Box 1. Box1 | Protocol to reconstitute immunodeficient mice with immunocompetent cells

From the following article

A method to enable the investigation of murine bronchial immune cells, their cytokines and mediators

Joachim H Maxeiner, Roman Karwot, Michael Hausding, Kerstin A Sauer, Petra Scholtes & Susetta Finotto

Nature Protocols 2, 105 - 112 (2007) Published online: 22 February 2007

doi:10.1038/nprot.2007.8

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SCID mice are organized in cages in accordance with the experimental adoptive transfer protocol

Day -1: The day before the adoptive cell transfer, aerosolize the mice with the antigen (e.g., 5% OVA in PBS) for 30 min (refs. 1, 2; Fig. 2e). The donor mice should already have undergone the standard sensitization and challenge protocol with antigen as described previously (ref. 3; Fig. 2e)

Day 0: Isolate spleen CD4+ T cells or other immunocompetent cells to be used in the adoptive transfer from donor mice and resuspend in Sterofundin at a concentration of approximately 2 times 106 cells in 200 mul. Inject 1 times 106 to 3 times 106 CD4+ T cells or other cells intraperitoneally into SCID mice (2 times 106 in 200 mul of Sterofundin)

Days 1–3: Expose SCID mice to OVA aerosol (5% in PBS)

Day 4: Perform non-invasive airways hyper-responsiveness (AHR) measurements, if desired

Day 5: Perform invasive measurements of AHR and BALF

Note that invasive AHR and BALF can be performed on day 4

Analysis of reconstituted immunodeficient mice (including the reconstitution itself) takes 5–7 days when the cells that are to be used for the adoptive transfer are available. The time required for generating the cells used for the adoptive transfer varies on the purpose of the experiment. For TH1 and TH2 clone generation, 7 days are required; for asthma protocols, 7–28 days are required (ref. 3; Fig. 1e)