IL-4-IRES-eGFP (4get) reporter mice: Monitor polarized immune response

Validation data

• Fig. 1) eGFP expression correlates with IL-4-expressing T cells and reflects IL-4 production

  ‍A: In contrast to Th1 conditions, the Th2-specific conditions on a 4get cell's eGFP- population, leads to a great increase expression of eGFP, reflecting IL-4 production.

  ‍Purified, naive CD4+ eGFP- homozygous 4get T cells were labeled with PKH26 red fluor. Cells were cultured under Th1, neutral, or Th2 conditions in the presence of antigen-presenting cells and analyzed on day 4, after gating on CD4+ cells.

  ‍B: Only under Th2 conditions did 4get cells specifically express IL-4.

  ‍Splenocytes from wild-type (+/+, open bars) and heterozygous (+/4get, hatched bars) or homozygous (4get/4get, dark bars) 4get mice were depleted of CD8+ cells by complement lysis and stimulated under Th1 or Th2 conditions for 5 days. Cells were washed and re-stimulated and supernatants were collected after 48 hours and analyzed for IL-4 and IFN-γ by ELISA. Mean and positive standard deviations of triplicate cultures are shown.

  

• Fig. 2) 4get cells retain the capacity to respond to N. brasiliensis infection in vivo by producing huge amount of IL-4

  In 4get mice infected, 4get cells produce IL-4 in response to N. brasiliensis infection.

  ‍Homozygous 4get mice were infected with N. brasiliensis. FACS analysis of lung, mesenteric lymph nodes (MLN), peripheral lymph nodes (PLN), and spleen (SPL) was performed after 10 days. Cells from the forward- and sidescatter lymphocyte gate were examined for CD4 and eGFP expression. Non-infected control mice were housed in the same facility as infected mice. Cells from three mice were pooled for each analysis. Numbers indicate percentages per quadrant.

  

• Validation data