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Article ; Online: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs.

Gromadzka, Beata / Chraniuk, Milena / Hovhannisyan, Lilit / Uranowska, Karolina / Szewczyk, Bogusław / Narajczyk, Magdalena / Panasiuk, Mirosława

Viruses

2022 Volume 14, Issue 4

Abstract: 1) Background: Avian influenza viruses (AIVs) promptly evade preexisting immunity by constantly altering the immunodominant neutralizing antibody epitopes (antigenic drift) or by procuring new envelope serotypes (antigenic shift). As a consequence, the ... ...

Abstract	(1) Background: Avian influenza viruses (AIVs) promptly evade preexisting immunity by constantly altering the immunodominant neutralizing antibody epitopes (antigenic drift) or by procuring new envelope serotypes (antigenic shift). As a consequence, the majority of antibodies elicited by infection or vaccination protect only against closely related strains. The immunodominance of the globular head of the main glycoprotein has been shown to mask the immunogenicity of the conserved regions located within the hemagglutinin (HA) protein. It has been shown that the broadly neutralizing universal antibodies recognize the HA2 domain in headless hemagglutinin (HA-stalk). Therefore, the HA-stalk is a highly conserved antigen, which makes it a good candidate to be used in universal vaccine development against AIVs. (2) Methods: Sf9 insect cells were used to produce triple H5N1/NA-HA-M1 influenza virus-like particles (VLPs) via co-expression of neuraminidase, hemagglutinin and matrix proteins from a tricistronic expression cassette. Purified influenza VLPs were used to immunize broiler hens. An in-depth characterization of the immune response was performed with an emphasis on the pool of elicited universal antibodies. (3) Results: Our findings suggest, that after vaccination with triple H5N1/NA-HA-M1 VLPs, hens generate a pool of broad-spectrum universal anti-HA-stalk antibodies. Furthermore, these universal antibodies are able to recognize the mammalian-derived HA-stalk recombinant proteins from homologous H5N1 and heterologous H7N9 AIVs as well as from the heterosubtypic human H1N1 influenza strain. (4) Conclusions: Our findings may suggest that highly pathogenic avian influenza H5 HA protein contain functional epitopes that are attractive targets for the generation of broad-spectrum antibodies against AIVs in their native hosts.
MeSH term(s)	Animals ; Antibodies, Viral ; Chickens ; Epitopes ; Female ; Hemagglutinin Glycoproteins, Influenza Virus/genetics ; Hemagglutinins ; Humans ; Immunity ; Influenza A Virus, H1N1 Subtype ; Influenza A Virus, H5N1 Subtype ; Influenza A Virus, H7N9 Subtype ; Influenza Vaccines ; Influenza in Birds/prevention & control ; Influenza, Human/prevention & control ; Mammals ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections ; Vaccination/veterinary
Chemical Substances	Antibodies, Viral ; Epitopes ; Hemagglutinin Glycoproteins, Influenza Virus ; Hemagglutinins ; Influenza Vaccines
Language	English
Publishing date	2022-03-30
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v14040730
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Functional Severe Acute Respiratory Syndrome Coronavirus 2 Virus-Like Particles From Insect Cells.

Naskalska, Antonina / Dabrowska, Agnieszka / Szczepanski, Artur / Jasik, Krzysztof P / Gromadzka, Beata / Pyrc, Krzysztof

Frontiers in microbiology

2021 Volume 12, Page(s) 732998

Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a major epidemic threat since the beginning of 2020. Efforts to combat the virus and the associated coronavirus disease 2019 (COVID-19) disease are being undertaken worldwide. To ... ...

Abstract	Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a major epidemic threat since the beginning of 2020. Efforts to combat the virus and the associated coronavirus disease 2019 (COVID-19) disease are being undertaken worldwide. To facilitate the research on the virus itself, a number of surrogate systems have been developed. Here, we report the efficient production of SARS-CoV-2 virus-like particles (VLPs) in insect cells. Contrary to widely used pseudovirus particles, where only one coronaviral protein is displayed within a heterologous scaffold, developed VLPs are structurally similar to the native virus and allow for more throughput studies on the biology of the infection. On the other hand, being devoid of the viral genome, VLPs are unable to replicate and thus safe to work with. Importantly, this is the first report showing that SARS-CoV-2 VLPs can be efficiently produced in insect cells and purified using scalable affinity chromatography.
Language	English
Publishing date	2021-10-20
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2021.732998
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Article ; Online: The Preliminary Assessment of New Biomaterials Necessitates a Comparison of Direct and Indirect Cytotoxicity Methodological Approaches.

