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. 2022 Mar 28;27(7):2195.
doi: 10.3390/molecules27072195.

Cell Differentiation and Proliferation in the Bone Marrow and Other Organs of 2D2 Mice during Spontaneous Development of EAE Leading to the Production of Abzymes

Kseniya S Aulova  1 , Andrey E Urusov  1 , Ludmila B Toporkova  2 , Sergey E Sedykh  1 , Juliya A Shevchenko  2 , Valeriy P Tereshchenko  2 , Sergei V Sennikov  2 , Irina A Orlovskaya  2 , Georgy A Nevinsky  1
Affiliations

Cell Differentiation and Proliferation in the Bone Marrow and Other Organs of 2D2 Mice during Spontaneous Development of EAE Leading to the Production of Abzymes

Kseniya S Aulova et al. Molecules. .

Abstract

The exact cellular and molecular mechanisms of multiple sclerosis and other autoimmune diseases have not been established. Autoimmune pathologies are known to be associated with faults in the immune system and changes in the differentiation profiles of bone marrow stem cells. This study analyzed various characteristics of experimental autoimmune encephalomyelitis (EAE) in 2D2 mice. Differentiation profiles of six hematopoietic stem cells of bone marrow were found to significantly differ in 2D2 male and female mice during the spontaneous development of EAE. In addition, we found various properties of B and T cells, CD4+ and CD8+ lymphocytes in blood and several organs (bone marrow, spleen, thymus, and lymph nodes) of 2D2 male and female mice to be considerably different. These changes in hematopoietic stem cells differentiation profiles and level of lymphocyte proliferation in various organs of 2D2 mice were found to induce the production of IgGs against DNA, myelin basic protein, and myelin oligodendrocyte glycoprotein, increasing the number of autoantibodies hydrolyzing these substrates. We compared the changes of these immunological and biochemical parameters in 2D2 mice with those of mice of two other lines (Th and C57BL/6), also prone to spontaneous development of EAE. Some noticeable and even extreme variations were found in the time-related development of parameters between male and female mice of 2D2, Th, and C57BL/6 lines. Despite some differences, mice of all three lines demonstrated the changes in hematopoietic stem cells profiles, lymphocyte content, and production of catalytic autoantibodies. Given that these changes are harmful to mice, we believe them to cause the development of experimental autoimmune encephalomyelitis.

Keywords: 2D2; Th and C57BL/6 EAE mice; catalytic antibodies; development of experimental autoimmune encephalomyelitis (EAE); hematopoietic stem cells differentiation; lymphocyte proliferation in different organs.

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Conflict of interest statement

The coauthors have no competing financial interests.

Figures

Figure 1
Figure 1
Relative changes in proteinuria during time-related development of EAE by 2D2 (black symbols), Th (red symbols) male and female, and C57BL/6 (green symbols) male mice. For comparison, the C57BL/6 [34,35,36,37] and Th [38,39] mice data were taken from our previously published articles. The duration of the experiment and the time of blood sampling for different strains of mice were different.
Figure 2
Figure 2
Changes during time-related development of spontaneous EAE in BFU-E (A), CFU-E (B), CFU-GM (C), and CFU-GEMM (D) cells as well as in BM relative amounts of B (E) and T lymphocytes (F) in 2D2 (black symbols), Th (red symbols), and C57BL/6 (green symbols) mice. The number of total cells is calculated for 15,000 bone marrow cells. All dependencies for all lines of males and females are marked in the Figure. For comparison, the C57BL/6 [34,35,36,37] and Th [38,39] mice data were taken from our previously published articles. The duration of the experiment and the time of blood sampling were different for different strains of mice.
Figure 3
Figure 3
Time-related dependencies in the changes of relative numbers of B cells in different organs of 2D2 (black symbols) and Th (red symbols) males and females. All dependencies for all organs of various males and females are marked in the figures (A–D). For comparison, the data for Th mice were taken from our previously published articles [38,39]. The duration of the experiment and the time of blood sampling for different strains of mice were different.
Figure 4
Figure 4
Time-related dependencies in the changes of relative numbers of T cells in various organs of male and female 2D2 (black symbols) and Th (red symbols) mice. All the dependencies for all organs of various males and females are given in (A–D) panels. For comparison, the data for Th mice were taken from our previously published articles [38,39]. The duration of the experiment and the time of blood sampling were different for different strains of mice.
Figure 5
Figure 5
Time-related dependencies in the relative numbers of total CD4 cells in various organs of male and female 2D2 (black symbols) and Th (red symbols) mice. All dependencies for all organs of different males and females are marked in the Figure (A–E). For comparison, the data for Th mice were taken from our previously published articles [38,39]. The duration of the experiment and the time of blood sampling for different strains of mice were different.
Figure 6
Figure 6
Time-related dependencies in the relative numbers of total CD8 cells in different organs of male and female 2D2 (black symbols) and Th (red symbols) mice. All dependencies for all organs of different males and females are marked in the Figure (A–E). For comparison, the data for Th mice were taken from our previously published articles [38,39]. The duration of the experiment and the time of blood sampling for different strains of mice were different.
Figure 7
Figure 7
The IgGmix (14 μg) homogeneity analysis by SDS-PAGE under non-reducing conditions in the absence of DTT (A); silver staining. Panel A demonstrates the position of IgGs. The relative activities (RA, %) in the hydrolysis of DNA (∆), MOG (□), MBP (●), and histones (o) were estimated using eluates of gel fragments (2–3 mm) (B). After substrates incubation for 24 h with eluates, complete hydrolysis of all substrates was taken for 100% (B). The errors of the relative activities estimation from two independent experiments did not exceed 7–10%.
Figure 8
Figure 8
Time-related changes in the relative concentration of IgGs against DNA, MBP, MOG, and histones in purified Abs (A–D). Dependencies corresponding to 2D2 (black symbols), Th (red signs), and C57BL/6 (green symbols) males and females are shown in (A–D) panels. For comparison, the C57BL/6 [34,35,36,37] and Th [38,39] mice data were taken from our earlier published articles. The duration of the experiment and the time of blood sampling for different strains of mice were different.
Figure 9
Figure 9
Time-related changes in the IgGs relative activities in the hydrolysis of DNA, MBP, and MOG (A–C). Dependencies corresponding to 2D2 (black symbols), Th (red signs), and C57BL/6 (green symbols) males and females are marked in the Figure. For comparison, the data for C57BL/6 [34,35,36,37] and Th [38,39] mice were taken from our earlier published articles. The duration of the experiment and the time of blood sampling for different strains of mice were different.
Figure 10
Figure 10
Time-related changes in the RA of IgGs in the hydrolysis of individual histones: H1 (A), H2A (B), H2B (C), H3 (D), and H4 (E). Dependencies corresponding to 2D2 male (■) and female (□) mice are marked in (A–E) panels.
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