Publications on the project |
110 Interaction of diphtheria toxin B subunit with sensitive and insensitive mammalian cells |
Authors: | A.J. Labyntsev, N.V. Korotkevich, A.A. Kaberniuk, S.I. Romaniuk, D.V. Kolibo, S.V. Komisarenko | |
Summary: | The recombinant fluorescent derivative of diphtheria toxin (EGFP-SbB) obtained by the replacement of toxin A subunit by enhanced green fluorescent protein (EGFP) has been used for visualization of the interaction of diphtheria toxin (DT) with sensitive and insensitive cells. It was shown that EGFP-SbB could interact with cell surface of both toxin-sensitive monkey cells (Vero cell line) and toxin-resistant mouse cells (3T3 cell line). The affinity of this protein for receptors of Vero cells was three times higher as compared with 3T3 cells. It was demonstrated that fluorescent derivate was able to interact with receptors of both cell lines and to internalize into these cells. Internalization of EGFP-SbB into the cells was inhibited by endocytosis inhibitor phenyl arsine oxide. We suppose that diverse sensitivity to DT of monkey and mouse cells can be explained not only by differences in their receptor affinity for DT but also by the processes that occur after internalization of the toxin into the cells. | |
Keywords: | fluorescent derivates, B subunit of diphtheria toxin, toxin-sensitive and toxinresistant cells, receptor binding, endocytosis | |
Edition: | | | | 2010,
65-75,Ukrainian |
110 Perspectives of application of recombinant diphtheria toxin derivatives |
Authors: | Romaniuk S.I., Kolibo D.V., Komisarenko S.V. | |
Summary: | Diphtheria toxin (DT) is a unique bacterial protein, which selectively kills certain cell populations due to strict functional specialization of domains that allows using this toxin in protein engineering for constructing recombinant derivatives with defined properties. The article covers structural and functional features of DT molecule, both fundamental and practical aspects of recombinant DT derivatives' applications in different fields. In particular, applications of recombinant DT derivatives as unique instruments for fundamental research of cell receptors' functions, mechanism of DT action and participation of different cell populations in biological processes are presented. Perspectives of recombinant DT derivatives practical applications for the development of vaccines, cytotoxins, HB-EGF blockers, diagnostic test-systems, serotherapeutic medications and constructions for drug delivery have been discussed. This review reflects recent advances and current problems in using recombinant DT derivatives for treatment and prophylaxis of oncologic, autoimmune, infectious and others diseases. | |
Keywords: | diphtheria toxin, fragments and domains of diphtheria toxin, recombinant proteins, heparin-binding epidermal growth factor-like growth factor (HB-EGF), vaccines, cytotoxins | |
Edition: | Bioorganicheskaia khimiia. 38(6) | | | 2012,
639-652,English |
110 Immunobiology of diphtheria. Recent approaches for the prevention, diagnosis, and treatment of the disease |
Authors: | D.V. Kolibo, A.J. Labyntsev, S.I. Romaniuk, A.A. Kaberniuk, E.S. Oliinyk, N.V. Korotkevich, S.V. Komisarenko | |
Summary: | Diphtheria is a highly contagious life-threatening disease caused by the toxi genic strains of Corynebacterium diphtheria, which are transformed by a bacteriophage carrying the toxin gene. Diphtheria causative agent and its major virulence factor diphtheria toxin are well studied, but outbreaks of disease still occur worldwide. Rapid development of new methods in immunology and molecular biology is currently leading to improvement of prophylaxis, diagnosis and treatment of diphtheria. This review highlights the microbiological, epidemiological and immunological aspects of diphtheria infection, role of diphtheria toxin and others virulence factors in diphtheria pathogenesis and role of humoral anti-toxic immunity in the protection against disease. Perspectives in development of new diagnostic tests, anti-diphtheria vaccines, immunobiological preparations and antidotes for prevention of diphtheria infection, and other anti-diphteria means was also discussed. | |
Keywords: | diphtheria, diphtheria toxin, immunity, diagnostic tests, vaccines, antidotes, recombinant proteins | |
