A validated real-time quantitative PCR approach shows a correlation between tumor burden and successful ex vivo purging in follicular lymphoma patients

M. Ladetto, S. Sametti, J. W. Donovan, D. Ferrero, M. Astolfi, M. Mitterer, I. Ricca, D. Drandi, P. Corradini, P. Coser, A. Pileri, J. G. Gribben, C. Tarella

Research output: Contribution to journalArticle

Abstract

Objective - Purging procedures are increasingly used to provide stem cell collections devoid of contaminating tumor cells. In follicle center lymphoma (FCL), most approaches eradicate polymerase chain reaction (PCR);-detectable disease in only a fraction of harvests undergoing ex vivo manipulation. In this study we evaluated whether there is a relationship between tumor burden of stem cell harvests and successful clearance of PCR-detectable disease following ex vivo manipulation. Materials and Methods - To address this issue, we developed a real-time PCR approach for quantitative measurement of tumor contamination using the bcl-2 rearrangement. Real-time PCR was used to evaluate the relationship between tumor burden of stem-cell harvests and purging effectiveness in PCR+ samples derived from 10 FCL patients. Ex vivo purging was performed using the MaxSep cell separator (Baxter Immunotherapy, Deerfield, IL, USA). Results - Our real-time PCR method proved effective, sensitive, accurate, and reproducible. Four collections were successfully cleared of minimal residual disease (MRD) whereas six remained PCR+. Real-time PCR showed that the four collections successfully cleared of MRD had a prepurging tumor burden significantly lower than those remaining PCR+ (p = 0.04). Conclusion - This study provides the first evidence that evaluation of tumor burden in stem-cell harvests by real-time PCR can predict the effectiveness of therapeutic intervention in non-Hodgkin's lymphoma. Based on these findings, we foresee a more widespread use of this technique to evaluate the impact of different therapeutic approaches in FCL.

Original languageEnglish
Pages (from-to)183-193
Number of pages11
JournalExperimental Hematology
Volume29
Issue number2
DOIs
Publication statusPublished - 2001

Keywords

  • bcl-2 rearrangement
  • Ex vivo purging
  • Follicle center lymphoma
  • Minimal residual disease
  • Real-time PCR

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

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    Ladetto, M., Sametti, S., Donovan, J. W., Ferrero, D., Astolfi, M., Mitterer, M., Ricca, I., Drandi, D., Corradini, P., Coser, P., Pileri, A., Gribben, J. G., & Tarella, C. (2001). A validated real-time quantitative PCR approach shows a correlation between tumor burden and successful ex vivo purging in follicular lymphoma patients. Experimental Hematology, 29(2), 183-193. https://doi.org/10.1016/S0301-472X(00)00651-2