Abstract

Type I interferons (IFNs) are essential regulators of antiviral defense, yet individual IFNα subtypes can elicit divergent immune outcomes. To systematically define these functional differences, we established an in vitro and in vivo workflow combining defined IFNα stimulations, T cell polarization assays, and viral infection models. Seven murine IFNα subtypes were compared for their capacity to modulate CD4⁺ T cell responses to Friend retrovirus. The protocol integrates co-culture systems with dendritic cells, metabolic and interferon-stimulated gene (ISG) profiling, and downstream functional readouts in infected mice. Using this approach, we found that IFNα11 drives Th1 polarization, augments metabolic activity, and induces a broad ISG signature, whereas IFNα2 promotes Th17 differentiation with limited metabolic and ISG responses. Functional partitioning revealed that antiproliferative effects require dendritic-cell IFNAR signaling, while cytokine modulation is intrinsic to CD4⁺ T cells.

In this webinar, participants will learn how to implement these assays, interpret subtype-specific IFNα effects, and apply integrated readouts of metabolism, ISG induction, and T cell differentiation. Attendees will gain practical insights into designing experiments to dissect immune signaling at the cellular and organismal level and understand how subtype-specific IFN signaling can inform therapeutic strategies in antiviral and immunotherapy contexts.

Highlights

- Comparative workflow: Parallel testing of multiple IFNα subtypes in matched cellular and in vivo assays.

- Integrated readouts: Combines metabolic profiling, ISG quantification, and functional T cell differentiation analyses.

- Mechanistic insight: Distinguishes dendritic-cell-dependent and T-cell-intrinsic IFNα effects.

- Practical outcomes: Attendees learn how to design, execute, and analyze experiments to study subtype-specific interferon responses for research and therapeutic applications.