斯氏艾美耳球虫重组表面抗原SAG13和SAG14对兔的免疫保护效果初步观察

doi:10.11843/j.issn.0366-6964.2022.03.021

Abstract

Abstract: This experiment was conducted to observe the immunoprotective effects of the prokaryotic expression products of surface antigens SAG13 and SAG14 of Eimeria stiedae in rabbits. We analyzed the transcription level of Es-SAGs in unsporulated oocyst stage, sporulated oocyst stage, schizogony stage and gametogony stage by relative fluorescence quantitative PCR, and expressed Es-SAG specifically expressed in the schizogony stage. Then forty 45-day-old coccidia-free rabbits were randomly divided into four groups:blank control group (B), parasite challenge control group (P), rEs-SAG13 immunization group (SAG13) and rEs-SAG14 immunization group (SAG14). Rabbits of group B, group P, group SAG13 and group SAG14 were subcutaneously injected with 1 mL PBS, 1 mL PBS, 1 mL 100 μg·mL^-1 rEs-SAG13 and 1 mL 100 μg·mL^-1 rEs-SAG14, respectively, then the same amount of booster immunity was given after 14 days. Two weeks after the second immunization, 1×10⁴ sporulated oocysts were infected orally in all groups except group B. Three weeks after infection, the differences of clinical symptoms, oocyst excretion, relative weight gain, feed conversion ratio, liver index and liver lesions were observed in each group. Results revealed that the transcription level of Es-SAGs at each stage is significantly different, and SAG13 and SAG14 are members that are specifically expressed in schizogony stage. The rEs-SAG13 and rEs-SAG14 have good reactogenicity. The typical symptoms of rabbit hepatic coccidiosis appeared in the control group, while the symptoms of the other groups were not obvious. The oocyst reduction rates of rEs-SAG13 immunization group and rEs-SAG14 immunization group were 82.8% and 51.9%, respectively. The average relative weight gain of immune group was significantly higher than that of parasite challenge control group. There were significant differences in liver index and liver lesion score between rEs-SAG13 immunization group and parasite challenge control group. However, there was no significant difference in liver index and liver lesion score between rEs-SAG14 immunization group and parasite challenge control group (P<0.05). These results indicated that rEs-SAG13 and rEs-SAG14 can reduce weight gain loss and oocyst output in varying degrees, alleviate liver lesions, but rEs-SAG13 is more effective.

Key words: Eimeria stiedae, surface antigens, surface antigen 13, surface antigen 14, prokaryotic expression, immunoprotective

CLC Number:

S852.723

LUO Yuejun, REN Yongjun, BAI Xin, CHEN Hao, PU Jiayan, ZHENG Ruoyu, XIAO Jie, YANG Guangyou. Preliminary Observation on the Immunoprotective Effects of Recombinant Surface Antigens SAG13 and SAG14 of Eimeria stiedae in Rabbits[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 883-893.

