Peptide immunogens provide an approach to focus antibody responses to specific neutralizing sites on the HIV envelope protein (Env) trimer or on other pathogens. However, the physical characteristics of peptide immunogens can limit their pharmacokinetic and immunological properties. Here, we have designed synthetic "star" nanoparticles based on biocompatible N-[(2-hydroxypropyl)methacrylamide] (HPMA)-based polymer arms extending from a poly(amidoamine) (PAMAM) dendrimer core. In mice, these star nanoparticles trafficked to lymph nodes (LNs) by 4 hours following vaccination, where they were taken up by subcapsular macrophages and then resident dendritic cells (DCs). Immunogenicity optimization studies revealed a correlation of immunogen density with antibody titers. Furthermore, the co-delivery of Env variable loop 3 (V3) and T-helper peptides induced titers that were 2 logs higher than if the peptides were given in separate nanoparticles. Finally, we performed a nonhuman primate (NHP) study using a V3 glycopeptide minimal immunogen that was structurally optimized to be recognized by Env V3/glycan broadly neutralizing antibodies (bnAbs). When administered with a potent Toll-like receptor (TLR) 7/8 agonist adjuvant, these nanoparticles elicited high antibody binding titers to the V3 site. Similar to human V3/glycan bnAbs, certain monoclonal antibodies (mAbs) elicited by this vaccine were glycan dependent or targeted the GDIR peptide motif. To improve affinity to native Env trimer affinity, nonhuman primates (NHPs) were boosted with various SOSIP Env proteins; however, significant neutralization was not observed. Taken together, this study provides a new vaccine platform for administration of glycopeptide immunogens for focusing immune responses to specific bnAb epitopes.

• MeSH
• epitopy imunologie   MeSH
• HIV infekce imunologie   MeSH
• HIV obalový protein gp120 chemie   MeSH
• HIV séropozitivita imunologie   MeSH
• HIV-1 imunologie   MeSH
• Macaca mulatta MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nanočástice chemie   terapeutické užití   MeSH
• neutralizující protilátky imunologie   MeSH
• peptidy MeSH
• primáti MeSH
• tvorba protilátek imunologie   MeSH
• vakcíny proti AIDS imunologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Intramural MeSH

Východisko: Glykozylácia konštantných oblastí protilátok zásadne ovplyvňuje ich interakčné schopnosti s bunkami imunitného systému. Jedná sa o modifikáciu, ktorá okrem biologickej aktivity protilátok zasahuje aj do ich konformácie, stability, rozpustnosti, sekrécie, farmakokinetiky a imunogénnosti. Pre ich správnu funkciu nie je podstatná len lokalizácia samotných glykozylácií na molekule protilátok, ale aj štruktúra jednotlivých glykánov. Zmeny glykozylačných profilov protilátok boli popísané u niektorých fyziologických procesov, akými sú tehotenstvo alebo starnutie, ale taktiež u mnohých patologických stavov ako reumatoidnej artritídy, či nádorov žalúdka, pľúc alebo prostaty. Stále existuje celé množstvo neobjasnených mechanizmov, ktoré riadia glykozyláciu protilátok alebo sú týmito modifikáciami naopak regulované. Viaceré zdroje popisujú význam niektorých špecifických glykozylácií ako potenciálnych biomarkerov. Cieľ: Cieľom tohto prehľadového článku je zhrnúť a priblížiť doterajšie poznatky o glykozylácii protilátok a upozorniť na ich vplyv na imunitné odpovede a ich úlohu v priebehu ochorenia. Ich dôležitosť podčiarkuje aj to, že väčšina vyvíjaných a využívaných terapeutických protilátok je modifikovaných glykozyláciou. Práve cielené vnesenie vhodných glykozylácií, ktoré podporujú aktivity akými sú napr. bunková cytotoxicita závislá na protilátkach, bunková fagocytóza závislá na protilátkach alebo cytotoxicita závislá na komplemente, viedlo k zlepšeniu schopností týchto protilátok likvidovať patogény alebo nádorové bunky. Preto je oblasti glykozylácie protilátok venovaná stále väčšia pozornosť. Získané znalosti môžu prispieť k ďalšiemu vývoju efektívnych nástrojov diagnostiky a terapie rôznych ochorení.

