Human immunodeficiency virus type 1 (HIV-1) vaccine immunogens capable of inducing broadly neutralizing antibodies (bNAbs) remain obscure. HIV-1 evades immune responses through enormous diversity and hides its conserved vulnerable epitopes on the envelope glycoprotein (Env) by displaying an extensive immunodominant glycan shield. In elite HIV-1 viremic controllers, glycan-dependent bNAbs targeting conserved Env epitopes have been isolated and are utilized as vaccine design templates. However, immunological tolerance mechanisms limit the development of these antibodies in the general population. The well characterized bNAbs monoclonal variants frequently exhibit extensive levels of somatic hypermutation, a long third heavy chain complementary determining region, or a short third light chain complementarity determining region, and some exhibit poly-reactivity to autoantigens. This review elaborates on the obstacles to engaging and manipulating the Env glycoprotein as an effective immunogen and describes an alternative reverse vaccinology approach to develop a novel category of bNAb-epitope-derived non-cognate immunogens for HIV-1 vaccine design.
- MeSH
- Epitopes immunology MeSH
- env Gene Products, Human Immunodeficiency Virus immunology MeSH
- HIV Infections immunology MeSH
- HIV Antibodies * immunology MeSH
- HIV-1 * immunology MeSH
- Humans MeSH
- Ligands MeSH
- Molecular Mimicry immunology MeSH
- Antibodies, Neutralizing * immunology MeSH
- Polysaccharides immunology MeSH
- AIDS Vaccines * immunology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Nicotinamide adenine dinucleotide (NAD) is a critical component of the cellular metabolism and also serves as an alternative 5' cap on various RNAs. However, the function of the NAD RNA cap is still under investigation. We studied NAD capping of RNAs in HIV-1-infected cells because HIV-1 is responsible for the depletion of the NAD/NADH cellular pool and causing intracellular pellagra. By applying the NAD captureSeq protocol to HIV-1-infected and uninfected cells, we revealed that four snRNAs (e.g., U1) and four snoRNAs lost their NAD cap when infected with HIV-1. Here, we provide evidence that the presence of the NAD cap decreases the stability of the U1/HIV-1 pre-mRNA duplex. Additionally, we demonstrate that reducing the quantity of NAD-capped RNA by overexpressing the NAD RNA decapping enzyme DXO results in an increase in HIV-1 infectivity. This suggests that NAD capping is unfavorable for HIV-1 and plays a role in its infectivity.
- MeSH
- HIV Infections * virology metabolism MeSH
- HIV-1 * MeSH
- Humans MeSH
- RNA, Small Nucleolar * metabolism genetics MeSH
- NAD * metabolism MeSH
- RNA Caps metabolism MeSH
- RNA, Small Nuclear * metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Anti-viral and anti-tumor vaccines aim to induce cytotoxic CD8+ T cells (CTL) and antibodies. Conserved protein antigens, such as p24 from human immunodeficiency virus, represent promising component for elicitation CTLs, nevertheless with suboptimal immunogenicity, if formulated as recombinant protein. To enhance immunogenicity and CTL response, recombinant proteins may be targeted to dendritic cells (DC) for cross presentation on MHCI, where mannose receptor and/or other lectin receptors could play an important role. Here, we constructed liposomal carrier-based vaccine composed of recombinant p24 antigen bound by metallochelating linkage onto surface of nanoliposomes with surface mannans coupled by aminooxy ligation. Generated mannosylated proteonanoliposomes were analyzed by dynamic light scattering, isothermal titration, and electron microscopy. Using murine DC line MutuDC and murine bone marrow derived DC (BMDC) we evaluated their immunogenicity and immunomodulatory activity. We show that p24 mannosylated proteonanoliposomes activate DC for enhanced MHCI, MHCII and CD40, CD80, and CD86 surface expression both on MutuDC and BMDC. p24 mannosylated liposomes were internalized by MutuDC with p24 intracellular localization within 1 to 3 h. The combination of metallochelating and aminooxy ligation could be used simultaneously to generate nanoliposomal adjuvanted recombinant protein-based vaccines versatile for combination of recombinant antigens relevant for antibody and CTL elicitation.
