淋巴细胞亚群、炎症因子和氨基末端B型钠尿肽原联合检测诊断川崎病

doi:10.3969/j.issn.1673-8640.2023.06.005

摘要/Abstract

摘要：

目的分析川崎病（KD）急性期患儿淋巴细胞亚群和相关炎症因子、心脏标志物的变化，探讨其在KD诊断中的临床价值。方法选取2016年10月—2019年3月上海市儿童医院222例KD急性期患儿（KD组）和177例感染发热的非KD患儿（感染发热组）。采用流式荧光法检测患儿外周血13种炎症因子[白细胞介素（IL）-8、IL-1β、IL-2、IL-6、IL-10、IL-4、IL-5、IL-12P70、IL-17A、IL-18、肿瘤坏死因子α（TNF-α）、γ-干扰素（INF-γ）、sCD25]水平，采用流式细胞术检测外周血淋巴细胞。比较KD组与感染发热组外周血炎症因子和淋巴细胞水平的差异，并比较KD患儿丙种球蛋白静脉治疗（IVIG）前后各炎症因子水平的变化。采用Spearman相关分析评价145例KD患儿心脏标志物[心肌肌钙蛋白I（cTnI）、肌红蛋白（MYO）、氨基末端B型钠尿肽原（NT-proBNP）]与淋巴细胞亚群和炎症因子的相关性。结果与感染发热组比较，除IL-2外，KD组其他炎症因子水平均升高（P<0.05 ）；外周血CD3⁺CD4⁺T细胞百分比、CD16⁺CD56⁺自然杀伤细胞百分比、CD4/CD8比值升高，CD3⁺CD8⁺ T细胞百分比下降（P<0.05 ）；CD16⁺CD56⁺自然杀伤细胞绝对值升高，CD3⁺CD8⁺ T细胞绝对值下降（P<0.05）。KD患儿治疗后血清IL-8、IL-1β、IL-2、IL-6、IL-10、IL-18、TNF-α、sCD25均明显降低（P<0.05 ）。KD患儿外周血cTnI 、MYO未见明显升高；有62%的患儿外周血NT-proBNP明显升高，与IL-6、IL-10、IL-17A、IL-18、TNF-α、INF-γ、sCD25、CD19⁺ B细胞百分比和绝对值、CD4/CD8比值呈正相关，与CD3⁺CD8⁺ T细胞百分比呈负相关。结论 KD急性期患儿存在炎症因子上调和淋巴细胞亚群比例失衡，NT-proBNP联合免疫细胞和炎症因子检测可更好地辅助临床诊疗。

关键词: 淋巴细胞亚群, 炎症因子, 氨基末端B型钠尿肽原, 川崎病

Abstract:

Objective To investigate the changes and roles of lymphocyte subsets，inflammatory factors and myocardial marker in children with acute Kawasaki disease（KD）. Methods From October 2016 to March 2019，222 acute KD children and 177 non-KD children with infection and fever were enrolled from Children's Hospital of Shanghai. Lymphocytes were determined by flow cytometry. The levels of 13 inflammatory factors [interleukin（IL）-8，IL-1β，IL-2，IL-6，IL-10，IL-4，IL-5，IL-12P70，IL-17A，IL-18，tumor necrosis factor-alpha（TNF-α），interferon-gamma （INF-γ）and sCD25] were determined flow cytometry and immunofluorescence technique Peripheral blood lymphocytes were determined by flow cytometry. The expressions of inflammatory cytokines and lymphocytes in peripheral blood of children with KD in acute phase and non-KD patients with fever-infected were compared. The expressions of inflammatory factors were compared before and after intravenous injection gamma globulin （IVIG） in children with KD. The changes of cardiac markers，including cardiac troponin I （cTnI），myoglobin （MYO） and N-terminal pro-B-type natriuretic peptide （NT-proBNP），in 145 children with KD in acute phase were determined，and the correlations were analyzed. Results Compared with children with infection and fever，other inflammatory factors except IL-2 were increased in children with KD（P<0.05）. Peripheral blood CD3⁺CD4⁺ T cell percentage，CD16⁺CD56⁺ natural killer cell percentage，CD4/CD8 were increased，and CD3⁺CD8⁺ T cell percentage was decreased （P<0.05）. The absolute value of CD16⁺CD56⁺ natural killer cells was increased，and the absolute value of CD3⁺CD8⁺ T cells was decreased （P<0.05）. After treatment，serum IL-8，IL-1β，IL-2，IL-6，IL-10，IL-18，TNF-α and sCD25 were decreased（P<0.05）. There was no significant increase of cTnI and MYO in peripheral blood of children with KD. There was a significant increase of NT-proBNP in peripheral blood of 62% children，which was positively correlated with IL-6，IL-10，IL-17A，IL-18，TNF-α，INF-γ，sCD25，the percentage and absolute value of CD19⁺ B cells，CD4/CD8，and which was negatively correlated with the percentage of CD3⁺CD8⁺ T cells. Conclusions There is up-regulation of inflammatory factors and imbalance of lymphocyte subsets in acute KD. The determination of NT-proBNP combined with immune cells and inflammatory factors can assist clinical diagnosis and treatment.

