Irradiated Chimeric Mouse Model
Objective: Demonstrate that TLR4 expression on radioresistant lung structural cells is required and sufficient for dendritic cell activation in the lung and for priming of effector T helper responses to house dust mite allergen
Gather these items before starting the experiment. Check off items as you prepare.
As an Amazon Associate, we earn from qualifying purchases. Product links help support this free resource.
Protocol Steps
Generate irradiated chimeric mice
Create chimeric mice through irradiation to establish models with differential TLR4 expression on lung structural cells versus hematopoietic cells
Note: These mice allow testing of whether TLR4 on radioresistant lung structural cells is required for DC activation and allergic responses
View evidence from paper
“Using irradiated chimeric mice, we demonstrate that TLR4 expression on radioresistant lung structural cells is required and sufficient for DC activation in the lung”
Expose mice to house dust mite allergen
Administer HDM allergen to induce allergic airway inflammation and assess TLR4 signaling requirements
Note: Used to test whether TLR4 on structural cells but not hematopoietic cells is necessary for allergic responses
View evidence from paper
“The absence of TLR4 on structural cells, but not on hematopoietic cells, abolished HDM driven allergic airway inflammation”
Measure dendritic cell activation
Assess DC activation in the lung following TLR4 triggering on structural cells
Note: DC activation is a key outcome measure for determining TLR4 signaling on structural cells
View evidence from paper
“TLR4 expression on radioresistant lung structural cells is required and sufficient for DC activation in the lung”
Measure innate proallergic cytokine production
Quantify production of thymic stromal lymphopoietin, granulocyte-macrophage colony stimulating factor, interleukin-25 and IL-33 from structural cells following TLR4 triggering
Note: These cytokines are produced by structural cells in response to TLR4 triggering and drive allergic inflammation
View evidence from paper
“TLR4 triggering on structural cells caused production of the innate proallergic cytokines thymic stromal lymphopoietin, granulocyte-macrophage colony stimulating factor, interleukin-25 and IL-33”
Assess T helper cell priming
Evaluate priming of effector T helper responses to HDM allergen
Note: Measures whether TLR4 on structural cells is sufficient for T cell responses
View evidence from paper
“TLR4 expression on radioresistant lung structural cells is required and sufficient for DC activation in the lung and for priming of effector T helper responses to HDM”
Administer TLR4 antagonist via inhalation
Deliver TLR4 antagonist through inhalation to target exposed epithelial cells
Note: Used to suppress asthma features including bronchial hyperreactivity
View evidence from paper
“Finally, inhalation of a TLR4 antagonist to target exposed epithelial cells suppressed the salient features of asthma including bronchial hyperreactivity”
Measure bronchial hyperreactivity
Assess bronchial hyperreactivity as a measure of asthma severity
Note: Key outcome measure for evaluating effectiveness of TLR4 antagonist treatment
View evidence from paper
“inhalation of a TLR4 antagonist to target exposed epithelial cells suppressed the salient features of asthma including bronchial hyperreactivity”