Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination

Area of Science:

• Immunology
• Molecular Biology
• Vaccinology

Background:

• Gene expression quantification is crucial for assessing vaccine-induced immune responses.
• Lack of established reference genes for mouse draining lymph node studies post-vaccination hinders reliable analysis.

Purpose of the Study:

• To identify and validate reliable reference genes for gene expression studies in mouse draining lymph nodes following vaccination.
• To establish a foundation for accurate immunological gene expression analysis in this context.

Main Methods:

• Evaluated seven candidate reference genes using TaqMan primer/probe mixes.
• Utilized RefFinder web-based program incorporating geNorm, NormFinder, BestKeeper, and delta-Ct methods for analysis.
• Validated candidate genes by applying them in relative expression calculations for immune response-related genes.

Main Results:

• The ubiquitin C gene (Ubc) emerged as the most reliable reference gene.
• Ubc demonstrated stability when validated against genes with low expression levels post-vaccination.
• Optimal sample collection time was found to be gene-dependent.

Conclusions:

• The ubiquitin C gene (Ubc) is identified as the most dependable reference gene for mouse vaccination studies.
• This research provides essential data for immunological gene expression studies in draining lymph nodes.