INFERTILITY TESTING OF RECOMBINANT SP-10 VACCINE

这项资助的主要目标是测试抗生育潜力 在雌性狒狒身上注射避孕疫苗。 主要成分的 在疫苗中是一种抗原，称为SP-10，存在于人身上， 狒狒精子头与顶体膜相连， 最近从人类睾丸中克隆出来的 疫苗将是一种融合 蛋白质构建体，含有与26 kD连接的SP-10的248个氨基酸 谷胱甘肽转移酶 疫苗将被注射 肌肉注射 为了实现这一主要目标，最初的目标将是基因工程 将SP-10多肽导入pGex表达载体， 通过亲和层析纯化纯融合蛋白， 作为免疫原和用于各种测定。 为了确保 人和狒狒SP-10之间同源性是可以理解的，狒狒SP-10将 利用cDNA文库从狒狒睾丸中克隆并测序 我们将用人SP-10核苷酸探针构建并筛选。 狒狒所有主要器官的信使RNA将被分离和探测 通过用人SP-10探针进行北方印迹以确定是否存在 狒狒睾丸以外的器官表达SP-10的mRNA。 这将 加强免疫组化和ELISA数据已经获得， 表明SP-10是一种组织特异性抗原， 自身免疫反应 该疫苗的生育试验将在15只雌性狒狒中进行 其中15只雌性接受了不含SP-10的疫苗成分 免疫原 免疫接种后，将对每只狒狒进行监测 9个月经周期内的妊娠证据和抗SP-10 血清和宫颈粘液中的抗体反应。 组织病理 狒狒器官的检查将在实验结束时进行。 study. 将使用狒狒血清中的SP-10抗体， 确定SP-10分子的免疫显性部分。 本提案中概述的研究目标遵循以下直接路径： 开发一种可能用于人类的避孕疫苗 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.