Edition: | Biotechnologia Acta – 2013. – Vol. 6. – N 4. | | | 2013,
43–62,English |
110 Recombinant fluorescent models for studying the diphtheria toxin |
Authors: | A.J. Labyntsev, N.V. Korotkevych, K.J. Manoilov, A.A. Kaberniuk, D.V. Kolybo, S.V. Komisarenko | |
Summary: | Diphtheria toxin is a major factor of the pathogenicity of the causative agent of diphtheria Corynebacterium. Due to a small size, it is of considerable interest as the basis for the development of synthetic protein molecules with a transport function, e.g., immunotoxins. In this work we describe the expression and characterized nontoxic recombinant fluorescent derivatives of the diphtheria toxin and its nontoxic CRM197 mutant. The proteins obtained can be used for studying receptor-binding and transport functions of the toxin in cells, evaluation of the expression level of the toxin proHB-EGF receptor membranes, immunization and generation of specific antibodies against the toxin, as well as for the development of diagnostic test-systems for the diphtheria toxin and antitoxic antibodies. | |
Keywords: | fluorescent proteins, molecular cloning, proHBEGF, recombinant nontoxic bacterial toxin analogues | |
Edition: | Russian Journal of Bioorganic Chemistry, 2014, Vol. 40, No. 4 | | | 2014,
pp. 401–409,English |
110 Cellobiose-coated poly(lactideco- glycolide) particles loaded with diphtheria toxoid for per os immunization |
Authors: | Tetiana Chudina, Andrii Labyntsev, Kyrylo Manoilov, Denys Kolybo, Serhiy Komisarenko | |
Summary: | Aim To evaluate the dose-dependent immunogenic properties of poly(lactide-co-glycolide) (PLGA) particles coated with cellobiose as antigen carriers for oral immunization.
Methods Two types of PLGA-cellobiose particles (PLGAcellobiose-1, ~ 0.8 ìm and PLGA-cellobiose-2, ~ 1.2 ìm) containing non-toxic recombinant subunit B (SbB) of diphtheria toxin fused with enhanced green fluorescent protein were characterized in vitro for their size, shape, antigen loading, and ability to induce phagocytosis. Different doses of antigen, immobilized on the particles (2.5 ìg, 25 ìg, 250 ìg, and 2500 ìg per 1 kg of body weight), were administered per os 3 times with intervals of 2 weeks to BALB/c female mice. The antigen-specific IgG and IgA antibodies were estimated in serum by ELISA.
Results After the first immunization, increase in concentration of blood antitoxic antibodies was detected. Antigen dose 250 ìg/kg was the most immunogenic for IgG antibodies induction for both types of PLGA-cellobiose particles. Antigen doses 25 ìg/kg and 2.5 ìg/kg were the most immunogenic for IgA antibodies induction by PLGA-cellobiose 1 and 2 particles, respectively. The second and the third treatment had no significant effect on the immune response or even reduced it, which could be explained by immune tolerance induction by the antigens delivered per os.
Conclusion Our results suggest that the correct dose of PLGA-cellobiose particles loaded with antigen could significantly increase the humoral immune response against the introduced antigen already after the first immunization. Thus, PLGA particles can be considered as a potent component of oral vaccines. | |
Keywords: | oral vaccination, immunity, microparticles, nanoparticles, PLGA, diphtheria toxoid | |
Edition: | Croat Med J. 2015;56 | | | ,
pp 85-93,English |
The events in the framework of the project |
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110 4.2. Physical and chemical basis of synthesis and formation properties of nanomaterials medical Purpose: Expected results:Other (to increase production, improve working conditions, environmental improvement, energy savings, material savings, reduced equipment wear, increase productivity, improve efficiency of diagnosis and treatment, etc.) Stage 1:Синтез колоїдних наночасток металів різного розміру та отримання їх комплексів з нетоксичними фрагментами дифтерійного токсину. Stage 2:Дослідження процесів взаємодії колоїдних наночасток з антиген-презентувальними клітинами in vitro. Stage 3:Дослідження цитотоксичних властивостей наночасток in vitro Stage 4:Оптимізація складу наночасток для пероральної імунізації Stage 5:Оцінка ефективності прояву імуногенних властивостей кон’югованого з наночастками антигену в залежності від типу наночасток.
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