Hajtó Tibor, Berki Timea, Boldizsár Ferenc, Németh Péter
Department of Immunology and Biotechnology, Faculty of Medicine, University of Pécs, Szigeti út 12, H-7643 Pécs, Hungary.
Immunol Lett. 2003 Mar 3;86(1):23-7. doi: 10.1016/s0165-2478(02)00265-1.
Galactoside-specific plant lectin, Viscum album agglutinin-I (VAA-I) has been shown to act as a biomodulator with proinflammatory and apoptosis-inducing effects, however its cellular targets and mechanism of immunobiological action in vivo are less well understood. Therefore, in the present work the short- and long-term in vivo effects of VAA-I on thymocyte subpopulations and peripheral T cells were tested using a murine (Balb/c) model. Cell surface CD4/CD8 staining and flow cytometry allowed us to follow the changes of thymocyte subpopulations: CD4-CD8- double negative (DN), CD4+CD8+ double positive (DP), CD4+ or CD8+ single positive (SP) and mature peripheral T cells after single or repeated injections with low doses of VAA-I. The apoptosis of the cells was detected by flow cytometry using propidium iodide (PI) and Annexin V staining. To detect the short-term effects of the lectin, the animals were investigated 24 h after a single injection of 1 or 30 ng/kg body weight (BW) VAA-I+/-1 mg/kg Dexamethasone (DX). The total number of mature CD8+ SP thymocytes increased significantly with an enhancement of the ratio of apoptotic cells. In contrast, in the blood samples an elevated CD4/CD8 ratio was found. In the next trial, Balb/c mice were treated twice weekly with 1 or 30 ng/kg VAA-I+/-1 mg/kg DX for 3 weeks. The total cell count of thymocytes showed significant increases after both doses of VAA-I, but an elevated percentage of apoptotic cells was found only after treatment with 30 ng/kg VAA-I. SP thymocytes revealed higher increases in lectin-induced apoptosis than DN or DP cells. In addition, both lectin doses significantly inhibited the DX-induced reduction of all thymocyte subpopulations investigated. In conclusion, our data suggest that VAA-I is able to modulate the maturation of thymocytes in vivo.
半乳糖苷特异性植物凝集素,欧洲槲寄生凝集素-I(VAA-I)已被证明可作为一种生物调节剂,具有促炎和诱导凋亡的作用,然而其在体内的细胞靶点和免疫生物学作用机制尚不清楚。因此,在本研究中,使用小鼠(Balb/c)模型测试了VAA-I对胸腺细胞亚群和外周T细胞的短期和长期体内效应。细胞表面CD4/CD8染色和流式细胞术使我们能够追踪胸腺细胞亚群的变化:在单次或重复注射低剂量VAA-I后,CD4-CD8-双阴性(DN)、CD4+CD8+双阳性(DP)、CD4+或CD8+单阳性(SP)以及成熟外周T细胞的变化。使用碘化丙啶(PI)和膜联蛋白V染色通过流式细胞术检测细胞凋亡。为了检测凝集素的短期效应,在单次注射1或30 ng/kg体重(BW)VAA-I±1 mg/kg地塞米松(DX)后24小时对动物进行研究。成熟CD8+SP胸腺细胞总数显著增加,凋亡细胞比例升高。相反,在血液样本中发现CD4/CD8比值升高。在接下来的试验中,Balb/c小鼠每周两次接受1或30 ng/kg VAA-I±1 mg/kg DX治疗3周。两种剂量的VAA-I处理后胸腺细胞总数均显著增加,但仅在30 ng/kg VAA-I处理后发现凋亡细胞百分比升高。SP胸腺细胞在凝集素诱导的凋亡中显示出比DN或DP细胞更高的增加。此外,两种凝集素剂量均显著抑制DX诱导的所有研究胸腺细胞亚群的减少。总之,我们的数据表明VAA-I能够在体内调节胸腺细胞的成熟。