Preliminary study on the changes of plymorphonucler myeloid-derived suppressor cells in the spleen of mice infected with Schistosoma japonicum

doi:10.12140/j.issn.1000-7423.2022.03.008

Abstract

Abstract:

Objective To explore the dynamic changes of the proportion, function and spleen histopathology of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in the spleen of mice infected with Schistosoma japonicum. Methods Thirty-six BALB/c mice aged 6-8 weeks were randomly assigned into the infected group and control group, with 18 mice in each group. Mice in the infection group were infected with S. japonicum cercariae (20 ± 1)/mouse. At 4, 6 and 8 weeks post-infection, the spleen from 6 mice, which were randomly selected from each group, were collected, followed by calculating the spleen coefficients after weighting. The spleen tissues were fixed, sliced and stained with hematoxylin-eosin (HE) to observe the pathological changes microscopically. Spleen single cell suspension was prepared for examining the dynamic changes of PMN-MDSCs proportion in splenic lymphocytes by flow cytometry. Fluorescent quantitative PCR was used to determine the mRNA relative expression expression level of PMN-MDSC related inflammation factors, including interleukin-6 (IL-6), S100 calcium binding protein A8 (S100A8）, S100A9, and the cell function factor arginase 1 (Arg1), nitric oxide synthase (iNOS), heme-binding membrane glycoprotein 91(gp91), transforming growth factor-β(TGF-β), and IL-10 in the spleen tissues. Results At 4, 6 and 8 weeks after infection, the spleen weight and spleen coefficient in infection group were(179 ± 10.19)mg, (350.3 ± 16.84)mg, (414.3 ± 18.98)mg, and (0.93 ± 0.03)%, (1.97 ± 0.10)%, (2.31 ± 0.08)%, respectively, which were all significantly higher than that in the control group[(108.2 ± 9.93)mg and (0.51 ± 0.04)%] (F = 101.3, 143.7, P < 0.01). HE staining showed that at 4-6 weeks after infection, the inflammatory cell band gradually increased, while the lymphoid follicles of the spleen decreased, and the germinal center decreased or even disappeared compared with the control group. At 6-8 weeks after infection, the inflammatory cell band decreased gradually, and the lymphoid follicles and germinal centre structure of the spleen proliferated gradually compared with the control group. Splenic PMN-MDSCs proportion in infection group were (1.53 ± 0.16)%, (28.40 ± 2.35)%, (38.67 ± 1.94)%, which were all significantly higher than that in the control group (0.80 ± 0.10)% (F = 326.5, P < 0.01). Real-time quantitative PCR showed that at 6 weeks post-infection, the relative level of mRNA expression of IL-6, S100A8, S100A9, gp91, Arg1, iNOS, IL-10 and TGF-β in splenic tissue were 2.74 ± 0.25, 51.4 ± 1.25, 39.20 ± 2.83, 2.15 ± 0.08, 2.33 ± 0.39, 1.57 ± 0.08, 2.20 ± 0.39 and 1.44 ± 0.05, respectively, which were all significantly higher than that in the control group [1.05 ± 0.10, 1.01 ± 0.11, 1.02 ± 0.07, 1.04 ± 0.09, 1.01 ± 0.06, 1.00 ± 0.05, 0.98 ± 0.20 and 1.00 ± 0.04(t = 6.367, 40.07, 13.50, 9.311, 3.315, 5.642, 2.764, 6.914, P < 0.05, P < 0.01)]. The relative iNOS mRNA expression in splenic PMN-MDSCs was higher than that in the control group (t = 0.6134, P < 0.01) and the relative of IL-10 mRNA expression level was no statistically significantly different between the infection group and the control group(t = 1.176, P > 0.05). At 8 weeks after infection, the relative IL-6, S100A8, S100A9, Arg1, iNOS and IL-10 mRNA expression levels in splenic tissue were all significantly higher than those in the control group (t = 2.496, 5.145, 9.518, 3.938, 4.819, 2.251, P < 0.05, P < 0.01). In the splenic of PMN-MDSCs, the relative iNOS mRNA expression level and IL-10 were 32.12 ± 2.30, and 2.64 ± 0.37, respectively, which were higher than that at 4 weeks after infection (1.08 ± 0.01, 1.14 ± 0.35), and also higher than that at 6 weeks after infection (12.06 ± 1.80, 1.50 ± 0.36), as well as the control group (1.02 ± 0.13, 1.06 ± 0.12) (F = 100.6, 5.471, P < 0.01). Conclusion At 4~6 weeks post S. japonicum infection, the spleen tissue presented strong inflammatory response, at 6~8 weeks after infection, the secretion of the inhibitory factor IL-10 of PMN-MDSC was significantly increased, and the spleen tissue structure was gradually proliferated.

