Primary characterization of platelet activation derived from mice infected by Babesia microti

doi:10.11853/j.issn.1003.8280.2018.01.009

Chines Journal of Vector Biology and Control ›› 2018, Vol. 29 ›› Issue (1): 34-37.DOI: 10.11853/j.issn.1003.8280.2018.01.009

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Primary characterization of platelet activation derived from mice infected by Babesia microti

PAN Zhi-hua^1,2, ZHENG Chen¹, CHEN Jia-xu³, ZHENG Kui-yang¹, LIU Xiang-ye¹

1 Jiangsu Key Laboratory of Immunity and Metabolism, Department of Pathogenic Biology and Immunology, Xuzhou Medical University, Xuzhou 221004, Jiangsu Province, China;
2 Experimental Teaching Center of Morphology, Xuzhou Medical University;
3 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, Ministry of Health of China, WHO Collaborating Centre for Malaria, Schistosomiasis and Filariasis

Received:2017-08-21 Online:2018-02-20 Published:2018-02-20
Supported by:
Supported by the China Postdoctoral Science Foundation (No. 2015M580472), Natural Science Foundation of Jiangsu Province of China (No. BK20150212) and Research Foundation of Xuzhou Medical University (No. D2015006)

田鼠巴贝西虫感染致宿主血小板活化初步研究

潘智华^1,2, 郑辰¹, 陈家旭³, 郑葵阳¹, 刘相叶¹

1 江苏省免疫与代谢重点实验室, 徐州医科大学病原生物学与免疫学教研室, 江苏徐州 221004;
2 徐州医科大学形态学教学实验中心, 江苏徐州 221004;
3 中国疾病预防控制中心寄生虫病预防控制所, 卫生部寄生虫病原与媒介生物学重点实验室, 世界卫生组织疟疾、血吸虫病和丝虫病合作中心, 上海 200025

通讯作者: 刘相叶,Email:liuxy83@xzhmu.edu.cn
作者简介:潘智华,女,硕士,实验师,主要从事寄生虫病致病机制研究,Email:562777067@qq.com
基金资助:
中国博士后科学基金（2015M580472）；江苏省自然科学基金（BK20150212）；徐州医科大学优秀人才科研启动基金（D2015006）

Abstract

Abstract: Objective To characterize platelet activation and its correlations with parasitemia in peripheral blood of mice infected by Babesia microti. Methods BALB/c mice were intraperitoneally inoculated with B. microti infected blood, which was collected, heparinized, and diluted with sterile saline to achieve a 30% RBC parasitic infection rate. Following the infection, anti-coagulated peripheral blood was collected from the mice at each day. The blood was stained with anti-CD61 and anti-CD62P antibodies, and then used to analyze the activation of platelet with flow cytometry. Furthermore, the percentage of parasitized red blood cells was determined via Giemsa-stained thin blood films. Finally, Pearson correlation analysis was used to evaluate the correlation of platelet activation and parasitemia. Results The percentage of platelet activation started to increase on the second day post infection, reached the peak on the third day[(15.50±0.17)%], and declined from the fourth day, but was absence of obvious regularities. The percentage of parasitized red blood cells gradually ascended post infection and reached the peak at the eighth day[(63.10±3.43)%], and then declined. However, there was no significant difference between the percentage of platelet activation and parasitized red blood cells, which was characterized by Pearson correlation analysis (P>0.05). Conclusion The platelet activation can be induced by B. microti infection, but it has no significant correlation with parasitemia.

Key words: Babesia microti, Platelet activation, Flow cytometry, Parasitemia

摘要： 目的探讨BALB/c小鼠感染田鼠巴贝西虫后外周血中血小板活化情况及其与红细胞染虫率的关系。方法利用田鼠巴贝西虫红细胞染虫率为30%的种鼠抗凝血经腹腔注射感染BALB/c小鼠，感染后第1天开始采集BALB/c小鼠外周抗凝血，利用流式细胞术分别以白细胞分化抗原CD61（整合蛋白β3）和CD62P（P选择素）抗体标记，检测血小板和活化血小板数量；同时利用吉姆萨染色检测红细胞染虫率。采用Pearson相关性检验分析血小板活化率与红细胞染虫率的相关性。结果 BALB/c小鼠感染田鼠巴贝西虫后第2天其外周血中血小板的活化率开始升高，第3天最高，达（15.50±0.17）%，第4天开始下降，但无明显规律性；BALB/c小鼠在感染田鼠巴贝西虫后外周血中红细胞染虫率逐渐升高，第8天最高，达（63.10±3.43）%，之后逐渐下降。血小板活化率与红细胞染虫率比较差异无统计学意义（P=0.768）。结论田鼠巴贝西虫感染可引起宿主外周血中血小板的活化率升高，但与红细胞染虫率无显著相关性。

关键词: 田鼠巴贝西虫, 血小板活化, 流式细胞术, 红细胞染虫率

CLC Number:

PAN Zhi-hua, ZHENG Chen, CHEN Jia-xu, ZHENG Kui-yang, LIU Xiang-ye. Primary characterization of platelet activation derived from mice infected by Babesia microti[J]. Chines Journal of Vector Biology and Control, 2018, 29(1): 34-37.

潘智华, 郑辰, 陈家旭, 郑葵阳, 刘相叶. 田鼠巴贝西虫感染致宿主血小板活化初步研究[J]. 中国媒介生物学及控制杂志, 2018, 29(1): 34-37.

