Экспрессия Toll-подобных рецепторов TLR2 и TLR4 типа на иммунных клетках и продукция про- и противовоспалительных цитокинов в трансгенной модели болезни Паркинсона у мышей

G. V. Idova; E. I. Alperina; S. Ya. Zhanaeva; M. A. Tikhonova; M. M. Gevorgyan

doi:10.25557/2310-0435.2022.03.38-43

G. V. Idova Research Institute of Neurosciences and Medicine, Novosibirsk, Russian Federation http://orcid.org/0000-0002-9090-514X
E. I. Alperina Research Institute of Neurosciences and Medicine, Novosibirsk, Russian Federation
S. Ya. Zhanaeva Research Institute of Neurosciences and Medicine, Novosibirsk, Russian Federation http://orcid.org/0000-0002-4798-054X
M. A. Tikhonova Research Institute of Neurosciences and Medicine, Novosibirsk, Russian Federation http://orcid.org/0000-0002-4265-8981
M. M. Gevorgyan Research Institute of Neurosciences and Medicine, Novosibirsk, Russian Federation

DOI: https://doi.org/10.25557/2310-0435.2022.03.38-43

Keywords: Parkinson's disease, A53T transgenic mice, monocytes, T- and B-cell subpopulations, ; toll-like receptors 2 and 4, proinflammatory and anti-inflammatory cytokines

Abstract

Current reports suggest that immuno-inflammatory disorders underlie the pathogenesis of Parkinson's disease (PD), a progressive neurodegenerative condition characterized by loss of dopaminergic (DA) neurons in the nigrostriatal system and accumulation of aggregated α-synuclein. There is currently no evidence of the role of these processes at early preclinical stages of PD.

The aim of the study was to assess the expression of TLR2 and TLR4 on circulating monocytes, T and B cells, as well as on the peripheral production of pro-inflammatory (IFNγ, IL-6, IL-17A) and anti-inflammatory (IL-4 and IL-10) cytokines in young A53T transgenic mice compared with the control WT mice.

Methods. Male mice of the B6.CG-Tg (Prnp-SNCA*A53T)23MKle/J strain (A53T, 2.0-2.5 months) expressing the A53T mutation of human α-synuclein were used as an animal model of PD. Locomotor activity was studied in the open field test using the Noldus automatic registration system (Noldus Information Technology, the Netherlands). Motor coordination and balance were assessed with the “Rotarod” hardware-software complex (Neurobotics Trading LLC, Moscow, Russia) at different cylinder rotation speeds. Levels of monocytes, T- and B-lymphocytes and their subpopulations in the peripheral blood, as well as the expression of TLR2 and TLR4 on these cells, were measured with a FACSCanto II (BD) flow cytometer. Spontaneous and mitogen-induced production of pro-inflammatory (IFNγ, IL-6, IL-17A) and anti-inflammatory (IL-4 and IL-10) cytokines in the culture supernatant of blood mononuclear cells (PBMCs) was measured using the multiplex analysis of proteins and nucleic acids (Milliplex Luminex 200, Merk Millipore).

Results. Young transgenic A53T mice did not show changes in the motor function. However, the numbers of circulating CD115⁺CD11b⁺ monocytes, CD3⁺ T cells, CD3⁺CD4⁺ T helpers (Th) were increased, while the content of CD3⁺CD4⁺CD25⁺ T regulatory (Treg) cells was reduced compared to control WT mice. А53Т mice also showed a higher expression of TLR2 and TLR4 only on Treg cells and a tendency to increased TLR4 expression on monocytes. An increase in spontaneous production of the pro-inflammatory cytokine IL-6 was associated with a decrease in spontaneous and stimulated production of the anti-inflammatory cytokine IL-10. The production of IFNγ, IL-17 and IL-4 did not differ significantly between A53T and WT mice.

Conclusion. Changes in immunity parameters of A53T mice overexpressing α-synuclein are observed at an early stage of parkinsonism, before the onset of motor disorders, indicating the development of inflammation.

Expression of TLR2 and TLR4 Toll-like receptors on immune cells and production of pro- and anti-inflammatory cytokines in a transgenic mouse model of Parkinson’s disease

Abstract