[母胎微嵌合体在活化人类白细胞抗原半相合动员外周血干细胞中对实体瘤治疗作用的初步研究]

[A preliminary study on the effects of feto-maternal microchimerism in activated human leukocyte antigen haploidentical mobilized peripheral blood stem cells on treatment of solid tumors].

作者信息

Cao Shui, Yu Jin-pu, Li Hui, An Xiu-mei, Xin Ning, Ren Xiu-bao

机构信息

Department of Biological Therapy, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China.

出版信息

Zhonghua Nei Ke Za Zhi. 2009 Oct;48(10):857-61.

PMID:20079231

Abstract

OBJECTIVE

To study the effect of feto-maternal microchimerism in the treatment of activated human leukocyte antigen (HLA) haploidentical mobilized peripheral blood cells against solid tumors.

METHODS

Genomic DNA samples of 25 pairs of HLA haploidentical donors and recipients were extracted. The donor-derived HLA-DRB loci were detected with nested PCR-sequence specific primer (SSP) typing. The mixed lymphocyte proliferation action between the patients and respective donors, the engraftment of donor's cells and the serum levels of Th1/Th2 type of cytokines were measured with MTT, FISH and ELISA method respectively. The survival time of patients with or without feto-maternal microchimerism were compared as well.

RESULTS

Using nested PCR-SSP typing, the positive rates of feto-maternal microchimerism in the 25 pairs of HLA haploidentical donors and recipients were 40% in the maternal/children pairs and 0 in the paternal/children pairs. The chimerism positive patients showed less proliferation activity when cocultured with respective donors as compared with unrelated ones (P = 0.03). Only one chimerism positive patient experienced the engraft of donor's cell 3 months after treatment as the donor derived XX chromosome was identified with FISH. When the data of chimerism positive patients were deleted, the serum levels of IFNgamma 1 month after treatment dropped dramatically from 171.4 (26.3 approximately 258.4) ng/L to 29.4 (1.2 approximately 39.9) ng/L. The survival time in chimerism positive patients of the maternal/children pairs was significantly longer than that in chimerism negative patients, which was (31.2 +/- 4.3) months and (11.1 +/- 3.3) months, respectively (P = 0.036).

CONCLUSION

Feto-maternal microchimerism might induce anergy in the HLA haploidentical donors, favor the engraftment of donor's progenitors and maintenance of positive microenvironment and prolong the survival time.

摘要

目的

研究母胎微嵌合体在治疗活化人类白细胞抗原（HLA）半相合动员外周血单个核细胞抗实体瘤中的作用。

方法

提取25对HLA半相合供受者的基因组DNA样本。采用巢式聚合酶链反应-序列特异性引物（PCR-SSP）分型法检测供者来源的HLA-DRB基因座。分别采用MTT法、荧光原位杂交（FISH）法和酶联免疫吸附测定（ELISA）法检测患者与各自供者之间的混合淋巴细胞增殖反应、供者细胞植入情况及Th1/Th2型细胞因子血清水平。同时比较有或无母胎微嵌合体患者的生存时间。

结果

采用巢式PCR-SSP分型法，25对HLA半相合供受者中，母-子对的母胎微嵌合体阳性率为40%，父-子对为0。与无关供者相比，嵌合体阳性患者与各自供者共培养时增殖活性较低（P = 0.03）。仅1例嵌合体阳性患者在治疗3个月后出现供者细胞植入，FISH检测鉴定为供者来源的XX染色体。剔除嵌合体阳性患者数据后，治疗1个月后干扰素γ（IFNγ）血清水平从171.4（26.3～258.4）ng/L大幅降至29.4（1.2～39.9）ng/L。母-子对嵌合体阳性患者的生存时间显著长于嵌合体阴性患者，分别为（31.2±4.3）个月和（11.1±3.3）个月（P = 0.036）。