High-throughput Gene Expression Analysis In Pigs As Model For Respiratory Infections

Abstract

Influenza A virus infections have great impact on human health and welfare and significant resources are linked to influenza epidemics due to excess hospitalizations and lost productivity. Up to 15% of the human population is affected when Influenza spreads around the world in seasonal epidemics (WHO). Animal models are essential in understanding the mechanisms involved in human infectious disease and for the development of effective prevention and treatment strategies. It is increasingly realized that large animal models like the pig are exceptionally human like and serve as an excellent model for disease and inflammation. Pigs are fully susceptible to human influenza, and have been demonstrated to be involved in influenza evolution and ecology. Pigs share many similarities with humans regarding lung physiology and innate immune cell infiltration of the respiratory system and thus seem to be an obvious large animal model for respiratory infections. This study aimed at providing a better understanding of the involvement of circulating non-coding RNA and innate immune factors in porcine blood leukocytes during influenza virus infection. By employing the pig as a model we were able to perform highly controlled experimental infections and to study changes of symptoms, viral titer, and expression of microRNAs/mRNAs as the influenza infection progresses in time, generating information that would be difficult to obtain from human patients. The Gram-negative bacterium Actinobacillus pleuropneumoniae causes pneumonia in pigs, a disease which is associated with high morbidity and mortality, as well as impaired animal welfare. The rapidly evolving pneumonia is characterized by large areas of lung necrosis resulting from the combined effect of tissue damage caused by the bacteria, and a strong proinflammatory immune response. To obtain in-depth understanding of this infection, concurrent gene expression of host and pathogen in lung samples collected from pigs experimentally infected with A. pleuropneumoniae was studied. We applied high-throughput RT-qPCR for the simultaneous analysis of host and pathogen gene expression. This parallel analysis was done in lung tissue samples as well as in the immediate surroundings of infection loci after laser capture microdissection. Regulation of gene expression of several immune factors was observed in agreement with protein levels of these factors in lung tissue, infection status and histopathological findings.