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1. Gene network landscape of mouse splenocytes reveals integrin complex as the A151 ODN-responsive hub molecule in the immune transcriptome

Author

Volkan Yazar, Ismail Cem Yilmaz, Artun Bulbul, Dennis M. Klinman, and Ihsan Gursel

Subjects
MT: Bioinformatics, A151 oligodeoxynucleotide, immune suppression, Itgam, Itga6, integrin, Therapeutics. Pharmacology, RM1-950
Abstract

Homeostatic restoration of an inflammatory response requires quenching of the immune system after pathogen threats vanish. A continued assault orchestrated by host defense results in tissue destruction or autoimmunity. A151 is the epitome of synthetic oligodeoxynucleotides (ODNs) that curb the immune response by a subset of white corpuscles through repetitive telomere-derived TTAGGG sequences. Currently, the genuine effect of A151 on the immune cell transcriptome remains unknown. Here, we leveraged an integrative approach where weighted gene co-expression network analysis (WGCNA), differential gene expression analysis, and gene set enrichment analysis (GSEA) of our in-house microarray datasets aided our understanding of how A151 ODN suppresses the immune response in mouse splenocytes. Our bioinformatics results, together with experimental validations, indicated that A151 ODN acts on components of integrin complexes, Itgam and Itga6, to interfere with immune cell adhesion and thereby suppresses the immune response in mice. Moreover, independent lines of evidence in this work converged on the observation that cell adhesion by integrin complexes serves as a focal point for cellular response to A151 ODN treatment in immune cells. Taken together, the outcome of this study sheds light on the molecular basis of immune suppression by a clinically useful DNA-based therapeutic agent.

Published
2023
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2. Development and preclinical evaluation of virus-like particle vaccine against COVID-19 infection

Author

Gulay Korukluoglu, İlayda Baydemir, Tugce Yildirim, Muzaffer Yildirim, Irem Evcili, Sefa Burak Cam, Ismail Selim Yildirim, Nese Guvencli, Ihsan Gursel, Naz Surucu Yilmaz, Yasemin Ceylan, İsmail Cem Yılmaz, Osman Erganiş, Basak Kayaoglu, Yagmur Aydin, Mayda Gursel, Hakan Enul, Emre Mert İpekoğlu, Fahriye Sarac, Asli Gulce Bartan, Merve Gizer, Gamze Aykut, Cumhur Adiay, Hasan Ersin Zeytin, Petek Korkusuz, Bilgi Gungor, Eda Çiftci Dede, Hivda Polat, Ihsan Cihan Ayanoglu, Şaban Tekin, Tugce Bildik, Berfu Saraydar, Hatice Asena Sanli, Nilsu Turay, Irem Abras, Artun Bulbul, Bilgehan Ibibik, Serdar Uzar, Yılmaz, İsmail Cem, Bülbül, Artun, Turay, Nilsu, Yıldırım, Muzaffer, Evcili, İrem, Güngör, Bilgi, Saraydar, Berfu, Bartan, Aslı Gülce, İbibik, Bilgehan, Bildik, Tuğçe, Ceylan, Yasemin, Yıldırım, Tuğçe, Abraş, İrem, Aykut, Gamze, and Gürsel, İhsan

Subjects
COVID-19 Vaccines, viruses, Immunology, Antibodies, Viral, complex mixtures, SARS‐CoV‐2, Immune system, Virus-like particle, Antigen, COVID‐19, vaccine, Immunology and Allergy, Animals, Humans, Vaccines, Virus-Like Particle, SARS-CoV-2, Immunogenicity, HEK 293 cells, fungi, virus diseases, COVID-19, Transfection, biochemical phenomena, metabolism, and nutrition, Virology, Antibodies, Neutralizing, Vaccination, HEK293 Cells, Allergen‐Specific Immunotherapy and Biologics, Humoral immunity, Original Article, ORIGINAL ARTICLES, Covid-19, Vaccine, virus‐like particle, CpG ODN Adjuvant
Abstract

Background Vaccines that incorporate multiple SARS‐CoV‐2 antigens can further broaden the breadth of virus‐specific cellular and humoral immunity. This study describes the development and immunogenicity of SARS‐CoV‐2 VLP vaccine that incorporates the four structural proteins of SARS‐CoV‐2. Methods VLPs were generated in transiently transfected HEK293 cells, purified by multimodal chromatography, and characterized by tunable‐resistive pulse sensing, AFM, SEM, and TEM. Immunoblotting studies verified the protein identities of VLPs. Cellular and humoral immune responses of immunized animals demonstrated the immune potency of the formulated VLP vaccine. Results Transiently transfected HEK293 cells reproducibly generated vesicular VLPs that were similar in size to and expressing all four structural proteins of SARS‐CoV‐2. Alum adsorbed, K3‐CpG ODN‐adjuvanted VLPs elicited high titer anti‐S, anti‐RBD, anti‐N IgG, triggered multifunctional Th1‐biased T‐cell responses, reduced virus load, and prevented lung pathology upon live virus challenge in vaccinated animals. Conclusion These data suggest that VLPs expressing all four structural protein antigens of SARS‐CoV‐2 are immunogenic and can protect animals from developing COVID‐19 infection following vaccination., SARS‐CoV‐2 VLP vaccine that incorporates the four structural proteins of SARS‐CoV‐2 is reproducibly produced in suspension adapted HEK293 cells. Alum adsorbed, K3‐CpG ODN‐adjuvanted VLPs elicit high titer anti‐S, anti‐RBD, anti‐N IgG, and neutralizing antibodies in mice, rats, and ferrets. The VLP vaccine supports multifunctional Th1‐biased T‐cell responses and demonstrate immunoprotective activity against live SARS‐CoV‐2 challenge in vaccinated mice.

