Enumeration of human lymphocyte subsets by monoclonal antibodies and flow cytometry: a comparative study using whole blood or mononuclear cells separated by density gradient centrifugation.

Author(s): De Paoli P, Reitano M, Battistin S, Castiglia C, Santini G

Publication: J Immunol Methods, 1984, Vol. 72, Page 349-53

PubMed ID: 6332151 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   Monoclonal antibodies recognizing T or B lymphocytes, or natural killer cells were used to quantitate lymphocyte subpopulations in whole blood or after cell separation via density gradient centrifugation.

   Summary of Findings:

   Cell separation resulted in significant changes in the percentage of large granular lymphocytes while T and B lymphocyte subpopulations did not change. Differences between the two separation methods were also observed. Natural killer cells (Leu7+ Leu11+) were selectively enriched at the Percoll interface I, while OKT8+, Leu7+, Leu11- cells were enriched at the bottom of the Ficoll tubes. Interestingly, the results suggest the Leu7+ subpopulation of lymphocytes can be further categorized based on reactivity with Leu11, natural killer activity, and density.

   Biospecimens

   • Bodily Fluid - Blood
   • Cell - White Cells

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Blood and blood products	Lymphocyte Whole blood
   Analyte Extraction and Purification	Analyte isolation method	Percoll separation Ficoll-Paque separation
   Flow cytometry Specific	Targeted peptide/protein	OKT3 OKT4 OKT8 B1 HLA-DR IgD Leu7 Leu11

   View more