Immunophenotyping of normal lymphocytes

Area of Science:

• Immunology
• Hematology
• Cell Biology

Background:

• Accurate enumeration of peripheral blood mononuclear cells is crucial for diagnosing and monitoring various health conditions.
• Establishing reliable reference ranges for immune cell populations is essential for clinical interpretation.

Purpose of the Study:

• To establish reproducible normal reference values for peripheral blood mononuclear cell populations.
• To validate flow cytometric analysis for enumerating T cells, B cells, natural killer (NK) cells, and myeloid cells.
• To assess the clinical utility of these reference values in both healthy individuals and patients with non-hematological disorders.

Main Methods:

• Direct and indirect immunofluorescence labeling of peripheral blood mononuclear cells.
• Flow cytometric analysis to define cell surface antigen expression.
• Use of polyclonal and monoclonal antibodies for T cells, T-cell subsets, B cells, NK cells, and myeloid cells.
• Comparison with established methods like whole blood lysis and E rosettes.

Main Results:

• Normal values for absolute numbers and percentages of mononuclear cell populations were established with high reproducibility.
• Flow cytometry results were comparable to other established immunological methods.
• Immune cell recovery was generally complete, indicating the reliability of the method.
• Results were consistent between normal controls and patients with non-hematological disorders.

Conclusions:

• The study successfully established clinically useful, reproducible reference values for peripheral blood mononuclear cell populations.
• Flow cytometry is a reliable method for enumerating immune cell subsets.
• These reference values serve as a crucial benchmark for lymphoid sub-populations in normal controls and can aid in clinical assessments.