Tomita Shuhei, Jiang Hou-Bo, Ueno Tomoo, Takagi Satoshi, Tohi Keiko, Maekawa Shin-ichi, Miyatake Akira, Furukawa Aizo, Gonzalez Frank J, Takeda Junji, Ichikawa Yoshiyuki, Takahama Yousuke
Department of Biochemistry, Kagawa Medical University, Kagawa, Japan.
J Immunol. 2003 Oct 15;171(8):4113-20. doi: 10.4049/jimmunol.171.8.4113.
The arylhydrocarbon receptor nuclear translocator (ARNT) is a member of the basic helix-loop-helix, PER-ARNT-SIM family of heterodimeric transcription factors, and serves as a dimerization partner for arylhydrocarbon receptor (AHR) and hypoxia-inducible factor-1alpha. To assess the function of ARNT in T cells, we disrupted the Arnt gene specifically in T cells of mice by conditional gene targeting using T cell-specific p56(lck)-Cre (Lck-Cre) transgenic Arnt-floxed mice. Thus generated, T cell-specific Arnt-disrupted mice (Lck-Cre;Arnt(flox/Delta) transgenic mice) exhibited complete loss of the expression of ARNT protein only in T cells, and were viable and appeared normal. The Arnt-disrupted T cells in the thymus were phenotypically and histologically normal. The Arnt-deficient T cells in the spleen were capable of responding to TCR stimulation in vitro. However, unlike normal mice in which exposure to the environmental pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), an AHR ligand, resulted in thymic involution, the thymus of Lck-Cre;Arnt(flox/Delta) mice were resistant to TCDD treatment in vivo. In contrast, benzo(a)pyrene, another AHR ligand, still caused thymic involution in Lck-Cre;Arnt(flox/Delta) mice. Finally, fetal thymus organ culture using Lck-Cre;Arnt(flox/Delta) and K5-Cre;Arnt(flox/Delta) (epithelial cell-specific Arnt-disrupted mice) showed that thymocytes rather than thymic epithelial cells are predominantly responsible for TCDD-induced thymic atrophy. Our results indicate that ARNT in T lineage cells is essential for TCDD-mediated thymic involution.
芳烃受体核转运蛋白(ARNT)是碱性螺旋-环-螺旋PER-ARNT-SIM异二聚体转录因子家族的成员,作为芳烃受体(AHR)和缺氧诱导因子-1α的二聚化伴侣。为了评估ARNT在T细胞中的功能,我们使用T细胞特异性p56(lck)-Cre(Lck-Cre)转基因Arnt-floxed小鼠,通过条件基因靶向技术在小鼠T细胞中特异性破坏Arnt基因。由此产生的T细胞特异性Arnt破坏小鼠(Lck-Cre;Arnt(flox/Delta)转基因小鼠)仅在T细胞中表现出ARNT蛋白表达完全缺失,并且存活且外观正常。胸腺中Arnt破坏的T细胞在表型和组织学上是正常的。脾脏中Arnt缺陷的T细胞能够在体外对TCR刺激作出反应。然而,与正常小鼠不同,正常小鼠暴露于环境污染物2,3,7,8-四氯二苯并对二恶英(TCDD)(一种AHR配体)会导致胸腺萎缩,而Lck-Cre;Arnt(flox/Delta)小鼠的胸腺在体内对TCDD治疗具有抗性。相比之下,另一种AHR配体苯并(a)芘仍会导致Lck-Cre;Arnt(flox/Delta)小鼠胸腺萎缩。最后,使用Lck-Cre;Arnt(flox/Delta)和K5-Cre;Arnt(flox/Delta)(上皮细胞特异性Arnt破坏小鼠)进行的胎儿胸腺器官培养表明,胸腺细胞而非胸腺上皮细胞是TCDD诱导胸腺萎缩的主要原因。我们的结果表明,T谱系细胞中的ARNT对于TCDD介导的胸腺萎缩至关重要。