Key words: S. japonicum, PMN-MDSC, Spleen, Cytokine, Inflammation

CLC Number:

R383.24

ZHANG Xiao-cheng, GAO Yuan, HU Yuan, CAO Jian-ping. Preliminary study on the changes of plymorphonucler myeloid-derived suppressor cells in the spleen of mice infected with Schistosoma japonicum[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(3): 330-336.

Figures/Tables 5

Table 1

Fig. 1

Fig. 2

Table 2

Table 3

References 26

[1]	LoVerde PT. Schistosomiasis[J]. Adv Exp Med Biol, 2019, 1154: 45-70. doi: 10.1007/978-3-030-18616-6_3 pmid: 31297759
[2]	Zhang LJ,, Xu ZM,, Yang F, et al. Endemic status of schistosomiasis in People’s Republic of China in 2020[J]. Chin J Schisto Control, 2021, 33(3): 225-233. (in Chinese)
	( 张利娟,, 徐志敏,, 杨帆, 等. 2020年全国血吸虫病疫情通报[J]. 中国血吸虫病防治杂志, 2021, 33(3): 225-233.)
[3]	Zhao J,, Chen Y,, Yu YR, et al. STAT3 promotes schistosome-induced liver injury by inflammation, oxidative stress, proliferation, and apoptosis signal pathway[J]. Infect Immun, 2021, 89(3): e00309-320.
[4]	Kong H,, He J,, Huang J, et al. Endothelin receptors promote schistosomiasis induced hepatic fibrosis via splenic B cells[J]. PLoS Pathog, 2020, 16(10): e1008947.
[5]	Zhou HH,, Lv NY,, Tong SJ, et al. Resveratrol regulates M1/M2 polarization of mice infected with Schistosoma japonicum through mitochondria[J]. Chin pharmacol Bull, 2021, 37(1):98-106. (in Chinese)
	( 周慧慧,, 吕年银,, 佟书娟, 等. 白藜芦醇通过线粒体调控血吸虫感染小鼠M1/M2极化[J]. 中国药理学通报, 2021, 37(1): 98-106.)
[6]	Zheng B,, Zhang JQ,, Chen H, et al. T lymphocyte mediated liver immunopathology of schistosomiasis[J]. Front Immunol, 2020, 11: 61. doi: 10.3389/fimmu.2020.00061 pmid: 32132991
[7]	Dorhoi A,, Glaría E,, Garcia-Tellez T, et al. MDSCs in infectious diseases: regulation roles, and readjustment[J]. Cancer Immunol Immun, 2019, 68(4): 673-685. doi: 10.1007/s00262-018-2277-y
[8]	Yang ZZ,, Guo JC,, Weng LL, et al. Myeloid-derived suppressor cells-new and exciting players in lung cancer[J]. J Hematol Oncol, 2020, 13(1): 10. doi: 10.1186/s13045-020-0843-1
[9]	Yin K,, Xia XL,, Rui K, et al. Myeloid-derived suppressor cells: a new and pivotal player in colorectal cancer progression[J]. Front Oncol, 2020, 10: 610104. doi: 10.3389/fonc.2020.610104
[10]	Veglia F,, Perego M,, Gabrilovich D. Myeloid-derived suppressor cells coming of age[J]. Nat Immunol, 2018, 19(2): 108-119. doi: 10.1038/s41590-017-0022-x
[11]	Yang SH,, Kim J,, Lee MJ, et al. Abnormalities of plasma cytokines and spleen in senile APP/PS₁/Tau transgenic mouse model[J]. Sci Rep, 2015, 5: 15703. doi: 10.1038/srep15703
[12]	Aiello I,, Finkielstein CV,, Paladino N, et al. Circadian disruption promotes tumor-immune micro environment remodeling favoring tumor cell proliferation[J]. Sci Adv, 2020, 6(42): eaaz4530.
[13]	Liu D,, Wu J,, Cyster JG, et al. Requirements for cDC2 positioning in blood-exposed regions of the neonatal and adult spleen[J]. J Exp Med, 2020, 217(11): e20192300.
