Abstract
Pigs are popular animal models in biomedical research. RNA-Seq is becoming the predominant tool to investigate transcriptional changes of the pig’s response to infection. The high sensitivity of this tool requires a strict control of the study design beginning with the selection of healthy animals to provide accurate interpretation of research data. Pigs chronically infected with Mycoplasma suis often show no obvious clinical signs, however the infection may affect the validity of animal research. The goal of this study was to investigate whether or not this silent infection is also silent at the host transcriptional level. Therefore, immunocompetent pigs were experimentally infected with M. suis and transcriptional profiles of whole blood, generated by RNA-Seq, were analyzed and compared to non-infected animals. RNA-Seq showed 55 differentially expressed (DE) genes in the M. suis infected pigs. Down-regulation of genes related to innate immunity (tlr8, chemokines, chemokines receptors) and genes containing IFN gamma-activated sequence (gbp1, gbp2, il15, cxcl10, casp1, cd274) suggests a general suppression of the immune response in the infected animals. Sixteen (29.09%) of the DE genes were involved in two protein interaction networks: one involving chemokines, chemokine receptors and interleukin-15 and another involving the complement cascade. Genes related to vascular permeability, blood coagulation, and endothelium integrity were also DE in infected pigs. These findings suggest that M. suis subclinical infection causes significant alterations in blood mRNA levels, which could impact data interpretation of research using pigs. Screening of pigs for M. suis infection before initiating animal studies is strongly recommended.
Highlights
The pig (Sus scrofa) is an important meat-production animal and the most commonly used large animal in research, as a mammalian model for the study of human diseases [1]
Aliquots collected from a specific pathogen free (SPF) pig inoculated with M. suis and at the peak of bacteremia [29], representing the second passage of this strain, were frozen at −80 °C and used in the experimental infection described
The raw RNA sequencing (RNA-Seq) data was submitted to the Sequence Read Archive (SRA) database at the National Center for Biotechnology Information (NCBI) under Bioproject Accession Number PRJNA321932
Summary
The pig (Sus scrofa) is an important meat-production animal and the most commonly used large animal in research, as a mammalian model for the study of human diseases [1]. Their size, anatomy, and physiology as well as the progression of many diseases, such as diabetes, atherosclerosis, and cardiovascular disease, are similar to humans [1]. RNA sequencing (RNA-Seq) using next-generation sequencing (NGS) technology is becoming the predominant tool for largescale gene expression analyses. The selection of healthy pigs is the first step toward the goal of generating valid research data that will provide results that are more predictive of human diseases
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