References 36

[1]	WEI W R, SHEN N X, XIAO J, et al. Expression analysis and serodiagnostic potential of microneme proteins 1 and 3 in Eimeria stiedai[J]. Genes, 2020, 11(7):725.
[2]	XIE Y, XIAO J, ZHOU X, et al. Global transcriptome landscape of the rabbit protozoan parasite Eimeria stiedae[J]. Parasites Vectors, 2021, 14(1):308
[3]	JING F, YIN G W, LIU X Y, et al. Large-scale survey of the prevalence of Eimeria infections in domestic rabbits in China[J]. Parasitol Res, 2012, 110(4):1495-1500.
[4]	YIN G W, GORAYA M U, HUANG J H, et al. Survey of coccidial infection of rabbits in Sichuan Province, Southwest China[J]. SpringerPlus, 2016, 5(1):870.
[5]	BACHENE M S, TEMIM S, AINBAZIZ H, et al. A vaccination trial with a precocious line of Eimeria magna in Algerian local rabbits Oryctolagus cuniculus[J]. Vet Parasitol, 2018, 261:73-76.
[6]	GU X L, LIU H B, LI C, et al. Selection and characterization of a precocious line of Eimeria media[J]. Parasitol Res, 2019, 118(10):3033-3041.
[7]	LI C, TAO G R, GU X L, et al. Selection and identification of a precocious line of Eimeria intestinalis with enlarged oocysts and deletion of one generation of schizogony[J]. Parasitol Res, 2019, 118(3):969-976.
[8]	FANG S F, GU X L, EL-ASHRAM S, et al. Immune protection provided by a precocious line trivalent vaccine against rabbit Eimeria[J]. Vet Parasitol, 2019, 275:108927.
[9]	DANFORTH H D. Development and use of hybridoma antibodies directed against Eimeria acervulina merozoites for cross-reactive and ferritin-labeling studies[J]. Avian Dis, 1987, 31(1):99-104.
[10]	JENKINS M C, AUGUSTINE P C, DANFORTH H D, et al. X-irradiation of Eimeria tenella oocysts provides direct evidence that sporozoite invasion and early schizont development induce a protective immune response (s)[J]. Infect Immun, 1991, 59(11):4042-4048.
[11]	LILLEHOJ H S. Immune response during coccidiosis in SC and FP chickens. I. In vitro assessment of T cell proliferation response to stage-specific parasite antigens[J]. Vet Immunol Immunopathol, 1986, 13(4):321-330.
[12]	CONSTANTINOIU C C, MOLLOY J B, JORGENSEN W K, et al. Antibody response against endogenous stages of an attenuated strain of Eimeria tenella[J]. Vet Parasitol, 2008, 154(3-4):193-204.
[13]	SONG X K, GAO Y L, XU L X, et al. Partial protection against four species of chicken coccidia induced by multivalent subunit vaccine[J]. Vet Parasitol, 2015, 212(3-4):80-85.
[14]	LIU T Q, HUANG J W, LI Y L, et al. Molecular characterisation and the protective immunity evaluation of Eimeria maxima surface antigen gene[J]. Parasites Vectors, 2018, 11(1):325.
[15]	ZHAO P F, LI Y C, ZHOU Y Q, et al. In vivo immunoprotective comparison between recombinant protein and DNA vaccine of Eimeria tenella surface antigen 4[J]. Vet Parasitol, 2020, 278:109032.
[16]	李祥瑞,汪志楷.斯氏艾美耳球虫生活史的研究[J].南京农业大学学报, 1989, 12(2):97-101.LI X R, WANG Z K. Study on life cycle of Eimeria stiedai[J]. Journal of Nanjing Agricultural University, 1989, 12(2):97-101.(in Chinese)
[17]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the method[J]. Methods, 2001, 25(4):402-408.
[18]	PEETERS J E, GEEROMS R. Efficacy of toltrazuril against intestinal and hepatic coccidiosis in rabbits[J]. Vet Parasitol, 1986, 22(1-2):21-35.
[19]	张龙现,林昆华.从病理变化角度探讨黄艾美耳球虫活虫苗不同免疫程序的免疫保护性[J].畜牧兽医学报, 1995, 26(5):426-434.ZHANG L X, LIN K H. Protection of Eimeria flavescens live virulent vaccine used indifference immune procedure by comparison of the intestinal mucosal surface deform[J]. Acta Veterinaria et Zootechnica Sinica, 1995, 26(5):426-434.(in Chinese)
[20]	LICOIS D, COUDERT P, DROUET-VIARD F, et al. Eimeria magna:pathogenicity, immunogenicity and selection of a precocious line[J]. Vet Parasitol, 1995, 60(1-2):27-35.
