Table 1.
Other asthma-related allergens contain disulfide bonds and thus may require GILT for optimal MHC class II-presentation.
Figure 1.
GILT−/− lymphocytes exhibit deficient recall proliferation to Der p 1.
A, Crystal structure of Der p 1 adapted from Chruszcz et al [14]. Disulfide bonds are shown in red; immunodominant epitopes are shown in yellow (p110), cyan (p78), and orange (p21). B and C, Recall proliferation analysis of lymphocytes isolated from Der p 1-challenged mice and incubated with increasing concentrations of purified Der p 1 (B) or immunodominant peptides (C). Data shown as mean ± SEM of triplicate wells and are representative of three independent experiments; * p<0.05, ** p<0.001. D, Recall proliferation analysis of lymphocytes from Der p 1-challenged mice that were depleted of CD8+ or CD4+ T cells prior to incubation with increasing concentrations of purified Der p 1 or p110 peptide. Data shown as mean ± SEM of triplicate wells and are representative of two independent experiments. E, Crystal structure of papain adapted from Kamphuls et al [19] and recall proliferation analysis of lymphocytes isolated from papain-challenged mice and incubated with increasing concentrations of papain. Data shown as mean ± SEM of triplicate wells and are representative of three independent experiments; p>0.05 for all points.
Figure 2.
Wild-type CD4+ T cells exhibit deficient proliferation following co-incubation with Der p 1- but not papain-pulsed Gilt−/− dendritic cells.
A, Proliferation of CD4+ T cells isolated from Der p 1-challenged wild-type mice following a three day co-incubation with wild-type or Gilt−/− bone marrow-derived dendritic cells that had been pulsed with increasing concentrations of purified Der p 1. B, Proliferation of CD4+ T cells isolated from papain-challenged wild-type mice following a three day co-incubation with wild-type or Gilt−/− bone marrow-derived dendritic cells that had been pulsed with increasing concentrations of papain. Data shown as mean ± SEM of triplicate wells and are representative of at least two independent experiments.
Figure 3.
Gilt-/- mice develop reduced bronchoalveolar lavage cellularity following intranasal Der p 1 challenge.
A, Schematic of the Der p 1/papain allergic asthma model. Mice were intranasally primed on day 0 and 1 with 0.1 µg LPS as Th2 adjuvant. Mice were rechallenged on day 12 and 13, then sacrificed 24 hours later for BAL analysis. B, BAL cellularity from age- and sex-matched wild-type and Gilt−/− mice challenged with PBS, Der p 1, or papain. Pooled data from three to six mice from three (papain) or four (Der p 1) experiments are shown as mean ± SEM. C. Percentage of total BAL cellularity represented by eosinophils and total eosinophilia in Der p 1-challenged or control wild-type and Gilt−/− mice. D. Relative expression of cytokines (IL-5, IL-13, IFN-γ) or MUC5AC in wild-type or Gilt−/− lung tissue following intranasal Der p 1 challenge. Expression is normalized to β-tubulin expression and control samples. Data shown as mean ± SEM of samples from two to four mice and are representative of two experiments. Asterisks denotes p value less than 0.05.
Figure 4.
Gilt−/− mice develop less bronchoalveolar lavage cellularity and histological symptoms following intranasal house dust mite (HDM) challenge.
A, Schematic of the HDM asthma model used for BAL and histological analysis. Mice were intranasally primed on day 0 and 1 with 100–300 µg HDM extract and rechallenged on day 12 and 13 (BAL), then sacrificed 24 hours later for analysis. B, BAL cellularity from age- and sex-matched wild-type and Gilt−/− mice. Pooled data from three to nine mice from two experiments are shown as mean ± SEM. C. Percentage of total BAL cellularity represented by eosinophils and total eosinophilia in HDM-challenged or control wild-type and Gilt−/− mice. D. Relative cytokine expression (IL-13, IFN-γ) or MUC5AC in wild-type or Gilt−/− lung tissue following intranasal HDM challenge. Expression is normalized to β-tubulin expression and control samples. Data shown as mean ± SEM of samples from two to nine mice and are averaged from two experiments.
Figure 5.
Gilt−/− lymphocytes exhibit deficient recall proliferation to the cockroach allergen Bla g 2 but not to the house dust mite allergen Der f 1.
Recall proliferation analysis of lymphocytes isolated from (A) Bla g 2- or (B) Der f 1-challenged mice and incubated with increasing concentrations of purified (A) Bla g 2 or (B) Der f 1. Data shown as mean ± SEM of triplicate wells and are representative of two independent experiments; * p<0.05, ** p<0.001.