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Updated: Aug 11, 2026
IP-FCM: Immunoprecipitation Detected by Flow Cytometry
Published on: December 2, 2010
Flow cytometry using the monoclonal antibody CD10-Pe/Cy5 is a useful tool to identify follicular lymphoma cells
1Departments of Hematology and Pathology. Hospital de la Santa Creu i Sant Pau, Barcelona, Spain.
Insights
Flow cytometry analysis for follicular lymphoma (FL) can be challenging. A novel CD10 antibody (CD10-Pe/Cy5) aids in identifying neoplastic cells with the t(14;18) translocation in FL and diffuse large cell lymphoma.
Area of Science:
- Hematology
- Immunophenotyping
- Flow Cytometry
Background:
- Follicular lymphoma (FL) diagnosis relies on characteristic histology, phenotype, and molecular markers.
- Traditional flow cytometry markers like CD19, CD10, and surface immunoglobulin (SIg) can be weakly expressed or negative in FL neoplastic cells.
- Accurate immunophenotyping is crucial for differentiating FL from other lymphoproliferative disorders.
Purpose of the Study:
- To establish the phenotypic pattern of neoplastic cells carrying the t(14;18)(q32;q21) chromosomal translocation using advanced flow cytometry techniques.
- To evaluate the utility of a novel CD10 antibody (CD10-Pe/Cy5) in identifying FL and related lymphomas.
- To correlate flow cytometry findings with histological diagnosis and molecular rearrangements.
Main Methods:
- Utilized triple antigen combinations with monoclonal antibodies (MoAbs) against CD10 (CD10-FITC and CD10-Pe/Cy5).
- Employed a long-distance polymerase chain reaction (PCR) approach to detect the bcl-2/JH rearrangement (t(14;18)(q32;q21)).
- Analyzed neoplastic cells for reactivity against CD20, CD19, CD22, SIg, CD5, CD11c, and CD10-FITC/Pe/Cy5.
Main Results:
- Neoplastic cells in FL exhibited stronger CD20 than CD19 reactivity, positivity for CD22 and SIg, and negativity for CD5, CD11c, and CD10-FITC.
- CD10-Pe/Cy5 was characteristically expressed in all samples with positive bcl-2/JH rearrangements.
- High correlation (96%) between histological diagnosis and CD10-Pe/Cy5 reactivity was observed in FL.
- CD10-Pe/Cy5 identified t(14;18)-positive cases in diffuse large cell lymphomas (DLCL) and Burkitt lymphomas.
Conclusions:
- CD10-Pe/Cy5 is a valuable antibody for identifying neoplastic cells carrying the t(14;18)(q32;q21) translocation in FL and DLCL.
- Combined with other MoAbs, anti-CD10 (HI10a, Cy-Chrome) helps define a characteristic FL immunophenotype.
- This approach improves the differential diagnosis of lymphoproliferative disorders.
Abstract:
Follicular lymphoma (FL) is a specific entity defined by characteristic histology, phenotype and molecular rearrangements. Classically, reactivity for CD19, CD10, and strong positivity for the surface light chain immunoglobulin (SIg) are considered to be phenotypic signs typically expressed in FL. In practice, this pattern is difficult to identify since most neoplastic cells analysed by flow cytometry (FC) show weak intensity for CD19-Pe/Cy5 and for SIg and negativity for CD10-FITC. We used triple antigen combinations including two monoclonal antibodies (MoAbs) against CD10 (CD10-FITC and CD10-Pe/Cy5) and a long-distance polymerase chain reaction (PCR) approach to establish the phenotypic pattern of neoplastic cells carrying t(14;18)(q32;q21). Neoplastic cells showed the following immunophenotype: stronger reactivity against CD20 than against CD19, positivity for CD22 and SIg and negativity for CD5, CD11c and CD10-FITC. Characteristically, CD10-Pe/Cy5 was expressed in all the samples with positive bcl-2/JH rearrangements. In FL, there was a high correlation between histologic diagnosis and reactivity against CD10-Pe/Cy5 (96% cases). In diffuse large cell lymphomas (DLCL), CD10-Pe/Cy5 identified positive cases with t(14;18)(q32;q21) chromosomal translocation, whereas Burkitt lymphomas showed all cases reactivity against CD10-Pe/Cy5. In conclusion, CD10-Pe/Cy5 is a useful antibody for identifying neoplastic cells carrying t(14;18)(q32;q21) in FL and DLCL. In combination with other MoAbs, anti-CD10 (HI10a, Cy-Chrome) can be used to identify a characteristic phenotypic profile of FL against other lymphoproliferative disorders.
