Enterovirus 71 capsid protein VP1 specifically recognizes and activates CD4+ T cell immune activation in children with hand, foot and mouth disease

doi:10.3877/cma.j.issn.1674-1358.2018.05.003

Abstract

Abstract:

Objective

To investigate the characteristics of interferon-γ(IFN-γ) response induced by enterovirus 71 (EV71) capsid protein VP1 in peripheral blood T lymphocytes of children with EV71 infection.

Methods

From September 2010 to December 2011, a total of 31 patients with hand, foot and mouth disease (HFMD) infected with EV71 were collected from the Second Affiliated Hospital of Medical College of Xi’an Jiaotong University and Xi’an Children’s Hospital. According to the clinical features, 31 cases were divided into three groups: acute infection group (16 cases), clinical convalescence group (9 cases) and one month after recovery of infection group (6 cases), in addition, 10 healthy children were selected as control group. Total of 36 overlapping peptides of EV71 capsid protein VP1 were synthesized by overlapping peptide technique as antigens. The immunological responses of EV71 VP1 specific CD4⁺T and CD8⁺T cells were detected by IFN-γenzyme-linked immunoblot assay (ELISPOT) and magnetic cell sorting (MACs) in vitro.

Results

In the acute infection group, 4 cases (25%) had T cell immune response to VP1 protein, 5 cases (56%) in the clinical convalescence group had T cell immune response to VP1 protein. The strength of T-cell responses in clinical convalescence group had the strongest, significantly higher than acute infection group (Z=-2.042,P= 0.041), and one month after recovery of infection group (Z=-2.105,P = 0.035) and control group (Z=-2.633,P = 0.008), with significant differences. The T cell immune response was not detected in the control group and one month after recovery of infection group. The results suggested that the frequency of VP1 protein recognition (the number of cases) disappeared with the disappearance of clinical symptoms and signs in children with EV71 infection. The cellular immune response induced by VP1 protein was dominated by specific CD4⁺T cells. Eight CD4⁺T cell epitopes of EV71 VP1 protein region were identified.

Conclusions

There was a specific T cell response to VP1 protein in children with EV71 infection, and the specific CD4⁺T cells were dominant. The CD4⁺T cell epitopes of VP1 protein were identified in EV71 infected children. It is helpful to elucidate the cellular immunopathogenesis of EV71 infection.

Key words: Enterovirus 71, T cell-mediated immunity, Capsid protein VP1, Enzyme-linked immunospot assays, T cell Epitope

Ning Gao, Mei Li, Yaping Li, Wenjun Wang, Juanjuan Shi, Xiaoli Jia, Shuangsuo Dang. Enterovirus 71 capsid protein VP1 specifically recognizes and activates CD4⁺ T cell immune activation in children with hand, foot and mouth disease[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2018, 12(05): 427-433.

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References 25

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临床特征	急性感染期组(16例)	临床恢复期组(9例)	感染恢复后1个月组(6例)
发病年龄(月)^a	23.63(8～49)	18.22(9～41)	28.17(13～43)
性别(女/男，例)	8/8	2/7	4/2
采血时间(发病天数)^a	3(2～4)	11(8～16)	30
病程(d)^a	11.56(8～20)	11.56(8～17)	10.17(8～11)
合并中枢神经系统症状[例(%)]	7(43.75)	1(11.11)	0(0.00)
轻型/重型(例)	6/10	7/2	6/0
hsCRP(mg/L)^a	2.80(0.8～4.7)	8.10(4.7～11.9)	—
最高体温(℃)^a	38.4(38.0～40.0)	38.5(38.0～39.2)	38.2(37.8～38.5)
白细胞计数(10⁹/L)^a	10.98(8.30～14.70)	11.06(6.70～19.90)	7.21(3.81～11.20)

临床特征	急性感染期组(16例)	临床恢复期组(9例)	感染恢复后1个月组(6例)
发病年龄(月)^a	23.63(8～49)	18.22(9～41)	28.17(13～43)
性别(女/男，例)	8/8	2/7	4/2
采血时间(发病天数)^a	3(2～4)	11(8～16)	30
病程(d)^a	11.56(8～20)	11.56(8～17)	10.17(8～11)
合并中枢神经系统症状[例(%)]	7(43.75)	1(11.11)	0(0.00)
轻型/重型(例)	6/10	7/2	6/0
hsCRP(mg/L)^a	2.80(0.8～4.7)	8.10(4.7～11.9)	—
最高体温(℃)^a	38.4(38.0～40.0)	38.5(38.0～39.2)	38.2(37.8～38.5)
白细胞计数(10⁹/L)^a	10.98(8.30～14.70)	11.06(6.70～19.90)	7.21(3.81～11.20)

组别	Z值	P值
急性感染期组vs.临床恢复期组	－2.042	0.041^a
急性感染期组 vs.感染恢复后1个月组	－1.315	0.189
急性感染期组 vs.对照组	－1.678	0.093
临床恢复期组 vs.感染恢复后1个月组	－2.105	0.035^a
临床恢复期组 vs.对照组	－2.633	0.008^a
感染恢复后1个月组 vs.对照组	0.000	1.000

组别	Z值	P值
急性感染期组vs.临床恢复期组	－2.042	0.041^a
急性感染期组 vs.感染恢复后1个月组	－1.315	0.189
急性感染期组 vs.对照组	－1.678	0.093
临床恢复期组 vs.感染恢复后1个月组	－2.105	0.035^a
临床恢复期组 vs.对照组	－2.633	0.008^a
感染恢复后1个月组 vs.对照组	0.000	1.000

患者编号	重叠肽段^a	氨基酸位置	应答强度^b(SFUs/10⁶cells)	肽段序列^c	CD4/CD8为主导
1	VP1-22	734～751	711	LAWQTATNPSVFVKLSDP	4
	VP1-23	742～759	736	PSVFVKLSDPPAQVSVPF	?
	VP1-25	758～775	658	PFMSPASAYQWFYDGYPT	?
2	VP1-19	710～727	40	EVVPQLLQYMFVPPGAPK	4
	VP1-28	782～799	46	EKDLEYGACPNNMMGTFS	?
	VP1-30	798～815	53	FSVRTVGSSKSKYPLVIR	?
	VP1-33	822～839	53	HVRAWIPRPMRNQNYLFK	?
	VP1-34	830～847	33	PMRNQNYLFKSNPNYAGD	?

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