Role of cyclooxygenase-2 in antibody responses to vaccination

描述（由申请人提供）：如果要保护人类免受潜在破坏性传染性病原体的侵害，则对疫苗接种的最佳体液（即抗体）反应是必不可少的。这些病毒包括引起天花的天花病毒。天花病毒和其他传染性病原体由于可能被用作针对平民和军队人口的生物恐怖主义武器而受到了极大的关注。B淋巴细胞是免疫系统的重要组成部分，负责合成抗体，在宿主防御包括病毒在内的感染性微生物方面发挥关键作用。本实验室已经证明，人和小鼠B淋巴细胞在激活时高表达前列腺素生成酶环氧化酶-2 （Cox-2）。重要的是，常用的非甾体抗炎药（NSAIDs），如双Cox-1/Cox-2抑制剂（如吲哚美辛）和较新的Cox-2选择性药物（如西乐brex），在多克隆和抗原特异性刺激下，显著降低了B细胞产生抗体的能力。基因缺乏Cox-2的小鼠对病毒样颗粒和牛痘病毒（用于天花疫苗接种）感染也会产生抗体滴度降低的反应。这些观察结果导致假设Cox-2是B淋巴细胞对刺激做出最佳反应并产生抗体所必需的。因此，非甾体抗炎药，尤其是针对Cox-2的非甾体抗炎药，可能会削弱体液免疫。如果得到证实，抗体对疫苗接种/感染的反应将会降低，特别是在免疫剂对B细胞免疫的刺激较弱的情况下。进一步的后果包括对免疫系统较弱的人的影响，如老年人和免疫功能受损的患者（艾滋病毒，癌症等），他们经常使用非甾体抗炎药。本文提出了两个特定的目的来研究Cox-2在牛痘抗体应答中的作用。目的1将确定Cox-2在小鼠对牛痘病毒感染的抗体反应中的作用。将使用两种互补的方法：一种是使用敲除Cox-2的小鼠的遗传策略，另一种是使用使用Cox-1/Cox-2或Cox-2小分子抑制剂治疗的正常小鼠的药理学策略。将评估牛痘感染的体液反应，包括免疫表型，并确定抗体反应需要Cox-2活性的关键时间框架。目的2将确定Cox-2在人B细胞产生牛痘抗体中的作用。献血者将包括1972年以前接种过天花疫苗的人和最近参加过评估天花疫苗临床试验的人。多克隆B细胞活化与非甾体抗炎药Cox-1/Cox-2或Cox-2抑制剂一致将决定这些药物是否抑制对牛痘的召回抗体反应。本研究的总体意义在于，在感染或接种疫苗后的关键时期，使用非甾体抗炎药或其他抑制Cox-2活性或表达的药物可能是禁忌的。这一发现将提高人类应对常规疫苗接种以及任何生物恐怖威胁的能力。对公共卫生的影响对疫苗接种（免疫）的最佳反应对于防止破坏性的传染性微生物至关重要。拟议的研究将研究广泛使用非甾体抗炎药（NSAIDs）（如西乐brex，布洛芬等）对免疫抗体反应的潜在负面影响。在疫苗供应不足、无法诱导保护性反应或接种者免疫系统较弱的情况下，这项研究尤为重要。本研究的总体意义在于，在感染或接种疫苗后的关键时期，使用非甾体抗炎药可能是禁忌。这一发现将提高我们应对常规疫苗接种以及任何生物恐怖威胁的能力。

DESCRIPTION (provided by applicant): Optimal humoral (i.e. antibody) responses to vaccination are essential if humans are to be protected against potentially devastating infectious agents. These include viruses such as variola that cause smallpox. Variola and other infectious agents have received significant attention due to their potential use as weapons for bioterrorism against both the civilian and military populations. B lymphocytes are crucial elements of the immune system responsible for the synthesis of antibodies that play key roles in host defense against infectious microorganisms including viruses. This laboratory has shown that human and mouse B lymphocytes highly express the prostaglandin-generating enzyme cyclooxygenase-2 (Cox-2) when activated. Importantly, commonly used non-steroidal anti-inflammatory drugs (NSAIDs) such as dual Cox-1/Cox-2 inhibitors (e.g. Indomethacin) and newer Cox-2 selective drugs (e.g. Celebrex) significantly blunt the ability of B cells to produce antibody in response to both polyclonal and antigen-specific stimulation. Mice genetically deficient in Cox-2 also respond with reduced antibody titers to virus-like particles and to infection with vaccinia virus (used in vaccination to smallpox). These observations led to the hypothesis that Cox-2 is required for B lymphocytes to optimally respond to stimulation and produce antibody. Thus NSAIDs, especially those that target Cox-2, may blunt humoral immunity. If proven, antibody responses to vaccination/infection would be reduced, especially where the immunizing agent weakly stimulates B cell immunity. Further ramifications include effects on humans with weakened immune systems such as the elderly and immuno-compromised patients (HIV, cancer, etc), who frequently use NSAIDs. Herein, two specific aims are proposed to study the role of Cox-2 in the antibody response to vaccinia. Aim 1 will determine the role of Cox-2 in the antibody response of mice to infection with vaccinia virus. Two complementary approaches will be used: a genetic strategy employing Cox-2 knock out mice and a pharmacologic one using normal mice treated with small molecule inhibitors of either Cox-1/Cox-2 or Cox-2. Humoral responses to vaccinia infection, including immune phenotyping will be evaluated and the critical time frame when Cox-2 activity is required for antibody responses will be determined. Aim 2 will determine the role of Cox-2 in antibody production to vaccinia for human B cells. Blood donors will consist of those vaccinated against smallpox prior to 1972 and those who participated in a recent clinical trial evaluating the smallpox vaccine. Polyclonal B cell activation in concert with NSAID inhibitors of either Cox-1/Cox-2 or Cox-2 will determine if these drugs dampen a recall antibody response to vaccinia. The overall significance of this research is that the use of NSAIDs or other drugs that inhibit Cox-2 activity or expression may be contraindicated during critical time periods after infection or vaccination. Such a finding will improve the ability of humans to respond to routine vaccination, as well as any bioterror threats. Impact on public health Optimal responses to vaccination (immunization) are essential to protect against devastating infectious microorganisms. The proposed research will study the potentially negative impact of the widespread use of non-steroidal anti-inflammatory drugs (NSAIDs) (e.g. Celebrex, Ibuprofen, etc.) on antibody responses to immunization. This research is especially important where vaccines are in short supply, are poor at inducing protective responses or the recipients have weakened immune systems. The overall significance of this research is that the use of NSAIDs may be contraindicated during critical time periods after infection or vaccination. Such a finding would improve our ability to respond to routine vaccination, as well as any bioterror threats.