Selection and transplantation of autologous CD34+ B-lineage negative cells in advanced-phase multiple myeloma patients: A pilot study

Roberto M. Lemoli, Giovanni Martinelli, Attilio Olivieri, Maria Rosa Motta, Simonetta Rizzi, Carolina Terragna, Giuliana Leopardi, Monica Benni, Sonia Ronconi, Isabella Cantori, Damiano Rondelli, Serena Mangianti, Pietro Leoni, Mauro Montanari, Michele Cavo, Sante Tura

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The feasibility of sequential positive and negative selection of mobilized CD34+ B-lineage negative cells to achieve tumour-free autografts in multiple myeloma (MM) patients was evaluated. Peripheral blood stem cells (PBSC) of 14 patients with advanced disease were mobilized. CD34+ cells were enriched in 12 of the patients by the avidin-biotin immunoabsorption technique. Subsequently, CD10+, CD19+, CD20+ and CD56+ cells (B-lin cells) were removed by immunomagnetic depletion. Minimal residual disease (MRD) was detected by flow cytometry and PCR-based molecular analysis of the patient specific IgH complementary-determining region III (CDRIII). Positive selection of stem cells produced a median recovery of 54.7% of the initial content of CD34+ cells (median purity 71.9%). Negative depletion of B- lineage cells reduced the number of CD34+ cells to 33.3% of the baseline value (median purity 72.7%). However, long-term culture assays showed the recovery of >60% of primitive haemopoietic progenitor cells after depletion of the B-lineage-positive cells. All evaluable patients had detectable disease in PBSC collections. The first step of positive selection of CD34+ cells resulted in >2 logs of tumour cell purging. However, molecular assessment showed the persistence of the disease in 6/7 cases. Immunofluorescence analysis demonstrated 1 additional log of B-cell purging by negative depletion. More importantly, molecular evaluation of IgH CDRIII region showed the disappearance of myeloma cells in 6/7 patients. 12 patients received a median of 3.9 x 106 CD34+ B-lin- cells/kg after conditioning with high-dose melphalan and showed a rapid reconstitution of haemopoiesis. These results were similar to two similar cohorts of patients who received either unmanipulated PBSC or positively selected CD34+ cells after the same conditioning regimen. Severe extrahaematological toxicity was limited to mucositis; no late infections were observed. We concluded that autotransplantation of purified CD34+ B-lin- cells was associated with a rapid and sustained recovery of haemopoiesis and low peritransplant morbidity. Sequential positive and negative enrichment of stem cells reduced tumour cell contamination in B-cell malignancies below the lower limit of detection of molecular analysis.

Original languageEnglish
Pages (from-to)419-428
Number of pages10
JournalBritish Journal of Haematology
Volume107
Issue number2
DOIs
Publication statusPublished - 1999

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Autologous Transplantation
Multiple Myeloma
B-Lymphocytes
Stem Cells
Neoplasms
Mucositis
Melphalan
Neoplastic Stem Cells
Avidin
Autografts
Residual Neoplasm
Biotin
Fluorescent Antibody Technique
Limit of Detection
Flow Cytometry

Keywords

  • Autologous transplantation
  • CD34 cells
  • Multiple myeloma
  • Positive/negative selection
  • Purging

ASJC Scopus subject areas

  • Hematology

Cite this

Selection and transplantation of autologous CD34+ B-lineage negative cells in advanced-phase multiple myeloma patients : A pilot study. / Lemoli, Roberto M.; Martinelli, Giovanni; Olivieri, Attilio; Motta, Maria Rosa; Rizzi, Simonetta; Terragna, Carolina; Leopardi, Giuliana; Benni, Monica; Ronconi, Sonia; Cantori, Isabella; Rondelli, Damiano; Mangianti, Serena; Leoni, Pietro; Montanari, Mauro; Cavo, Michele; Tura, Sante.

In: British Journal of Haematology, Vol. 107, No. 2, 1999, p. 419-428.

Research output: Contribution to journalArticle

Lemoli, RM, Martinelli, G, Olivieri, A, