@misc{MTMT:35067737, title = {LNP-mRNS vakcinák pszeudoallergiás és proinflammatorikus hatásainak vizsgálata sertés hiperszenzitivitási modellben}, url = {https://m2.mtmt.hu/api/publication/35067737}, author = {Dézsi, László and Kökény, Gábor and Révész, Csaba and Mészáros, Tamás and Bakos, Tamás and Kozma, Gergely Tibor and Dobos, Attila Balázs and Merkely, Béla Péter and Radovits, Tamás and Szebeni, János}, unique-id = {35067737}, abstract = {Bevezetés Korábban kimutattuk, hogy COVID-19 védőoltásként alkalmazott LNP-mRNS vakcinák sertés komplement-aktivációval kapcsolatos pszeudoallergiás (CARPA) modellben - humán adáshoz hasonló - korai hiperszenzitivitási reakcióhoz vezetnek. A jelenlegi vizsgálatban a fentiek mellett az LNP-mRNS vakcinák korai és késői proinflammatorikus mellékhatásait teszteltük. Anyag és módszer Kísérleteinkben a Pfizer SARS-CoV-2 elleni monovalens Comirnaty (CMT) és a bivalens Comirnaty/Omikron vakcinát altatott sertéseknek (n=10) 5x humán dózisban, 3-szori ismétléssel i.v. adtuk. Ezen kívül pegilált üres liposzóma (doxebo) előkezelés immunogén hatását, valamint az ismételt vakcinaadás általi tachyphylaxis (saját-tolerancia) kialakulását is vizsgáltuk. A vakcinabeadások előtt és után vérmintákat vettünk, melyekből perifériás mononukleáris sejteket (PBMC) izoláltunk. A kísérletek végén több szervből (pl. szív, vese, máj, lép, agy) szövetmintákat vettünk. A PBMC-ben, ill. a szövetekben a vakcinabeadást követő gyulladásos citokinek és a tüskefehérje (SP) expresszióját az őket kódoló mRNS szekvenciák qPCR módszerrel történő kimutatása alapján elemeztük. A paraméterek változását akut és krónikus (6-8 hét) kísérletekben (n=6) is követtük. Eredmények Az akut pszeudoallergiás reakció létrejöttét mindkét vakcina adásakor a sertés CARPA modell több (pl. hemodinamikai, hematológiai) markere jelezte. Doxebo előkezelés az anaphylaxiás sokk gyakoriságát növelte. Ismételt CMT adáskor esetenként tachypylaxist láttunk, a többi állatban mindhárom vakcinabeadás CARPA reakciót váltott ki. CMT adása PBMC-ben már a 15. percben gyulladásos citokinek mRNS expresszióját indukálta. Az mRNS szintek időbeli változása követte a 3-szori vakcinabeadást, és a kísérlet végén (6 óránál) volt a legmagasabb. CMT adása emellett a PBMC-ben tüskefehérje (SP1) mRNS expresszióhoz vezetett, ami 6 óra után a szívből, veséből, májból, lépből és az agyból vett szövetminták egy részében is kimutatható volt. Krónikus kísérleteinkben CMT adása után akár 2 hónappal az állatok szívében és veséjében kismértékű, de egyértelmű SP1 mRNS expressziót találtunk. Egyes állatokban immunfestéssel SP1-pozitív vesetubulusokat is kimutattunk. Következtetés Eredményeink szerint monovalens és bivalens CMT vakcina is pszeudolallergiás reakciót váltott ki. Emellett PBMC-ben a vakcinabeadás után pár perccel, a szövetmintákban 6 óra múlva citokin, ill. SP mRNS expressziót láttunk, ami akár 2 hónap után is fennállt. Ezáltal sertéseken a CMT adást kísérő gyulladásos mellékhatásokat sikeresen tudtuk modellezni.