[21]	崔平,方素芳,顾小龙,等.大型艾美耳球虫早熟株的选育及18S rDNA系统发育分析[J].中国兽医学报, 2018, 38(7):1327-1331.CUI P, FANG S F, GU X L, et al. Selection and phylogenetic analysis of 18S rDNA from precocious line of Eimeria magna[J]. Chinese Journal of Veterinary Science, 2018, 38(7):1327-1331.(in Chinese)
[22]	LICOIS D, COUDERT P, DROUET-VIARD F, et al. Eimeria media:selection and characterization of a precocious line[J]. Parasitol Res, 1994, 80(1):48-52.
[23]	PAKANDL M, JELÍNKOVÁ A. The rabbit coccidium Eimeria piriformis:selection of a precocious line and life-cycle study[J]. Vet Parasitol, 2006, 137(3-4):351-354.
[24]	PAKANDL M. Selection of a precocious line of the rabbit coccidium Eimeria flavescens Marotel and Guilhon (1941) and characterisation of its endogenous cycle[J]. Parasitol Res, 2005, 97(2):150-155.
[25]	孟庆玲,乔军,才学鹏,等.斯氏艾美耳球虫MIC-5与兔IL-15基因在真核细胞中的共表达[J].中国农业科学, 2011, 44(19):4096-4101.MENG Q L, QIAO J, CAI X P, et al. Co-expression of MIC-5 Gene of Eimeria stiedai and rabbit IL-15 in Eucaryotic cell line[J]. Scientia Agricultura Sinica, 2011, 44(19):4096-4101.(in Chinese)
[26]	孟庆玲,乔军,才学鹏,等.携带兔斯氏艾美耳球虫MIC-5基因减毒鼠伤寒沙门菌活疫苗的安全性、稳定性与免疫原性[J].中国兽医学报, 2011, 31(5):659-662.MENG Q L, QIAO J, CAI X P, et al. Safety, stability and immunogenicity of attenuated Salmonella typhimurium carrying MIC-5 gene of Eimeria stiedae[J]. Chinese Journal of Veterinary Science, 2011, 31(5):659-662.(in Chinese)
[27]	LICOIS D, COUDERT P, BOIVIN M, et al. Selection and characterization of a precocious line of Eimeria intestinalis, an intestinal rabbit coccidium[J]. Parasitol Res, 1990, 76(3):192-198.
[28]	OMATA Y, SUEDA M, KOYAMA T, et al. Identification and the role of soluble antigens detected in bile from Eimeria stiedai-infected rabbits[J]. J Parasitol, 2001, 87(2):287-291.
[29]	HANADA S, UMEMOTO Y, OMATA Y, et al. Eimeria stiedai merozoite 49-kDa soluble antigen induces protection against infection[J]. J Parasitol, 2003, 89(3):613-617.
[30]	HANADA S, OMATA Y, UMEMOTO Y, et al. Relationship between liver disorders and protection against Eimeria stiedai infection in rabbits immunized with soluble antigens from the bile of infected rabbits[J]. Vet Parasitol, 2003, 111(2-3):261-266.
[31]	GRIMWOOD J, SMITH J E. Toxoplasma gondii:the role of a 30-kDa surface protein in host cell invasion[J]. Exp Parasitol, 1992, 74(1):106-111.
[32]	LI M X, AO Y S Q, GUO J Y, et al. Surface antigen 1 is a crucial secreted protein that mediates Babesia microti invasion into host cells[J]. Front Microbiol, 2020, 10:3046.
[33]	COELHO C, VIEIRA-PINTO M, VILARES A, et al. PCR Detection of Toxoplasma gondii in European wild rabbit (Oryctolagus cuniculus) from Portugal[J]. Microorganisms, 2020, 8(12):1926.
[34]	梁凯,李艳文,王贝贝,等.刚地弓形虫重组蛋白表面抗原1与4诱导小鼠免疫保护性的比较研究[J].中国寄生虫学与寄生虫病杂志, 2019, 37(2):155-160.LIANG K, LI Y W, WANG B B, et al. Protective immunity induced by recombinant surface protein-1 and surface protein-4 against Toxoplasma gondii infection in mice[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2019, 37(2):155-160.(in Chinese)
[35]	TABARÉS E, FERGUSON D, CLARK J, et al. Eimeria tenella sporozoites and merozoites differentially express glycosylphosphatidylinositol-anchored variant surface proteins[J]. Mol Biochem Parasitol, 2004, 135(1):123-132.
[36]	潘晓亮,丁熙成,蒋金书. TA4和EtMIC-2表达产物免疫后对感染柔嫩艾美耳球虫(Eimeria tenella)鸡增重和盲肠卵囊数的影响[J].黑龙江畜牧兽医, 2002(8):6-7.PAN X L, DING X C, JIANG J S. Protection of chickens against Eimeria tenella by three expressed proteins and plasmids DNA[J]. Heilongjiang Journal of Animal Science and Veterinary Medicine, 2002(8):6-7.(in Chinese)

Preliminary Observation on the Immunoprotective Effects of Recombinant Surface Antigens SAG13 and SAG14 of Eimeria stiedae in Rabbits

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