transcripts could be determined based on BLASTX searches and anno tated with DAVID or by searching against the GenBank database. Among these, 135 unique genes were grouped into 39 categories based on biological pro cess Gene Ontology terms or according to their potential Biology Process Classification sellckchem by referring to recent publications. Unsurprisingly, the majority of genes were related to the immune response, Transcription, Transport, material and energy metabo lism, etc. Validation of microarray data by quantitative real time PCR The qPCR was performed to validate the expression pat terns during infection for specific genes identified in the microarray assay. In order to validate the differential expression of various identified genes, 16 up regulated genes, with the increase ranging from 2.
0 fold to 18. 6 fold, and 3 down regulated genes, with the decrease ran ging from 2. 5 fold to 5. 9 fold, were selected for qPCR analysis. All the selected down regulated genes could be amplified from the control samples but failed to achieve significant detectable signs from WT infected spleens, except for ALOX15 which showed 3. 2 fold down regu lated expression. All selected up regulated genes showed higher expression in WT infected samples than in the control samples. Though variation in fold changes could be observed between qPCR and microar ray, the differential expression patterns were coincident between the results of the two techniques, which indicated the reliability of the microarray analysis.
Induction of inflammasomes and acute phase proteins by SS2 infection Highly pathogenic SS2 infection could cause up regu lated expression of a large set of genes involved in the inflammatory response and acute phase proteins by microarray analysis. IL 1B, IL 6 and IL 8 could be induced by foreign pathogens and play essential roles in controlling infections. However, they may also cause pathology when these productions are excessive or uncontrolled. Ye et al. also found that signifi cantly high level of cytokines could be induced by highly pathogenic SS2 strain and play important roles in sepsis, which is in coincidence with ours. In addition, quite a few genes related to inflammatory response were found up regulated, such as S100 family proteins, Pentraxin 3 and Resistin.
They play important roles in med iating inflammatory responses, recruiting inflammatory cells to sites of tissue damage or contributing to resist ing the invasion of various pathogens. Acute phase proteins, Batimastat such as Lactotransferrin, Haptoglobin, Serum amyloid A 2 and coagulation factor XIII, were involved in physiologic reactions initiated early in the inflammatory process, and could be a response to S. suis infection. CEBPD belonging to the CCAAT enhancer binding pro tein family which is crucial in the regulation of genes involved in immunity and inflammation. These up regulated genes are the representative of host acute response struggling selleck inhibitor to eliminate invading pathogens. Induction of g