Humanized Mouse Model of HIV Infection Leontyev, D. S.; Glazkova, D. V.; Bezborodova, O. A. ...
Bulletin of experimental biology and medicine,
05/2023, Letnik:
175, Številka:
1
Journal Article
Recenzirano
The development of new drugs for the treatment of HIV infection requires testing of their efficacy in a relevant animal model, such as humanized mice, which, unfortunately, are not yet available in ...Russia. In the present study, we have developed conditions for the humanization of immunodeficient NSG mice with human hematopoietic stem cells. Humanized animals generated during the study showed a high degree of chimerism and harbored repopulation of the entire range of human lymphocytes required for HIV replication in the blood and organs. Inoculation of these mice with HIV-1 virus led to stable viremia, which was confirmed by the presence of viral RNA in blood plasma throughout the entire period of observation and proviral DNA in the organs of animals 4 weeks after HIV infection.
•Novel single-tube real-time PCR to determine a ratio of wild type and rMVA in plaques.•Rapid generation of new pure rMVA in 11 days.•Plaque purification is less effective then end-point dilution as ...the last purification step.•Various marker gene selected plaques differ greatly in proportion of rMVA.
Obtaining a pure recombinant Modified Vaccinia Ankara (MVA) virus is a multistage, time-consuming procedure. We describe a novel single-tube real-time PCR which enables determination of the amount of wild type and recombinant viruses and their ratio in plaques. Use of the real-time PCR significantly reduce the time and efforts needed to obtain purified recombinant MVA. The new approach has been applied to generate recombinant MVAs encoding different SARS-COV-2 antigens.
Influenza virus is one of the most rapidly evolving human pathogens and causes significant morbidity and mortality worldwide. This feature enables the virus to avoid natural or vaccine-induced ...immunity. For this reason, there is an intensive search for new approaches to create a universal influenza vaccine. Here, we propose pipelines based on modern prediction algorithms that allowed us to select 10 B-cell epitopes, 10 CD8+ T-cell epitopes and 6 CD4+ T-cell epitopes from influenza viruses that were characterized by high conservation and antigenicity. These epitopes could be used to create universal vaccines against influenza viruses. In addition, the scripts used in these pipelines are universal and can be used to select epitopes from other pathogens.
Influenza, Universal vaccine, T- and B-cell epitopes, Computational approaches.
The chimeric protein TRIM5α-HRH is a promising antiviral factor for HIV-1 gene therapy. This protein is able to protect cells from HIV-1 by blocking the virus in the cytoplasm. We are developing ...protocol of HIV-1 gene therapy, which involves the delivery of the
TRIM5
α
-HRH
gene into CD4
+
T-lymphocytes by lentiviral vectors (LVs). However, LVs containing
TRIM5
α
-HRH
have a low infectious titer, which prevents effective T cell modification. Here, we found that the expression of
TRIM5
α
-HRH
during pseudoviral particle production in HEK293 T cells, as well as the presence of the Ef1α promoter in our construction are responsible for titer reduction. These results allow us to determine the directions for further optimization of LV with the
TRIM5
α
-HRH
gene to improve its infectious titer.
One of the most important steps in the development of drugs and vaccines against a new coronavirus infection is their testing on a relevant animal model. The laboratory mouse, with well-studied ...immunology, is the preferred mammalian model in experimental medicine. However, mice are not susceptible to infection with SARS-CoV-2 due to the lack of human angiotensin-converting enzyme (hACE2), which is the cell receptor of SARS-CoV-2 and necessary for the entry of the virus into the cell. In present work, it was shown that intranasal administration of the adeno-associated vectors AAV9 and AAV-DJ encoding the hACE2 provided a high level of expression of
ACE2
gene in the lungs of mice. In contrast, the introduction of the AAV6 vector led to a low level
ACE2
expression. Infection with SARS-CoV-2 of mice expressing hACE2 in the lungs led to virus replication and development of bronchopneumonia on the 7th day after infection. Thus, a simple method for delivering the human
ACE2
gene to mouse lungs by intranasal administration of the AAV vector has been proposed. This approach enabled rapid generation of mouse model for studying coronavirus infection.
The mechanisms for the protection of the human body from viral or bacterial agents are extremely diverse. In one such mechanism, an important role belongs to the cytidine deaminase APOBEC3 family, ...which is the factor of congenital immunity and protects the organism from numerous viral agents. One of the proteins of this family, APOBEC3G, is able to protect against Human Immunodeficiency Virus type 1 in the absence of viral protein Vif. In turn, Vif opposes APOBEC3G action, causing polyubiquity of the protein and degradation in the proteasome. The review describes possible ways to increase the anti-HIV activity of APOBEC3G, giving it resistance to viral protein Vif, as well as potential approaches to the use of modified APOBEC3G in gene therapy for HIV.
The chimeric protein TRIM5α-HRH is a promising antiviral factor for HIV-1 gene therapy. This protein is able to protect cells from HIV-1 by blocking the virus in the cytoplasm. We are developing ...protocol of HIV-1 gene therapy, which involves the delivery of the TRIM5α-HRH gene into CD4^(+) T-lymphocytes by lentiviral vectors (LVs). However, LVs containing TRIM5α-HRH have a low infectious titer, which prevents effective T cell modification. Here, we found that the expression of TRIM5α-HRH during pseudoviral particle production in HEK293 T cells, as well as the presence of the Eflα promoter in our construction are responsible for titer reduction. These results allow us to determine the directions for further optimization of LV with the TRIM5α-HRH gene to improve its infectious titer.
Abstract
The work is devoted to improving clonal micropropagation methods of the genus
Rubus
representatives. When cultivating
R. arcticus
L. cultivars, the optimal concentration of ...6-benzylaminopurine (6-BAP) was 0.3 mg L
−1
. Significant effect of FeEDTA concentration in the nutrient medium on multiplication of
R. idaeus
L. cultivars was established. The interaction of auxin type and plant genotype was revealed during rooting of blackberry cultivars and raspberry-blackberry hybrids. The nutrient medium supplemented with 0.5 mg L-1 indoleacetic acid contributed to the highest percentage of rhizogenesis in
R. arcticus
representatives (82%). The optimal growth regulator for conservation of raspberry explants was 6-BAP at a concentration of 0.3 mg L-1. One of the most representative
in vitro
collections of
Rubus
cultivars has been created.
—
It is commonly known that the antiviral activity of the TRIM5α protein, the intracellular retrovirus restriction factor, underlies the resistance of the Old World monkeys to HIV-1. This fact ...suggests that TRIM5α can potentially be used to cure HIV-1 infection in humans. The present review considers the mechanisms of HIV-1 replication inhibition by the TRIM5α protein and the prospects for using it in gene therapy of HIV infection.
It is commonly known that the antiviral activity of the TRIM5α protein, the intracellular retrovirus restriction factor, underlies the resistance of the Old World monkeys to HIV-1. This fact suggests ...that TRIM5α can potentially be used to cure HIV-1 infection in humans. The present review considers the mechanisms of HIV-1 replication inhibition by the TRIM5a protein and the prospects for using it in gene therapy of HIV infection.
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