Article

Eosinophil adoptive transfer system to directly evaluate pulmonary eosinophil trafficking in vivo

Division of Allergy and Immunology, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH 45229.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 03/2013; 110(15). DOI: 10.1073/pnas.1220572110
Source: PubMed

ABSTRACT

Most in vivo studies of granulocytes draw conclusions about their trafficking based on examination of their steady-state tissue/blood levels, which result from a combination of tissue homing, survival, and egress, rather than direct examination of cellular trafficking. Herein, we developed a unique cell transfer system involving the adoptive transfer of a genetically labeled, bone-marrow-derived unique granulocyte population (eosinophils) into an elicited inflammatory site, the allergic lung. A dual polychromatic FACS-based biomarker-labeling system based on the IL4-eGFP transgene (4get) or Cd45.1 allele was used to track i.v. transferred eosinophils into the airway following allergen or TH2-associated stimuli in the lung in multiple mouse strains. The system was amenable to reverse tagging of recipients, thus allowing transfer of nonlabeled eosinophils and competitive tracking of multiple populations of eosinophils in vivo. The half-life of eosinophils in the blood was 3 h, and migration to the lung was dependent upon the dosage of transferred eosinophils, sensitive to pertussis toxin pretreatment, peaked at ∼24 h after adoptive transfer, and revealed a greater than 8-d eosinophil half-life in the lung. Eosinophil migration to the lung was dependent upon recipient IL-5 and IL-13 receptor α1 and donor eosinophil C-C chemokine receptor type 3 (CCR3) and interleukin 1 receptor-like 1 (ST2) in vivo. Taken together, this unique eosinophil transfer system provides an unprecedented opportunity to examine airway eosinophil migration without the need for extensive efforts to acquire donor source and time-consuming genetic crossing and has already been used to identify a long eosinophil half-life in the allergic lung and a definite role for ST2 in regulating eosinophil trafficking.

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    • "To test directly whether eosinophil trafficking was reduced in HFD fed mice, we measured the number of fluorescently labelled bone marrow-derived eosinophils in the intestine 3–4 days after intravenous injection. However, as has previously been reported using this method in naïve, unchallenged mice, we found minimal trafficking to the target tissue (<0.1percent of cells) of normal chow or HFD animals confounding definitive interpreta- tion[33]Intestinal eosinophil depletion is caused by the fat content of the diet rather than obesity To determine whether the high fat content of the diet or obesity itself was the cause of the eosinophil deficiency, we analyzed lamina propria eosinophils in Ob/Ob mice, since these mice become obese on chow diets. In contrast to the HFD mice, there was no detectable differencein eosinophil content in the intestine of Ob/Ob mice (Fig 4A). "
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    • "on has led to the generation of CD45 . 1 mutant mice strains , which is now – similar to eGFP – a crucial tool in the experimental design in immunology ( Deak et al . , 2010 ) . Although both transgenes , eGFP and CD45 . 1 , are commonly used for cells tracing , e . g . upon adoptive transfer of cells or when generating bone marrow chimeric mice ( Wen et al . , 2013 ) , insights on the sensitivity of the cell detection level are still elusive ."
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    • "on has led to the generation of CD45 . 1 mutant mice strains , which is now – similar to eGFP – a crucial tool in the experimental design in immunology ( Deak et al . , 2010 ) . Although both transgenes , eGFP and CD45 . 1 , are commonly used for cells tracing , e . g . upon adoptive transfer of cells or when generating bone marrow chimeric mice ( Wen et al . , 2013 ) , insights on the sensitivity of the cell detection level are still elusive ."

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