Background: The glycosylation of constant regions of antibodies significantly affects their interaction capabilities with immune cells. It is a modification that, in addition to the biological activity of antibodies, has an impact on their conformation, stability, solubility, secretion, pharmacokinetics, and immunogenicity. The location of glycosylations on the molecule is essential for the proper function of the antibody, as is the structure of the individual glycans. Changes in the glycosylation profiles of antibodies have been described in some physiological processes like pregnancy or ageing, but also in many pathological conditions such as rheumatoid arthritis or gastric, lung and prostate tumours. There are still several unexplained mechanisms that control the glycosylation of antibodies or immune responses, which in turn are regulated by these modifications. Multiple sources describe the importance of some specific glycosylations as potential biomarkers. Purpose: The aim of this review is to summarise and present the knowledge of the glycosylation of antibodies and to highlight their influence on immune responses and their role during disease. Their importance is also underlined by the fact that the most of these therapeutic antibodies used and developed are modified by glycosylation. The targeted introduction of appropriate glycosylations, which can promote activities such as antibody-dependent cellular cytotoxicity, antibody-dependent cellular phagocytosis or complement-dependent cytotoxicity, have improved the ability of these antibodies to kill pathogens or tumour cells. Therefore, more attention is being paid to this area. In the future, more effective tools for diagnosing and treating certain diseases can be created with better knowledge.

• MeSH
• farmakoterapie metody   MeSH
• glykosylace MeSH
• imunitní systém MeSH
• lidé MeSH
• protilátky * imunologie   terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy 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
• epitopy MeSH
• genové produkty env - virus lidské imunodeficience MeSH
• HIV infekce * MeSH
• HIV protilátky MeSH
• HIV-1 * MeSH
• imunoglobulin A MeSH
• imunoglobulin G MeSH
• lidé MeSH
• myši MeSH
• neutralizující protilátky MeSH
• non-progresoři MeSH
• polysacharidy MeSH
• široce neutralizující protilátky MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem 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
• epitopy MeSH
• HIV obalový protein gp120 MeSH
• HIV protilátky * MeSH
• HIV-1 * MeSH
• myši MeSH
• neutralizující protilátky MeSH
• polysacharidy MeSH
• široce neutralizující protilátky MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Human immunodeficiency virus type 1 (HIV-1) entry is mediated by the interaction between a variably glycosylated envelope glycoprotein (gp120) and host-cell receptors. Approximately half of the molecular mass of gp120 is contributed by N-glycans, which serve as potential epitopes and may shield gp120 from immune recognition. The role of gp120 glycans in the host immune response to HIV-1 has not been comprehensively studied at the molecular level. We developed a new approach to characterize cell-specific gp120 glycosylation, the regulation of glycosylation, and the effect of variable glycosylation on antibody reactivity. A model oligomeric gp120 was expressed in different cell types, including cell lines that represent host-infected cells or cells used to produce gp120 for vaccination purposes. N-Glycosylation of gp120 varied, depending on the cell type used for its expression and the metabolic manipulation during expression. The resultant glycosylation included changes in the ratio of high-mannose to complex N-glycans, terminal decoration, and branching. Differential glycosylation of gp120 affected envelope recognition by polyclonal antibodies from the sera of HIV-1-infected subjects. These results indicate that gp120 glycans contribute to antibody reactivity and should be considered in HIV-1 vaccine design.

• MeSH
• AIDS imunologie   MeSH
• buněčné linie MeSH
• buňky Hep G2 metabolismus   MeSH
• ELISA metody   MeSH
• glykosidhydrolasy metabolismus   MeSH
• glykosylace MeSH
• HIV obalový protein gp120 genetika   imunologie   izolace a purifikace   metabolismus   MeSH
• HIV séropozitivita imunologie   metabolismus   MeSH
• HIV-1 imunologie   metabolismus   MeSH
• hmotnostní spektrometrie MeSH
• Jurkat buňky metabolismus   MeSH
• komplementární DNA genetika   MeSH
• lektin vázající mannosu genetika   MeSH
• lidé MeSH
• mannosa metabolismus   MeSH
• nádorové buněčné linie MeSH
• oligosacharidy chemie   izolace a purifikace   MeSH
• plazmidy MeSH
• polysacharidy chemie   izolace a purifikace   MeSH
• protilátky imunologie   MeSH
• specificita protilátek MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

• MeSH
• biomarkery farmakologické MeSH
• biotechnologie metody   MeSH
• preklinické hodnocení léčiv MeSH
• protilátky farmakologie   terapeutické užití   MeSH
• Publikační typ
• příručky MeSH

• Konspekt
• Farmacie. Farmakologie
• NLK Obory
• farmacie a farmakologie
• alergologie a imunologie
• NLK Publikační typ
• kolektivní monografie

