The identification and analysis of allergen-specific CD4+ T cells is critical for understanding how these cells contribute to atopic disease and how to subvert their behavior through immune therapy. The advent of fluorescently labeled soluble tetramers of peptide:MHCII complexes (pMHCII tetramers) has provided investigators with an invaluable means to achieve this goal. Although pMHCII tetramers were first developed over two decades ago, their widespread use has been limited by the technical difficulty of generating these reagents. However, the adoption of various technical innovations from several labs over time has contributed greatly to the increased success in tetramer generation today. Here, we describe a comprehensive protocol for generating pMHCII tetramers using as an example a Derp1:I-Ab tetramer used to study allergen-specific CD4+ T cell responses in murine models of airway inflammation and allergic disease.
Keywords: Allergy; CD4+ T cell; House dust mite; Peptide:MHC complex; Tetramer.