多发性骨髓瘤患者淋巴细胞来源微泡的检测及其临床意义

doi:10.11958/20231100

摘要/Abstract

摘要：

目的探讨多发性骨髓瘤（MM）患者淋巴细胞来源微泡（LMP）的表达及其临床意义。方法选取65例初诊MM患者（初诊组）及30例健康体检志愿者为对照组，初诊组经4疗程化疗后8例在3个月内死亡，余57例纳入化疗后组。流式细胞仪检测3组外周血LMP的表达及初诊组外周血淋巴细胞亚群及骨髓MM细胞免疫表型表达；比较化疗后不同疗效组间LMP；受试者工作特征（ROC）曲线确定各LMP预测死亡的截断值，LMP≥截断值为High（H）组，<截断值的为Low（L）组，进行Kaplan-Meier生存分析；将Kaplan-Meier分析中P<0.05的变量纳入Cox回归分析，分析患者死亡的影响因素；比较不同LMP组间淋巴细胞亚群及骨髓瘤细胞免疫表型差异。结果初诊组LMP、CD3⁺LMP、CD3⁺CD8⁺LMP比例低于对照组，NKLMP比例、CD4⁺/CD8⁺LMP高于对照组（P<0.05）。化疗后组CD3⁺CD8⁺LMP比例高于初诊组，CD3⁺CD4⁺LMP比例、CD4⁺/CD8⁺LMP低于初诊组（P<0.05）。完全缓解（CR）+非常好的部分缓解（VGPR）组CD3⁺LMP、CD3⁺CD8⁺LMP比例高于部分缓解（PR）+微小缓解（MR）+疾病进展（PD）组（P<0.01），而NKLMP比例、CD4⁺/CD8⁺LMP则低于PR+MR+PD组（P<0.05）。Kaplan-Meier分析显示，LLMP组中位生存时间（OS）较HLMP组缩短（P<0.01）；LNKTLMP组中位OS较HNKTLMP组缩短（P<0.05）。Cox回归分析显示LLMP、LNKTLMP是患者死亡的独立危险因素（HR分别为4.620、2.706，P<0.05）。LLMP组CD3⁺CD4⁺T比例、CD4⁺/CD8⁺T高于HLMP组（P<0.05）。LLMP组MM细胞CD117⁺比例高于HLMP组（P<0.05）。结论 MM患者存在LMP分泌紊乱，LMP、NKTLMP与MM预后密切相关，未来靶向调节LMP分泌或可延长生存，改善预后。

关键词: 多发性骨髓瘤, 流式细胞术, 预后, B淋巴细胞, T淋巴细胞, 淋巴细胞来源微泡

Abstract:

Objective To investigate the expression and clinical significance of lymphocyte-derived microvesicles (LMP) in patients with multiple myeloma (MM). Methods A total of 65 newly diagnosed MM patients were used as the initial diagnosis group, and 30 health examination volunteers were selected as the control group. After 4 courses of chemotherapy, 8 patients in the initial diagnosis group died within 3 months, and the remaining 57 patients were included in the post chemotherapy group. Flow cytometry was used to detect the expression of LMP in peripheral blood of three groups, as well as the immune phenotype expression of peripheral blood lymphocyte subsets and bone marrow MM cells in the initial diagnosis group. Values of LMP between different therapeutic groups after chemotherapy were compared. The receiver operating characteristic (ROC) curve was used to determine the cutoff value for predicting death for each LMP. For the high (H) group with LMP ≥ cut-off value and the low (L) group with LMP<cut-off value, Kaplan Meier survival analysis was performed. Variables with a P value<0.05 in Kaplan Meier analysis were included into Cox regression analysis to analyze influencing factors of patient death. The differences in lymphocyte subpopulations and myeloma cell immunophenotypes were compared between different LMP groups. Results The proportions of LMP, CD3⁺LMP and CD3⁺CD8⁺LMP were lower in the initial diagnosis group than those in the control group, while the proportions of NKLMP and CD4⁺/CD8⁺LMP were higher than those in the control group (P<0.05). The proportion of CD3⁺CD8⁺LMP was higher in the post chemotherapy group than that in the initial diagnosis group, while the proportion of CD3⁺CD4⁺LMP and the ratio of CD4⁺/CD8⁺LMP were lower than those in the initial diagnosis group (P<0.05). The proportion of CD3⁺LMP and ratio of CD3⁺CD8⁺LMP were higher in the CR+VGPR group than those in the PR+MR+PD group (P<0.01), while the proportion of NKLMP and ratio of CD4⁺/CD8⁺LMP were lower than those in the PR+MR+PD group (P<0.05). Kaplan Meier analysis showed that the median survival time (OS) was shorter in the LLMP group than that of the HLMP group (P<0.01). The median OS was shorter in the LNKTLMP group than that in the HNKTLMP group (P<0.05). Cox regression analysis showed that LLMP and LNKTLMP were independent risk factors for patient mortality, with HR 4.620 and 2.706 (P<0.05). The proportions of CD3⁺CD4⁺T and CD4⁺/CD8⁺T were higher in the LLMP group than those in the HLMP group (P<0.05). The proportion of CD117⁺ in MM cells was higher in the LLMP group than that in the HLMP group (P<0.05). Conclusion Patients with MM have abnormal secretion of LMP. The LMP and NKTLMP are closely correlated with prognosis of MM. Targeted regulation of LMP secretion may improve the survival and prognosis of MM.

