U UREALYTICUM ANTIGEN

描述（改编自申请人摘要）：U。解脲支原体是一种 它是泌尿生殖道的常见疾病，然而，它是导致 绒毛膜羊膜感染与早产儿发病率和死亡率 对人类免疫反应中重要的抗原知之甚少。 有14个公认的血清型，可分为两个生物型。 据推测，只有某些血清型或单一血清型的成员， 生物型可能能够引起侵袭性疾病， 生物型特异性抗体可能是重要的危险因素。 本 补助金申请是第一次提交的补助金的竞争性更新， 对RFA 88-AI-12的反应，其目标是开发单克隆抗体， 抗体（mAb）用于表位作图以鉴定血清型或 血清群特异性抗原。 研究人员和同事此前 鉴定了Uu上的抗原复合物，命名为MB，这表明， 血清型和/或血清群特异性是在表位而不是 全抗原水平。 在此期间，他们展示了 所有14种血清型均存在MB;该抗原含有两种血清型 和交叉反应性表位; MB不仅在体外产生， 在体内; MB是Uu期间识别的最主要抗原之一 MB的mAb可以保护人类免受 疾病 通过抗体反应性肽扫描（Pepscan），他们已经表明， MB血清型3和血清型 特异性映射到羧基区域。 通过Pepscan分析， 建立了一种合成肽酶免疫测定法（EIA）， 以确定抗体对大多数抗原表位的反应性 在支原体感染患者的血清中。 重要的是，他们有 确定羧基结构域的独特结构允许 导致MB抗原大小变化的突变。 他们已经表明 羧基末端由多联体6个氨基酸重复组成 在不同的临床分离株中以少至7个拷贝和多至 共42个副本。 研究者指出，所有迹象表明， 变异可能是疾病产生的关键决定因素。 他们 已经开发了PCR引物，其允许检测大小变异， 已经开发了PCR技术，作为检测培养物是可靠的 患者标本中的Uu。 本补助金申请的具体目的 （1）从剩余的13个Uu血清型中克隆和测序MB， 方法建立在目前的赠款期;（2）发展PCR方法， 在体内检测血清型和MB大小变体;和（3）通过Pepscan建立 其余13种血清型和生物型的特异性表位 血清型;并使用基于合成肽的EIA来确定血清型， B组和总MB抗体（包括同种型和亚类）应答 在有记录的侵袭性Uu感染的患者中。 抗体谱将 与受影响者中存在的血清型和大小变体进行比较 通过PCR确定的位点。 这些研究将建立、表征和 验证研究者认为应该证明有用的试剂， 阐明不同血清型和生物型在生产 血清型特异性抗体在Uu感染中的作用 保护

DESCRIPTION (Adapted from applicant's abstract): U. urealyticum is a common commensal of the urogenital tract, yet, it is an important cause of chorioamnion infection and morbidity and mortality in premature infants. Antigens important in the human immune response are poorly understood. There are 14 recognized serovars which can be divided into two biotypes. It has been postulated that only certain serovars or members of a single biotype may be able to cause invasive disease and that lack of serovar or biotype specific antibody may be an important risk factor. The present grant application is the first competitive renewal of a grant submitted in response to RFA 88-AI-12 the goal of which was to develop monoclonal antibodies (mAbs) to be used in epitope mapping to identify serovar or serogroup specific antigens. The investigator and colleagues previously identified an antigen complex on Uu, designated MB, which suggested that serovar- and/or serogroup specificity is at the epitope rather than the whole antigen level. During this grant period, they have shown the existence of MB on all 14 serovars; that this antigen contains both serovar and cross-reactive epitopes; that MB is produced not only in vitro but also in vivo; MB is one of the most predominant antigens recognized during Uu infection of humans; and mAbs to MB can protect against development of disease. By antibody-reactive peptide scanning (Pepscan), they have shown that the major immunoreactive site of MB serovar 3 as well as serovar specificity maps to the carboxy region. Using Pepscan analysis they have established a synthetic peptide enzyme immunoassay (EIA) which will allow them to determine the antibody reactivity to the most antigenic epitope(s) in sera of ureaplasma infected patients. Importantly, they have established that the unique structure of the carboxy domain allows mutations that result in size variation of the MB antigen. They have shown that the carboxy terminus is composed of concatemeric 6 amino acid repeats occurring in different clinical isolates in as few as 7 copies and as many as 42 copies. The investigator states that all indications are that this variation may be a critical determinant in production of disease. They have developed PCR primers which allow detection of size variants plus they have developed PCR techniques which are reliable as culture for detection of Uu in patient specimens. Specific aims of the present grant application are to: (1) clone and sequence MB from the remaining 13 Uu serovars using methods established in the present grant period; (2) develop PCR methods to detect serovars and MB size variants in vivo; and (3) establish by Pepscan the serovar and biotype specific epitopes of each of the remaining 13 serovars; and use a synthetic peptide-based EIA to determine the serovar, biogroup, and total MB antibody (including isotype and subclass) response in patients with documented invasive Uu infection. Antibody profiles will be compared to the serovar(s) and size variant(s) present in the affected site as determined by PCR. These studies will establish, characterize, and validate reagents which the investigators feel should prove useful in elucidating the role of different serovars and biotypes in production of invasive Uu disease and the role of serovar specific antibody in protection.