title: Blood_Control_CD44low [GRA769A4]
organism: Mus musculus
attributes.tag: source_name
attributes.value: Blood_Control_CD44low
attributes.tag: mouse model
attributes.value: House dust mite (HDM) allergy
attributes.tag: mouse strain
attributes.value: C57BL/6/J
attributes.tag: condition
attributes.value: Control
attributes.tag: infection protocol
attributes.value: Female C57BL/6/J mice (MRC NIMR) were sensitized with 10 mg HDM (Greer) and 2 mg Imject Alum (Thermo Scientific) in 200 ml PBS or Alum alone as control by i.p. injections on days 0 and 14, followed by i.t. challenge with 10 mg HDM in 20 ml of PBS or PBS on days 21 and 24 as described6. Whole blood and lung samples were collected from individual HDM and PBS control treated mice on day 25. For FACS sorting of T cells, pooled blood from 4-5 individual HDM or PBS treated mice was collected into heparin sodium (Wockhardt) at 10 to 30 international units per ml of blood. Peripheral blood mononuclear cells were isolated by density separation with Lympholyte®-Mammal (Cedarlane). Lung CD4+ T cells were enriched by positive selection (Miltenyi Biotech) from a corresponding pool of lungs. Blood and lung cells were stained with CD3 (145-2C11) APC, CD4 (RM4-5) eFluor450 and CD44 (IM7) PE (all from eBioscience) and CD3+CD4+, CD3+CD4+ CD44low and CD44high cells were sorted on MoFlo XDP (Beckman Coulter) and BD FACSAria™ Fusion (Beckton Dickinson) flow cytometers and 15,000 per population collected into TRI-Reagent LS (Sigma-Aldrich).
attributes.tag: cell type
attributes.value: CD3+CD4+ CD44lo
attributes.tag: mouse id
attributes.value: MS0818
attributes.tag: tissue
attributes.value: Blood