Lee H R, Ho W Z, Douglas S D
Division of Allergy, Immunology and Infectious Diseases, Children's hospital of Philadelphia, PA 19104, USA
Clin Diagn Lab Immunol. 1994 Jul;1(4):419-23. doi: 10.1128/cdli.1.4.419-423.1994.
Substance P (SP) is an undecapeptide that has the amino sequence Arg-Pro-Lys-Pro-Gin-Gln-Phe-Phe-Gly-Leu-Met-NH2 and that belongs to a family of structurally related peptides known as tachykinins, the latter are widely distributed in the central nervous system. SP is involved in the biological activities of cells in the immune system, including the induction of cytokines in immune cells. We have investigated the effects of SP on constitutive and/or lipopolysaccharide (LPS)-induced expression of tumor necrosis factor (TNF) in cultured blood monocyte-derived macrophages (MDM). Cells cultured in vitro for 14 days were treated with SP at various concentrations (10(-10) to 10(-6M) in the presence of LPS before culture supernatants were harvested. TNF bioactivity in culture supernatants was measured with L929 cell line MDM from 10 of 12 donors treated with a SP alone showed increased TNF production. SP and LPS also interacted in a synergistic fashion in upregulating TNF production in MDM from responders. The stimulatory effect of SP was inhibited by two SP antagonists, spantide ([D-Arg-1-D-Trp-7-D-Trp-7-D-Trp-9-leu-11]-SP) and CP-96,345 (a nonpeptide antagonist of the SP receptor). In addition, an anti SP polyclonal antibody blocked the SP effect on TNF production in cultured MDM, further indicating the specificity of these effects. These results demonstrate that SP is an important regulator of monokine production by human monocytes/macrophages.
P物质(SP)是一种十一肽,其氨基酸序列为Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2,属于一类结构相关的肽家族,即速激肽,后者广泛分布于中枢神经系统。SP参与免疫系统中细胞的生物活性,包括诱导免疫细胞中的细胞因子。我们研究了SP对培养的血液单核细胞衍生巨噬细胞(MDM)中肿瘤坏死因子(TNF)的组成性和/或脂多糖(LPS)诱导表达的影响。在收获培养上清液之前,将体外培养14天的细胞在LPS存在下用各种浓度(10^-10至10^-6M)的SP处理。用L929细胞系测量培养上清液中的TNF生物活性,12名供体中有10名单独用SP处理的MDM显示TNF产生增加。SP和LPS在上调反应者MDM中的TNF产生方面也以协同方式相互作用。SP的刺激作用被两种SP拮抗剂,即spantide([D-Arg-1-D-Trp-7-D-Trp-7-D-Trp-9-leu-11]-SP)和CP-96,345(SP受体的非肽拮抗剂)抑制。此外,抗SP多克隆抗体阻断了SP对培养的MDM中TNF产生的影响,进一步表明了这些作用的特异性。这些结果表明,SP是人类单核细胞/巨噬细胞产生单核因子的重要调节因子。