Chraniuk, Milena / Panasiuk, Mirosława / Hovhannisyan, Lilit / Żołędowska, Sabina / Nidzworski, Dawid / Ciołek, Lidia / Woźniak, Anna / Jaegermann, Zbigniew / Biernat, Monika / Gromadzka, Beata

Polymers

2022 Volume 14, Issue 21

Abstract: Background: Cytotoxicity testing is a primary method to establish the safety of biomaterials, e.g., biocomposites. Biomaterials involve a wide range of medical materials, which are usually solid materials and are used in bone regeneration, cardiology, ... ...

Abstract	Background: Cytotoxicity testing is a primary method to establish the safety of biomaterials, e.g., biocomposites. Biomaterials involve a wide range of medical materials, which are usually solid materials and are used in bone regeneration, cardiology, or dermatology. Current advancements in science and technology provide several standard cytotoxicity testing methods that are sufficiently sensitive to detect various levels of cellular toxicity, i.e., from low to high. The aim was to compare the direct and indirect methodology described in the ISO guidelines UNE-EN ISO 10993-5:2009 Part 5. Methods: Cell proliferation was measured using WST-1 assay, and cytotoxicity was measured using LDH test kit. Results: The results indicate that the molecular surface of biomaterials have impact on the cytotoxicity and proliferation profile. Based on these results, we confirm that the indirect method does not provide a clear picture of the cell condition after the exposure to the surface, and moreover, cannot provide complete results about the effects of the material. Conclusions: Comparison of both methods shows that it is pivotal to investigate biomaterials at the very early stages using both indirect and direct methods to access the influence of the released toxins and surface of the material on the cell condition.
Language	English
Publishing date	2022-10-25
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527146-5
ISSN	2073-4360 ; 2073-4360
ISSN (online)	2073-4360
ISSN	2073-4360
DOI	10.3390/polym14214522
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Article ; Online: Effect of Selected Crosslinking and Stabilization Methods on the Properties of Porous Chitosan Composites Dedicated for Medical Applications.

Biernat, Monika / Woźniak, Anna / Chraniuk, Milena / Panasiuk, Mirosława / Tymowicz-Grzyb, Paulina / Pagacz, Joanna / Antosik, Agnieszka / Ciołek, Lidia / Gromadzka, Beata / Jaegermann, Zbigniew

Polymers

2023 Volume 15, Issue 11

Abstract: Chitosan is one of the most commonly employed natural polymers for biomedical applications. However, in order to obtain stable chitosan biomaterials with appropriate strength properties, it is necessary to subject it to crosslinking or stabilization. ... ...

Abstract	Chitosan is one of the most commonly employed natural polymers for biomedical applications. However, in order to obtain stable chitosan biomaterials with appropriate strength properties, it is necessary to subject it to crosslinking or stabilization. Composites based on chitosan and bioglass were prepared using the lyophilization method. In the experimental design, six different methods were used to obtain stable, porous chitosan/bioglass biocomposite materials. This study compared the crosslinking/stabilization of chitosan/bioglass composites with ethanol, thermal dehydration, sodium tripolyphosphate, vanillin, genipin, and sodium β-glycerophosphate. The physicochemical, mechanical, and biological properties of the obtained materials were compared. The results showed that all the selected crosslinking methods allow the production of stable, non-cytotoxic porous composites of chitosan/bioglass. The composite with genipin stood out with the best of the compared properties, taking into account biological and mechanical characteristics. The composite stabilized with ethanol is distinct in terms of its thermal properties and swelling stability, and it also promotes cell proliferation. Regarding the specific surface area, the highest value exposes the composite stabilized by the thermal dehydration method.
Language	English
Publishing date	2023-05-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527146-5
ISSN	2073-4360 ; 2073-4360
ISSN (online)	2073-4360
ISSN	2073-4360
DOI	10.3390/polym15112507
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Article: Characterization of surface-exposed structural loops as insertion sites for foreign antigen delivery in calicivirus-derived VLP platform.