}, year = {2024}, pages = {4}, orcid-numbers = {Dézsi, László/0000-0002-4190-9793; Kökény, Gábor/0000-0002-0345-6914; Révész, Csaba/0000-0001-6016-526X; Bakos, Tamás/0000-0002-0569-8343; Merkely, Béla Péter/0000-0001-6514-0723} } @article{MTMT:34877474, title = {Comirnaty-induced cardiopulmonary distress and other symptoms of complement-mediated pseudo-anaphylaxis in a hyperimmune pig model: Causal role of anti-PEG antibodies}, url = {https://m2.mtmt.hu/api/publication/34877474}, author = {Barta, Bálint András and Radovits, Tamás and Dobos, Attila Balázs and Kozma, Gergely Tibor and Mészáros, Tamás and Berényi, Petra and Facskó, Réka and Fulop, Tamas and Merkely, Béla Péter and Szebeni, János}, doi = {10.1016/j.jvacx.2024.100497}, journal-iso = {VACCINE X}, journal = {VACCINE: X}, volume = {19}, unique-id = {34877474}, issn = {2590-1362}, year = {2024}, eissn = {2590-1362}, orcid-numbers = {Facskó, Réka/0000-0002-5611-0597; Merkely, Béla Péter/0000-0001-6514-0723} } @article{MTMT:34801936, title = {Flow Cytometry-Based Assay to Detect Alpha Galactosidase Enzymatic Activity at the Cellular Level}, url = {https://m2.mtmt.hu/api/publication/34801936}, author = {Fekete, Nóra and Li, Luca Kamilla and Kozma, Gergely Tibor and Fekete, György and Pállinger, Éva and Kovács, Árpád Ferenc}, doi = {10.3390/cells13080706}, journal-iso = {CELLS-BASEL}, journal = {CELLS}, volume = {13}, unique-id = {34801936}, abstract = {Background: Fabry disease is a progressive, X chromosome-linked lysosomal storage disorder with multiple organ dysfunction. Due to the absence or reduced activity of alpha-galactosidase A (AGAL), glycosphingolipids, primarily globotriaosyl-ceramide (Gb3), concentrate in cells. In heterozygous women, symptomatology is heterogenous and currently routinely used fluorometry-based assays measuring mean activity mostly fail to uncover AGAL dysfunction. The aim was the development of a flow cytometry assay to measure AGAL activity in individual cells. Methods: Conventional and multispectral imaging flow cytometry was used to detect AGAL activity. Specificity was validated using the GLA knockout (KO) Jurkat cell line and AGAL inhibitor 1-deoxygalactonojirimycin. The GLA KO cell line was generated via CRISPR-Cas9-based transfection, validated with exome sequencing, gene expression and substrate accumulation. Results: Flow cytometric detection of specific AGAL activity is feasible with fluorescently labelled Gb3. In the case of Jurkat cells, a substrate concentration of 2.83 nmol/mL and 6 h of incubation are required. Quenching of the aspecific exofacial binding of Gb3 with 20% trypan blue solution is necessary for the specific detection of lysosomal substrate accumulation. Conclusion: A flow cytometry-based assay was developed for the quantitative detection of AGAL activity at the single-cell level, which may contribute to the diagnosis of Fabry patients.}, year = {2024}, eissn = {2073-4409}, orcid-numbers = {Fekete, György/0000-0002-5312-2541; Pállinger, Éva/0000-0002-5789-0951; Kovács, Árpád Ferenc/0000-0002-7742-160X} } @article{MTMT:34754128, title = {mRNA-LNP COVID-19 Vaccine Lipids Induce Complement Activation and Production of Proinflammatory Cytokines: Mechanisms, Effects of Complement Inhibitors, and Relevance to Adverse Reactions}, url = {https://m2.mtmt.hu/api/publication/34754128}, author = {Bakos, Tamás and Mészáros, Tamás and Kozma, Gergely Tibor and Berényi, Petra and Facskó, Réka and Farkas, Henriette and Dézsi, László and Heirman, Carlo and de Koker, Stefaan and Schiffelers, Raymond and Glatter, Kathryn Anne and Radovits, Tamás and Szénási, Gábor and Szebeni, János}, doi = {10.3390/ijms25073595}, journal-iso = {INT J MOL SCI}, journal = {INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES}, volume = {25}, unique-id = {34754128}, issn = {1661-6596}, abstract = {A small fraction of people vaccinated with mRNA–lipid nanoparticle (mRNA-LNP)-based COVID-19 vaccines display acute or subacute inflammatory