- MeSH
- Antigens MeSH
- Dendritic Cells MeSH
- HIV-1 * MeSH
- Humans MeSH
- Liposomes metabolism MeSH
- Mannans metabolism MeSH
- Mice MeSH
- Recombinant Proteins metabolism MeSH
- AIDS Vaccines * immunology MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Léčba infekce HIV modifikovala původně smrtelnou infekci do typicky chronického onemocnění s potřebou celoživotní léčby. U léčených pacientů však nedochází ke kompletní normalizaci imunitní aktivace, známek zánětu a protrombotického stavu. Tento stav je důsledkem mnoha faktorů, za hlavní příčinu je ale považována reziduální produkce RNA HIV-1 a virových proteinů infikovanými buňkami v buněčných rezervoárech. Perzistence imunitní aktivace/zánětu/protrombotického stavu vede k patofyziologii „sterilního zánětu“ a tzv. non -AIDS onemocněním, která se u infikovaných manifestují o jednu až dvě dekády dříve. Přes veškerá úskalí a nežádoucí sekundární projevy antiretrovirotik dokázala léčba infekce HIV zvrátit trajektorii fatální pandemie a umožnila přistoupit k terapeutickým modalitám, které byly ještě před několika lety absolutně nepředstavitelné. Transplantace solidních orgánů je pro pacienty s infekcí HIV dnes zcela legitimní terapeutická metoda a vysoce supresivní léčba umožňuje dokonce transplantaci od dárce s infekcí HIV. Níže uvedený text předkládá stručný přehled základních úskalí, ale i úspěchů současné vysoce supresivní léčby infekce HIV.
Treatment of HIV infection has modified the initially fatal infection into a typically chronic disease requiring lifelong treatment. However, there is no complete normalization of immune activation, signs of inflammation and prothrombotic state in treated patients. This condition is the result of many factors, but the main cause is thought to be the residual production of HIV-1 RNA and viral proteins by infected cells in cellular reservoirs. Persistence of immune activation/inflammation/prothrombotic state leads to the pathophysiology of "sterile inflammation" and so-called non-AIDS diseases, which manifest one to two decades earlier in those infected. Despite all the pitfalls and unwanted secondary manifestations of antiretroviral drugs, the treatment of HIV infection has managed to reverse the trajectory of a fatal pandemic and has made it possible to approach therapeutic modalities that were absolutely unimaginable just a few years ago. Solid organ transplantation is now a completely legitimate therapeutic method for patients living with HIV, and highly suppressive treatment even allows transplantation from an HIV-infected donor. The text below presents a brief overview of the basic pitfalls, but also of the successes, of the current highly suppressive treatment of HIV infection.
- MeSH
- Anti-Retroviral Agents * therapeutic use MeSH
- HIV Infections * drug therapy immunology complications MeSH
- HIV-1 genetics MeSH
- Homeostasis MeSH
- Humans MeSH
- Organ Transplantation MeSH
- Inflammation drug therapy MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
The secondary structure of nucleic acids containing quartets of guanines, termed G-quadruplexes, is known to regulate the transcription of many genes. Several G-quadruplexes can be formed in the HIV-1 long terminal repeat promoter region and their stabilization results in the inhibition of HIV-1 replication. Here, we identified helquat-based compounds as a new class of anti-HIV-1 inhibitors that inhibit HIV-1 replication at the stage of reverse transcription and provirus expression. Using Taq polymerase stop and FRET melting assays, we have demonstrated their ability to stabilize G-quadruplexes in the HIV-1 long-terminal repeat sequence. Moreover, these compounds were not binding to the general G-rich region, but rather to G-quadruplex-forming regions. Finally, docking and molecular dynamics calculations indicate that the structure of the helquat core greatly affects the binding mode to the individual G-quadruplexes. Our findings can provide useful information for the further rational design of inhibitors targeting G-quadruplexes in HIV-1.
