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Immunology JoVE (Open Access) Citable · DOI

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

DOI: 10.3791/57554-v
What you'll learn
  • ✓ Perform tuberculin skin test (TST) intradermal injection technique
  • ✓ Evaluate and measure cutaneous recall responses at 48 hours
  • ✓ Harvest and collect blister fluid for immune cell analysis
  • ✓ Apply suction blister model to study human T-cell responses
Protocol

Here, we provide a demonstration of the suction blister cutaneous recall model. The model allows a simple access to study human in vivo adaptive immune responses, for instance in the context of vaccine development.

Difficulty
intermediate
Total time
~48–72 hours (48-hour observation period post-TST injection)
Biosafety
BSL-2

Steps

1
Administer tuberculin skin test intradermal injection

Inject tuberculin purified protein derivative (PPD) intradermally into the forearm skin using standard TST protocol. This initiates a delayed-type hypersensitivity reaction in previously sensitized individuals.

▶ 01:31
2
Evaluate skin induration and reaction size

At 48 hours post-injection, visually assess and measure the diameter of skin induration using a ruler or caliper. Document erythema and any systemic reaction signs.

▶ 03:56
3
Harvest blister fluid using suction device

Apply a sterile suction blister apparatus to the TST reaction site to create a fluid-filled blister. Carefully collect the accumulated blister fluid without contamination for downstream immune analysis.

▶ 07:24
4
Analyze blister fluid immune cell composition

Process harvested blister fluid to quantify T cells and characterize immune responses, including phenotyping and functional assays. Results demonstrate enrichment of antigen-specific memory T cells at the recall site.

▶ 08:35
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