Conti P, Pang X, Boucher W, Letourneau R, Reale M, Barbacane R C, Thibault J, Theoharides T C
Immunology Division, University of Chieti School of Medicine, Italy.
J Pathol. 1997 Nov;183(3):352-8. doi: 10.1002/(SICI)1096-9896(199711)183:3<352::AID-PATH938>3.0.CO;2-L.
RANTES (regulated upon activation normal T expressed and secreted) is another member of the intercrine beta subfamily which acts as a selective chemoattractant for human monocytes and CD4-positive lymphocytes and increases the adherence of monocytes to endothelial cells. In this work, the effect of RANTES was studied on rat skin injection sites. Rats were intradermally injected with 50 microliters of RANTES, at different concentrations, fMet-Leu-Phe (FMLP), or LPS (positive controls) or PBS vehicle (negative control). The animals were then injected with 0.6 ml of Evans' blue in the tail vein in order to obtain a blue colour in the areas where the compounds were injected. After 4 h the rats were killed and the maximum diameter of the blue extravasation area was measured. The coloured areas were then excised and optical and electron microscopic studies were performed. In addition, in some of the excised tissue, a Northern blot analysis for histidine decarboxylase (HDC) mRNA was performed along with an estimation of the amount of histamine generated in the tissue injection sites. In these studies it was found that intradermal injections of 5, 2.5, and 1.25 x 10(-5) M RANTES produced a strong inflammatory response with the accumulation of a great number of basophil cells compared with the PBS (50 microliters) negative control, or FMLP (10(-6) M/50 microliters) or LPS (10 ng/50 microliters) positive control, after 4 h. Moreover, 5, 2.5, 1.25 x 10(-5) M RANTES produced a dose-response stimulation of HDC mRNA in the tissues of skin injection sites. The increasing number of basophils in the RANTES inflamed tissues led to augmentation of histamine content, compared with the PBS control. In conclusion, the pro-inflammatory chemokine RANTES stimulates the generation of HDC mRNA in skin injection sites.
调节激活正常T细胞表达和分泌因子(RANTES)是白细胞介素β亚家族的另一个成员,它作为人单核细胞和CD4阳性淋巴细胞的选择性趋化因子,增加单核细胞与内皮细胞的黏附。在本研究中,对RANTES在大鼠皮肤注射部位的作用进行了研究。将不同浓度的RANTES、甲酰甲硫氨酰-亮氨酰-苯丙氨酸(FMLP)或脂多糖(LPS,阳性对照)或磷酸盐缓冲液(PBS,阴性对照)50微升皮内注射到大鼠体内。然后通过尾静脉给动物注射0.6毫升伊文思蓝,以便在注射化合物的部位呈现蓝色。4小时后处死大鼠,测量蓝色外渗区域的最大直径。然后切除染色区域并进行光学和电子显微镜研究。此外,在一些切除的组织中,进行了组胺脱羧酶(HDC)mRNA的Northern印迹分析,并估计了组织注射部位产生的组胺量。在这些研究中发现,与PBS(50微升)阴性对照、FMLP(10⁻⁶ M/50微升)或LPS(10纳克/50微升)阳性对照相比,皮内注射5×10⁻⁵ M、2.5×10⁻⁵ M和1.25×10⁻⁵ M的RANTES在4小时后产生了强烈的炎症反应,有大量嗜碱性粒细胞聚集。此外,5×10⁻⁵ M、2.5×10⁻⁵ M、1.25×10⁻⁵ M的RANTES对皮肤注射部位组织中的HDC mRNA产生了剂量反应性刺激。与PBS对照相比,RANTES炎症组织中嗜碱性粒细胞数量增加导致组胺含量增加。总之,促炎性趋化因子RANTES刺激皮肤注射部位HDC mRNA的产生。