A potent malaria transmission blocking vaccine based on codon harmonized full length Pfs48/45 expressed in Escherichia coli.
A potent malaria transmission blocking vaccine based on codon harmonized full length Pfs48/45 expressed in Escherichia coli.
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DOI：
10.1371/journal.pone.0006352
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发表时间：
2009-07-22
期刊：
影响因子：
3.7
通讯作者：
Kumar N
中科院分区：
文献类型：
--
作者：
Chowdhury DR;Angov E;Kariuki T;Kumar N
由恶性疟原虫引起的疟疾每年造成近100万人死亡。尽管最近取得了很大进展，但研制安全、有效和负担得起的疟疾疫苗仍然是一项挑战。针对疟原虫有性阶段的疫苗不仅可以减少雌性按蚊的疟疾传播，而且还可以减少能够逃避针对红细胞前和红细胞无性阶段疫苗引起的免疫的寄生虫的传播。我们将研究重点放在Pfs48/45上，这是一种在感染者体内发育的性阶段表达的蛋白质，也是最有希望的传播阻断疫苗靶点之一。Pfs48/45蛋白的功能性免疫原性需要适当的二硫键形成，因此迄今为止，重组全长Pfs48/45蛋白的免疫原性评估一直受到表达适当折叠蛋白困难的阻碍。在这里，我们提出了一种涉及密码子协调的策略，以成功地在大肠杆菌中表达全长Pfs48/45。纯化后的蛋白，被命名为CH-rPfs48/45，被单克隆抗体识别，该单克隆抗体针对天然蛋白中的还原敏感构象表位。小鼠免疫原性评价显示，在三种不同佐剂（明矾、蒙塔尼德ISA-51和完全弗氏佐剂）配制的ach - rpfs48 /45抗血清的膜喂养试验中，抗血清具有强大的传播阻断活性。更重要的是，CH-rPfs48/45与Montanide ISA-51配制后用于非人灵长类动物（橄榄狒狒，Papio anubis）时，在所有五种动物中均产生一致的高抗体反应（ELISA滴度为200万）。在单次免疫后，这些动物的血清在膜喂养试验中显示出超过93%的阻断活性，在疫苗加强剂量后达到几乎完全阻断。有效的传播阻断抗体在小鼠和非人灵长类动物中相对容易表达和诱导，这为开发基于CH-rPfs48/45的疟疾传播阻断疫苗提供了令人信服的理由和基础。
Malaria caused by Plasmodium falciparum is responsible for nearly 1 million deaths annually. Although much progress has been made in the recent past, the development of a safe, effective and affordable malaria vaccine has remained a challenge. A vaccine targeting sexual stages of the parasite will not only reduce malaria transmission by female Anopheles mosquitoes, but also reduce the spread of parasites able to evade immunity elicited by vaccines targeting pre-erythrocytic and erythrocytic asexual stages. We focused our studies on Pfs48/45, a protein expressed in the sexual stages developing within an infected person and one of the most promising transmission-blocking vaccine targets. Functional immunogenicity of Pfs48/45 protein requires proper disulfide bond formation, consequently evaluation of the immunogenicity of recombinant full-length Pfs48/45 has been hampered by difficulties in expressing properly folded protein to date. Here we present a strategy involving harmonization of codons for successful recombinant expression of full length Pfs48/45 in Escherichia coli. The purified protein, designated CH-rPfs48/45, was recognized by monoclonal antibodies directed against reduction-sensitive conformational epitopes in the native protein. Immunogenicity evaluation in mice revealed potent transmission blocking activity in membrane feeding assays of antisera elicited by CH-rPfs48/45 formulated in three different adjuvants, i.e. Alum, Montanide ISA-51 and complete Freund's adjuvant. More importantly, CH-rPfs48/45 formulated with Montanide ISA-51 when administered to nonhuman primates (Olive baboons, Papio anubis) resulted in uniformly high antibody responses (ELISA titers >2 million) in all five animals. Sera from these animals displayed greater than 93% blocking activity in membrane feeding assays after a single immunization, reaching nearly complete blocking after a booster dose of the vaccine. The relative ease of expression and induction of potent transmission blocking antibodies in mice and nonhuman primates provide a compelling rationale and basis for development of a CH-rPfs48/45 based malaria transmission blocking vaccine.
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影响因子：
1.5
作者：
KUMAR, N;CARTER, R
通讯作者：
CARTER, R
影响因子：
15.8
作者：
Guerra, Carlos A.;Gikandi, Priscilla W.;Tatem, Andrew J.;Noor, Abdisalan M.;Smith, Dave L.;Hay, Simon I.;Snow, Robert W.
通讯作者：
Snow, Robert W.
影响因子：
158.5
作者：
Abdulla, Salim;Oberholzer, Rolf;Tanner, Marcel
通讯作者：
Tanner, Marcel
影响因子：
3
作者：
Gosling, Roly D.;Ghani, Azra C.;Chandramohan, Daniel
通讯作者：
Chandramohan, Daniel
DOI：
10.4269/ajtmh.1995.52.60
发表时间：
1995-01-01
影响因子：
3.3
作者：
ROEFFEN, W;LENSEN, T;BECKERS, PJA
通讯作者：
BECKERS, PJA