ABSTRACT
We report that basophils in peripheral blood can be stained using histamine immunocytochemistry. The staining is based on the fixation of leucocytes with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (CDI) and the subsequent incubation of these cells with antisera raised against histamine conjugated to different carrier proteins using CDI. The staining appears to be specific for basophils and stained cells can be examined using both fluorescence microscopy and flow cytometry. In addition, histamine immunocytochemistry can be combined with conventional immunocytochemistry by incubating leucocytes with antibodies to cell surface antigens prior to or following fixation of the cells with CDI. Thus, histamine immunocytochemistry may be a valuable tool in future studies of human basophils.
Subject(s)
Basophils/immunology , Basophils/metabolism , Histamine/metabolism , Immunohistochemistry/methods , Alcian Blue , Antibodies, Monoclonal , Antigens, CD/metabolism , Coloring Agents , Ethyldimethylaminopropyl Carbodiimide , Evaluation Studies as Topic , Flow Cytometry , Histamine/immunology , Humans , Leukocytes/immunology , Leukocytes/metabolism , Microscopy, Fluorescence , Staining and Labeling/methodsABSTRACT
We report that NF-AT1 and NF-AT4 are expressed cytoplasmically in resting eosinophils, whereas NF-AT2 and NF-AT3 have not been seen. Likewise, NF-AT1 mRNA and NF-AT4 mRNA have been detected in resting eosinophils, and their levels can be significantly up-regulated by the Th2-associated cytokines IL-4 and IL-5. There is no detectable NF-AT protein expression in the nuclei of resting eosinophils. However NF-ATs appear in the nuclei of IL-4-, IL-5-, or ionomycin-stimulated eosinophils. Only NF-AT1 and NF-AT4, but not NF-AT2 and NF-AT3, have translocated into the nuclei in IL-4- or IL-5-stimulated eosinophils. These findings delineate a novel pathway in the cytokine network in which Th2 lymphocytes "control" eosinophils via the release of IL-4 and IL-5, and activation of NF-AT in eosinophils. The findings also suggest that a later feedback "talking" may exist between eosinophils and Th2 lymphocytes.
Subject(s)
DNA-Binding Proteins/biosynthesis , Eosinophils/metabolism , Interleukin-4/physiology , Interleukin-5/physiology , Lymphocyte Activation , Nuclear Proteins , T-Lymphocytes/metabolism , Transcription Factors/biosynthesis , DNA-Binding Proteins/blood , DNA-Binding Proteins/genetics , DNA-Binding Proteins/immunology , Eosinophils/immunology , Humans , Multigene Family/immunology , NFATC Transcription Factors , RNA, Messenger/biosynthesis , T-Lymphocytes/immunology , Th2 Cells/metabolism , Transcription Factors/blood , Transcription Factors/genetics , Transcription Factors/immunologyABSTRACT
Pharmacy Service of the Department of Veteran Affairs Medical Center in Lincoln, Nebraska recently implemented a computerized Baxter ATC-212 Unit Dose System. During implementation of this system, pharmacy technicians completed an accuracy comparison with manual filling. The Baxter ATC-212 System was 99.98% accurate and manual filling was 92.62% accurate. The accuracy of the combination of manually and ATC-212 filled drawers was 98.77%. The technicians had a median error rate of three errors per day with an average 352 doses filled per day.