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In wild-type FVB mice, leukocyte recruitment to lipopolysaccharide was sexually dimorphic, with a greater number of leukocytes recruited in females. In male beta(2)-adrenergic receptor knock out mice (bred on a congenic FVB background) the number of leukocytes recruited was increased approximately 4-fold, while in females there was no change, eliminating sexual dimorphism in leukocyte migration. While there were significantly fewer recruited CD62L(+) and CD11a(+) leukocytes in wild-type males, only in male beta-adrenergic receptor knock out mice was there an increase in the number of recruited CD11a(+) leukocytes, again eliminating sexual dimorphism. Thus, leukocyte migration and CD11a(+) adhesion molecule expression in male, but not in female, leukocytes is beta-adrenergic receptor-dependent. Our findings provide support for a role of beta(2)-adrenergic receptor mechanisms in the inflammatory response, and suggest that beta(2)-adrenergic receptor on male leukocytes contributes to sexual dimorphism in the effect of stress on inflammatory diseases.
Figure 1. In vivo effect of LPS on leukocyte recruitment in the air pouch
Total…
Figure 1. In vivo effect of LPS on leukocyte recruitment in the air pouch
Total number of leukocytes in pouch lavage was determined by flow cytometry, as described in Materials and Methods. LPS elicits leukocyte recruitment in vivo. Sterile PBS (saline), alone, or containing 100 ng/ml LPS was injected into the 6-day murine air pouches of β2-adrenergic receptor knockout and control female and male animals. After 4 hours, exudates were collected and the total number of leukocytes was enumerated by flow cytometry as described in Materials and Methods. Number within bars indicates n.
Figure 2. Neutrophil accumulation in air pouch…
Figure 2. Neutrophil accumulation in air pouch exudates in response to LPS
Cells were immunostained…
Figure 2. Neutrophil accumulation in air pouch exudates in response to LPS
Cells were immunostained with a FITC anti-mouse neutrophil and with staining for T helper lymphocytes, phycoerythrin (PE)-CD4. The four panel part of figure shows typical flow cytometry dot plots; the abscissa shows fluorescence intensity for PE-labeled anti-CD4 and the ordinate shows the fluorescence intensity for FITC-labeled anti-neutrophil antibody. An FSC/SSC plot (bottom panel) shows the PE-labeled CD4 cell population; FSC and SSC were adjusted to include only live and nucleated cells (using propidium iodide and Hoeschst (R1 and R2), respectively). The plot shows PE positive (CD4) cells as red dots, with an SSC indicating a lymphocyte population separated from a population of larger and more dense monocytes and granulocytes.
Figure 3. Number of leukocytes expressing adhesion…
Figure 3. Number of leukocytes expressing adhesion molecules CD62L and CD11a recruited into the air…
Figure 3. Number of leukocytes expressing adhesion molecules CD62L and CD11a recruited into the air pouch by LPS
CD62L and CD11a expression on leukocytes found in the air pouch after LPS was injected into the pouch of male and female wild-type or knockout mice (n=6 for each group) ; * indicates P
Figure 4. CD11a and CD62L adhesion molecule…
Figure 4. CD11a and CD62L adhesion molecule density as measured by fluorescent intensity on peripheral…
Figure 4. CD11a and CD62L adhesion molecule density as measured by fluorescent intensity on peripheral blood leukocyte
Mean fluorescence intensity significantly less in CD11a knock out males, while there was no change in females. Fluorescence intensity for CD62L was unchanged in knock out male and female mice (n=6 for each group); * indicates P
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