[14]	Lewis SM,, Williams A,, Eisenbarth SC. Structure and function of the immune system in the spleen[J]. Sci Immunol, 2019, 4(33): eaau6085.
[15]	Wang YJ,, Shen YJ,, Xu YX, et al. Observation on destroyed architecture of splenic lymphoid follicles in mice infected with Schistosoma japonicum by immunohistochemistry[J]. Chin J Schisto Control, 2017, 29(4): 468-470. (in Chinese)
	( 王燕娟,, 沈玉娟,, 徐馀信, 等. 免疫组化法观察日本血吸虫感染破坏小鼠脾脏淋巴滤泡结构[J]. 中国血吸虫病防治杂志, 2017, 29(4): 468-470.)
[16]	Tayukcuoglu E,, Horzum U,, Esendagli G, et al. Human splenic polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) are strategically located immune regulatory cells in cancer[J]. Eur J Immunol, 2020, 50(12): 2067-2074. doi: 10.1002/eji.202048666
[17]	Corzo CA,, Cotter MJ,, Cheng PY, et al. Mechanism regulating reactive oxygen species in tumor-induced myeloid-derived suppressor cells[J]. J Immunol, 2009, 182(9): 5693-5701. doi: 10.4049/jimmunol.0900092
[18]	Weber R,, Groth C,, Lasser S, et al. IL-6 as a major regulator of MDSC activity and possible target for cancer immunotherapy[J]. Cell Immunol, 2021, 359: 104254.
[19]	Wang S,, Song R,, Ma J, et al. S100A8/A9 in inflammation[J]. Front Immunol, 2018, 9: 1298. doi: 10.3389/fimmu.2018.01298
[20]	Ohl K,, Tenbrock K. Reactive oxygen species as regulators of MDSC-mediated immune suppression[J]. Front Immunol, 2018, 9: 2499. doi: 10.3389/fimmu.2018.02499
[21]	Voronov E,, Dotan S,, Apte RN. IL-1-induced inflammation promotes development of leishmaniasis in susceptible BALB/c mice[J]. Int Immunol, 2010, 22(4): 245-257. doi: 10.1093/intimm/dxq006 pmid: 20181656
[22]	Van Ginderachter JA, Beschin A,, Raes G, et al. Myeloid-derived suppressor cells in parasitic infections[J]. Eur J Immunol, 2010, 40(11): 2976-2985. doi: 10.1002/eji.201040911 pmid: 21061431
[23]	Yang Q,, Qiu H,, Huang J, et al. A Schistosoma japonicum infection promotes the expansion of myeloid-derived suppressor cells by activating the JAK/STAT3 pathway[J]. J Immunol, 2017, 198(12): 4716-4727. doi: 10.4049/jimmunol.1601860
[24]	Gao YQ,, Li L,, Hu L, et al. Immuno-supressive function of myeloid derived suppressor cells to T cells in BALB/c mice infected with Schistosoma japonicum[J]. Chin J Immunol, 2019, 35(3): 269-273. (in Chinese)
	( 高勇强,, 黎丽,, 胡丽, 等. 日本血吸虫感染小鼠的髓源抑制细胞对T细胞的免疫抑制[J]. 中国免疫学杂志, 2019, 35(3): 269-273.)
[25]	Rasta JL,, Green WR. Myeloid-derived suppressor cells in murine AIDS inhibit B-cell responses in part via soluble mediators including reactive oxygen and nitrogen species, and TGF-β[J]. Virology, 2016, 499: 9-22. doi: 10.1016/j.virol.2016.08.031
[26]	Schwacha MG,, Scroggins SR,, Cap AP, et al. Bum injury is associated with an infiltration of the wound site with myeloid-derived suppressor cells[J]. Cell Immunol, 2019, 338: 21-26. doi: S0008-8749(18)30555-0 pmid: 30902343

