Open AccessEnhancing the specific T cell immune response against micro- and nanoparticle immobilized antigen
Author(s) -
Р. Г. Сахабеев,
D.S. Polyakov,
Arina Goshina,
A. A. Vishnya,
I. V. Kudryavtsev,
E. S. Sinitcina,
Viktor Korzhikov-Vlakh,
Tatiana Tennikova,
M. M. Shavlovsky
Publication year - 2020
Publication title -
infekciâ i immunitet
Language(s) - English
Resource type - Journals
eISSN - 2313-7398
pISSN - 2220-7619
DOI - 10.15789/2220-7619-ets-1374
Subject(s) - polyethylene glycol , recombinant dna , conjugate , chemistry , peg ratio , immune system , virus like particle , polylactic acid , antigen , biophysics , microbiology and biotechnology , polymer , biochemistry , biology , immunology , organic chemistry , mathematical analysis , mathematics , finance , economics , gene
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.