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Clone-specific anti-CD45 blocking factor in patient plasma.

Samuel R Wellhausen¹, Stephen P Slone, James J Miller

¹Flow Cytometry Laboratory, James Graham Brown Cancer Center, University of Louisville, School of Medicine, Louisville, Kentucky 40202, USA. samuel.wellhausen@louisville.edu

Cytometry. Part B, Clinical Cytometry

|February 22, 2007

Summary

A plasma factor inhibited anti-CD45 staining in blood samples, but not other immune cell markers. This specific blocking activity was not identified as a heterophilic antibody.

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Area of Science:

Immunology
Hematology
Clinical Pathology

Background:

Flow cytometry is a crucial technique for analyzing immune cells.
Accurate staining of cell surface markers like CD45 is essential for reliable results.

Observation:

An unexplained inhibition of anti-CD45 staining was observed in a clinical blood specimen.
This inhibition affected anti-CD45 staining but not anti-CD3, -CD4, or -CD8 staining.

Findings:

A plasma factor was identified that specifically blocked anti-CD45 (clone 2D1) staining on lymphocytes.
The blocking factor's activity was specific to the anti-CD45 clone and not a general blocking agent.
Immunoabsorption and immunofluorescence studies ruled out heterophilic antibodies as the cause of the inhibition.

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Implications:

This finding highlights a potential interference in CD45 flow cytometry analysis due to unknown plasma factors.
Further investigation is needed to biochemically identify the blocking factor.
Understanding such interferences is critical for accurate clinical diagnostics and research.