引物名称 Primer name	引物序列（5′→3′） Primer sequence (5′→3′)
白介素-6 IL-6	F: TACCACTTCACAAGTCGGAGGC R: CTGCAAGTGCATCATCGTTGTTC
S100钙结合蛋白A8 S100A8	F: TGCCCTCTACAAGAATGACT R: AAGCTCTGCTACTCCTTGTG
S100钙结合蛋白A9 S100A9	F: CCCTGACACCCTGAGCAAGAAG R: TTTCCCAGAACAAAGGCCATTGAG
血红素结合膜糖蛋白91 gp91	F: TGGCGATCTCAGCAAAAGGTGG R: GTACTGTCCCACCTCCATCTTG
精氨酸酶1 Arg1	F: AAGAATGGAAGAGTCAGTGTGG R: GGGAGTGTTGATGTCAGTGTG
一氧化氮合成酶 iNOS	F: GAGACAGGGAAGTCTGAAGCAC R: CCAGCAGTAGTTGCTCCTCTTC
白介素-10 IL-10	F: CGGGAAGACAATAACTGCACCC R: CGGTTAGCAGTATGTTGTCCAGC
转换生长因子-β TGF-β	F: TGATACGCCTGAGTGGCTGTCT R: CACAAGAGCAGTGAGCGCTGAA
β-肌动蛋白 β-actin	F: CATTGCTGACAGGATGCAGAAGG R: TGCTGGAAGGTGGACAGTGAGG

脾组织相关因子 Splenic tissue related factor	对照组 Control group	感染组Infection group
脾组织相关因子 Splenic tissue related factor	对照组 Control group	感染后4周 4 weeks after infection	感染后6周 6 weeks after infection	感染后8周 8 weeks after infection
白细胞介素-6 IL-6	1.05 ± 0.10	0.92 ± 0.12	2.74 ± 0.25	1.73 ± 0.25
S100钙结合蛋白A8 S100A8	1.01 ± 0.11	0.01 ± 0.01	51.4 ± 1.25	4.67 ± 0.70
S100钙结合蛋白A9 S100A9	1.02 ± 0.07	0.02 ± 0.01	39.20 ± 2.83	5.99 ± 0.52
血红素结合膜糖蛋白91 gp91	1.04 ± 0.09	0.55 ± 0.01	2.15 ± 0.08	1.12 ± 0.14
精氨酸酶1 Arg1	1.01 ± 0.06	0.75 ± 0.13	2.33 ± 0.39	3.44 ± 0.61
一氧化氮合酶 iNOS	1.00 ± 0.05	1.24 ± 0.35	1.57 ± 0.08	3.12 ± 0.44
白细胞介素-10 IL-10	0.98 ± 0.20	1.33 ± 0.35	2.20 ± 0.39	4.59 ± 1.59
转换生长因子-β TGF-β	1.00 ± 0.04	0.62 ± 0.17	1.44 ± 0.05	0.74 ± 0.06

PMN-MDSC相关因子 PMN-MDSC related factor	对照组 Control group	感染组 Infection group
PMN-MDSC相关因子 PMN-MDSC related factor	对照组 Control group	感染后4周 4 weeks after infection	感染后6周 6 weeks after infection	感染后8周 8 weeks after infection
一氧化氮合酶 iNOS	1.02 ± 0.13	1.08 ± 0.01	12.06 ± 1.80	32.12 ± 2.30
白细胞介素-10 IL-10	1.06 ± 0.12	1.14 ± 0.35	1.50 ± 0.36	2.64 ± 0.37