IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUS
The aim of the study was to investigate immunogenic properties of mosaic recombinant proteins constructed on the data of hepatitis C virus NS4A and NS4B antigens. Four mosaic recombinant proteins, containing the T and B epitopes of the NS4A and NS4B antigens, were created by genetic engineering meth...
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| Format: | Article |
| Language: | English |
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Central Research Institute for Epidemiology
2018-06-01
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| Series: | Вопросы вирусологии |
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| Online Access: | https://virusjour.crie.ru/jour/article/viewFile/198/108 |
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| author | V. V. Koupriyanov L. I. Nikolaeva A. A. Zykova P. I. Makhnovskiy R. Y. Kotlyarov A. V. Vasilyev N. V. Ravin |
| author_facet | V. V. Koupriyanov L. I. Nikolaeva A. A. Zykova P. I. Makhnovskiy R. Y. Kotlyarov A. V. Vasilyev N. V. Ravin |
| author_sort | V. V. Koupriyanov |
| collection | DOAJ |
| description | The aim of the study was to investigate immunogenic properties of mosaic recombinant proteins constructed on the data of hepatitis C virus NS4A and NS4B antigens. Four mosaic recombinant proteins, containing the T and B epitopes of the NS4A and NS4B antigens, were created by genetic engineering methods in the E. coli system. To enhance the immune response they were linked in different variations to the nucleotide sequences of murine interleukin-2 (IL-2), the Neisseria meningiditis lipopeptide, and the T helper epitope of the core protein of hepatitis C virus. The immunogenic properties of these recombinant proteins were analyzed by immunoblotting, ELISA and ELISpot using sera from immunized mice and patients infected with hepatitis C virus. Recombinant proteins specifically reacted with the sera of immunized mice and infected patients in immunoblotting. According to the ELISA data, the predominant formation of antibodies to NS4B was observed when mice were immunized with the recombinant proteins containing both antigens. Analysis of gamma-interferon production by T-lymphocytes upon contact with activated dendritic cells showed in ELISpot that the maximum production of this cytokine was detected when adjuvant components were located at the N- and C-ends of the recombinant protein. The highest level of gamma-interferon production during stimulation with this drug was detected in lymphocytes from the bone marrow and lymph nodes. The recombinant protein containing the T and B epitopes of NS4A and NS4B, murine IL-2 and the lipopeptide Neisseria meningiditis had the greatest immunostimulate effect among the four constructions. This recombinant protein formed nanoparticles of 100-120 nm in size. |
| first_indexed | 2024-03-13T00:08:27Z |
| format | Article |
| id | doaj.art-29132cb7aeea4773a8396c90e5ff0154 |
| institution | Directory Open Access Journal |
| issn | 0507-4088 2411-2097 |
| language | English |
| last_indexed | 2024-03-13T00:08:27Z |
| publishDate | 2018-06-01 |
| publisher | Central Research Institute for Epidemiology |
| record_format | Article |
| series | Вопросы вирусологии |
| spelling | doaj.art-29132cb7aeea4773a8396c90e5ff01542023-07-12T20:00:24ZengCentral Research Institute for EpidemiologyВопросы вирусологии0507-40882411-20972018-06-0163313814310.18821/0507-4088-2018-63-3-138-143178IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUSV. V. Koupriyanov0L. I. Nikolaeva1A. A. Zykova2P. I. Makhnovskiy3R. Y. Kotlyarov4A. V. Vasilyev5N. V. Ravin6Federal Research Centre «Fundamentals of Biotechnology»D.I. Ivanovsky Institute of Virology, «National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya»Federal Research Centre «Fundamentals of Biotechnology»D.I. Ivanovsky Institute of Virology, «National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya»Federal Research Centre «Fundamentals of Biotechnology»D.I. Ivanovsky Institute of Virology, «National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya»Federal Research Centre «Fundamentals of Biotechnology»The aim of the study was to investigate immunogenic properties of mosaic recombinant proteins constructed on the data of hepatitis C virus NS4A and NS4B antigens. Four mosaic recombinant proteins, containing the T and B epitopes of the NS4A and NS4B antigens, were created by genetic engineering methods in the E. coli system. To enhance the immune response they were linked in different variations to the nucleotide sequences of murine interleukin-2 (IL-2), the Neisseria meningiditis lipopeptide, and the T helper epitope of the core protein of hepatitis C virus. The immunogenic properties of these recombinant proteins were analyzed by immunoblotting, ELISA and ELISpot using sera from immunized mice and patients infected with hepatitis C virus. Recombinant proteins specifically reacted with the sera of immunized mice and infected patients in immunoblotting. According to the ELISA data, the predominant formation of antibodies to NS4B was observed when mice were immunized with the recombinant proteins containing both antigens. Analysis of gamma-interferon production by T-lymphocytes upon contact with activated dendritic cells showed in ELISpot that the maximum production of this cytokine was detected when adjuvant components were located at the N- and C-ends of the recombinant protein. The highest level of gamma-interferon production during stimulation with this drug was detected in lymphocytes from the bone marrow and lymph nodes. The recombinant protein containing the T and B epitopes of NS4A and NS4B, murine IL-2 and the lipopeptide Neisseria meningiditis had the greatest immunostimulate effect among the four constructions. This recombinant protein formed nanoparticles of 100-120 nm in size.https://virusjour.crie.ru/jour/article/viewFile/198/108hepatitis c virus antigensrecombinant mosaic proteinsadjuvant components |
| spellingShingle | V. V. Koupriyanov L. I. Nikolaeva A. A. Zykova P. I. Makhnovskiy R. Y. Kotlyarov A. V. Vasilyev N. V. Ravin IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUS Вопросы вирусологии hepatitis c virus antigens recombinant mosaic proteins adjuvant components |
| title | IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUS |
| title_full | IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUS |
| title_fullStr | IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUS |
| title_full_unstemmed | IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUS |
| title_short | IMMUNOGENIC PROPERTIES OF RECOMBINANT MOZAIC PROTEINS BASED ON ANTIGENS NS4A AND NS4B OF HEPATITIS C VIRUS |
| title_sort | immunogenic properties of recombinant mozaic proteins based on antigens ns4a and ns4b of hepatitis c virus |
| topic | hepatitis c virus antigens recombinant mosaic proteins adjuvant components |
| url | https://virusjour.crie.ru/jour/article/viewFile/198/108 |
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