symptoms whose mechanism has not been clarified to date. To better understand the molecular mechanism of these adverse events (AEs), here, we analyzed in vitro the vaccine-induced induction and interrelations of the following two major inflammatory processes: complement (C) activation and release of proinflammatory cytokines. Incubation of Pfizer-BioNTech’s Comirnaty and Moderna’s Spikevax with 75% human serum led to significant increases in C5a, sC5b-9, and Bb but not C4d, indicating C activation mainly via the alternative pathway. Control PEGylated liposomes (Doxebo) also induced C activation, but, on a weight basis, it was ~5 times less effective than that of Comirnaty. Viral or synthetic naked mRNAs had no C-activating effects. In peripheral blood mononuclear cell (PBMC) cultures supplemented with 20% autologous serum, besides C activation, Comirnaty induced the secretion of proinflammatory cytokines in the following order: IL-1α < IFN-γ < IL-1β < TNF-α < IL-6 < IL-8. Heat-inactivation of C in serum prevented a rise in IL-1α, IL-1β, and TNF-α, suggesting C-dependence of these cytokines’ induction, although the C5 blocker Soliris and C1 inhibitor Berinert, which effectively inhibited C activation in both systems, did not suppress the release of any cytokines. These findings suggest that the inflammatory AEs of mRNA-LNP vaccines are due, at least in part, to stimulation of both arms of the innate immune system, whereupon C activation may be causally involved in the induction of some, but not all, inflammatory cytokines. Thus, the pharmacological attenuation of inflammatory AEs may not be achieved via monotherapy with the tested C inhibitors; efficacy may require combination therapy with different C inhibitors and/or other anti-inflammatory agents.}, year = {2024}, eissn = {1422-0067}, orcid-numbers = {Bakos, Tamás/0000-0002-0569-8343; Facskó, Réka/0000-0002-5611-0597; Farkas, Henriette/0000-0003-2929-1721; Dézsi, László/0000-0002-4190-9793; Szénási, Gábor/0000-0002-7350-6091} } @misc{MTMT:34521339, title = {mRNA-LNP COVID-19 vaccine lipids induce low level complement activation and production of proinflammatory cytokines: Mechanisms, effects of complement inhibitors, and relevance to adverse reactions}, url = {https://m2.mtmt.hu/api/publication/34521339}, author = {Bakos, Tamás and Mészáros, Tamás and Kozma, Gergely Tibor and Petra, Berényi and Facskó, Réka and Henriette, Farkas and Dézsi, László and Carlo, Heirman and Stefaan, de Koker and Raymond, Schiffelers and Kathryn, Anne Glatter and Radovits, Tamás and Szénási, Gábor and Szebeni, János}, unique-id = {34521339}, abstract = {Messenger RNA-containing lipid nanoparticles (mRNA-LNPs) enabled widespread COVID-19 vaccination with a small fraction of vaccine recipients displaying acute or sub-acute inflammatory symptoms. The molecular mechanism of these adverse events (AEs) remains undetermined. Here we report that the mRNA-LNP vaccine, Comirnaty, triggers low-level complement (C) activation and production of inflammatory cytokines, which may be key underlying processes of inflammatory AEs. In serum, Comirnaty and the control PEGylated liposome (Doxebo) caused different rises of C split products, C5a, sC5b-9, Bb and C4d, indicating stimulation of the classical pathway of C activation mainly by the liposomes, while a stronger stimulation of the alternative pathway was equal with the vaccine and the liposomes. Spikevax had similar C activation as Comirnaty, but viral or synthetic mRNAs had