Panasiuk, Mirosława / Chraniuk, Milena / Zimmer, Karolina / Hovhannisyan, Lilit / Krapchev, Vasil / Peszyńska-Sularz, Grażyna / Narajczyk, Magdalena / Węsławski, Jan / Konopacka, Agnieszka / Gromadzka, Beata

Frontiers in microbiology

2023 Volume 14, Page(s) 1111947

Abstract: Chimeric virus-like particles (cVLPs) show great potential in improving public health as they are safe and effective vaccine candidates. The capsid protein of caliciviruses has been described previously as a self-assembling, highly immunogenic delivery ... ...

Abstract	Chimeric virus-like particles (cVLPs) show great potential in improving public health as they are safe and effective vaccine candidates. The capsid protein of caliciviruses has been described previously as a self-assembling, highly immunogenic delivery platform. The ability to significantly induce cellular and humoral immunity can be used to boost the immune response to low immunogenic foreign antigens displayed on the surface of VLPs. Capsid proteins of caliciviruses despite sequence differences share similar architecture with structural loops that can be genetically modified to present foreign epitopes on the surface of cVLPs. Here, based on the VP1 protein of norovirus (NoV), we investigated the impact of the localization of the epitope in different structural loops of the P domain on the immunogenicity of the presented epitope. In this study, three distinct loops of NoV VP1 protein were genetically modified to present a multivalent influenza virus epitope consisting of a tandem repeat of M2/NP epitopes. cVLPs presenting influenza virus-conserved epitopes in different localizations were produced in the insect cells and used to immunize BALB/c mice. Specific reaction to influenza epitopes was compared in sera from vaccinated mice to determine whether the localization of the foreign epitope has an impact on the immunogenicity.
Language	English
Publishing date	2023-02-27
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2023.1111947
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Article ; Online: Low-volume label-free SARS-CoV-2 detection with the microcavity-based optical fiber sensor.

Janik, Monika / Gabler, Tomasz / Koba, Marcin / Panasiuk, Mirosława / Dashkevich, Yanina / Łęga, Tomasz / Dąbrowska, Agnieszka / Naskalska, Antonina / Żołędowska, Sabina / Nidzworski, Dawid / Pyrć, Krzysztof / Gromadzka, Beata / Śmietana, Mateusz

Scientific reports

2023 Volume 13, Issue 1, Page(s) 1512

Abstract: Accurate and fast detection of viruses is crucial for controlling outbreaks of many diseases; therefore, to date, numerous sensing systems for their detection have been studied. On top of the performance of these sensing systems, the availability of ... ...

Abstract	Accurate and fast detection of viruses is crucial for controlling outbreaks of many diseases; therefore, to date, numerous sensing systems for their detection have been studied. On top of the performance of these sensing systems, the availability of biorecognition elements specific to especially the new etiological agents is an additional fundamental challenge. Therefore, besides high sensitivity and selectivity, such advantages as the size of the sensor and possibly low volume of analyzed samples are also important, especially at the stage of evaluating the receptor-target interactions in the case of new etiological agents when typically, only tiny amounts of the receptor are available for testing. This work introduces a real-time, highly miniaturized sensing solution based on microcavity in-line Mach-Zehnder interferometer (μIMZI) induced in optical fiber for SARS-CoV-2 virus-like particles detection. The assay is designed to detect conserved regions of the SARS-CoV-2 viral particles in a sample with a volume as small as hundreds of picoliters, reaching the detection limit at the single ng per mL level.
MeSH term(s)	Humans ; Optical Fibers ; SARS-CoV-2 ; Interferometry ; Biosensing Techniques ; COVID-19/diagnosis
Language	English
Publishing date	2023-01-27
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-28790-y
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Article ; Online: Assessment of the Toxicity of Biocompatible Materials Supporting Bone Regeneration: Impact of the Type of Assay and Used Controls.