Cieľ: Cieľom štúdie bolo popísať výskyt HIV-1 subtypov a HIV-1 kmeňov rezistentných na antiretrovírusovú liečbu (ART) u HIV pozitívnych osôb novo diagnostikovaných na Slovensku v rokoch 2019–2021. Materiál a metódy: Študijnú skupinu tvorilo 184 HIV pozitívnych naivných pacientov novo diagnostikovaných na Slovensku v rokoch 2019–2021. Vírusová HIV-1 RNA bola izolovaná z plazmy pomocou QIAamp Viral RNA Mini Kit (QIAGEN, Nemecko). Pre RT-PCR a sekvenovanie oblasti HIV pol sme použili interné postupy podľa protokolu ANRS AC11 pre RT (reverzná transkriptáza), PRO (proteáza) a IN (integráza) (ANRS AC11 Resistance Study Group, 2015). Analýzu sekvencií sme uskutočnili pomocou softvéru Sequencing Analysis Software v5.3 (Applied Biosystems®). HIV sekvencie boli manuálne upravené pomocou BioEdit (verzia 7.2.5), porovnané s konsenzuálnymi HIV-1 sekvenciami v Los Alamos Sequence Database (URL 2), zarovnané pomocou CLUSTAL W (Labarga et al., 2007) a softvérových balíkov BioEdit (verzia 7.2 .5) (Hall, 1999). Na vyhodnotenie sekvencie sme použili algoritmus HIVDB (verzia 9.0) Stanfordskej databázy HIV liekovej rezistencie (URL 1.). Na analýzu HIV-1 subtypov sme použili nástroj REGA HIV-1 Subtyping Tool (De Oliviera et al., 2005) a fylogenetickú analýzu sme vypracovali pomocou programu MEGA X (Kumar et al., 2018). Výsledky: Fylogenetickú analýzu sme vykonali zo vzoriek 184 osôb, kde sme odhalili najrozšírenejší subtyp B (129/184, 70,11 %) prevládajúci v populácii u mužov, ktorí majú sex s mužmi (MSM) (96/129 74,42 %). Čo sa týka non-B subtypov (55/184, 29,89 %), najrozšírenejší bol subtyp A (48/184, 26,09 %) v porovnaní so subtypom F (F1) (3; 1,63 %), C (1; 0,54 %) a cirkulujúcimi rekombinantnými formami CRF02_AG (2; 1,09 %), CRF01_AE (1; 0,54 %). U 9,24 % (17/184) vzoriek sme zistili prítomnosť 25 mutácií asociovaných s HIV rezistenciou na ART, z toho 7,07 % (13/184) na inhibítory reverznej transkriptázy, 1,66 % (3/181) na inhibítory proteázy a 1,32 % (2/151) na inhibítory integrázy. Okrem toho u 1,63 % (3/184) pacientov bola prítomná viactriedna rezistencia. Mutácie asociované s rezistenciou HIV na ART sa našli u 9,30 % osôb infikovaných podtypom B. Záver: Naša štúdia potvrdila pretrvávajúci najvyšší výskyt subtypu B s mierne klesajúcou tendenciou v porovnaní s minulými rokmi. Detekcia mutácií vytvárajúcich rezistenciu HIV na ART podčiarkuje potrebu testovania rezistencie u naivných pacientov ešte pred začatím ART na Slovensku.
Aim: The aim of the study was to describe the prevalence of HIV-1 subtypes and HIV-1 strains resistant to antiretroviral therapy (ART) in HIV-positive persons newly diagnosed in Slovakia in 2019–2021. Materials and Methods: The study group consisted of 184 HIV-positive na?ve patients newly diagnosed in Slovakia from 2019 to 2021. The viral HIV-1 RNA was isolated from plasma by the QIAamp Viral RNA Mini Kit (QIAGEN, Germany). For RT-PCR and sequencing of the HIV pol region, in-house procedures were used according to the ANRS AC11 protocol for RT (reverse transcriptase), PRO (protease), and IN (integrase) [ANRS AC11 Resistance Study Group, 2015]. Analysis of sequences was performed using Sequencing Analysis Software v5.3 (Applied Biosystems®). HIV sequences were manually edited using BioEdit (version 7.2.5), compared with consensus HIV-1 sequences in the Los Alamos Sequence Database (URL 2), aligned using CLUSTAL W [Labarga et al., 2007] and BioEdit software packages (version 7.2 .5) [Hall, 1999]. HIVDB Algorithm (version 9.0) of the Stanford HIV Drug resistance database (URL 1.) was used for sequence evaluation. For HIV-1 subtype analysis, the REGA HIV-1 Subtyping Tool [De Oliviera et al., 2005] and phylogenetic analysis MEGA X [Kumar et al., 2018] were used. Results: Phylogenetic analyses performed in samples of 184 persons revealed the most prevalent subtype B (129/184, 70.11%), detected to the greatest extent in the population of men who have sex with men (MSM) (96/129 74.42%). Concerning non-B subtypes (55/184, 29.89%), subtype A was found with the highest prevalence (48/184, 26.09%) compared to subtype F (F1) (3; 1.63%), C (1; 0.54%) and circulating recombinant forms CRF02_AG (2; 1.09%), CRF01_AE (1; 0.54%). In 9.24% (17/184) of samples, 25 mutations clinically relevant and associated with HIV resistance ART were detected, of which 7.07% (13/184) to reverse transcriptase inhibitors, 1.66% (3/181) to protease inhibitors and 1.32% (2/151) to integrase inhibitors. In addition, multiclass resistance was present in 1.63% (3/184) of patients. Mutations associated with HIV resistance to ART were found in 9.30 % of persons infected with subtype B. Conclusion: Our study confirmed ongoing highest prevalence of subtype B with a slightly decreasing trend compared to last years. Detection of mutations causing HIV resistance to ART underlines the need for resistance testing in na?ve patients even before the initiation of ART in Slovakia.
