TY - JOUR
T1 - Using oligonucleotide aptamer probes for immunostaining of formalin-fixed and paraffin-embedded tissues
AU - Zeng, Zihua
AU - Zhang, Peng
AU - Zhao, Nianxi
AU - Sheehan, Andrea M.
AU - Tung, Ching Hsuan
AU - Chang, Chung Che
AU - Zu, Youli
N1 - Funding Information:
This study was supported in part by a clinical translation study award from the Methodist Hospital Research Institute (CTSA) and grants from the National Cancer Institutes (K22 CA113493, R43 CA148735, and developmental research project award of P50 CA126752).
PY - 2010/12
Y1 - 2010/12
N2 - For tissue immunostaining, antibodies are currently the only clinically validated and commercially available probes. Aptamers, which belong to a class of small molecule ligands composed of short single-stranded oligonucleotides, have emerged as probes over the last several decades; however, their potential clinical value has not yet been fully explored. Using cultured cells and an RNA-based CD30 aptamer, we recently demonstrated that the synthetic aptamer is useful as a specific probe for flow cytometric detection of CD30-expressing lymphoma cells. In this study, we further validated the use of this aptamer probe for immunostaining of formalin-fixed and paraffin-embedded lymphoma tissues. Using CD30 antibody as a standard control, we demonstrated that the synthetic CD30 aptamer specifically recognized and immunostained tumor cells of classical Hodgkin lymphoma and anaplastic large cell lymphoma, but did not react with background cells within tumor sites. Notably, the CD30 aptamer probe optimally immunostained lymphoma cells with lower temperature antigen retrieval (37 vs 96°C for antibody) and shorter probing reaction times (20 vs 90 min for antibody) than typical antibody immunostaining protocols. In addition, the CD30 aptamer probe showed no nonspecific background staining of cell debris in necrotic tissue and exhibited no cross-reaction to tissues that do not express CD30, as confirmed by a standard CD30 antibody staining. Therefore, our findings indicate that the synthetic oligonucleotide CD30 aptamer can be used as a probe for immunostaining of fixed tissue sections for disease diagnosis.
AB - For tissue immunostaining, antibodies are currently the only clinically validated and commercially available probes. Aptamers, which belong to a class of small molecule ligands composed of short single-stranded oligonucleotides, have emerged as probes over the last several decades; however, their potential clinical value has not yet been fully explored. Using cultured cells and an RNA-based CD30 aptamer, we recently demonstrated that the synthetic aptamer is useful as a specific probe for flow cytometric detection of CD30-expressing lymphoma cells. In this study, we further validated the use of this aptamer probe for immunostaining of formalin-fixed and paraffin-embedded lymphoma tissues. Using CD30 antibody as a standard control, we demonstrated that the synthetic CD30 aptamer specifically recognized and immunostained tumor cells of classical Hodgkin lymphoma and anaplastic large cell lymphoma, but did not react with background cells within tumor sites. Notably, the CD30 aptamer probe optimally immunostained lymphoma cells with lower temperature antigen retrieval (37 vs 96°C for antibody) and shorter probing reaction times (20 vs 90 min for antibody) than typical antibody immunostaining protocols. In addition, the CD30 aptamer probe showed no nonspecific background staining of cell debris in necrotic tissue and exhibited no cross-reaction to tissues that do not express CD30, as confirmed by a standard CD30 antibody staining. Therefore, our findings indicate that the synthetic oligonucleotide CD30 aptamer can be used as a probe for immunostaining of fixed tissue sections for disease diagnosis.
KW - anaplastic large cell lymphoma
KW - CD30 expression
KW - classical Hodgkin lymphoma
KW - immunohistochemistry
KW - immunostaining
KW - oligonucleotide aptamer probe
KW - paraffin-embedded tissue staining
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U2 - 10.1038/modpathol.2010.151
DO - 10.1038/modpathol.2010.151
M3 - Article
C2 - 20693984
AN - SCOPUS:78649703580
SN - 0893-3952
VL - 23
SP - 1553
EP - 1558
JO - Modern Pathology
JF - Modern Pathology
IS - 12
ER -