Immune reactivity and host modulatory roles of two novel Haemonchus contortus cathepsin B-like proteases.
Immune reactivity and host modulatory roles of two novel Haemonchus contortus cathepsin B-like proteases.
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DOI：
10.1186/s13071-021-05010-y
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发表时间：
2021-11-19
影响因子：
3.2
通讯作者：
Zarlenga D
中科院分区：
文献类型：
--
作者：
Bakshi M;Tuo W;Aroian RV;Zarlenga D
捻转血矛线虫（Haemonchuscontortus）是一种食血胃肠道线虫（GIN），其对全世界的小型反刍动物产业造成重大经济损失。尽管进行了广泛的努力，我们对GIN逃避宿主免疫反应的分子机制的理解是有限的。组织蛋白酶B样蛋白（CBPs）是半胱氨酸蛋白酶家族的成员，参与寄生虫入侵，因此提供了可行的疫苗候选物。使用计算机比较分析来鉴定进化枝V寄生线虫子集中的保守蛋白质，重点是吸血蠕虫，其中CBP表现突出。我们鉴定并表征了两种新的CBP，命名为Hc-CBP-1和Hc-CBP-2。用兔抗重组体（r）Hc-CBP-1和rHc-CBP-2抗体检测成虫排泄分泌产物（ESP）和虫体组织中天然蛋白的存在。扭曲用同源和异源抗血清以及地塞米松处理（Dex+）和未处理（Dex−）H的血清筛选rHc-CBP-1和rHc-CBP-2的肽阵列。感染的动物，以确定关键的免疫原性肽。Hc-cbp-1和Hc-cbp-2基因在H.用Dex+处理的感染旋毛虫的动物。最后，使用成熟的重组蛋白来评估它们调节细胞功能的能力。免疫组化结果显示Hc-CBP-1和Hc-CBP-2均存在于小肠刷状缘; Hc-CBP-2也存在于体壁的皮下组织中。用兔抗rHc-CBP-1和抗rHc-CBP-2筛选的肽展示揭示了蛋白质内占优势和重叠表位的区域。ELISA结果与H.扭曲的成年ESP H.来自Dex+动物的扭转显示Hc-cbp-2转录物增加三倍，而Hc-cbp-1表达没有变化。相反，rHc-CBP-1和rHc-CBP-2肽阵列与Dex+和Dex−动物血清的免疫反应性比较主要显示Hc-CBP-1结合的变化。最后，rHc-CBP-1抑制牛外周血单核细胞细胞因子/活化标志物（包括TNFα、IL-1、IL-6和CD 86）的mRNA表达。这些结果表明，作为分泌和隐蔽蛋白，Hc-CBP-1和Hc-CBP-2分别影响细胞和免疫活性，在感染过程中具有有趣的动力学，并可能提供可行的免疫相关的目标减毒H。扭体病传染性在线版本包含补充材料，可通过10.1186/s13071-021-05010-y获得。
Haemonchus contortus is a blood-feeding, gastrointestinal nematode (GIN) that causes significant economic losses to the small ruminant industry worldwide. Despite extensive efforts, our understanding of the molecular mechanisms used by GIN to evade host immune responses is limited. Cathepsin B-like proteins (CBPs) are members of the cysteine protease family and are involved in parasite invasion and thus provide viable vaccine candidates. In silico comparative analysis was used to identify conserved proteins among a subset of clade V parasitic nematodes with emphasis on blood-feeding worms, among which CBPs appeared prominently. We identified and characterized two novel CBPs designated Hc-CBP-1 and Hc-CBP-2. Rabbit anti-recombinant (r) Hc-CBP-1 and rHc-CBP-2 were used to detect the presence of native proteins in the excretory secretory products (ESP) and in worm tissues of adult H. contortus. Peptide arrays of rHc-CBP-1 and rHc-CBP-2 were screened with the homologous and heterologous anti-sera and with sera from dexamethasone-treated (Dex+) and non-treated (Dex−) H. contortus-infected animals to identify key immunogenic peptides. Gene transcription of Hc-cbp-1 and Hc-cbp-2 was also performed on H. contortus-infected animals treated with Dex+. Finally, the mature recombinant proteins were used to assess their abilities to modulate cell functions. Immunohistochemistry showed that both Hc-CBP-1 and Hc-CBP-2 are present on the brush borders of the intestine; Hc-CBP-2 was also present in the hypodermis of the body wall. Peptide displays screened with rabbit anti-rHc-CBP-1 and anti-rHc-CBP-2 revealed regions within the proteins where dominant and overlapping epitopes prevailed. ELISA results were consistent with only Hc-CBP-1 being present in H. contortus adult ESPs. H. contortus from Dex+ animals exhibited a threefold increase in Hc-cbp-2 transcript while Hc-cbp-1 expression did not change. In contrast, comparisons of immunoreactivities of rHc-CBP-1 and rHc-CBP-2 peptide arrays to sera from Dex+ and Dex− animals primarily showed changes in Hc-CBP-1 binding. Lastly, rHc-CBP-1 suppressed mRNA expression of bovine peripheral blood mononuclear cell cytokines/activation markers, including TNFα, IL-1, IL-6 and CD86. These results suggest that as secreted and cryptic proteins, respectively, Hc-CBP-1 and Hc-CBP-2 influence cellular and immunological activities that have interesting dynamics during infection and may provide viable immune-related targets for attenuating H. contortus infectivity. The online version contains supplementary material available at 10.1186/s13071-021-05010-y.
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影响因子：
20.3
作者：
Brossart, P;Grünebach, F;Brugger, WR
通讯作者：
Brugger, WR
影响因子：
5.1
作者：
Cathers, BE;Barrett, C;Rydzewski, RM
通讯作者：
Rydzewski, RM
影响因子：
15.3
作者：
Alderson, M R;Armitage, R J;Tough, T W;Strockbine, L;Fanslow, W C;Spriggs, M K
通讯作者：
Spriggs, M K
影响因子：
14.9
作者：
Benkert P;Künzli M;Schwede T
通讯作者：
Schwede T
DOI：
10.1093/bioinformatics/btq662
发表时间：
2011-02-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Benkert P;Biasini M;Schwede T
通讯作者：
Schwede T