Antibodies specific for the Hia adhesion proteins of nontypeable Haemophilus influenzae mediate opsonophagocytic activity.
Antibodies specific for the Hia adhesion proteins of nontypeable Haemophilus influenzae mediate opsonophagocytic activity.
复制标题
对不可分型流感嗜血杆菌的 Hia 粘附蛋白具有特异性的抗体介导调理吞噬活性。
DOI：
10.1128/cvi.00090-09
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Barenkamp,StephenJ
中科院分区：
文献类型：
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作者：
Winter,LindaE;Barenkamp,StephenJ
Hia自转运蛋白是由不可分型流感嗜血杆菌（NTHI）表达的高度免疫原性表面粘附素。本研究的目的是评估抗hia抗体对同源和异源NTHI的调理噬细胞活性。编码h的表面暴露部分的基因片段。将流感菌株11hia蛋白克隆到pGEMEX-2表达载体中。将构建的pGEMEX-Hia BstEII del重组质粒转化大肠杆菌jm101，获得重组融合蛋白。免疫血清抗重组GEMEX-Hia （rGEMEX-Hia）介导的同源NTHI菌株11以1:160滴度和5个异种表达hia的菌株滴度≥1:40。免疫血清不能介导基因失活的两株hia敲除菌株的杀伤，但在用含有该基因的质粒转化一株基因敲除菌株后，可以介导高滴度的杀伤。免疫血清不介导表达HMW1/ hmw2的NTHI菌株的杀伤，这些菌株不表达Hia粘附素。然而，当两个具有代表性的表达HMW1/ hmw2的菌株用含有hiagene的质粒转化时，它们表达了丰富的Hia，并且容易被免疫血清杀死。免疫血清不能介导无基因质粒转化的表达HMW1/ hmw2的菌株的杀伤。我们的研究结果表明，NTHI的Hia蛋白是抗噬细胞抗体的靶标，并且这些抗体识别的共享表位存在于无关的NTHI菌株的Hia蛋白上。这些数据支持继续研究Hia蛋白作为预防NTHI疾病的候选疫苗。
The Hia autotransporter proteins are highly immunogenic surface adhesins expressed by nontypeableHaemophilus influenzae(NTHI). The objective of our study was to assess the opsonophagocytic activity of anti-Hia antibodies against homologous and heterologous NTHI. A segment of thehiagene that encodes a surface-exposed portion of theH. influenzaestrain 11 Hia protein was cloned into a pGEMEX-2 expression vector.Escherichia coliJM101 was transformed with the resulting pGEMEX-Hia BstEII del recombinant plasmid, and recombinant fusion protein was recovered. An immune serum against recombinant GEMEX-Hia (rGEMEX-Hia)-mediated killing of the homologous NTHI strain 11 at a 1:160 titer and five heterologous Hia-expressing strains at titers of ≥1:40. Immune serum did not mediate killing of two Hia-knockout strains whosehiagenes were inactivated but did mediate killing of one knockout strain at a high titer after the strain was transformed with a plasmid containing thehiagene. Immune serum did not mediate killing of HMW1/HMW2-expressing NTHI strains, which do not express the Hia adhesin. However, when two representative HMW1/HMW2-expressing strains were transformed with the plasmid containing thehiagene, they expressed abundant Hia and were susceptible to killing by the immune serum. Immune serum did not mediate killing of HMW1/HMW2-expressing strains transformed with the plasmid without thehiagene. Our results demonstrate that the Hia proteins of NTHI are targets of opsonophagocytic antibodies and that shared epitopes recognized by such antibodies are present on the Hia proteins of unrelated NTHI strains. These data argue for the continued investigation of the Hia proteins as vaccine candidates for the prevention of NTHI disease.