Expanded clinical and experimental use of SOX11 - using a monoclonal antibody.
BACKGROUND
The transcription factor SOX11 is of diagnostic and prognostic importance in mantle cell lymphoma (MCL) and epithelial ovarian cancer (EOC), respectively. Thus, there is an unmet clinical and experimental need for SOX11-targeting assays with low background, high specificity and robust performance in multiple applications, including immunohistochemistry (IHC-P) and flow cytometry, which until now has been lacking.


METHODS
We have developed SOX11-C1, a monoclonal mouse antibody targeting SOX11, and successfully evaluated its performance in western blots (WB), IHC-P, fluorescence microscopy and flow cytometry.


RESULTS
We confirm the importance of SOX11 as a diagnostic antigen in MCL as 100% of tissue micro array (TMA) cases show bright nuclear staining, using the SOX11-C1 antibody in IHC-P. We also show that previous reports of weak SOX11 immunostaining in a fraction of hairy cell leukemias (HCL) are not confirmed using SOX11-C1, which is consistent with the lack of transcription. Thus, high sensitivity and improved specificity are demonstrated using the monoclonal SOX11-C1 antibody. Furthermore, we show for the first time that flow cytometry can be used to separate SOX11 positive and negative cell lines and primary tumors. Of note, SOX11-C1 shows no nonspecific binding to primary B or T cells in blood and thus, can be used for analysis of B and T cell lymphomas from complex clinical samples. Dilution experiments showed that low frequencies of malignant cells (~1%) are detectable above background using SOX11 as a discriminant antigen in flow cytometry.


CONCLUSIONS
The novel monoclonal SOX11-specific antibody offers high sensitivity and improved specificity in IHC-P based detection of MCL and its expanded use in flow cytometry analysis of blood and tissue samples may allow a convenient approach to early diagnosis and follow-up of MCL patients.
Department of Immunotechnology, Lund University, BMCD13, Lund, SE-221 84, Sweden.
Lena
L
Nordström
Ulrika
U
Andréasson
Mats
M
Jerkeman
Michael
M
Dictor
Carl
C
Borrebaeck
Sara
S
Ek
D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry* / Q000276:immunology; D015153:Blotting, Western / Q000379:methods; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007943:Leukemia, Hairy Cell / Q000175:diagnosis / Q000378:metabolism; D020522:Lymphoma, Mantle-Cell / Q000175:diagnosis / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D008856:Microscopy, Fluorescence / Q000379:methods; D015203:Reproducibility of Results; D055751:SOXC Transcription Factors / Q000032:analysis* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity; D014183:Transplantation, Heterologous
D058846:Antibodies, Monoclonal, Murine-Derived; D055751:SOXC Transcription Factors