no such effect. In autologous serum-supplemented peripheral blood mononuclear cell (PBMC) cultures, Comirnaty caused increases in the levels of sC5b-9 and proinflammatory cytokines in the following order: IL-1α < IFN-γ < IL-1β < TNF-α < IL-6 < IL-8, whereas heatinactivation of serum prevented the rises of IL-1α, IL-1β, and TNF-α. Clinical C inhibitors, Soliris and Berinert, suppressed vaccine-induced C activation in serum but did not affect cytokine production when applied individually. These findings suggest that the PEGylated lipid coating of mRNA-LNP nanoparticles can trigger C activation mainly via the alternative pathway, which may be causally related to the induction of some, but not all inflammatory cytokines. While innate immune stimulation is essential for the vaccine’s efficacy, concurrent production of C- and PBMC-derived inflammatory mediators may contribute to some of the AEs. Pharmacological attenuation of harmful cytokine production using C inhibitors likely requires blocking the C cascade at multiple points.}, year = {2024}, pages = {&}, orcid-numbers = {Bakos, Tamás/0000-0002-0569-8343; Facskó, Réka/0000-0002-5611-0597; Dézsi, László/0000-0002-4190-9793; Szénási, Gábor/0000-0002-7350-6091} } @article{MTMT:34095902, title = {Eculizumab suppresses zymosan-induced release of inflammatory cytokines IL-1α, IL-1β, IFN-γ and IL-2 in autologous serum-substituted PBMC cultures: Relevance to cytokine storm in Covid-19}, url = {https://m2.mtmt.hu/api/publication/34095902}, author = {Bakos, Tamás and Kozma, Gergely Tibor and Szebeni, János and Szénási, Gábor}, doi = {10.1016/j.biopha.2023.115294}, journal-iso = {BIOMED PHARMACOTHER}, journal = {BIOMEDICINE & PHARMACOTHERAPY}, volume = {166}, unique-id = {34095902}, issn = {0753-3322}, year = {2023}, eissn = {1950-6007}, orcid-numbers = {Bakos, Tamás/0000-0002-0569-8343; Szénási, Gábor/0000-0002-7350-6091} } @article{MTMT:34039197, title = {Role of anti-polyethylene glycol (PEG) antibodies in the allergic reactions to PEG-containing Covid-19 vaccines: Evidence for immunogenicity of PEG}, url = {https://m2.mtmt.hu/api/publication/34039197}, author = {Kozma, Gergely Tibor and Mészáros, Tamás and Berényi, Petra and Facskó, Réka and Patkó, Zsófia Panna and Oláh, Csaba Zsolt and Nagy, Adrienne and Fülöp, Tamás Gyula and Glatter, Kathryn Anne and Radovits, Tamás and Merkely, Béla Péter and Szebeni, János}, doi = {10.1016/j.vaccine.2023.06.009}, journal-iso = {VACCINE}, journal = {VACCINE}, volume = {41}, unique-id = {34039197}, issn = {0264-410X}, year = {2023}, eissn = {1873-2518}, pages = {4561-4570}, orcid-numbers = {Facskó, Réka/0000-0002-5611-0597; Merkely, Béla Péter/0000-0001-6514-0723} } @article{MTMT:33548672, title = {Zymosan Particle-Induced Hemodynamic, Cytokine and Blood Cell Changes in Pigs: An Innate Immune Stimulation Model with Relevance to Cytokine Storm Syndrome and Severe COVID-19}, url = {https://m2.mtmt.hu/api/publication/33548672}, author = {Kökény, Gábor and Bakos, Tamás and Barta, Bálint András and Nagy, Georgina Viktória and Mészáros, Tamás and Kozma, Gergely Tibor and Szabó, András and Szebeni, János and Merkely, Béla Péter and Radovits, Tamás}, doi = {10.3390/ijms24021138}, journal-iso = {INT J MOL SCI}, journal = {INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES}, volume = {24}, unique-id = {33548672}, issn = {1661-6596}, abstract = {Hemodynamic disturbance, a rise in neutrophil-to-lymphocyte ratio (NLR) and release of inflammatory cytokines into blood, is