Published
2021

3. Development and Preclinical Evaluation of Virus Like Particle Vaccine Against COVID-19 Infection

Author

Ismail Selim Yildirim, Cumhur Adiay, Şaban Tekin, Hakan Enul, Eda Çiftci Dede, Muzaffer Yildirim, Serdar Uzar, Bilgehan Ibibik, Sefa Burak Cam, Tugce Yildirim, Emre Mert İpekoğlu, Nilsu Turay, Tugce Bildik, Artun Bulbul, Asli Gulce Bartan, Mayda Gursel, Hatice Asena Sanli, Ihsan Gursel, Naz Surucu Yilmaz, Irem Abras, Fahriye Sarac, Ismail Yilmaz C, Berfu Saraydar, Bilgi Gungor, Hivda Polat, Yasemin Ceylan, Basak Kayaoglu, Ihsan Cihan Ayanoglu, Irem Evcili, Osman Erganiş, Nese Guvencli, Yagmur Aydin, Merve Gizer, Gamze Aykut, Hasan Ersin Zeytin, İlayda Baydemir, and Petek Korkusuz

Subjects
Chemistry, viruses, Immunogenicity, T cell, fungi, virus diseases, Transfection, biochemical phenomena, metabolism, and nutrition, complex mixtures, Virology, Vaccination, Immune system, medicine.anatomical_structure, Antigen, Virus-like particle, Humoral immunity, medicine
Abstract

Background Vaccines that incorporate multiple SARS-CoV-2 antigens can further broaden the breadth of virus-specific cellular and humoral immunity. This study describes the development and immunogenicity of SARS-CoV-2 VLP vaccine that incorporates the 4 structural proteins of SARS-CoV-2. Methods VLPs were generated in transiently transfected HEK293 cells, purified by multimodal chromatography and characterized by tunable resistive pulse sensing, AFM, SEM, and TEM. Immunoblotting studies verified the protein identities of VLPs. Cellular and humoral immune responses of immunized animals demonstrated the immune potency of the formulated VLP vaccine. Results Transiently transfected HEK293 cells reproducibly generated vesicular VLPs that were similar in size to and expressing all four structural proteins of SARS-CoV-2. Alum adsorbed, K3-CpG ODN adjuvanted VLPs elicited high titer anti-S, anti-RBD, anti-N IgG, triggered multifunctional Th1 biased T cell responses, reduced virus load and prevented lung pathology upon live virus challenge in vaccinated animals. Conclusion These data suggest that VLPs expressing all four structural protein antigens of SARS-CoV-2 are immunogenic and can protect animals from developing COVID-19 infection following vaccination.

Published
2021
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4. Development and Preclinical Evaluation of Virus Like Particle Vaccine Against COVID-19 Infection

Author

Ismail Yilmaz C, Emre Ipekoglu, Muzaffer Yildirim, Irem Evcili, Naz Yilmaz, Artun Bulbul, Nilsu Turay, Yagmur Aydin, Nese Guvencli, Bilgi Gungor, Berfu Saraydar, Asli Gulce Bartan, Bilgehan Ibibik, Tugce Bildik, İlayda Baydemir, Sefa Burak Cam, Eda Ciftci Dede, Merve Gizer, Osman Erganis, Fahriye Sarac, Serdar Uzar, Hakan Enul, Cumhur Adiay, Gamze Aykut, Hivda Polat, Saban Tekin, Hasan E. Zeytin, Petek Korkusuz, Ihsan Gursel, and Mayda Gursel

Subjects
viruses, fungi, virus diseases, biochemical phenomena, metabolism, and nutrition, complex mixtures
Abstract

Background Vaccines that incorporate multiple SARS-CoV-2 antigens can further broaden the breadth of virus-specific cellular and humoral immunity. This study describes the development and immunogenicity of SARS-CoV-2 VLP vaccine that incorporates the 4 structural proteins of SARS-CoV-2. Methods VLPs were generated in transiently transfected HEK293 cells, purified by multimodal chromatography and characterized by tunable resistive pulse sensing, AFM, SEM, and TEM. Immunoblotting studies verified the protein identities of VLPs. Cellular and humoral immune responses of immunized animals demonstrated the immune potency of the formulated VLP vaccine. Results Transiently transfected HEK293 cells reproducibly generated vesicular VLPs that were similar in size to and expressing all four structural proteins of SARS-CoV-2. Alum adsorbed, K3-CpG ODN adjuvanted VLPs elicited high titer anti-S, anti-RBD, anti-N IgG, triggered multifunctional Th1 biased T cell responses, reduced virus load and prevented lung pathology upon live virus challenge in vaccinated animals. Conclusion These data suggest that VLPs expressing all four structural protein antigens of SARS-CoV-2 are immunogenic and can protect animals from developing COVID-19 infection following vaccination.

Published
2021
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