ELISPOT ASSAY TO EVALUATE EFFECTS OF ANTI-HIV THERAPY

药物在治疗艾滋病毒方面的有益效果尤其明显。 临床上难以确定感染儿童和无症状儿童 成年人。目前可用的实验室测试来评估一种药物的 有效性，如血清p24定量，往往是负的。 这两个人群，或者说检测结果在治疗过程中可能不会改变， 例如，从血白细胞中分离病毒。在纵向研究中 建议感染艾滋病毒的儿童和无症状的成年人进入AZT 治疗将通过免疫分析-ELISPOT进行监测。这个化验， 在我们实验室进行了修改，以计数外周血淋巴细胞 分泌HIV特异性抗体，已在大约 四分之三受感染的儿童或无症状的成年人。在一个 初步横断面调查，13名接受AZT的成年人中有12人 发现埃利斯波特对艾滋病毒呈阴性。由于这一数字 抗体分泌细胞(ASC)与正在进行的 抗原刺激，有待检验的假设是，作为AZT 抑制病毒复制，抗原性刺激将减少， 循环中的Asc对HIV的相应减少。我们还观察到 最近，循环中的B细胞总数分泌 许多同种异型的免疫球蛋白无症状地显著升高。 受感染的成年人与未受感染的人相比。既然是这样 这一发现很可能与艾滋病病毒的直接或间接影响有关 非特异性B细胞激活，病毒复制的减少将是 预计将影响总发行量 免疫球蛋白分泌细胞(IgSC)。在拟议的纵向研究中， 患者将充当自己的控制者。循环中HIV-ASC的数量 或在给药前的两个不同时间测定的 将药物与不同时间间隔获得的结果进行比较 开始治疗。之后进行的测量结果的比较 停止(并可能随后重新建立)治疗应 也有助于评估药物的效果。虽然不是 这项研究的主要目标，与其他实验室测试和 临床表现可以提供更多关于 ELISPOT方法论在确定医疗保险有效性中的作用 有潜在价值的药物治疗儿童艾滋病毒感染和 成年人。

The beneficial effects of drugs in the treatment of HIV are particularly difficult to ascertain clinically in infected children and asymptomatic adults. Currently available laboratory tests to evaluate a drug's effectiveness, such as serum p24 quantitation, are very often negative in these two populations, or the test results may not change during therapy, e.g. isolation of virus from blood leukocytes. In the longitudinal study proposed here, HIV infected children and asymptomatic adults entering AZT therapy will be monitored with an immunoassay - ELISPOT. This assay, modified in our laboratory to enumerate peripheral blood lymphocytes which secrete HIV specific antibodies, has yielded positive results in about three-quarters of infected children or asymptomatic adults. In a preliminary cross-sectional survey, 12 of 13 adults receiving AZT were found to be ELISPOT negative for HIV. Since the number of antibody-secreting cells (ASC) is related to the extent of ongoing antigenic stimulation, the hypothesis to be tested is that, as AZT suppresses virus replication, the antigenic stimulus would be reduced, with a corresponding decrease in circulating ASC to HIV. We have also observed recently that the total number of circulating B cells secreting immunoglobulin of many isotypes is greatly elevated in asymptomatically infected adults, as compared to non-infected individuals. Since this finding is likely to be related to a direct or indirect effect of HIV in non-specific B cell activation, a reduction in virus replication would be expected to affect the total number of circulating immunoglobulin-secreting-cells (IgSC). In the proposed longitudinal study, patients will serve as their own control. The number of circulating HIV-ASC or IgSC determined at two different times prior to administration of the drug will be compared to the results obtained at various intervals after initiation of therapy. Comparisons of measurements made after discontinuation (and possibly subsequent reinstitution) of treatment should also be helpful in assessing the effects of the drug. Although not the primary objective of this study, comparisons to other laboratory tests and to clinical manifestations may provide additional information on the usefulness of the ELISPOT methodology in determining the effectiveness of drugs of potential value for the treatment of HIV infection in children and adults.