Monoclonal antibodies to native HIV type 1 reverse transcriptase and their interaction with enzymes from different subtypes.
Monoclonal antibodies to native HIV type 1 reverse transcriptase and their interaction with enzymes from different subtypes.
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针对天然 HIV 1 型逆转录酶的单克隆抗体及其与不同亚型酶的相互作用。
DOI：
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发表时间：
2000
影响因子：
1.5
通讯作者：
C. Källander
中科院分区：
文献类型：
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作者：
A. Rytting;L. Åkerblom;J. Albert;T. Unge;E. Björling;L. Al‐Khalili;J. Gronowitz;C. Källander
用重组HIV-1亚型B逆转录酶（RT）制备小鼠抗RT单克隆抗体（MAb）。通过将RT与ISCOM基质形成佐剂皂苷（Quil A）混合来进行免疫。两种不同的测定法，均基于天然RT和抗体的相互作用，用于监测小鼠中的免疫应答，并用于筛选、选择和表征MAb。第一个测定测量抗体抑制RT的聚合酶活性的能力，第二个测定测量抗体捕获酶活性RT的能力。利用MAb组来评估代表9种不同HIV 1亚型的18种不同RT的免疫学特性。RT-抑制单克隆抗体可以分为两组，根据其对不同的HIV-1 RT的交叉反应模式。具有RT捕获能力的单克隆抗体之间记录的多样性程度较大。鉴定了至少7组具有不同交叉反应性模式的单克隆抗体。因此，同工酶特异性的程度变化很大，从对亚型B RT非常特异的MAb到能够捕获除两个O组分离株之一以外的所有HIV-1分离株的RT的MAb B。总之，我们的研究表明，存在令人惊讶的大免疫学差异RT从不同的HIV-1亚型以及从相同的亚型。
Recombinant reverse transcriptase (RT) from HIV-1 subtype B was used to produce mouse anti-RT monoclonal antibodies (MAbs). Immunization was done by mixing RT with the ISCOM matrix-forming adjuvant saponin (Quil A). Two different assays, both based on the interaction of native RT and antibodies, were used to monitor the immune response in mice and for screening, selection, and characterization of the MAbs. The first assay measures the capacity of antibodies to inhibit the polymerase activity of the RT and the second assay measures the ability of antibodies to capture enzymatically active RT. Twelve clones with the capacity to inhibit at least 50% of the RT activity and 34 clones with high RT-capturing capacity were found. The MAb panel was utilized to evaluate the immunological properties of 18 different RTs representing 9 different HIV1 subtypes. The RT-inhibitory MAbs could be divided into two groups based on their pattern of cross-reactivity toward the different HIV-1 RTs. The degree of diversity recorded among MAbs with RT-capturing capacity was larger. At least seven groups of MAbs with distinct cross-reactivity patterns were identified. Thus, the degree of isoenzyme specificity varied greatly, from MAbs that were quite specific for subtype B RT to one MAb that was able to capture the RTs from all HIV-1 isolates tested except one of the two group O isolates. In conclusion, our study revealed that there exist surprisingly large immunological differences between RTs from different HIV-1 subtypes as well as from the same subtype.
影响因子：
56.9
作者：
KOHLSTAEDT, LA;WANG, J;STEITZ, TA
通讯作者：
STEITZ, TA