References

[1] Vannier E, Krause PJ. Human babesiosis[J]. N Engl J Med, 2012,366(25):2397-2407.
[2] Vannier E, Krause PJ. Babesiosis in China, an emerging threat[J]. Lancet Infect Dis,2015,15(2):137-139.
[3] Fang LQ, Liu K, Li XL, et al. Emerging tick-borne infections in mainland China:an increasing public health threat[J]. Lancet Infect Dis,2015,15(12):1467-1479.
[4] Lemieux JE,Tran AD,Freimark L,et al. A global map of genetic diversity in Babesia microti reveals strong population structure and identifies variants associated with clinical relapse[J]. Nat Microbiol,2016,1(7):16079.
[5] Obiegala A, Pfeffer M, Pfister K, et al. Molecular examinations of Babesia microti in rodents and rodent-attached ticks from urban and sylvatic habitats in Germany[J]. Ticks Tick Borne Dis,2015,6(4):445-449.
[6] Bullard JM, Ahsanuddin AN, Perry AM, et al. The first case of locally acquired tick-borne Babesia microti infection in Canada[J]. Can J Infect Dis Med Microbiol,2014,25(6):e87-89.
[7] Tuvshintulga B, Sivakumar T, Battsetseg B, et al. The PCR detection and phylogenetic characterization of Babesia microti in questing ticks in Mongolia[J]. Parasitol Int,2015,64(6):527-532.
[8] Smyth SS, McEver RP, Weyrich AS, et al. Platelet functions beyond hemostasis[J]. J Thromb Haemost,2009,7(11):1759-1766.
[9] Czapiga M, Kirk AD, Lekstrom-Himes J, et al. Platelets deliver costimulatory signals to antigen-presenting cells:a potential bridge between injury and immune activation[J]. Exp Hematol, 2004,32(2):135-139.
[10] Sharma J, Eickhoff CS, Hoft DF, et al. Absence of calcium-independent phospholipase A2β impairs platelet-activating factor production and inflammatory cell recruitment in Trypanosoma cruzi-infected endothelial cells[J]. Physiol Rep, 2014,2(1):e00196.
[11] Goncalves R,Zhang X,Cohen H,et al. Platelet activation attracts a subpopulation of effector monocytes to sites of Leishmania major infection[J]. J Exp Med,2011,208(6):1253-1265.
[12] McMorran BJ, Wieczorski L, Drysdale KE, et al. Platelet factor 4 and Duffy antigen required for platelet killing of Plasmodium falciparum[J]. Science,2012,338(6112):1348-1351.
[13] McMorran BJ, Marshall VM, de Graaf C, et al. Platelets kill intraerythrocytic malarial parasites and mediate survival to infection[J]. Science,2009,323(5915):797-800.
[14] Aggrey AA, Srivastava K, Ture S, et al. Platelet induction of the acute-phase response is protective in murine experimental cerebral malaria[J]. J Immunol,2013,190(9):4685-4691.
[15] Goddard A,Leisewitz AL,Kristensen AT,et al. Platelet activation and platelet-leukocyte interaction in dogs naturally infected with Babesia rossi[J]. Vet J,2015,205(3):387-392.
[16] Michelson AD, Barnard MR, Krueger LA, et al. Circulating monocyte-platelet aggregates are a more sensitive marker of in vivo platelet activation than platelet surface P-selection:studies in baboons, human coronary intervention, and human acute myocardial infarction[J]. Circulation,2001,104(13):1533-1537.
[17] Moritz A, Walcheck BK, Weiss DJ. Flow cytometric detection of activated platelets in the dog[J]. Vet Clin Pathol,2003,32(1):6-12.
[18] 张加,司晨晨, 蔡玉春,等. 田鼠巴贝虫可溶性抗原组分分析及其初步应用[J]. 中国人兽共患病学报,2014,30(5):469-478.
[19] Zhou X,Xia S,Huang JL, et al. Human babesiosis, an emerging tick-borne disease in the people's republic of China[J]. Parasit Vectors,2014,7:509.
[20] Emuchay CI, Usanga EA. Increased platelet factor 3 activity in Plasmodium falciparum malaria[J]. East Afr Med J,1997,74(8):527-529.
[21] Pain A,Ferguson DJP, Kai O, et al. Platelet-mediated clumping of Plasmodium falciparum-infected erythrocytes is a common adhesive phenotype and is associated with severe malaria[J]. Proc Natl Acad Sci USA,2001,98(4):1805-1810.
[22] Chotivanich K, Sritabal J, Udomsangpetch R, et al. Platelet-induced autoagglutination of Plasmodium falciparum-infected red blood cells and disease severity in Thailand[J]. J Infect Dis, 2004,189(6):1052-1055.
[23] 魏金龙,周勇志,龚海燕,等. 田鼠巴贝斯虫感染对宿主凝血系统的影响[J]. 中国兽医科学,2015,45(4):390-394.
[24] Piguet PF, Kan CD, Vesin C. Thrombocytopenia in an animal model of malaria is associated with an increased caspase-mediated death of thrombocytes[J]. Apoptosis,2002,7(2):91-98.
[25] Gramaglia I,Sahlin H,Nolan JP,et al. Cell-rather than antibody-mediated immunity leads to the development of profound thrombocytopenia during experimental Plasmodium berghei malaria[J]. J Immunol,2005,175(11):7699-7707.
[26] Evelyn ME, Ezeiruaku F, Ukaji DC. Experiential relationship between malaria parasite density and some haematological parameters in malaria infected male subjects in Port Harcourt, Nigeria[J]. Glob J Health Sci,2012,4(4):139-148.

Primary characterization of platelet activation derived from mice infected by Babesia microti

田鼠巴贝西虫感染致宿主血小板活化初步研究

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