Key words: Lymphocyte subset, Inflammatory factor, N-terminal pro-B-type natriuretic peptide, Kawasaki disease

中图分类号:

R446.1

刘亚楠, 夏敏, 胡韶华, 郑月, 张泓. 淋巴细胞亚群、炎症因子和氨基末端B型钠尿肽原联合检测诊断川崎病[J]. 检验医学, 2023, 38(6): 532-537.

LIU Yanan, XIA Min, HU Shaohua, ZHENG Yue, ZHANG Hong. Roles of lymphocyte subsets，inflammatory factors and NT-proBNP in diagnosis of Kawasaki disease[J]. Laboratory Medicine, 2023, 38(6): 532-537.

图/表 6

参考文献 17

[1]	ROWLEY A H, SHULMAN S T. The epidemiology and pathogenesis of Kawasaki disease[J]. Front Pediatr, 2018, 6:374. DOI PMID
[2]	RIFE E, GEDALIA A. Kawasaki disease:an update[J]. Curr Rheumatol Rep, 2020, 22(10):75. DOI
[3]	JCS Joint Working Group. Guidelines for diagnosis and management of cardiovascular sequelae in Kawasaki disease(JCS 2013). Digest version[J]. Circ J, 2014, 78(10):2521-2562. DOI URL
[4]	NAGATA S. Causes of Kawasaki disease-from past to present[J]. Front Pediatr, 2019, 7:18. DOI PMID
[5]	DEL PRINCIPE D, PIETRAFORTE D, GAMBARDELLA L, et al. Pathogenetic determinants in Kawasaki disease:the haematological point of view[J]. J Cell Mol Med, 2017, 21(4):632-639. DOI URL
[6]	SWANSON K V, DENG M, TING J P. The NLRP3 inflammasome:molecular activation and regulation to therapeutics[J]. Nat Rev Immunol, 2019, 19(8):477-489. DOI
[7]	KIM K Y, KIM D S. Recent advances in Kawasaki disease[J]. Yonsei Med J, 2016, 57(1):15-21. DOI PMID
[8]	WENG K P, HSIEH K S, HUANG S H, et al. Interleukin-18 and coronary artery lesions in patients with Kawasaki disease[J]. J Chin Med Assoc, 2013, 76(8):438-445. DOI URL
[9]	TERAURA H, KOTANI K, MINAMI T, et al. The serum concentration of soluble interleukin-2 receptor in patients with Kawasaki disease[J]. Ann Clin Biochem, 2017, 54(2):209-213. DOI PMID
[10]	WU L, CHEN Y, ZHONG S, et al. Blood N-terminal pro-brain natriuretic peptide and interleukin-17 for distinguishing incomplete Kawasaki disease from infectious diseases[J]. Indian Pediatr, 2015, 52(6):477-480. PMID
[11]	丁艳, 尹薇, 王瑞耕, 等. 川崎病患儿淋巴细胞亚群及免疫球蛋白的变化[J]. 临床儿科杂志, 2016, 34(1):29-32.
[12]	汪希珂, 刘晓英, 吴悦, 等. T细胞免疫及细胞因子在川崎病不同时期表达变化的研究[J]. 贵州医药, 2018, 42(3):276-280.
[13]	祝绚, 王雪梅, 刘成桂, 等. 川崎病患儿外周血淋巴细胞亚群变化及临床意义[J]. 国际检验医学杂志, 2016, 37(10):1393-1395.
[14]	LINDQUIST M E, HICAR M D. B cells and antibodies in Kawasaki disease[J]. Int J Mol Sci, 2019, 20(8):1834. DOI URL
[15]	ZHENG X, ZHANG Y, LIU L, et al. N-terminal pro-brain natriuretic peptide as a biomarker for predicting coronary artery lesion of Kawasaki disease[J]. Sci Rep, 2020, 10(1):5130. DOI PMID
[16]	DIONNE A, DAHDAH N. A decade of NT-proBNP in acute Kawasaki disease,from physiological response to clinical relevance[J]. Children(Basel), 2018, 5(10):141.
[17]	YANAGISAWA D, AYUSAWA M, KATO M, et al. Factors affecting N-terminal pro-brain natriuretic peptide elevation in the acute phase of Kawasaki disease[J]. Pediatr Int, 2016, 58(11):1105-1111. DOI PMID

组别	例数	IL-8									IL-1β				IL-2
组别	例数	P₅₀		P₇₅	P₉₀				P₉₅		P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
KD组	222	0.10		20.40	323.56				574.34		0.10	0.10	2.74	6.88	0.10	0.10	0.10	2.75
感染发热组	177	0.10		0.10	18.08				101.09		0.10	0.10	0.10	0.10	0.10	0.10	0.10	0.10
Z值		-5.347									-3.812				-1.893
P值		<0.001									<0.001				0.058
组别	IL-6				IL-10						IL-4				IL-5
组别	P₅₀	P₇₅	P₉₀	P₉₅	P₅₀			P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
KD组	0.10	25.11	14.91	357.13	0.10			7.64	29.48	60.92	0.10	0.10	0.10	5.89	0.10	0.10	0.10	5.89
感染发热组	0.10	0.10	0.10	0.29	0.10			0.10	3.02	15.41	0.10	0.10	0.10	0.10	0.10	0.10	0.10	0.10
Z值	-7.314				-6.687						-3.008				-3.239
P值	<0.001				<0.001						<0.001				0.001
组别	IL-12P70										IL-17A				IL-18
组别	P₅₀		P₇₅			P₉₀			P₉₅		P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
感染发热组	0.10		0.10			0.10			22.85		0.10	0.10	0.10	0.60	0.10	7.95	136.64	191.66
KD组	0.10		0.10			0.10			27.85		0.10	0.10	0.91	1.70	18.90	75.51	148.66	252.16
Z值	-2.856										-4.178				-5.950
P值	0.004										<0.001				<0.001
组别	TNF-α										INF-γ				sCD25
组别	P₅₀		P₇₅				P₉₀		P₉₅		P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
KD组	0.10		0.10				3.01		5.75		0.23	2.77	20.06	112.48	27 214.81	47 688.36	92 148.49	126 074.00
感染发热组	0.10		0.10				0.10		0.10		0.10	1.00	26.21	47.45	15 896.00	23 829.98	32 245.89	36 403.40
Z值	-5.420										-3.628				-7.521
P值	<0.001										<0.001				<0.001

组别	例数	IL-8									IL-1β				IL-2
组别	例数	P₅₀		P₇₅	P₉₀				P₉₅		P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
KD组	222	0.10		20.40	323.56				574.34		0.10	0.10	2.74	6.88	0.10	0.10	0.10	2.75
感染发热组	177	0.10		0.10	18.08				101.09		0.10	0.10	0.10	0.10	0.10	0.10	0.10	0.10
Z值		-5.347									-3.812				-1.893
P值		<0.001									<0.001				0.058
组别	IL-6				IL-10						IL-4				IL-5
组别	P₅₀	P₇₅	P₉₀	P₉₅	P₅₀			P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
KD组	0.10	25.11	14.91	357.13	0.10			7.64	29.48	60.92	0.10	0.10	0.10	5.89	0.10	0.10	0.10	5.89
感染发热组	0.10	0.10	0.10	0.29	0.10			0.10	3.02	15.41	0.10	0.10	0.10	0.10	0.10	0.10	0.10	0.10
Z值	-7.314				-6.687						-3.008				-3.239
P值	<0.001				<0.001						<0.001				0.001
组别	IL-12P70										IL-17A				IL-18
组别	P₅₀		P₇₅			P₉₀			P₉₅		P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
感染发热组	0.10		0.10			0.10			22.85		0.10	0.10	0.10	0.60	0.10	7.95	136.64	191.66
KD组	0.10		0.10			0.10			27.85		0.10	0.10	0.91	1.70	18.90	75.51	148.66	252.16
Z值	-2.856										-4.178				-5.950
P值	0.004										<0.001				<0.001
组别	TNF-α										INF-γ				sCD25
组别	P₅₀		P₇₅				P₉₀		P₉₅		P₅₀	P₇₅	P₉₀	P₉₅	P₅₀	P₇₅	P₉₀	P₉₅
KD组	0.10		0.10				3.01		5.75		0.23	2.77	20.06	112.48	27 214.81	47 688.36	92 148.49	126 074.00
感染发热组	0.10		0.10				0.10		0.10		0.10	1.00	26.21	47.45	15 896.00	23 829.98	32 245.89	36 403.40
Z值	-5.420										-3.628				-7.521
P值	<0.001										<0.001				<0.001

统计值		IL-8	IL-1β	IL-2	IL-6	IL-10	IL-4	IL-5	IL-12P70	IL-17A	IL-18	TNF-α	INF-γ	sCD25
r值		-0.028	0.100	-0.153	0.416	0.407	-0.047	0.100	0.116	0.259	0.256	0.308	0.238	0.531
P值		0.738	0.231	0.066	<0.001	<0.001	0.574	0.230	0.164	0.002	0.002	<0.001	0.004	<0.001

统计值		IL-8	IL-1β	IL-2	IL-6	IL-10	IL-4	IL-5	IL-12P70	IL-17A	IL-18	TNF-α	INF-γ	sCD25
r值		-0.028	0.100	-0.153	0.416	0.407	-0.047	0.100	0.116	0.259	0.256	0.308	0.238	0.531
P值		0.738	0.231	0.066	<0.001	<0.001	0.574	0.230	0.164	0.002	0.002	<0.001	0.004	<0.001

统计值	CD3⁺ T细胞		CD3⁺CD8⁺ T细胞		CD16⁺CD56⁺自然杀伤细胞		CD19⁺ B细胞		CD4/CD8比值
统计值	百分比	绝对值	百分比	绝对值	百分比	绝对值	百分比	绝对值	CD4/CD8比值
r值	-0.116	0.075	-0.241	-0.032	-0.147	-0.088	0.218	0.190	0.197
P值	0.165	0.368	0.003	0.702	0.077	0.291	0.008	0.022	0.017