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A Non-invasive and Technically Non-intensive Method for Induction and Phenotyping of Experimental Bacterial Pneumonia in Mice
Published on: September 28, 2016
Differential Gene Expression in Porcine Lung Compartments after Experimental Infection with Mycoplasma hyopneumoniae
Rubén S Rosales1, David Risco2, Obdulio García-Nicolás3,4
1Instituto Universitario de Sanidad Animal y Seguridad Alimentaria (IUSA), Veterinary Faculty, University of Las Palmas de Gran Canaria, Trasmontaña s/n, 35416 Arucas, Spain.
Abstract:
Mycoplasma hyopneumoniae (Mhyo) is the causative agent of porcine enzootic pneumonia (EP), as well as one of the main pathogens involved in the porcine respiratory disease complex. The host-pathogen interaction between Mhyo and infected pigs is complex and not completely understood; however, improving the understanding of these intricacies is essential for the development of effective control strategies of EP. In order to improve our knowledge about this interaction, laser-capture microdissection was used to collect bronchi, bronchi-associated lymphoid tissue, and lung parenchyma from animals infected with different strains of Mhyo, and mRNA expression levels of different molecules involved in Mhyo infection (ICAM1, IL-8, IL-10, IL-23, IFN-α, IFN-γ, TGF-β, and TNF-α) were analyzed by qPCR. In addition, the quantification of Mhyo load in the different lung compartments and the scoring of macroscopic and microscopic lung lesions were also performed. Strain-associated differences in virulence were observed, as well as the presence of significant differences in expression levels of cytokines among lung compartments. IL-8 and IL-10 presented the highest upregulation, with limited differences between strains and lung compartments. IFN-α was strongly downregulated in BALT, implying a relevant role for this cytokine in the immunomodulation associated with Mhyo infections. IL-23 was also upregulated in all lung compartments, suggesting the potential involvement of a Th17-mediated immune response in Mhyo infections. Our findings highlight the relevance of Th1 and Th2 immune response in cases of EP, shedding light on the gene expression levels of key cytokines in the lung of pigs at a microscopic level.
Insights
This study reveals how Mycoplasma hyopneumoniae (Mhyo) affects pig lungs by analyzing cytokine gene expression. Findings illuminate host-pathogen interactions crucial for developing better porcine enzootic pneumonia (EP) control strategies.
Area of Science:
- Veterinary immunology
- Swine respiratory diseases
- Molecular pathology
Background:
- Mycoplasma hyopneumoniae (Mhyo) causes porcine enzootic pneumonia (EP), a significant swine respiratory disease.
- Understanding Mhyo's host-pathogen interactions is vital for effective EP control strategies.
- Current knowledge of Mhyo's molecular impact on swine lung immunity remains incomplete.
Purpose of the Study:
- To investigate the host-pathogen interactions between Mhyo and pigs at a molecular level.
- To analyze the expression of key immune-related genes in different lung compartments following Mhyo infection.
- To identify strain-specific virulence differences and their impact on immune responses.
Main Methods:
- Laser-capture microdissection was employed to isolate specific lung tissues (bronchi, BALT, parenchyma).
- Quantitative PCR (qPCR) was used to measure mRNA expression levels of cytokines (ICAM1, IL-8, IL-10, IL-23, IFN-α, IFN-γ, TGF-β, TNF-α).
- Mhyo load, and macroscopic/microscopic lung lesions were quantified and scored.
Main Results:
- Significant differences in cytokine expression were observed across lung compartments and between Mhyo strains.
- IL-8 and IL-10 showed the highest upregulation, with minimal strain or compartment variation.
- IFN-α was downregulated in BALT, while IL-23 was upregulated in all compartments, suggesting Th17 involvement.
Conclusions:
- The study highlights the importance of Th1 and Th2 immune responses in porcine enzootic pneumonia.
- Gene expression analysis provides a microscopic view of immune responses to Mhyo infection in pigs.
- Findings offer insights into the complex interplay between Mhyo strains and the host immune system.

