Effects of Cannabinoids on the Immune Response to Murine Systemic Candidiasis

大麻素对小鼠系统性念珠菌病免疫反应的影响

• 批准号: 8248208
• 负责人: Nancy Elizabeth Buckley-Nieves
• 金额: $ 10.54万
• 依托单位: CALIFORNIA STATE POLY U POMONA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8248208
• 关键词: 2-arachidonylglycerol; Acquired Immunodeficiency Syndrome; Acute; Address; Animals; Anorexia; Antibody Formation; B-Cell Development; Bacterial Infections; Biomedical Research; British; Cancer Patient; Candida; Candida albicans; Cannabinoids; Cells; Cellular biology; Chemotaxis; Colitis; Collaborations; Colony-forming units; Data; Development; Disseminated candidiasis; Dose; Educational process of instructing; Endocannabinoids; Enzyme-Linked Immunosorbent Assay; Faculty; Funding; Future; Goals; Host resistance; Housing; Immune; Immune response; Immune system; Immunity; Immunology; Individual; Indwelling Catheter; Infection; Institution; Interferons; Interleukin-12; Interleukin-4; Interleukin-6; Investigational Therapies; Journals; Kidney; Knock-out; Knockout Mice; Laboratories; Learning; Legionnaires' Disease; Literature; Marijuana; Marijuana Smoking; Memory; Messenger RNA; Molecular Biology Techniques; Monitor; Morbidity - disease rate; Multiple Sclerosis; Mus; Mycoses; Necrosis; Neuraxis; Oral; Pain; Paper; Parasitic infection; Patients; Peer Review; Peripheral; Pharmacology; Phase; Postdoctoral Fellow; Process; Production; Progress Reports; Proteins; Psychotropic Drugs; Publications; Publishing; Reading; Research; Research Personnel; Role; Serum; Spleen; Splenocyte; Students; T-Cell Proliferation; T-Lymphocyte; T-Lymphocyte Subsets; TNF gene; Tetrahydrocannabinol; Time; TimeLine; Tissues; Training; Transplant Recipients; United States National Institutes of Health; Virus Diseases; Vomiting; Weight; Wild Type Mouse; Work; Writing; Yeasts; bacterial resistance; cannabinoid receptor; combat; cytokine; graduate student; in vitro testing; in vivo; macrophage; meetings; member; posters; programs; public health relevance; research study; response; tool; tumor; undergraduate student

DESCRIPTION (provided by applicant): The main psychoactive compound of marijuana, (-9-tetrahydrocannabinol (THC) has been shown to suppress host resistance to bacterial, viral and parasitic infections, presumably through the central or peripheral cannabinoid receptors, CB1R or CB2R, respectively. While CB1R is abundant in the central nervous system, CB2R is mainly expressed in immune cells. The effect of THC on Legionella pneumophila (L.p.) infection has been thoroughly studied by others. They found that THC suppressed T helper1 (Th1) cytokines (i.e interferon-3 (IFN- 3)) while increasing Th2 cytokines (i.e. interleukin-4 (IL-4)). However the effect of cannabinoids on fungal infections is unknown. The goal of the present proposal is to investigate the effects of cannabinoids (THC and 2-arachidonoylglycerol (2-AG)) and CB2R on a systemic Candida albicans (C. albicans) infection. C. albicans is the most common cause of systemic candidiasis, an infection that occurs frequently in immune compromised individuals (i.e., patients with AIDS), who may use marijuana to combat emesis and anorexia. Systemic candidiasis may also occur in immune competent individuals (i.e. patients with in-dwelling catheters). In the first aim of this proposal, we will investigate the role of cannabinoids and CB2R on the innate immune response to Candida infection. In the second aim, we will examine the effects of cannabinoids and CB2R on the adaptive immune response to the infection. To address the first aim, we will treat each wild type (WT) or CB2R knockout (CB2R-/-) mouse with vehicle or cannabinoids and 18h later will inject the mice with PBS or C. albicans (1x107 yeast). We will then collect serum, kidneys and spleens 2h, 8h and 1 day after the yeast infection. Others have shown that the acute phase cytokines (i.e. interleukin-6 (IL-6), tumor necrosis- 1 (TNF- 1) and IFN- 3) are elevated in the serum of C. albicans infected mice. We will compare serum cytokine and splenic macrophages cytokine mRNA levels from cannabinoid and vehicle treated mice using enzyme linked immunosorbent assay and real time RT-PCR, respectively. To address the second aim of this study, WT or CB2R-/- mice will receive vehicle or cannabinoids and 18h later will be injected with PBS or C. albicans (0.75- 1x106 yeast). To study the primary immune response, some of the mice will be sacrificed 1 day, 3 days and 7 days later to determine serum IFN-3 and IL-4 levels. Fifteen days after the 1st C. albicans dose, the remaining mice will receive a higher dose of C. albicans (1-2x107 yeast). Half of these mice will be housed for up to 14 days to investigate the effect of cannabinoids on the memory immune response. The remaining mice will be sacrificed 2h, 8h and 1 day after the 2nd yeast challenge to determine serum and splenic T cell IFN-3 and IL-4 protein and mRNA levels, respectively. Weight and morbidity will be monitored daily for all mice. Kidneys or spleens will be collected at the different time points to evaluate the level of infection by counting the yeast colony forming units. We expect that cannabinoids will alter the innate and adaptive immune responses to the fungal infection. If cannabinoids act via CB2R, cannabinoids will not alter the fungal infection in CB2R-/- mice. Public Health Relevance: Systemic Candida albicans yeast infections are common among AIDS, cancer and transplant patients, individuals who may use marijuana and related compounds, such as Marinol (delta-9-tetrahydrocannabinol (THC)), to combat emesis and anorexia. THC is known to suppress immunity to bacterial, viral and parasitic infections, but its effect on yeast infections is essentially unknown. The present proposal will investigate the effects of THC, its related compound 2-arachidonoylglycerol and the peripheral cannabinoid receptor on the immune responses to a systemic Candida albicans infection in mice.

