Metwali A, Elliott D, Blum A M, Li J, Sandor M, Weinstock J V
Department of Internal Medicine, University of Iowa, Iowa City 52242, USA.
J Immunol. 1996 Jul 1;157(1):265-70.
Granulomas form in the liver and intestines of mice infected with the parasite Schistosoma mansoni. Vasoactive intestinal peptide (VIP) is a neurokine that can modulate aspects of the immune response by acting through receptors within the granuloma. Cloned are two novel VIP receptor (VIPR) mRNAs (VIPR1 and VIPR2) that also bind a second neurokine called pituitary adenylated cyclase-activating polypeptide (PACAP). The objective of this study was to determine if granulomas express either VIPR1 or VIPR2. Using a radioligand-binding assay, it was established that PACAP is as effective as VIP at displacing radiolabeled VIP from splenocytes and granuloma cells, and that most if not all VIPRs in the spleen and granulomas bind PACAP. PCR amplification of reverse transcribed RNA determined that granulomas express both VIPR1 and VIPR2 mRNAs. Gel electrophoresis and nucleotide sequencing confirmed the authenticity of the PCR products. Also, both receptor subtypes were amplified from several granuloma CD4+ T cell lines; yet reverse transcribed RNA from T cell-depleted, dispersed granuloma cells had only VIPR1 RNA. It is notable that reverse transcriptase-PCR detected only VIPR1 in the thymus and spleen, which are organs rich in T lymphocytes. Thus, the granulomas and spleens from mice with schistosomiasis contain cells that display authentic VIP/PACAP receptors. Moreover, these data suggest that T cells in different compartments vary in VIPR subtype expression. VIPR1 and VIPR2 may have different physiologic roles in inflammation.
在感染曼氏血吸虫的小鼠肝脏和肠道中会形成肉芽肿。血管活性肠肽(VIP)是一种神经因子,可通过作用于肉芽肿内的受体来调节免疫反应的各个方面。克隆出了两种新型VIP受体(VIPR)mRNA(VIPR1和VIPR2),它们也能结合另一种称为垂体腺苷酸环化酶激活多肽(PACAP)的神经因子。本研究的目的是确定肉芽肿是否表达VIPR1或VIPR2。通过放射性配体结合试验确定,PACAP在从脾细胞和肉芽肿细胞中置换放射性标记的VIP方面与VIP一样有效,并且脾脏和肉芽肿中大多数(如果不是全部)VIP受体都能结合PACAP。对逆转录RNA进行PCR扩增确定肉芽肿表达VIPR1和VIPR2 mRNA。凝胶电泳和核苷酸测序证实了PCR产物的真实性。此外,从几个肉芽肿CD4 + T细胞系中扩增出了两种受体亚型;然而,来自去除T细胞的分散肉芽肿细胞的逆转录RNA仅含有VIPR1 RNA。值得注意的是,逆转录酶PCR在富含T淋巴细胞的胸腺和脾脏中仅检测到VIPR1。因此,血吸虫病小鼠的肉芽肿和脾脏含有表达真实VIP/PACAP受体的细胞。此外,这些数据表明不同区室中的T细胞在VIPR亚型表达上存在差异。VIPR1和VIPR2在炎症中可能具有不同的生理作用。