Cell Biochemistry Martinsried
1. Goal: Identification of juvenile asthma patients from flow cytometric and clinical chemistry parameters.
2. Experimental Data: Fourty nine clinical chemistry parameters as well as 103 percent cell frequency and fluorescence intensity values of CD4/CD8, CD3/CD19, CD3/HLA-DR, CD3/CD16+56, CD25/CD3, CD71/CD3, CD4/CD45RA, CD45RO/CD4, CD62L/CD4, CD4/CD29, CD57/CD8, CD8/CD11b, CD5/CD19, CD21/CD19, CD62L/CD20, IgG1/IgG2 two colour immunophenotypes as well as FCS-list mode files of: CD45/CD14, CD4/CD29, CD4/CD8, CD56/CD8, CD3/CD56, CD25/CD3, CD3/HLA-DR, CD71/CD3, CD3/CD19, CD5/CD19, IgG1/IgG2 immunophenotypes were available for analysis.
Data Pattern Classification:
The clinical chemistry and the flow cytometric data were
separately and jointly classified.
- Clinical Chemistry: Predictive values of 60.0% and 100.0% for healthy and asthmatic children in the learning set and of 57.1% and 77.7% for the unknown test set of children were obtained from the data pattern: thrombocyte and eosinophil counts, aspartate aminotransferase (ASAT), thyroid-stimulating hormone (TSH), ferritin, IgE, beta-globulin.
- Flow Cytometric Cell Frequency and Fluorescence Intensity: Predictive values of 100.0% and 100.0% for healthy and asthmatic children in the learning and unknown test sets were obtained from a lymphocyte data pattern of: CD45RA, CD45RO, CD4, CD8 antigen expression and % CD4neg/CD45RApos, % CD4neg/CD45RAneg, % CD5pos/CD19pos lymphocyte frequency.
- Flow Cytometric List Mode Analysis: Similar predictive values of 100.0% and 100.0% for healthy and asthmatic children were obtained from the exhaustive list mode analysis with parameters: CD45, HLA-DR, CD4, CD3, CD4 antigen expression and: % CD3pos/HLA-DRpos, % CD71pos lymphocytes, % CD25pos, % CD5pos/CD19neg granulocyte frequency , CD56/CD3 and CD19/CD5 antigen ratios on granulocytes.
The flow cytometric determination of leukocyte immunophenotypes from
peripheral blood discriminates healthy and asthmatic
children provides predictive values of 100.0% for the prospective classification
of unknown patients. This is superior to predictive values
between 57.1% and 77.7% for the prospective classification
with the most discriminating clinical chemistry parameters.
- A longterm goal of the studies consist in the early identification of children prior to disease outbreak e.g. from risk families. In this case, disease manifestation may be retarded or avoided by therapeutic intervention. Careful screening of patient leukocyte immunophenotypes seems a promising approach to this challenge in clinical cytomics.
L1 G.Valet, H.Kahle, F.Otto, E.Bräutigam, L.Kestens (2001) Prediction and precise diagnosis of diseases by data pattern analysis in multiparameter flow cytometry: Melanoma, Juvenile Asthma, HIV Infection. in: Cytometry (3rd edition) eds: Z Darzynkiewicz, JP Robinson, HA Crissman, Academic Press, San Diego, Methods Cell Biology 64:487-508
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