- MeSH
- Anti-Retroviral Agents therapeutic use MeSH
- Genetic Techniques MeSH
- Genotype MeSH
- HIV Infections * epidemiology drug therapy transmission MeSH
- HIV-1 genetics drug effects MeSH
- Clinical Studies as Topic MeSH
- Humans MeSH
- Mutation genetics MeSH
- Drug Resistance, Viral MeSH
- Check Tag
- Humans MeSH
- Geographicals
- Slovakia MeSH
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) represent cornerstones of current regimens for treatment of human immunodeficiency virus type 1 (HIV-1) infections. However, NNRTIs usually suffer from low aqueous solubility and the emergence of resistant viral strains. In the present work, novel bicyclic NNRTIs derived from etravirine (ETV) and rilpivirine (RPV), bearing modified purine, tetrahydropteridine, and pyrimidodiazepine cores, were designed and prepared. Compounds 2, 4, and 6 carrying the acrylonitrile moiety displayed single-digit nanomolar activities against the wild-type (WT) virus (EC50 = 2.5, 2.7, and 3.0 nM, respectively), where the low nanomolar activity was retained against HXB2 (EC50 = 2.2-2.8 nM) and the K103N and Y181C mutated strains (fold change, 1.2-6.7×). Most importantly, compound 2 exhibited significantly improved phosphate-buffered saline solubility (10.4 μM) compared to ETV and RPV (≪1 μM). Additionally, the binding modes of compounds 2, 4, and 6 to the reverse transcriptase were studied by X-ray crystallography.
- MeSH
- HIV Infections * drug therapy MeSH
- HIV Reverse Transcriptase metabolism MeSH
- HIV-1 * metabolism MeSH
- Reverse Transcriptase Inhibitors MeSH
- Anti-HIV Agents * chemistry MeSH
- Humans MeSH
- Drug Design MeSH
- Rilpivirine therapeutic use MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
INTRODUCTION: Imprinting broadly neutralizing antibody (bNAb) paratopes by shape complementary protein mimotopes represents a potential alternative for developing vaccine immunogens. This approach, designated as a Non-Cognate Ligand Strategy (NCLS), has recently been used for the identification of protein variants mimicking CD4 binding region epitope or membrane proximal external region (MPER) epitope of HIV-1 envelope (Env) glycoprotein. However, the potential of small binding proteins to mimic viral glycan-containing epitopes has not yet been verified. METHODS: In this work, we employed a highly complex combinatorial Myomedin scaffold library to identify variants recognizing paratopes of super candidate bNAbs, PGT121 and PGT126, specific for HIV-1 V3 loop epitopes. RESULTS: In the collection of Myomedins called MLD variants targeted to PGT121, three candidates competed with gp120 for binding to this bNAb in ELISA, thus suggesting an overlapping binding site and epitope-mimicking potential. Myomedins targeted to PGT126 designated MLB also provided variants that competed with gp120. Immunization of mice with MLB or MLD binders resulted in the production of anti-gp120 and -Env serum antibodies. Mouse hyper-immune sera elicited with MLB036, MLB041, MLB049, and MLD108 moderately neutralized 8-to-10 of 22 tested HIV-1-pseudotyped viruses of A, B, and C clades in vitro. DISCUSSION: Our data demonstrate that Myomedin-derived variants can mimic particular V3 glycan epitopes of prominent anti-HIV-1 bNAbs, ascertain the potential of particular glycans controlling neutralizing sensitivity of individual HIV-1 pseudoviruses, and represent promising prophylactic candidates for HIV-1 vaccine development.
