Zhang Tao, Sun Bing-zhong, Chen Xie-qun, Zhu Hua-feng, Qiao Qing-da
Department of Hematology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 May;22(3):353-5, 359.
To investigate the effects of both Th1/Th2 imbalance and its re-attainment on in vitro expansion and hematopoiesis of CD34+ cells from a severe aplastic anemia (sAA) patient.
A preliminary-diagnosed sAA patient was studied. (1) Bone marrow mononuclear cells (BMMNC) were isolated and CD34+ and CD4+ cells were enriched by magnetic beads respectively. (2) The Th1/Th2 cell ratio within CD4+ subset was detected by flow cytometry (FCM). (3) Enriched-CD34+ cells were expanded and re-enriched for enough cells to be divided into four groups: control, Th cell treatment, Th cell and IFN-gamma treatment, and Th cell and IL-4 treatment, respectively. (4) After expansion for 10 d, colony-forming unit assay was performed on cells in each group. (5) Patient's Th1/Th2 ratio was followed up by FCM after immunotherapy.
(1) Symptom remission was achieved after therapy for 5 months. (2) Th1/Th2 cell ratio of the patient before and after remission was 22.47 and 12.27, respectively, while that of healthy controls was 8.98+/-4.45. (3) The CD34+ cell expansion rates as well as CFU numbers, from high to low, were ranked as control, Th cell and IL-4 contained group, Th cell contained, and Th cell and IFN-gamma contained group.
Predominant Th1 cells seem to directly inhibit the self-renewal, proliferation and lineage differentiation of CD34+ cells in vitro, which can be counteracted by IL-4, probably mediated by switching Th1/Th2 balance.
探讨Th1/Th2失衡及其恢复对重型再生障碍性贫血(sAA)患者CD34+细胞体外扩增及造血的影响。
研究一名初诊为sAA的患者。(1)分离骨髓单个核细胞(BMMNC),分别通过磁珠富集CD34+细胞和CD4+细胞。(2)采用流式细胞术(FCM)检测CD4+亚群内的Th1/Th2细胞比例。(3)对富集的CD34+细胞进行扩增并再次富集,获得足够数量的细胞后分为四组:对照组、Th细胞处理组、Th细胞与干扰素-γ处理组、Th细胞与白细胞介素-4处理组。(4)扩增10 d后,对每组细胞进行集落形成单位测定。(5)免疫治疗后通过FCM对患者的Th1/Th2比例进行随访。
(1)治疗5个月后症状缓解。(2)缓解前后患者的Th1/Th2细胞比例分别为22.47和12.27,而健康对照者为8.98±4.45。(3)CD34+细胞扩增率及集落形成单位数量从高到低依次为对照组、含Th细胞与白细胞介素-4组、含Th细胞组、含Th细胞与干扰素-γ组。
优势Th1细胞似乎直接抑制CD34+细胞在体外的自我更新、增殖及谱系分化,白细胞介素-4可能通过转换Th1/Th2平衡介导来抵消这种抑制作用。