描述(申请人提供)：大麻的主要精神活性化合物-9-四氢大麻酚(THC)已被证明分别通过中枢或外周大麻素受体CB1R或CB2R抑制宿主对细菌、病毒和寄生虫感染的抵抗力。虽然CB1R在中枢神经系统中含量丰富，但CB2R主要在免疫细胞中表达。四氢呋喃对嗜肺军团菌的影响其他人已经对感染进行了彻底的研究。他们发现，THC抑制Th1(Th1)细胞因子(即干扰素-3)，而增加Th2细胞因子(即IL-4)。然而，大麻素对真菌感染的影响尚不清楚。本提案的目的是研究大麻素(THC和2-花生四烯酸甘油(2-AG))和CB2R对系统性白色念珠菌感染的影响。白念珠菌是系统性念珠菌病的最常见原因，这种感染经常发生在免疫功能受损的人(即艾滋病患者)中，他们可能会使用大麻来对抗呕吐和厌食症。系统性念珠菌病也可能发生在有免疫能力的个人(即有室内导管的患者)中。在这项建议的第一个目标，我们将调查大麻素和CB2R在念珠菌感染的先天性免疫反应中的作用。在第二个目标中，我们将研究大麻素和CB2R对感染的适应性免疫反应的影响。为了达到第一个目标，我们将给每个野生型(WT)或CB2R基因敲除(CB2R-/-)小鼠注射赋形剂或大麻类化合物，18小时后将给小鼠注射PBS或白色念珠菌(1x107酵母)。在酵母菌感染后2小时、8小时和1天分别采集血清、肾脏和脾组织。另有研究表明，感染白念珠菌的小鼠血清中的急性期细胞因子(即白介素6(IL-6)、肿瘤坏死因子-1(TNF-1)和干扰素-3)升高。我们将使用酶联免疫吸附试验和实时定量RT-PCR分别比较大麻素和赋形剂处理的小鼠的血清细胞因子和脾巨噬细胞细胞因子的mRNA水平。为了达到这项研究的第二个目的，WT或CB2R-/-小鼠将接受载体或大麻类药物，18小时后将注射PBS或白色念珠菌(0.75-1x106酵母)。为研究初次免疫应答，分别于1天、3天和7天后处死部分小鼠，测定血清中干扰素-3和白介素4水平。在第一剂白念珠菌接种15天后，其余的小鼠将接受更高剂量的白色念珠菌(1-2x107酵母)。其中一半的小鼠将被关押长达14天，以研究大麻类化合物对记忆免疫反应的影响。其余小鼠分别于第2次酵母菌攻击后2小时、8小时和1天处死，分别测定血清和脾T细胞干扰素-3、白介素4蛋白和mRNA水平。将每天监测所有小鼠的体重和发病率。在不同的时间点采集肾脏或脾，通过计数酵母菌集落形成单位来评估感染程度。我们预计，大麻素将改变对真菌感染的先天和获得性免疫反应。如果大麻素通过CB2R起作用，大麻素不会改变CB2R-/-小鼠的真菌感染。 公共卫生相关性：系统性白念珠菌酵母菌感染在艾滋病、癌症和移植患者中很常见，这些患者可能使用大麻和相关化合物，如Marinol(Delta-9-四氢大麻酚(THC))来对抗呕吐和厌食症。众所周知，THC可以抑制对细菌、病毒和寄生虫感染的免疫力，但它对酵母感染的影响基本上是未知的。本提案将研究THC、其相关化合物2-花生四烯基甘油和外周大麻素受体对小鼠系统性白念珠菌感染的免疫反应的影响。