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Lipopolysaccharide and Concanavalin A Differentially Induce the Expression of Immune Response Genes in Caprine
Vishakh Walia1, Rohit Kumar, Abhijit Mitra
1a Genome Analysis Laboratory, Animal Genetics Division , Indian Veterinary Research Institute , Izatnagar , India.
Insights
Concanavalin A (ConA) more effectively stimulates immune response genes in goat macrophages than lipopolysaccharide (LPS). This finding aids in selecting optimal methods for studying caprine cellular immunity and pathogen challenges.
Area of Science:
- Veterinary Immunology
- Cellular Immunology
- Molecular Biology
Background:
- Monocyte-derived macrophages (MDMs) are crucial in vitro models for pathogen challenge studies.
- Lipopolysaccharide (LPS) and concanavalin A (ConA) are common inducers for assaying cellular immunity.
- Goat-specific immune response studies using these inducers are limited.
Purpose of the Study:
- To compare the efficacy of LPS and ConA in initiating immune responses in caprine MDMs.
- To analyze the expression profiles of key immune response (IR) genes in goat macrophages.
- To determine the optimal inducer for studying general immune responses in caprine MDMs.
Main Methods:
- Caprine MDMs were induced with LPS and ConA.
- Real-Time quantitative PCR (RT-qPCR) was used to analyze gene expression.
- Expression levels of TNFA, IFNG, IL2, GMCSF, IL10, TGFB, NRAMP1, NOS2, and CASP1 were quantified.
Main Results:
- Both LPS and ConA upregulated GMCSF, TGFB1, IL10, and IFNG, and downregulated NRAMP1.
- ConA upregulated TNFA and IL2, while LPS upregulated NOS2.
- CASP1 expression remained unaltered by either inducer.
- ConA demonstrated a more pronounced effect on IR gene regulation.
Conclusions:
- ConA is a more potent inducer for studying general immune responses in caprine MDMs compared to LPS.
- This study provides valuable insights for selecting appropriate methods in caprine immunology research.
- Findings contribute to a better understanding of goat macrophage activation and immune gene expression patterns.
Abstract:
Monocyte derived macrophages (MDMs), as an in vitro model in pathogen challenge studies, are generally induced with lipopolysaccharide (LPS) and concanavalin A (ConA) to assay cellular immunity. General immune responses to LPS and ConA have been studied in a wide range of species, but similar studies are limited to goats. In the present study, caprine MDMs were induced with LPS and ConA and the expression profile of immune response (IR) genes, namely, Tumor Necrosis Factor Alpha (TNFA), Interferon Gamma (IFNG), Interleukin 2 (IL2), Granulocyte Macrophage Colony Stimulating Factor (GMCSF), Interleukin 10 (IL10), Transforming Growth Factor Beta (TGFB), Natural Resistance-Associated Macrophage Protein-1 (NRAMP1), inducible nitric oxide synthase (NOS2), and caspase1 (CASP1) were studied to compare the potential of LPS and ConA in initiating immune responses in goat macrophages. Real Time quantitative PCR (RT-qPCR) analysis revealed that both LPS and ConA caused an upregulation (p < 0.05) of GMCSF, TGFB1, IL10, and IFNG and down-regulation of NRAMP1. TNFA and IL2, and NOS2 were upregulated (p < 0.05) by ConA and LPS, respectively. Whereas, the expression of CASP1 remain unaltered. Comparatively, the effect of ConA was more pronounced (p < 0.05) in regulating the expression of IR genes suggesting its suitability for studying the general immune responses in caprine MDM.
