牛支原体体外肺精准切片感染模型的建立

doi:10.11843/j.issn.0366-6964.2024.10.035

Abstract

Abstract:

Mycoplasma bovis (M. bovis) is an important pathogen of bovine respiratory disease complex (BRDC), mainly causing pneumonia, mastitis, arthritis, keratoconjunctivitis, otitis, and genital disorders, leading to high economic losses in dairy and beef cattle production. Lack of small animals' models have greatly hindered the progress of effective vaccines and drugs for prevention and control in Mycoplasma bovis. In this study, we have established an in vitro infection model of M. bovis in Precision-cut lung tissue slices (PCLS). We collected bovine lungs from apparently healthy two-months cattle after slaughter, filled the lung airway with low melting point agarose, transferred it to the vibratome tissue slicer and then made the 250 μm-thickness precision cut lung slices. After M. bovis wildtype strain HB0801(P1) and its attenuated strain P150 at a MOI of 10⁸ CFU infected, quantitative real-time PCR, lactate dehydrogenase cytotoxicity assay, 3D fluorescence microscope panoramic scanning, immunohistochemistry analysis were performed. The results showed that cultured PCLS remained 60% cell viability for at least 72 h and maintained normal structural integrity including edge sharpness and alveolar structure well in uninfected PCLS. After infected with wildtype and attenuated M. bovis strains respectively, we found that both M. bovis could survive and proliferate in PCLS, tropism to alveolar and bronchial space, and showed cytotoxic reactions until 18 h post-infection. We also observed that after 36 h incubation with M. bovis wildtype strain P1 and its attenuated strain P150, cultured PCLS produced pro-inflammatory factors IL-1β and IL-8 in PCLS, and M. bovis wildtype strain HB0801 could induce significantly higher level than P150 (P < 0.05). In conclusion, this study provides a model and method to investigate M. bovis infection in vitro which can mimic M. bovis infection in vivo. It is expected to provide reference for subsequent research on host-pathogen interaction in M. bovis.

Key words: Mycoplasma bovis, precision-cut lung tissue slices, infected in vitro, inflammatory response

CLC Number:

S852.62

Hui ZHANG, Doukun LU, Yiqiu ZHANG, Gang ZHAO, Yingyu CHEN, Xi CHEN, Changmin HU, Aizhen GUO. Establishment of Cattle Precision-Cut Lung Tissue Slices (PCLS) Models Infected with Mycoplasma bovis in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4638-4645.

Figures/Tables 5

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References 17

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Establishment of Cattle Precision-Cut Lung Tissue Slices (PCLS) Models Infected with Mycoplasma bovis in vitro

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