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表达马红球菌vapA基因的DNA疫苗诱导成年马的回忆性免疫反应及新生动物的初次免疫反应分析。

Analysis of anamnestic immune responses in adult horses and priming in neonates induced by a DNA vaccine expressing the vapA gene of Rhodococcus equi.

作者信息

Lopez A Marianela, Hines Melissa T, Palmer Guy H, Knowles Donald P, Alperin Debra C, Hines Stephen A

机构信息

Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA 99164-7040, USA.

出版信息

Vaccine. 2003 Sep 8;21(25-26):3815-25. doi: 10.1016/s0264-410x(03)00329-3.

Abstract

Rhodococcus equi remains one of the most important pathogens of early life in horses, yet conventional vaccines to prevent rhodococcal pneumonia have not been successful. DNA vaccination offers an alternative to conventional vaccines with specific advantages for immunization of neonates. We developed a DNA vaccine expressing the vapA gene (pVR1055vapA) that induced an anamnestic response characterized by virulence associated protein A (VapA)-specific IgG antibodies in sera and bronchoalveolar lavage fluid (BALF) as well as VapA-specific proliferation of pulmonary lymphocytes when tested in adult ponies. In contrast, none of the adults receiving the control plasmid responded. To determine if pVR1055vapA induced VapA-specific responses in the foal, the targeted age group for vaccination against R. equi, 10 naïve foals were randomly assigned at birth to two groups of five. At 8-15 days of age (day 1), foals were vaccinated by intranasal and intradermal (i.d.) routes with either pVR1055vapA or the negative control pVR1055vapA_rev. All foals were DNA boosted at day 14 and protein boosted at day 30 with either recombinant VapA or recombinant CAT (control group). Prior to the protein boost, neither group developed VapA-specific immune responses. However, at day 45, two of the VR1055vapA-vaccinated foals had increased titers of VapA-specific IgGb, IgM and IgGa in the sera, and IgG in the BALF. The induction of the opsonizing isotypes IgGa and IgGb has been previously shown to be associated with protection against R. equi. No VapA-specific immune responses were detected in the control group. This study indicates that the DNA vaccine effectively stimulates anamnestic systemic and pulmonary immune responses in adult horses. The results in foals suggest that the DNA vaccine also primed a subset of immunized neonates. These data support further development and modification to produce a DNA vaccine to more effectively prime neonatal foals.

摘要

马红球菌仍然是马早期生命中最重要的病原体之一,然而预防红球菌性肺炎的传统疫苗并不成功。DNA疫苗为传统疫苗提供了一种替代方案,对新生儿免疫具有特定优势。我们开发了一种表达vapA基因的DNA疫苗(pVR1055vapA),在成年小马中进行测试时,它能诱导一种以血清和支气管肺泡灌洗液(BALF)中与毒力相关蛋白A(VapA)特异性IgG抗体以及肺淋巴细胞VapA特异性增殖为特征的回忆反应。相比之下,接受对照质粒的成年马均无反应。为了确定pVR1055vapA是否能在马驹(针对马红球菌疫苗接种的目标年龄组)中诱导VapA特异性反应,10匹新生马驹在出生时被随机分为两组,每组5匹。在8 - 15日龄(第1天)时,马驹通过鼻内和皮内(i.d.)途径接种pVR1055vapA或阴性对照pVR1055vapA_rev。所有马驹在第14天接受DNA加强免疫,在第30天接受蛋白加强免疫,加强免疫所用的分别是重组VapA或重组CAT(对照组)。在蛋白加强免疫之前,两组均未产生VapA特异性免疫反应。然而,在第45天,接种VR1055vapA的两匹马驹血清中VapA特异性IgGb、IgM和IgGa以及BALF中IgG的滴度升高。此前已证明调理同型抗体IgGa和IgGb的诱导与抗马红球菌感染的保护作用相关。对照组未检测到VapA特异性免疫反应。这项研究表明,DNA疫苗能有效刺激成年马的回忆性全身和肺部免疫反应。马驹的结果表明,DNA疫苗也能使一部分免疫的新生儿产生免疫初免。这些数据支持进一步开发和改进,以生产一种能更有效地使新生马驹产生免疫初免的DNA疫苗。

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