Chraniuk, Milena / Panasiuk, Mirosława / Hovhannisyan, Lilit / Żołędowska, Sabina / Nidzworski, Dawid / Ciołek, Lidia / Woźniak, Anna / Kubiś, Agnieszka / Karska, Natalia / Jaegermann, Zbigniew / Rodziewicz-Motowidło, Sylwia / Biernat, Monika / Gromadzka, Beata

Toxics

2022 Volume 10, Issue 1

Abstract: Assessing the toxicity of new biomaterials dedicated to bone regeneration can be difficult. Many reports focus only on a single toxicity parameter, which may be insufficient for a detailed evaluation of the new material. Moreover, published data ... ...

Abstract	Assessing the toxicity of new biomaterials dedicated to bone regeneration can be difficult. Many reports focus only on a single toxicity parameter, which may be insufficient for a detailed evaluation of the new material. Moreover, published data frequently do not include control cells exposed to the environment without composite or its extract. Here we present the results of two assays used in the toxicological assessment of materials' extracts (the integrity of the cellular membrane and the mitochondrial activity/proliferation), and the influence of different types of controls used on the obtained results. Results obtained in the cellular membrane integrity assay showed a lack of toxic effects of all tested extracts, and no statistical differences between them were present. Control cells, cells incubated with chitosan extract or chitosan-bioglass extract were used as a reference in proliferation calculations to highlight the impact of controls used on the result of the experiment. The use of different baseline controls caused variability between obtained proliferation results, and influenced the outcome of statistical analysis. Our findings confirm the thesis that the type of control used in an experiment can change the final results, and it may affect the toxicological assessment of biomaterial.
Language	English
Publishing date	2022-01-06
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2733883-6
ISSN	2305-6304 ; 2305-6304
ISSN (online)	2305-6304
ISSN	2305-6304
DOI	10.3390/toxics10010020
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Article ; Online: Chimeric virus-like particles presenting tumour-associated MUC1 epitope result in high titers of specific IgG antibodies in the presence of squalene oil-in-water adjuvant: towards safe cancer immunotherapy.

Panasiuk, Mirosława / Zimmer, Karolina / Czarnota, Anna / Narajczyk, Magdalena / Peszyńska-Sularz, Grażyna / Chraniuk, Milena / Hovhannisyan, Lilit / Żołędowska, Sabina / Nidzworski, Dawid / Żaczek, Anna J / Gromadzka, Beata

Journal of nanobiotechnology

2022 Volume 20, Issue 1, Page(s) 160

Abstract: Background: Immunotherapy is emerging as a powerful treatment approach for several types of cancers. Modulating the immune system to specifically target cancer cells while sparing healthy cells, is a very promising approach for safer therapies and ... ...

Abstract	Background: Immunotherapy is emerging as a powerful treatment approach for several types of cancers. Modulating the immune system to specifically target cancer cells while sparing healthy cells, is a very promising approach for safer therapies and increased survival of cancer patients. Tumour-associated antigens are favorable targets for cancer immunotherapy, as they are exclusively expressed by the cancer cells, minimizing the risk of an autoimmune reaction. The ability to initiate the activation of the immune system can be achieved by virus-like particles (VLPs) which are safe and potent delivery tools. VLP-based vaccines have evolved dramatically over the last few decades and showed great potential in preventing infectious diseases. Immunogenic potency of engineered VLPs as a platform for the development of effective therapeutic cancer vaccines has been studied extensively. This study involves recombinant VLPs presenting multiple copies of tumour-specific mucin 1 (MUC1) epitope as a potentially powerful tool for future immunotherapy. Results: In this report VLPs based on the structural protein of Norovirus (NoV VP1) were genetically modified to present multiple copies of tumour-specific MUC1 epitope on their surface. Chimeric MUC1 particles were produced in the eukaryotic Leishmania tarentolae expression system and used in combination with squalene oil-in-water emulsion MF59 adjuvant to immunize BALB/c mice. Sera from vaccinated mice demonstrated high titers of IgG and IgM antibodies which were specifically recognizing MUC1 antigen. Conclusions: The obtained results show that immunization with recombinant chimeric NoV VP1- MUC1 VLPs result in high titers of MUC1 specific IgG antibodies and show great therapeutic potential as a platform to present tumour-associated antigens.
MeSH term(s)	Animals ; Epitopes ; Humans ; Immunization ; Immunoglobulin G ; Mice ; Mucin-1 ; Neoplasms/therapy ; Squalene ; Water
Chemical Substances	Epitopes ; Immunoglobulin G ; MUC1 protein, human ; Mucin-1 ; Water (059QF0KO0R) ; Squalene (7QWM220FJH)
Language	English
Publishing date	2022-03-27
Publishing country	England
Document type	Journal Article
ZDB-ID	2100022-0
ISSN	1477-3155 ; 1477-3155
ISSN (online)	1477-3155
ISSN	1477-3155
DOI	10.1186/s12951-022-01357-1
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Article ; Online: A multiplex real-time PCR assay for detection of oseltamivir-resistant strains of influenza virus

