LAB ADAPTATION IN HIV-1

该提案的目标包括阐明CD 4亲和力在细胞内的作用。 HIV-1 gp 120在确定实验室适应或主要表型中起作用， 产生将支持T细胞嗜性的生长的细胞系， 分离株，并建立定量感染性测定， 分析嗜巨噬细胞的HIV-1分离株。建议解剖 当HIV-1经历了一系列的突变后， 实验室适应性，并比较感染特性，和gp 120 生物化学属性，如CD 4亲和力，对那些父母 初级分离物，从而深入了解如何培养初级分离物。 能够支持原代分离株生长的T细胞系将被 通过使用逆转录病毒载体引入大量的CD 4 白血病T细胞系，从而使其更容易为主要分离物 附着并造成感染定量感染性测定 对于嗜巨噬细胞的HIV-1分离株， 粘附细胞系如HeLa CD 4或CEM-SS中的CD 44 S。最 敏感品系将用于感染性试验。可溶性CD 4和 gp 120亚单位疫苗（基于实验室改造的 菌株）未能消除HIV- 1的主要菌株。 了解原代分离株的传染性与 实验室适应的分离物的那些将有助于细胞培养的发展 允许HIV-1的原代菌株繁殖的系统， 开发疫苗和其他抗病毒策略。

Goals of this proposal include elucidating the role that CD4 affinity of HIV-1 gp120 plays in determining lab adapted or primary phenotype, generating cell lines which will support the growth of T-cell tropic isolates, and establishing quantitative infectivity assays with which to analyze macrophage-tropic isolates of HIV-1. It is proposed to dissect out distinct mutational changes that occur in gp120 as HIV-1 undergoes lab adaptation, and to compare infectious properties, and gp120 biochemical attributes such as CD4 affinity, to those of the parental primary isolate, thus gaining insight into ways to grow primary isolates. T-cell lines capable of supporting primary isolate growth will be generated by using a retroviral vector to introduce high amounts of CD4 into leukemic T-cell lines, thus making it easier for primary isolates to attach to and establish an infection. A quantitative infectivity assay for macrophage-tropic isolates of HIV-1 will be established by expressing CD44S in adherent cell lines such as HeLa CD4 or CEM-SS. The most susceptible line will be used for infectivity assays. Soluble CD4 and gp120 subunit vaccines (both based and initially tested on lab adapted strains) have failed to inactivate primary strains of HIV- 1. Understanding how infectious properties of primary isolates differ from those of lab adapted isolates will aid in the development of cell culture systems allowing propagation of primary strains of HIV-1 for use in the development of vaccines and other antiviral strategies.