Key words: multiple myeloma, flow cytometry, prognosis, B-lymphocytes, T-lymphocytes, lymphocyte-derived

中图分类号:

R733.3

图/表 11

参考文献 32

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组别	n	LMP/%	CD3⁺LMP/%	CD3⁺CD4⁺LMP/%	CD3⁺CD8⁺LMP/%
对照组	30	7.20（6.98，8.10）	76.90（72.20，79.13）	35.71（33.68，39.59）	25.39（22.62，32.18）
初诊组	65	1.22（0.49，2.56）	63.01（55.81，73.38）	36.28（27.91，39.51）	21.42（13.35，27.52）
Z		7.346^＊＊	4.864^＊＊	0.865	3.047^＊＊
组别	CD4⁺/CD8⁺LMP		NKLMP/%	NKTLMP/%	BLMP/%
对照组	1.28（1.23，1.56）		12.19（9.16，18.03）	8.42（7.16，10.22）	9.96（9.06，10.66）
初诊组	1.63（1.12，2.31）		21.32（15.18，28.67）	9.76（5.73，14.78）	6.35（3.52，11.78）
Z	2.042^＊		4.472^＊＊	1.173	1.934

组别	n	LMP/%	CD3⁺LMP/%	CD3⁺CD4⁺LMP/%	CD3⁺CD8⁺LMP/%
对照组	30	7.20（6.98，8.10）	76.90（72.20，79.13）	35.71（33.68，39.59）	25.39（22.62，32.18）
初诊组	65	1.22（0.49，2.56）	63.01（55.81，73.38）	36.28（27.91，39.51）	21.42（13.35，27.52）
Z		7.346^＊＊	4.864^＊＊	0.865	3.047^＊＊
组别	CD4⁺/CD8⁺LMP		NKLMP/%	NKTLMP/%	BLMP/%
对照组	1.28（1.23，1.56）		12.19（9.16，18.03）	8.42（7.16，10.22）	9.96（9.06，10.66）
初诊组	1.63（1.12，2.31）		21.32（15.18，28.67）	9.76（5.73，14.78）	6.35（3.52，11.78）
Z	2.042^＊		4.472^＊＊	1.173	1.934

组别	n	LMP/%	CD3⁺LMP/%	CD3⁺CD4⁺LMP/%	CD3⁺CD8⁺LMP/%
初诊组	65	1.22（0.49，2.56）	63.01（55.81，73.38）	36.28（27.91，39.51）	21.42（13.35，27.52）
化疗后组	57	1.10（0.28，2.86）	68.67（53.23，79.91）	30.56（23.20，34.25）	28.00（18.61，34.29）
Z		0.338	1.208	2.497^＊	2.540^＊
组别	CD4⁺/CD8⁺LM		NKLMP/%	NKTLMP/%	BLMP/%
初诊组	1.63（1.12，2.31）		21.32（15.18，28.67）	9.76（5.73，14.78）	6.35（3.52，11.78）
化疗后组	0.99（0.93，1.33）		16.75（10.52，28.85）	9.39（4.88，13.07）	11.13（3.32，18.60）
Z	3.677^＊＊		1.950	0.834	1.634

组别	n	LMP/%	CD3⁺LMP/%	CD3⁺CD4⁺LMP/%	CD3⁺CD8⁺LMP/%
初诊组	65	1.22（0.49，2.56）	63.01（55.81，73.38）	36.28（27.91，39.51）	21.42（13.35，27.52）
化疗后组	57	1.10（0.28，2.86）	68.67（53.23，79.91）	30.56（23.20，34.25）	28.00（18.61，34.29）
Z		0.338	1.208	2.497^＊	2.540^＊
组别	CD4⁺/CD8⁺LM		NKLMP/%	NKTLMP/%	BLMP/%
初诊组	1.63（1.12，2.31）		21.32（15.18，28.67）	9.76（5.73，14.78）	6.35（3.52，11.78）
化疗后组	0.99（0.93，1.33）		16.75（10.52，28.85）	9.39（4.88，13.07）	11.13（3.32，18.60）
Z	3.677^＊＊		1.950	0.834	1.634

组别	n	LMP/%	CD3⁺LMP/%	CD3⁺CD4⁺LMP/%	CD3⁺CD8⁺LMP/%
CR+VGPR组	31	1.25（0.31，3.77）	72.58（66.54，81.79）	31.89（26.58，35.53）	33.15（26.33，37.03）
PR+MR+PD组	26	0.93（0.08，1.85）	56.12（46.31，76.69）	27.38（21.71，32.11）	20.66（12.61，30.90）
Z		1.626	2.948^＊＊	1.738	3.020^＊＊
组别	CD4⁺/CD8⁺LMP		NKLMP/%	NKTLMP/%	BLMP/%
CR+VGPR组	0.97（0.92，1.02）		15.61（8.86，19.17）	9.84（5.96，13.08）	9.71（3.43，15.59）
PR+MR+PD组	1.23（0.96，1.51）		23.01（12.66，33.75）	7.44（4.42，12.71）	13.36（3.21，22.32）
Z	2.275^＊		2.243^＊	0.713	1.330