Enhancing the specific T cell immune response against micro- and nanoparticle immobilized antigen

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The current study was a part of the project on generating viral particle traps occurring due to covalent immobilization on the interface of recombinant virus-specific polymer-based nano- and microparticles. It is assumed that protein-particle conjugates could be able to bind virions followed by engulfment by immune cells. The study was aimed to examine the effect of polylactic acid (PLA) and PLA block-copolymer with polyethylene glycol (PLA-PEG)-based micro- and nanoparticles on the cellular immune response against polymeric particle-bound antigen. Materials and methods. A recombinant chimeric protein beta-2-microglobulin — green fluorescent protein (β2M-sfGFP) was obtained by affine chromatography. The recombinant protein was immobilized onto the polymer particles, which were further used for mice immunization. Female F1 hybrid mice (CBA x C57BL) in experimental and control groups consisted of 4–6-month-old 15 animals (weighted 20–25 g). Intracellular cytokine staining was used to evaluate the cellular immune response. Results and discussion. It was shown that the nanoparticles of PLA block-copolymer with polyethylene glycol (PLA-PEG) were able to bind 10 microgram protein per 1 mg polymer. The polylactic acid nanoparticles were able to bind 2,3 microgram protein per 1 mg polymer. In experiment, mice in group 1 were immunized with 100 nm PLA-PEG particle-β2M-sfGFP conjugate, in group 2 — with same particles together with soluble β2M-sfGFP. In group 3, mice were immunized with 1400 nm PLA particles-β2M-sfGFP conjugate, and in group 4 — with same particles together with soluble protein. The spleens isolated 2 weeks after the four-time intraperitoneal immunization. Comparison of immune response between groups was assessed by nonparametric Kruskal–Wallis criterion with Tukey correction. It was shown that the number of antigen-specific CD4+ T cells produced to model protein was significantly higher after immunization with particle-β2M-sfGFP conjugate, as compared to control groups, wherein immunization was performed with a mixture of protein and unmodified particles (p < 0.001). It was found that the number of antigen-specific CD8+ T cells formed against β2m-sfGFP did not differ between all groups examined.

About the authors

R. G. Sakhabeev

Institute of Experimental Medicine

Author for correspondence.
Email: helm505@mail.ru
ORCID iD: 0000-0002-4367-4924

Rodion G. Sakhabeev,  Junior Researcher, Department of Molecular Genetics 

197376, St. Petersburg, Academician Pavlov str., 12

Phone: +7 953 152-68-07 

Russian Federation

D. S. Polyakov

Institute of Experimental Medicine

Email: ravendoctor@mail.ru

PhD (Medicine), Researcher, Department of Molecular Genetics 

St. Petersburg 

A. D. Goshina

St. Petersburg State University

Email: arina8goshina@gmail.com

4th-year Student

St. Petersburg 

A. A. Vishnya

Herzen State Pedagogical University of Russia

Email: tlou000@yandex.ru

2th-year Master Student

St. Petersburg 

I. V. Kudryavtsev

Institute of Experimental Medicine

Email: igorek1981@yandex.ru

PhD (Biology), Head of the Laboratory of Immunoregulation, Department of Immunology

St. Petersburg 

E. S. Sinitcina

Institute of Chemistry, St. Petersburg State University; Institute of Macromolecular Compounds of the Russian Academy of Sciences

Email: kat_sinitsyna@mail.ru

PhD (Chemistry), Senior Researcher, Interdisciplinary Laboratory of Biomedical Chemistry; Researcher, Laboratory of Polymer Sorbents and Carriers for Biotechnology

St. Petersburg 

V. А. Korzhikov-Vlakh

Institute of Chemistry, St. Petersburg State University

Email: v_korzhikov@mail.ru

PhD (Chemistry), Associate Professor, Interdisciplinary Laboratory of Biomedical Chemistry

St. Petersburg 

T. B. Tennikova

St. Petersburg State University; Institute of Chemistry, St. Petersburg State University

Email: tennikova@mail.ru

PhD, MD (Chemistry), Professor, Leading Researcher and Head of the Interdisciplinary Laboratory of Biomedical Chemistry

St. Petersburg 

M. M. Shavlovsky

Institute of Experimental Medicine; Pavlov First St. Petersburg State Medical University

Email: mmsch@rambler.ru

PhD, MD (Medicine), Head of the Molecular Genetics Laboratory; Professor

St. Petersburg 


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Copyright (c) 2020 Sakhabeev R.G., Polyakov D.S., Goshina A.D., Vishnya A.A., Kudryavtsev I.V., Sinitcina E.S., Korzhikov-Vlakh V.А., Tennikova T.B., Shavlovsky M.M.

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