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首页> 外文期刊>Infection and immunity >Monoclonal Antibody against the Plasmodium falciparum Chitinase, PfCHT1, Recognizes a Malaria Transmission-Blocking Epitope in Plasmodium gallinaceum Ookinetes Unrelated to the Chitinase PgCHT1
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Monoclonal Antibody against the Plasmodium falciparum Chitinase, PfCHT1, Recognizes a Malaria Transmission-Blocking Epitope in Plasmodium gallinaceum Ookinetes Unrelated to the Chitinase PgCHT1

机译：抗恶性疟原虫几丁质酶PfCHT1的单克隆抗体识别与几丁质酶PgCHT1不相关的疟原虫在Ookinetes中阻断疟疾传播的表位。

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摘要

To initiate invasion of the mosquito midgut, Plasmodium ookinetes secrete chitinases that are necessary to cross the chitin-containing peritrophic matrix en route to invading the epithelial cell surface. To investigate chitinases as potential immunological targets of blocking malaria parasite transmission to mosquitoes, a monoclonal antibody (MAb) was identified that neutralized the enzymatic activity of the sole chitinase of Plasmodium falciparum, PfCHT1, identified to date. This MAb, designated 1C3, previously shown to react with an apical structure of P. falciparum ookinetes, also reacts with a discrete apical structure of P. gallinaceum ookinetes. In membrane feeding assays, MAb 1C3 markedly inhibited P. gallinaceum oocyst development in mosquito midguts. MAb 1C3 affinity isolated an ～210-kDa antigen which, under reducing conditions, became a 35-kDa antigen. This isolated 35-kDa protein cross-reacted with an antiserum raised against a synthetic peptide derived from the P. gallinaceum chitinase active site, PgCHT1, even though MAb 1C3 did not recognize native or recombinant PgCHT1 on Western blot. Therefore, this affinity-purified 35-kDa antigen appears similar to a previously identified protein, PgCHT2, a putative second chitinase of P. gallinaceum. Epitope mapping indicated MAb 1C3 recognized a region of PfCHT1 that diverges from a homologous amino acid sequence conserved within sequenced chitinases of P. berghei, P. yoelii, and P. gallinaceum (PgCHT1). A synthetic peptide derived from the mapped 1C3 epitope may be useful as a component of a subunit transmission-blocking vaccine.
机译：为了引发蚊子中肠的入侵， Plasmodium ookinetes分泌几丁质酶，这些酶必须穿过含几丁质的营养层基质才能侵入上皮细胞表面。为了研究几丁质酶作为阻止疟原虫向蚊子传播的潜在免疫学靶标，我们确定了一种单克隆抗体（MAb），该抗体中和了迄今为止确定的恶性疟原虫（PfCHT1）的唯一几丁质酶的酶活性。先前显示为标为1C3的MAb与 P的顶端结构反应。恶性疟原虫的人，也与 P的离散的顶端结构反应。鸡油菌。在膜进料分析中，MAb 1C3显着抑制 P。鸡中肠的鸡胆卵囊发育。 MAb 1C3亲和力分离出〜210 kDa的抗原，该抗原在还原条件下变为35 kDa的抗原。该分离的35kDa蛋白与针对源自 P的合成肽产生的抗血清交叉反应。 Gallinaceum几丁质酶活性位点PgCHT1，即使MAb 1C3在Western blot上不能识别天然或重组PgCHT1。因此，这种亲和纯化的35-kDa抗原看起来类似于先前鉴定的蛋白质PgCHT2，一种假定的 P第二种几丁质酶。鸡胸草。表位作图表明，MAb 1C3识别了PfCHT1区域，该区域与 P的几丁质酶序列中保守的同源氨基酸序列不同。 berghei，P。yoelii 和 P。鸡胆草（PgCHT1）。来源于作图的1C3表位的合成肽可用作亚基传递阻断疫苗的组分。

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