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Published on: October 16, 2013
Comparison of a lysed whole blood method to purified cell preparations for lymphocyte immunophenotyping: differences
K R Tamul1, M R O'Gorman, M Donovan
1Clinical Microbiology-Immunology Laboratory, University of North Carolina Hospitals, Chapel Hill 27514.
Insights
Comparing lysed whole blood (LWB) and peripheral blood mononuclear cells (PBMC) methods for HIV+ patient samples reveals significant differences. Results indicate these immunophenotyping methods are not interchangeable for accurate cell quantitation in clinical research.
Area of Science:
- Immunology
- Virology
- Clinical Laboratory Science
Background:
- Clinical laboratories commonly use lysed whole blood (LWB) for routine immunophenotyping.
- Researchers may require purified cell preparations, like peripheral blood mononuclear cells (PBMC), for specific studies involving human immunodeficiency virus (HIV+) patients.
Purpose of the Study:
- To compare the effects of LWB and PBMC sample preparation methods on immunophenotyping results.
- To assess the impact of these methods on specimens from both healthy donors and HIV+ patients.
- To determine the equivalence of immunophenotyping data obtained from LWB versus PBMC preparations.
Main Methods:
- Peripheral blood specimens from normal donors and HIV+ patients were analyzed.
- Both LWB and PBMC preparation methods were employed for comparison.
- Reproducibility and holding studies (pre- and post-processing) were conducted.
Main Results:
- Sample preparation methods demonstrated differential effects on abnormal patient specimens compared to healthy controls.
- Immunophenotyping outcomes varied significantly between LWB and PBMC methods.
- The observed discrepancies suggest method-specific alterations in cell populations.
Conclusions:
- LWB and PBMC sample preparation methods yield non-equivalent immunophenotyping results.
- Direct comparison of quantitative data derived from LWB and PBMC methods is not advisable.
- Method selection is critical for accurate cell quantitation in HIV+ research.
Abstract:
Although the majority of clinical laboratories now use a lysed whole blood (LWB) method for routine immunophenotyping, researchers wishing to perform other types of studies with lymphocytes from HIV+ patients may still need to use purified cell preparations, such as peripheral blood mononuclear cells (PBMC). A comparison study of the two methods was performed, using peripheral blood specimens from normal donors and from patients infected with the human immunodeficiency virus (HIV+). Reproducibility studies and several types of holding studies (both before and after specimen processing) were also performed. The results suggest that the two different methods of sample preparation have different effects upon abnormal patient specimens than those observed in healthy controls. Immunophenotyping results derived from the two different methods cannot be considered equivalent for the purposes of quantitating the presence of a particular type of cell.
