A Suction Blister Protocol to Study Human T-cell Recall Responses In Vivo

Area of Science:

• Immunology
• Cell-mediated immunity
• Vaccine development

Background:

• Cutaneous antigen-recall models are crucial for studying human memory T-cell responses in vivo.
• Existing methods may have limitations in accessing sufficient antigen-specific T-cells and their microenvironment.
• Suction blister (SB) induction offers a potential avenue for enhanced accessibility to these cells.

Purpose of the Study:

• To describe a practical, safe, and minimally invasive procedure for cutaneous antigen recall using skin suction blisters.
• To demonstrate the isolation and analysis of antigen-specific T-cells and cytokine microenvironments from SB specimens.
• To highlight the utility of this method for studying cell-mediated immunity and human memory responses, particularly for vaccine research.

Main Methods:

• Antigenic recall using the tuberculin skin test in individuals vaccinated with Bacillus Calmette-Guérin.
• Induction of skin suction blisters (SB) for sample collection.
• Harvest of antigen-specific T-cells and analysis of cytokine microenvironments using multiplex and flow cytometry.

Main Results:

• The SB method successfully facilitated the isolation of antigen-specific T-cells.
• Analysis revealed high fractions of antigen-specific polyfunctional CD4+ T-cells in SB specimens.
• These fractions were notably higher compared to cells isolated from peripheral blood.

Conclusions:

• The described cutaneous recall with SB induction is a safe and effective method for studying in vivo human immune memory.
• This technique provides superior access to antigen-specific T-cells and their local cytokine milieu compared to blood sampling.
• The method holds significant promise for advancing research in cell-mediated immunity, human memory, and vaccine development.