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
• epitopy imunologie   MeSH
• genové produkty env - virus lidské imunodeficience imunologie   MeSH
• HIV infekce imunologie   MeSH
• HIV protilátky * imunologie   MeSH
• HIV-1 * imunologie   MeSH
• lidé MeSH
• ligandy MeSH
• molekulární mimikry imunologie   MeSH
• neutralizující protilátky * imunologie   MeSH
• polysacharidy imunologie   MeSH
• vakcíny proti AIDS * imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Glycosylation is a topic of intense current interest in the development of biopharmaceuticals because it is related to drug safety and efficacy. This work describes results of an interlaboratory study on the glycosylation of the Primary Sample (PS) of NISTmAb, a monoclonal antibody reference material. Seventy-six laboratories from industry, university, research, government, and hospital sectors in Europe, North America, Asia, and Australia submitted a total of 103 reports on glycan distributions. The principal objective of this study was to report and compare results for the full range of analytical methods presently used in the glycosylation analysis of mAbs. Therefore, participation was unrestricted, with laboratories choosing their own measurement techniques. Protein glycosylation was determined in various ways, including at the level of intact mAb, protein fragments, glycopeptides, or released glycans, using a wide variety of methods for derivatization, separation, identification, and quantification. Consequently, the diversity of results was enormous, with the number of glycan compositions identified by each laboratory ranging from 4 to 48. In total, one hundred sixteen glycan compositions were reported, of which 57 compositions could be assigned consensus abundance values. These consensus medians provide community-derived values for NISTmAb PS. Agreement with the consensus medians did not depend on the specific method or laboratory type. The study provides a view of the current state-of-the-art for biologic glycosylation measurement and suggests a clear need for harmonization of glycosylation analysis methods.

• MeSH
• biofarmacie metody   MeSH
• biologické přípravky * MeSH
• glykomika metody   MeSH
• glykopeptidy metabolismus   MeSH
• glykosylace MeSH
• laboratoře MeSH
• lidé MeSH
• monoklonální protilátky chemie   metabolismus   MeSH
• polysacharidy metabolismus   MeSH
• posttranslační úpravy proteinů MeSH
• proteomika metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Natural antibodies (Abs) can target host glycans on the surface of pathogens. We studied the evolution of glycan-reactive B cells of rhesus macaques and humans using glycosylated HIV-1 envelope (Env) as a model antigen. 2G12 is a broadly neutralizing Ab (bnAb) that targets a conserved glycan patch on Env of geographically diverse HIV-1 strains using a unique heavy-chain (VH) domain-swapped architecture that results in fragment antigen-binding (Fab) dimerization. Here, we describe HIV-1 Env Fab-dimerized glycan (FDG)-reactive bnAbs without VH-swapped domains from simian-human immunodeficiency virus (SHIV)-infected macaques. FDG Abs also recognized cell-surface glycans on diverse pathogens, including yeast and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike. FDG precursors were expanded by glycan-bearing immunogens in macaques and were abundant in HIV-1-naive humans. Moreover, FDG precursors were predominately mutated IgM+IgD+CD27+, thus suggesting that they originated from a pool of antigen-experienced IgM+ or marginal zone B cells.

• MeSH
• B-lymfocyty imunologie   MeSH
• COVID-19 imunologie   MeSH
• dimerizace MeSH
• epitopy imunologie   MeSH
• genové produkty env - virus lidské imunodeficience chemie   genetika   imunologie   MeSH
• glykoprotein S, koronavirus imunologie   MeSH
• glykosylace MeSH
• HIV infekce imunologie   MeSH
• HIV protilátky imunologie   MeSH
• HIV-1 imunologie   MeSH
• imunoglobuliny - Fab fragmenty chemie   imunologie   MeSH
• lidé MeSH
• Macaca mulatta MeSH
• neutralizující protilátky imunologie   MeSH
• polysacharidy chemie   imunologie   MeSH
• receptory antigenů B-buněk chemie   MeSH
• SARS-CoV-2 imunologie   MeSH
• široce neutralizující protilátky imunologie   MeSH
• vakcíny imunologie   MeSH
• virus opičí imunodeficience genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Studies of molecular and cellular interactions involved in the pathogenesis of IgA nephropathy have revealed the autoimmune nature of this most common primary glomerulonephritis. In patients with this disease, altered glycan structures in the unique hinge region of the heavy chains of IgA1 molecules lead to the exposure of antigenic determinants, which are recognized by naturally occurring antiglycan antibodies of the IgG and/or IgA1 isotype. As a result, nephritogenic immune complexes form in the circulation and deposit in the glomerular mesangium. Deposited immune complexes induce proliferation of resident mesangial cells, increased production of extracellular matrix proteins and cytokines, and ultimately loss of glomerular function. Structural elucidation of the nature of these immune complexes and their biological activity should provide a rational basis for an effective, immunologically mediated inhibition of the formation of nephritogenic immune complexes that could be used as a disease-specific therapeutic approach.

• MeSH
• epitopy imunologie   metabolismus   MeSH
• glomerulární mesangium imunologie   metabolismus   MeSH
• IgA nefropatie * imunologie   patologie   MeSH
• imunoglobulin A imunologie   metabolismus   MeSH
• imunokomplex imunologie   metabolismus   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• sekvence aminokyselin MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH