儿童异基因造血干细胞移植后细胞免疫重建的临床研究

doi:10.12372/jcp.2022.21e1384

摘要/Abstract

摘要：

目的探讨儿童异基因造血干细胞移植（allo-HSCT）后免疫重建的临床特性。方法选取2019年7月至2020年10月接受allo-HSCT且临床资料完整的患儿为研究对象。动态监测患儿移植后第0、14、21、28、60及100 d外周血淋巴细胞亚群（CD3⁺ T细胞、CD4⁺ T细胞、CD8⁺ T细胞、CD16⁺ CD56⁺ NK细胞、CD19⁺ B细胞、CD4⁺ CD25⁺ Treg细胞）绝对计数;将患儿按有无急性移植物抗宿主病（aGVHD）分为non-aGVHD组和aGVHD组,比较两组间淋巴细胞亚群的差异。结果共75例患儿接受allo-HSCT,其中移植后满100 d、临床资料完整的42例患儿纳入本研究,男20例、女22例,中位年龄5.4（2.8~9.3）岁。移植前后不同时间点之间患儿CD3⁺ T细胞、CD4⁺ T细胞、CD8⁺ T细胞、CD16⁺ CD56⁺ NK细胞、CD19⁺ B细胞、CD4⁺CD25⁺ Treg细胞水平以及CD4⁺/CD8⁺比值差异均有统计学意义（P<0.05）。其中CD16⁺ CD56⁺ NK细胞恢复最快,在移植后21 d基本可达到移植前水平;CD3⁺ T细胞在移植后60 d基本可以达到移植前水平,CD8⁺ T 细胞到移植后60 d已超过移植前水平。42例患儿中发生aGVHD 16例,aGVHD 发生的中位时间为20.0（6.0~36.0）d。移植后28 d和60 d,aGVHD 组CD16⁺ CD56⁺ NK细胞绝对计数均低于non-aGVHD 组,差异有统计学意义（P<0.05）。结论儿童allo-HSCT后淋巴细胞亚群中重建最早的为CD16⁺ CD56⁺ NK细胞。NK细胞重建与aGVHD 的发生密切相关,或有助于早期识别aGVHD 患儿。

关键词: 异基造血干细胞移植, 淋巴细胞亚群, 急性移植物抗宿主病, 儿童

Abstract:

Objective To investigate the clinical characteristics of immune reconstitution after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children. Methods Children with complete clinical data who received allo-HSCT in the department of hematology and oncology from July 2019 to October 2020 were selected as the study subjects. The absolute count of peripheral blood lymphocyte subsets (CD3⁺ T cells, CD4⁺ T cells, CD8⁺ T cells, CD16⁺ CD56⁺ NK cells, CD19⁺ B cells, CD4⁺ CD25⁺ Treg cells) was dynamically monitored on day 0, 14, 21, 28, 60 and 100 after transplantation. The children were divided into non-aGVHD group and aGVHD group according to the presence or absence of acute graft-versus-host disease (aGVHD), and the differences of lymphocyte subsets between the two groups were compared. Results A total of 75 children received allo-HSCT, and 42 of them with complete clinical data after 100 days of transplantation were included in this study. There were 20 boys and 22 girls with a median age of 5.4 (2.8~9.3) years. There were statistically significant differences in the levels of CD3⁺ T cells, CD4⁺ T cells, CD8⁺ T cells, CD16⁺ CD56⁺ NK cells, CD19⁺B cells, CD4⁺CD25⁺ Treg cells and CD4⁺/CD8⁺ ratio at different time points before and after transplantation (P<0.05). CD16⁺CD56⁺ NK cells recovered the earliest and reached the level of pre-transplantation at 21 days post-transplantation. CD3⁺ T cells almost reached the pre-transplantation level at 60 days after transplantation, while CD8⁺ T cells exceeded the pre-transplantation level at 60 days after transplantation. The aGVHD occurred in 16 of 42 children, and the median time of aGVHD occurrence was 20.0 (6.0-36.0) days. At 28d and 60d after transplantation, the absolute count of CD16⁺ CD56⁺ NK cells in aGVHD group was lower than that in non-aGVHD group, the difference was statistically significant (P<0.05). Conclusions CD16⁺ CD56⁺ NK cells were the earliest lymphocyte subsets reconstructed after allo-HSCT in children. NK cell reconstitution is closely related to the occurrence of aGVHD after allo-PBSCT, which may be used as an effective predictor of prognosis in patients with aGVHD.

Key words: allogeneic peripheral blood stem cell transplantation, lymphocyte subsets, acute graft versus host disease, child

宋娜, 孙鸣, 祁闪闪, 王卓, 杨李, 卢文婕, 吴旻, 夏维, 陈燕, 熊昊. 儿童异基因造血干细胞移植后细胞免疫重建的临床研究[J]. 临床儿科杂志, 2022, 40(5): 388-394.

SONG Na, SUN Ming, QI Shanshan, WANG Zhuo, YANG Li, LU Wenjie, WU Min, XIA Wei, CHEN Yan, XIONG Hao. Clinical research on cellular immune reconstitution after allogeneic hematopoietic stem cell transplantation in children[J]. Journal of Clinical Pediatrics, 2022, 40(5): 388-394.

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参考文献 20

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时间	组别	CD16⁺ CD56⁺ NK细胞		CD3⁺ T细胞		CD4⁺ T细胞
时间	组别	M(P₂₅~P₇₅)	统计分析	M(P₂₅~P₇₅)	统计分析	M(P₂₅~P₇₅)	统计分析
移植前	non-aGVHD(n=26)	56.0(27.5~121.3)	Z=0.00	925.0(436.0~1 687.5)	Z=0.09	406.5(197.3~1119.5)	Z=1.55
移植前	aGVHD(n=16)	60.0(19.8~181.8)	P=1.000	1 046.5(243.0~1 974.3)	P=0.928	230.0(108.3~726.5)	P=0.120
0 d	non-aGVHD(n=26)	0.0(0.0~0.0)	Z=0.31	1.0(0.0~8.0)	Z=1.93	0.0(0.0~6.0)	Z=2.11
0 d	aGVHD(n=16)	0.0(0.0~0.0)	P=0.758	10.0(1.0~17.0)	P=0.053	7.0(1.0~13.0)	P=0.035
14 d	non-aGVHD(n=26)	3.0(0.0~14.0)	Z=0.17	4.0(0.0~119.0)	Z=1.65	2.0(0.0~65.0)	Z=1.15
14 d	aGVHD(n=16)	2.5(0.3~13.3)	P=0.862	47.5(6.5~124.5)	P=0.099	16.5(1.5~34.5)	P=0.249
21 d	non-aGVHD(n=26)	46.5(7.5~154.0)	Z=1.10	130.0(20.3~285.3)	Z=1.17	42.0(13.3~73.5)	Z=1.30
21 d	aGVHD(n=16)	25.0(8.0~55.0)	P=0.273	275.0(40.0~565.0)	P=0.242	70.0(13.0~117.0)	P=0.194
28 d	non-aGVHD(n=26)	186.0(77.3~389.8)	Z=2.28	301.0(124.8~939.8)	Z=0.98	81.5(28.5~141.3)	Z=0.76
28 d	aGVHD(n=16)	53.0(22.0~170.0)	P=0.023	333.0(80.0~589.0)	P=0.326	54.0(21.5~151.0)	P=0.447
60 d	non-aGVHD(n=26)	314.0(143.5~883.0)	Z=2.61	1 074.0(338.0~2 319.0)	Z=1.69	171.0(100.0~303.5)	Z=1.88
60 d	aGVHD(n=16)	152.0(54.0~218.0)	P=0.009	450.5(190.3~1351.0)	P=0.092	107.0(67.3~129.5)	P=0.060
100 d	non-aGVHD(n=26)	366.0(246.0~1 024.0)	Z=1.90	1 239.0(804.0~2 406.0)	Z=1.58	206.0(148.5~357.5)	Z=2.19
100 d	aGVHD(n=16)	269.0(155.0~387.0)	P=0.057	562.0(240.0~1921.0)	P=0.114	106.0(59.0~240.0)	P=0.028 （个/µL）

时间	组别	CD8⁺ T细胞		CD19⁺ B细胞		CD4⁺CD25⁺ Treg细胞
时间	组别	M(P₂₅~P₇₅)	统计分析	M(P₂₅~P₇₅)	统计分析	M(P₂₅~P₇₅)	统计分析
移植前	non-aGVHD(n=26)	502.5(213.3~712.8)	Z=0.18	48.5(1.0~150.0)	Z=0.83	67.0(38.8~121.5)	Z=0.84
移植前	aGVHD(n=16)	437.5(143.8~1 038.0)	P=0.856	22.5(1.0~86.8)	P=0.406	61.5(24.8~100)	P=0.400
0 d	non-aGVHD(n=26)	0.0(0.0~2.0)	Z=1.37	0.0(0.0~0.0)	Z=1.16	0.0(0.0~2.0)	Z=1.28
0 d	aGVHD(n=16)	1.0(0.0~3.0)	P=0.172	0.0(0.0~0.0)	P=0.247	1.0(0.0~6.0)	P=0.200
14 d	non-aGVHD(n=26)	1.0(0.0~40.0)	Z=1.89	0.0(0.0~1.0)	Z=0.93	0.0(0.0~4.0)	Z=0.08
14 d	aGVHD(n=16)	22.0(1.8~107.5)	P=0.059	0.0(0.0~0.0)	P=0.353	0.0(0.0~6.5)	P=0.938
21 d	non-aGVHD(n=26)	77.5(9.5~264.3)	Z=1.31	0.0(0.0~1.0)	Z=0.26	6.0(1.0~15.8)	Z=0.55
21 d	aGVHD(n=16)	119.0(27.0~474.0)	P=0.189	0.0(0.0~1.0)	P=0.799	2.0(1.0~14.0)	P=0.581
28 d	non-aGVHD(n=26)	216.0(63.3~676.5)	Z=0.86	0.0(0.0~3.0)	Z=1.46	10.0(2.8~26.5)	Z=1.26
28 d	aGVHD(n=16)	196.0(51.0~411.5)	P=0.388	0.0(0.0~0.5)	P=0.145	1.0(1.0~15.0)	P=0.209
60 d	non-aGVHD(n=26)	922.0(265.5~1 767.5)	Z=1.56	1.0(0.0~95.5)	Z=1.50	19.0(5.5~44.5)	Z=2.66
60 d	aGVHD(n=16)	320.5(112.8~1 273.5)	P=0.119	0.5(0.0~2.5)	P=0.134	2.0(0.3~17.0)	P=0.008
100 d	non-aGVHD(n=26)	957.0(674.5~2 120.0)	Z=1.66	37.0(0.0~95.5)	Z=1.82	29.0(18.0~46.5)	Z=1.36
100 d	aGVHD(n=16)	446.0(224.0~1 605.0)	P=0.096	2.0(0.0~25.0)	P=0.069	19.0(4.0~38.0)	P=0.175