ELISPOT ASSAY TO EVALUATE EFFECTS OF ANTI-HIV THERAPY

ELISPOT 测定法评估抗 HIV 治疗的效果

• 批准号: 3422628
• 负责人: FRANCIS K LEE
• 金额: $ 6.67万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-30 至 1992-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3422628
• 关键词: AIDS; AIDS therapy; B lymphocyte; adult human (21+); child (0-11); human immunodeficiency virus 1; human subject; human therapy evaluation; human tissue; immunologic techniques; longitudinal human study; monocyte; secretion; zidovudine

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.

药物在治疗艾滋病毒方面的有益效果特别明显， 临床上难以确定感染儿童和无症状儿童 成年人了目前可用的实验室测试，以评估药物的 有效性，如血清p24定量， 这两个群体，或者测试结果在治疗期间可能不会改变， 例如从血液白细胞中分离病毒。在纵向研究中， 在此建议，艾滋病毒感染儿童和无症状成人服用AZT， 将用免疫测定- ELISPOT监测治疗。该测定， 在我们的实验室修改，以计数外周血淋巴细胞， 分泌艾滋病毒特异性抗体，已产生积极的结果， 四分之三的受感染儿童或无症状成人。中 初步横断面调查显示，13名接受AZT治疗的成年人中有12人 发现ELISPOT对HIV呈阴性。自号 抗体分泌细胞（ASC）与正在进行的 抗原刺激，要测试的假设是，作为AZT 抑制病毒复制，抗原刺激将减少， 相应地减少了循环ASC到HIV。我们还注意到 最近，循环B细胞分泌的 许多同种型的免疫球蛋白在无症状地 感染者与未感染者相比。由于这 这一发现可能与艾滋病毒在以下方面的直接或间接影响有关： 非特异性B细胞活化，病毒复制减少， 预计将影响流通总量 免疫球蛋白分泌细胞（IgSC）。在拟议的纵向研究中， 患者将作为自己的对照。传播中的艾滋病毒非典型肺炎数量 或IgSC，在施用所述化合物之前的两个不同时间测定。 将药物与在不同时间间隔获得的结果进行比较， 开始治疗。测量结果的比较 停止治疗（可能随后重新开始治疗）应 也有助于评估药物的效果。虽然不是 本研究的主要目的是与其他实验室检查进行比较， 临床表现可能会提供更多的信息， ELISPOT方法在确定 对治疗儿童艾滋病毒感染具有潜在价值的药物， 成年人了

项目成果

Diagnosis of human immunodeficiency virus infection by enzyme-linked immunospot assays in a prospectively followed cohort of infants of human immunodeficiency virus-seropositive women.

在人类免疫缺陷病毒血清阳性女性婴儿的前瞻性随访队列中，通过酶联免疫斑点测定诊断人类免疫缺陷病毒感染。

• 发表时间: 1992
• 期刊: The Pediatric infectious disease journal
• 作者: Nesheim,S;Lee,F;Sawyer,M;Jones,D;Lindsay,M;Slade,B;Shaffer,N;Holmes,R;Ashby,R;Grimes,V
• 通讯作者: Grimes,V