OPIATE RECEPTOR MEDIATED EFFECTS OF STRESS ON IMMUNITY

阿片受体（DORs）调节大量的免疫功能，如T细胞增殖、IL-2的产生和趋化因子介导的趋化。最近，三角洲阿片激动剂被报道通过DOR-转染的T细胞株（DOR- ju）抑制HIV- lp24抗原的表达。1)。胸腺和脾淋巴细胞诱导脑啡肽的表达强烈提示DORs对T细胞的调节是生理性的。虽然几个实验室已经在啮齿动物和灵长类动物的单核细胞中检测到DOR mRNA，但对DOR mRNA的调控表达知之甚少。这些研究将通过在存在和不存在抗cd28共刺激的情况下交联T细胞受体（TCR）来评估其体外诱导作用。在未刺激的培养和通过TCR，将确定表达DOR mRNA的淋巴细胞的表型以及活化的蛋白激酶C对DOR诱导的影响。进一步的研究将集中于使用天然超抗原葡萄球菌肠毒素B （SEB）在体内诱导DOR mRNA。介导DORs免疫调节作用的细胞内信号尚未确定。虽然有丝分裂原活化蛋白激酶（MAPKs）， ERKs 1,2在DOR- ju， 1细胞中被DORs激活，但DOR激动剂抑制脾脏T细胞中tcr依赖性的ERKs磷酸化和活化。在特定目标#2中，磷酸酶（例如SHP-1、- 2、MKP 1-3、PAC-1）在DORs抑制erk1、2磷酸化中的作用将通过描述DOR激动剂对T细胞磷酸酶的表达、磷酸化和活性的影响来评估。其他研究将评估DORs对TCR依赖性ERKs 1、2激活所需的早期事件的影响，例如TCR复合物关键亚基的磷酸化（例如CD3-zeta）。具体目标#3将通过继续我们对hiv - 1表达的研究来解决dor依赖性免疫调节在人外周血CD4+ T细胞中的临床意义。在药理学研究中，两种DOR激动剂SNC-80和DADLE对hiv - 1原代分离株感染的正常CD4+ T细胞中hiv - 1 p24抗原表达的影响将与明尼苏达菌株进行比较。机制研究将这些激动剂的功效与激活的人CD4+ T细胞DOR mRNA的表达联系起来。还将评估参与hiv - 1复制反激活的核因子kappaB的结合活性，以及趋化因子受体CXCR4和CCR5的表达和磷酸化，这两种受体是hiv - 1摄取的共受体。总之，这些研究将有助于阐明我们对T细胞DORs的表达和功能的理解，以及它们在调节T细胞对HIV- 1的反应中的潜在免疫药理学作用。

Delta opioid receptors (DORs) modulate a myriad of immune functions such as T cell proliferation, IL-2 production and chemokine- mediated chemotaxis. Recently, delta opiate agonists have been reported to inhibit the expression of HIV- l p24 antigen by a DOR- transfected T cell line (DOR-Ju. 1). The inducible expression of enkephalins by thymic and splenic lymphocytes strongly suggests that the modulation of T cells by DORs is physiological. Although several laboratories have detected DOR mRNA in mononuclear cells from rodents and primates, little is known about the regulated expression of DOR mRNA. These investigations will evaluate its induction in vitro by cross-linking the T cell receptor (TCR) in the presence and absence of anti-CD28 costimulation. The phenotype(s) of the lymphocytes expressing DOR mRNA and the effects of activated protein kinase C on the induction of DOR in unstimulated cultures and through the TCR will be determined. Additional studies will focus on the induction of DOR mRNA in vivo, using the naturally occurring superantigen, staphylococcal enterotoxin B (SEB). The intracellular signals mediating the immunomodulatory effects of DORs have not been identified. Although the mitogen activated protein kinases (MAPKs), ERKs l, 2, are activated by the DORs in DOR-Ju,l cells, DOR agonists suppress the TCR-dependent phosphorylation and activation of ERKs in splenic T cells. In specific aim #2, the role of phosphatases (e.g. SHP-1, - 2, MKP 1-3, PAC-1) in the suppression of ERK l, 2 phosphorylation by DORs will be evaluated by characterizing the effects of DOR agonists on the expression, phosphorylation and activity of T cell phosphatases. Additional studies will assess the effects of DORs on early events required for the TCR-dependent activation of ERKs l, 2, such as the phosphorylation of key subunits of the TCR complex (e.g. CD3-zeta). Specific aim #3 will address the clinical significance of DOR-dependent immunomodulation in human peripheral blood CD4+ T cells by continuing our studies of HIV-l expression. In pharmacological studies, the effects of two DOR agonists, SNC-80 and DADLE, on HIV-l p24 antigen expression by normal CD4+ T cells infected with primary isolates of HIV-l will be compared to the Minnesota strain. Mechanistic studies will correlate the efficacy of these agonists with the expression of DOR mRNA by activated human CD4+ T cells. The binding activity of nuclear factor kappaB, which is involved in transactivation of HIV-l replication, and the expression and phosphorylation of the chemokine receptors CXCR4 and CCR5, which are co-receptors for HIV-l uptake, will also be evaluated. In summary, these investigations will help clarify our understanding of the expression and function of T cell DORs and their potential immunopharmacological role in modulating the T cell response to HIV- 1.