Nidzworski Dawid / Dobkowska Joanna / Hołysz Marcin / Gromadzka Beata / Szewczyk Bogusław

Open Life Sciences, Vol 9, Iss 6, Pp 628-

2014 Volume 633

Keywords	influenza ; detection ; oseltamivir ; h275y ; real-time pcr ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2014-06-01T00:00:00Z
Publisher	De Gruyter
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Immunization with Leishmania tarentolae-derived norovirus virus-like particles elicits high humoral response and stimulates the production of neutralizing antibodies.

Panasiuk, Mirosława / Zimmer, Karolina / Czarnota, Anna / Grzyb, Katarzyna / Narajczyk, Magdalena / Peszyńska-Sularz, Grażyna / Żołędowska, Sabina / Nidzworski, Dawid / Hovhannisyan, Lilit / Gromadzka, Beata

Microbial cell factories

2021 Volume 20, Issue 1, Page(s) 186

Abstract: Background: Noroviruses are a major cause of epidemic and sporadic acute non-bacterial gastroenteritis worldwide. Unfortunately, the development of an effective norovirus vaccine has proven difficult and no prophylactic vaccine is currently available. ... ...

Abstract	Background: Noroviruses are a major cause of epidemic and sporadic acute non-bacterial gastroenteritis worldwide. Unfortunately, the development of an effective norovirus vaccine has proven difficult and no prophylactic vaccine is currently available. Further research on norovirus vaccine development should be considered an absolute priority and novel vaccine candidates are needed. One of the recent approaches in safe vaccine development is the use of virus-like particles (VLPs). VLP-based vaccines show great immunogenic potential as they mimic the morphology and structure of viral particles without the presence of the virus genome. Results: This study is the first report showing successful production of norovirus VLPs in the protozoan Leishmania tarentolae (L. tarentolae) expression system. Protozoan derived vaccine candidate is highly immunogenic and able to not only induce a strong immune response (antibody titer reached 10 Conclusions: Norovirus VLPs produced in L. tarentolae could be relevant for the development of the norovirus vaccine.
MeSH term(s)	Animals ; Antibodies, Neutralizing/blood ; Immunization ; Immunoglobulin G/blood ; Leishmania/genetics ; Leishmania/immunology ; Leishmania/virology ; Male ; Mice ; Mice, Inbred BALB C ; Norovirus/genetics ; Norovirus/immunology ; Vaccine Development ; Vaccines, Virus-Like Particle/administration & dosage ; Vaccines, Virus-Like Particle/immunology ; Viral Vaccines/administration & dosage ; Viral Vaccines/genetics ; Viral Vaccines/immunology
Chemical Substances	Antibodies, Neutralizing ; Immunoglobulin G ; Vaccines, Virus-Like Particle ; Viral Vaccines
Language	English
Publishing date	2021-09-24
Publishing country	England
Document type	Journal Article
ISSN	1475-2859
ISSN (online)	1475-2859
DOI	10.1186/s12934-021-01677-1
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