Araujo Ricardo N, Padilha Terezinha, Zarlenga Dante, Sonstegard Tad, Connor Erin E, Van Tassel Curt, Lima Walter S, Nascimento Evaldo, Gasbarre Louis C
USDA, ARS, Bovine Functional Genomics Laboratory, Beltsville, MD, USA.
Vet Parasitol. 2009 May 26;162(1-2):106-15. doi: 10.1016/j.vetpar.2008.12.017. Epub 2008 Dec 25.
In the present study, we use microarray technology to investigate the expression patterns of 381 genes with known association to host immune responses. Hybridization targets were derived from previously characterized bovine cDNAs. A total of 576 reporters (473 sequence-validated cDNAs and 77 controls) were spotted onto glass slides in two sets of four replicates. Two color, comparative hybridizations across both mesenteric lymph node (MLN) and small intestine mucosa (SIM) RNA samples were done between animals with previously demonstrated phenotypic differences based on natural exposure to gastrointestinal (GI) nematodes over a 6-month exposure period. A total of 138 significant hybridization differences were detected by mixed model analysis of variance. A subset of these significant differences was validated by quantitative, real-time RT-PCR to assay transcript levels for 18 genes. These results confirmed that in the SIM, susceptible animals showed significantly higher levels in the genes encoding IGHG1, CD3E, ACTB, IRF1, CCL5 and C3, while in the MLN of resistant animals, higher levels of expression were confirmed for PTPRC, CD1D and ITGA4. Combined, the results indicate that immune responses against GI nematode infections involve multiple response pathways. Higher levels of expression for IgE receptor, integrins, complement, monocyte/macrophage and tissue factors are related to resistance. In contrast, higher levels of expression for immunoglobulin chains and TCRs are related to susceptibility. Identification of these genes provides a framework to better understand the genetic variation underlying parasite resistance.
在本研究中，我们使用微阵列技术来研究381个与宿主免疫反应已知相关基因的表达模式。杂交靶标源自先前鉴定的牛cDNA。总共576个报告基因（473个经序列验证的cDNA和77个对照）以两组四个重复的方式点样到载玻片上。在经过6个月暴露期自然感染胃肠道（GI）线虫后具有先前证明的表型差异的动物之间，对肠系膜淋巴结（MLN）和小肠黏膜（SIM）RNA样本进行了双色、比较杂交。通过方差混合模型分析检测到总共138个显著的杂交差异。通过定量实时RT-PCR对其中一部分显著差异进行验证，以测定18个基因的转录水平。这些结果证实，在SIM中，易感动物在编码IGHG1、CD3E、ACTB、IRF1、CCL5和C3的基因中表达水平显著更高，而在抗性动物的MLN中，PTPRC、CD1D和ITGA4的表达水平更高。综合来看，结果表明针对GI线虫感染的免疫反应涉及多种反应途径。IgE受体、整合素、补体、单核细胞/巨噬细胞和组织因子的较高表达水平与抗性相关。相反，免疫球蛋白链和TCR的较高表达水平与易感性相关。这些基因的鉴定为更好地理解寄生虫抗性背后的遗传变异提供了一个框架。
