The Expression Dynamics of Key Immune-Related Genes in Response to Mannheimia Haemolytica in Sheep Alveolar Macrophages In Vitro

Abstract

Alveolar macrophages (AMs) respond to these infections as the first line of defense and trigger the lung's immune response. Knowing the expression dynamics of key immune-related genes in alveolar macrophages against M. haemolytica challenge will help deepen the understanding of disease immunopathogenesis. For this purpose, first time an in vitro obtained from bronchoalveolar lavage (BAL) fluid and treated with three doses (T1: 1800, T2: 2700, T3: 5400 CFU) of M. haemolytica inoculum, while keeping one untreated as a control. Then, total RNA was extracted, and cDNA was synthesized. The real-time quantification data indicated that the expression of IL-1β, IL-6, IL-8, IL-10, and NF-κB genes were significantly changed as compared to control. Our study revealed that the exposure to M. haemolytica stimulates the immune response in the sheep alveolar macrophages in a dose and time-dependent manner.

Keywords

• Alveolar Macrophages
• Immune related Genes
• Gene Expression
• M. haemolytica
• Pneumonia

Koyun Alveoler Makrofajlarında Mannheimia Haemolytica'ya Yanıt Olarak Bağışıklık ile İlişkili Genlerin İn Vitro mRNA Ekspresyon Dinamikleri

Abstract

Alveoler makrofajlar (AM) enfeksiyonlara ilk savunma hattı olarak yanıt verir ve akciğerin bağışıklık tepkisini tetikler. Alveoler makrofajlarda Mannheimia haemolytica mücadelesine karşı anahtar bağışıklık ile ilişkili genlerin ekspresyon dinamiklerini bilmek, hastalık immünopatogenezinin anlaşılmasını derinleştirmeye yardımcı olacaktır. Bu amaçla, ilk kez interlökin-1β, IL-6, IL-8, IL-10 ve NF-κB'nin ekspresyon desenini doz ve zamana bağlı bir şekilde araştırmak için in vitro deneysel bir enfeksiyon modeli kullanıldı. Bronkoalveoler lavajdan (BAL) sıvıdan toplam %95 saf alveoler makrofaj kültür plakaları elde edildi ve kontrol grubu ile M. haemolytica inokulumun üç dozu (T1: 1800, T2: 2700, T3: 5400 CFU) analiz edildi. Daha sonra toplam RNA ekstrakte edilerek ve cDNA sentezlendi. Gerçek zamanlı PCR verileri, IL-1β, IL-6, IL-8, IL-10 ve NF-κB genlerinin ekspresyonunun, kontrole kıyasla önemli ölçüde değiştiğini gösterdi. Çalışmamız, M. haemolytica'ya maruz kalmanın, koyun alveoler makrofajlarındaki bağışıklık tepkisini dozda ve zamana bağlı bir şekilde uyardığını ortaya koymuştur.

Keywords

• Alveolar makrofaj
• Bağışıklıkla ilişkili genler
• Gen ifadesi
• M. haemolytica
• pnömoni

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