- MeSH
- Epitopes MeSH
- HIV Envelope Protein gp120 MeSH
- HIV Antibodies * MeSH
- HIV-1 * MeSH
- Mice MeSH
- Antibodies, Neutralizing MeSH
- Polysaccharides MeSH
- Broadly Neutralizing Antibodies MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Decrypting the B cell ontogeny of HIV-1 broadly neutralizing antibodies (bNAbs) is paramount for vaccine design. Here, we characterized IgA and IgG bNAbs of three distinct B cell lineages in a viremic controller, two of which comprised only IgG+ or IgA+ blood memory B cells; the third combined both IgG and IgA clonal variants. 7-269 bNAb in the IgA-only lineage displayed the highest neutralizing capacity despite limited somatic mutation, and delayed viral rebound in humanized mice. bNAbs in all three lineages targeted the N332 glycan supersite. The 2.8-Å resolution cryo-EM structure of 7-269-BG505 SOSIP.664 complex showed a similar pose as 2G12, on an epitope mainly composed of sugar residues comprising the N332 and N295 glycans. Binding and cryo-EM structural analyses showed that antibodies from the two other lineages interact mostly with glycans N332 and N386. Hence, multiple B cell lineages of IgG and IgA bNAbs focused on a unique HIV-1 site of vulnerability can codevelop in HIV-1 viremic controllers.
- MeSH
- Epitopes MeSH
- env Gene Products, Human Immunodeficiency Virus MeSH
- HIV Infections * MeSH
- HIV Antibodies MeSH
- HIV-1 * MeSH
- Immunoglobulin A MeSH
- Immunoglobulin G MeSH
- Humans MeSH
- Mice MeSH
- Antibodies, Neutralizing MeSH
- Elite Controllers MeSH
- Polysaccharides MeSH
- Broadly Neutralizing Antibodies MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Fullerene derivatives with hydrophilic substituents have been shown to exhibit a range of biological activities, including antiviral ones. For a long time, the anti-HIV activity of fullerene derivatives was believed to be due to their binding into the hydrophobic pocket of HIV-1 protease, thereby blocking its activity. Recent work, however, brought new evidence of a novel, protease-independent mechanism of fullerene derivatives' action. We studied in more detail the mechanism of the anti-HIV-1 activity of N,N-dimethyl[70]fulleropyrrolidinium iodide fullerene derivatives. By using a combination of in vitro and cell-based approaches, we showed that these C70 derivatives inhibited neither HIV-1 protease nor HIV-1 maturation. Instead, our data indicate effects of fullerene C70 derivatives on viral genomic RNA packaging and HIV-1 cDNA synthesis during reverse transcription-without impairing reverse transcriptase activity though. Molecularly, this could be explained by a strong binding affinity of these fullerene derivatives to HIV-1 nucleocapsid domain, preventing its proper interaction with viral genomic RNA, thereby blocking reverse transcription and HIV-1 infectivity. Moreover, the fullerene derivatives' oxidative activity and fluorescence quenching, which could be one of the reasons for the inconsistency among reported anti-HIV-1 mechanisms, are discussed herein.
- MeSH
- Fullerenes metabolism pharmacology MeSH
- Genome, Viral drug effects MeSH
- gag Gene Products, Human Immunodeficiency Virus metabolism MeSH
- HEK293 Cells MeSH
- HIV-1 drug effects genetics metabolism physiology MeSH
- Anti-HIV Agents metabolism pharmacology MeSH
- Humans MeSH
- Nucleocapsid Proteins metabolism MeSH
- Reverse Transcription MeSH
- RNA, Viral metabolism MeSH
- Virus Uncoating drug effects MeSH
- Protein Binding MeSH
- Virion metabolism MeSH
- Viral Genome Packaging drug effects MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