a bad prognostic indicator in severe COVID-19 and other diseases involving cytokine storm syndrome (CSS). The purpose of this study was to explore if zymosan, a known stimulator of the innate immune system, could reproduce these changes in pigs. Pigs were instrumented for hemodynamic analysis and, after i.v. administration of zymosan, serial blood samples were taken to measure blood cell changes, cytokine gene transcription in PBMC and blood levels of inflammatory cytokines, using qPCR and ELISA. Zymosan bolus (0.1 mg/kg) elicited transient hemodynamic disturbance within minutes without detectable cytokine or blood cell changes. In contrast, infusion of 1 mg/kg zymosan triggered maximal pulmonary hypertension with tachycardia, lasting for 30 min. This was followed by a transient granulopenia and then, up to 6 h, major granulocytosis, resulting in a 3–4-fold increase in NLR. These changes were paralleled by massive transcription and/or rise in IL-6, TNF-alpha, CCL-2, CXCL-10, and IL-1RA in blood. There was significant correlation between lymphopenia and IL-6 gene expression. We conclude that the presented model may enable mechanistic studies on late-stage COVID-19 and CSS, as well as streamlined drug testing against these conditions.}, year = {2023}, eissn = {1422-0067}, orcid-numbers = {Kökény, Gábor/0000-0002-0345-6914; Bakos, Tamás/0000-0002-0569-8343; Szabó, András/0000-0001-8320-4946; Merkely, Béla Péter/0000-0001-6514-0723} } @misc{MTMT:33209367, title = {Role of anti-polyethylene glycol (PEG) antibodies in the allergic reactions and immunogenicity of PEG-containing Covid-19 vaccines}, url = {https://m2.mtmt.hu/api/publication/33209367}, author = {Kozma, Gergely Tibor and Mészáros, Tamás and Petra, Berényi and Facskó, Réka and Patkó, Zsófia Panna and Oláh, Csaba Zsolt and Adrienne, Nagy and Tamás, Gyula Fülöp and Kathryn, Anne Glatter and Radovits, Tamás and Merkely, Béla Péter and Szebeni, János}, unique-id = {33209367}, abstract = {The polyethylene-glycol (PEG)-containing Covid-19 vaccines can cause hypersensitivity reactions (HSRs), or rarely, life-threatening anaphylaxis. A causal role of anti-PEG antibodies (Abs) has been proposed, but not yet proven in humans. The 191 blood donors in this study included 10 women and 5 men who displayed HSRs to Comirnaty or Spikevax Covid-19 vaccines with 3 anaphylaxis. 118 donors had pre-vaccination anti-PEG IgG/IgM values as measured by ELISA, of which >98% were over background regardless of age, indicating the presence of these Abs in almost everyone. Their values varied over 2-3 orders of magnitude and displayed strong left-skewed distribution with 3-4% of subjects having >15-30-fold higher values than the respective median. First, or booster injections with both vaccines led to significant rises of anti-PEG IgG/IgM with >10-fold rises in about ∼10% of Comirnaty, and all Spikevax recipients, measured at different times after the injections. The anti-PEG Ab levels measured within 4-months after the HSRs were significantly higher than those in nonreactors. Serial testing of plasma (n=361 tests) showed the SARS-CoV-2 neutralization IgG to vary over a broad range, with a trend for biphasic dose dependence on anti-PEG Abs. The highest prevalence of anti-PEG Ab positivity in human blood reported to date represents new information which can most easily be rationalized by daily exposure to common PEG-containing medications and/or household items. The significantly higher, HSR-non-coincidental blood