð»ÑÑÐµÐ½Ð½ÑÐµ ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÑ Ñ ÑÐ¾ÑÐºÐ¸ Ð·ÑÐµÐ½Ð¸Ñ Ð¿ÑÐ°ÐºÑÐ¸ÑÐµÑÐºÐ¾Ð¹ Ð¿Ð¾Ð»ÑÐ·Ñ Ð¼Ð¾Ð³ÑÑ Ð±ÑÑÑ ÑÐ°ÑÑÐ¼Ð¾ÑÑÐµÐ½Ñ ÐºÐ°Ðº ÑÑÐ°ÑÑÐ¾Ð²ÑÐµ Ð¿ÑÐ¸ ÑÐ°ÑÑÐ¸ÑÐµÐ½Ð¸Ð¸ ÑÐ¿Ð¸ÑÐºÐ° Ð¸ÑÑÐ»ÐµÐ´ÑÐµÐ¼ÑÑ ÑÑÐ°Ð¼Ð¼Ð¾Ð² Ð²Ð¸ÑÑÑÐ° Ð³ÑÐ¸Ð¿Ð¿Ð°, Ð° ÑÐ°ÐºÐ¶Ðµ Ð¿ÑÐ¸ Ð¿ÐµÑÐµÑÐ¾Ð´Ðµ Ðº ÑÐ¸ÑÑÐµÐ¼Ð°Ð¼ in vivo Ñ Ð¿Ð¾ÑÑÐµÐ¿ÐµÐ½Ð½ÑÐ¼ ÑÐ¾Ð²ÐµÑÑÐµÐ½ÑÑÐ²Ð¾Ð²Ð°Ð½Ð¸ÐµÐ¼ Ð¸Ñ Ð¼Ð¾Ð´ÐµÐ»ÐµÐ¹ Ð¸ ÑÑÐ»Ð¾Ð¶Ð½ÐµÐ½Ð¸ÐµÐ¼ ÑÐµÐ°ÐºÑÐ¸Ð¹ Ð¸Ð¼Ð¼ÑÐ½Ð½Ð¾Ð³Ð¾ Ð¾ÑÐ²ÐµÑÐ°.
description: The search for antiviral drugs that are active against different strains of the influenza virus is an extremely urgent task. While mRNA vaccines approved for emergency use during COVID-19 have demonstrated safety and high efficacy, the therapeutic approach based on mRNA technology represents a promising alternative strategy. The given work is devoted to in vitro study of the preventive and treatment potential of mRNAs encoding Fab-fragments (2/3-Fab) of non-neutralizing antibodies to the internal conserved protein of influenza B virus â nucleoprotein. While studying the main properties of influenza B/Austria/1359417/2021 virus, its titer was obtained in units of 50% cytopathic activity by hemagglutination assay (HA) and enzyme-linked immunosorbent assay (ELISA). Fluorescence microscopy and luciferase assay confirmed the ability of 2X3-DOPE liposomes to successfully transfect MDCK cells and deliver model mRNAs of green fluorescent protein eGFP and firefly luciferase (FLuc) respectively. The intracellular production of 2/3-Fab was established and the dynamics of their accumulation was described. The effect of 2X3-DOPE liposome and mRNA (FLuc) complexes on infection of MDCK cells with influenza B/Austria/1359417/2021 virus was characterized. As a consequence, mRNA-2/3-Fab delivery conditions were optimized under which the transport of lipoplex structures is neutral with respect to infection, i.e. neither pro-viral nor antiviral in nature. Based on the selected time parameters, an experiment was conducted to determine the antiviral effect of therapeutic mRNA-2/3-Fab, which confirmed a significant suppression of viral replication. mRNA-2/3-Fab noticeably reduced the titer and resulted in rates comparable to the effect of zanamivir. Considering practical benefit, the obtained results can be a starting point for expanding the list of influenza virus strains under study, as well as for the transition to in vivo systems with gradual improvement of their models and complication of immune response reactions.