INFERTILITY TESTING OF RECOMBINANT SP-10 VACCINE

这笔赠款的主要目的是测试抗生育潜力。 一种避孕疫苗在雌性狒狒身上的应用。主要成分 在疫苗中有一种抗原，称为SP-10，存在于人类和 狒狒精子头与顶体膜和顶体膜的关系 最近从人类睾丸中克隆出来的。疫苗将是一种融合 蛋白质结构，包含248个氨基酸的SP-10，与26kD相连 谷胱甘肽转移酶。疫苗将被注射 肌肉内。 朝着这个首要目标，最初的目标将是基因工程。 将SP-10多肽插入pGex表达载体，构建成重组表达载体pGex 亲和层析法纯化服务用纯融合蛋白 作为免疫原，用于各种检测。以确保准确的 人类和狒狒SP-10之间的同源性已被理解，狒狒SP-10将 利用cDNA文库从狒狒睾丸中克隆和测序 我们将构建并使用人类SP-10核苷酸探针进行筛选。 将从所有主要的狒狒器官中分离和探测信使RNA 通过用人SP-10探针进行Northern杂交来确定是否有 除了睾丸以外的狒狒器官表达SP-10的mRNAs。此日期将 重新加强免疫组织化学和ELISA法已获得的数据 提示SP-10是一种组织特异性抗原，不太可能引发 自身免疫反应。 该疫苗的生育试验将在15只雌性狒狒身上进行。 有15名女性接种了减去SP-10的疫苗成分 免疫基因。在接种疫苗后，将对每只狒狒进行监测 通过9个月经周期提供受孕证据和抗SP-10 血清和宫颈粘液中的抗体反应。组织病理学 在会议结束时，将对狒狒器官进行检查。 学习。针对SP-10的恒河猴血清抗体将用于 确定SP-10分子的免疫优势部分。 本提案中概述的研究目标遵循以下直接路径 一种具有人类潜在用途的避孕疫苗的研制 女性。

The primary goal of this grant is to test the anti-fertility potential of a contraceptive vaccine in female baboons. The principal ingredient in the vaccine is an antigen, termed SP-10, existing on the human and baboon sperm head in association with the acrosomal membranes and recently cloned from the human testis. The vaccine will be a fusion protein construct, containing 248 amino acids of SP-10 linked to 26 kD of glutathione transferase. The vaccine will be injected intramuscularly. Toward this primary goal, an initial aim will be the genetic engineering of the SP-10 polypeptide into the pGex expression vector and the purification by affinity chromatography of pure fusion protein to serve as the immunogen and for various assays. To insure that the exact homology between human and baboon SP-10 is understood, baboon SP-10 will be cloned and sequenced from baboon testis utilizing cDNA libraries which we will construct and screen with a human SP-10 nucleotide probe. Messenger RNA from all major baboon organs will be isolated and probed by Northern blotting with the human SP-10 probe to determine if any baboon organ other than testis expresses mRNAs for SP-10. This dat will re-inforce immunohistochemical and ELISA data already obtained which indicate SP-10 is a tissue specific antigen unlikely to elicit autoimmune reactions. Fertility trials of the vaccine will be conducted in 15 female baboons with 15 females receiving the vaccine ingredients minus the SP-10 immunogen. Following immunization, each baboon will be monitored through 9 menstrual cycles for evidence of conception and for anti-SP-10 antibody responses in serum and cervial mucus. Histopathological examination of baboon organs will be conducted at the conclusion of the study. The baboon serum antibodies to SP-10 will be employed to determine the immunodominant portions of the SP-10 molecule. The research aims outlined in this proposal follow the direct path to development of a contraceptive vaccine with potential use in human females.