level of anti-PEG Abs in hypersensitivity reactor vs. non-reactors, taken together with relevant clinical and experimental data in the literature, suggest that anti-PEG Ab supercarrier people might be at increased risk for HSRs to PEG-containing vaccines, which themselves can induce these Abs via bystander immunogenicity. Our data also raise the possibility that anti-PEG Abs might also contribute to the reduction of these vaccines’ virus neutralization efficacy. Thus, screening for anti-PEG Ab supercarriers may identify people at risk for HSRs or reduced vaccine effectiveness.}, year = {2022}, orcid-numbers = {Facskó, Réka/0000-0002-5611-0597; Merkely, Béla Péter/0000-0001-6514-0723} } @article{MTMT:32666145, title = {A naturally hypersensitive porcine model may help understand the mechanism of COVID-19 mRNA vaccine-induced rare (pseudo) allergic reactions: complement activation as a possible contributing factor}, url = {https://m2.mtmt.hu/api/publication/32666145}, author = {Dézsi, László and Mészáros, Tamás and Kozma, Gergely Tibor and H-Velkei, Mária and Oláh, Csaba Zsolt and Szabó, Miklós and Patkó, Zsófia Panna and Fülöp, Tamás and Hennies, Mark and Szebeni, Miklós and Barta, Bálint András and Merkely, Béla Péter and Radovits, Tamás and Szebeni, János}, doi = {10.1007/s11357-021-00495-y}, journal-iso = {GEROSCIENCE}, journal = {GEROSCIENCE: OFFICIAL JOURNAL OF THE AMERICAN AGING ASSOCIATION (AGE)}, volume = {44}, unique-id = {32666145}, issn = {2509-2715}, abstract = {A tiny fraction of people immunized with lipid nanoparticle (LNP)-enclosed mRNA (LNP-mRNA) vaccines develop allergic symptoms following their first or subsequent vaccinations, including anaphylaxis. These reactions resemble complement (C) activation-related pseudoallergy (CARPA) to i.v. administered liposomes, for which pigs provide a naturally oversensitive model. Using this model, we injected i.v. the human vaccination dose (HVD) of BNT162b2 (Comirnaty, CMT) or its 2-fold (2x) or 5-fold (5x) amounts and measured the hemodynamic changes and other parameters of CARPA. We observed in 6 of 14 pigs transient pulmonary hypertension along with thromboxane A2 release into the blood and other hemodynamic and blood cell changes, including hypertension, granulocytosis, lymphopenia, and thrombocytopenia. One pig injected with 5x CMT developed an anaphylactic shock requiring resuscitation, while a repeat dose failed to induce the reaction, implying tachyphylaxis. These typical CARPA symptoms could not be linked to animal age, sex, prior immune stimulation with zymosan, immunization of animals with Comirnaty i.v., or i.m. 2 weeks before the vaccine challenge, and anti-PEG IgM levels in Comirnaty-immunized pigs. Nevertheless, IgM binding to the whole vaccine, used as antigen in an ELISA, was significantly higher in reactive animals compared to non-reactive ones. Incubation of Comirnaty with pig serum in vitro showed significant elevations of C3a anaphylatoxin and sC5b-9, the C-terminal complex. These data raise the possibility that C activation plays a causal or contributing role in the rare HSRs to Comirnaty and other vaccines with similar side effects. Further studies are needed to uncover the factors controlling these vaccine reactions in pigs and to understand their translational value to humans.}, year = {2022}, eissn = {2509-2723}, pages = {597-618}, orcid-numbers = {Dézsi, László/0000-0002-4190-9793; Fülöp, Tamás/0000-0003-2889-1370; Merkely, Béla Péter/0000-0001-6514-0723} }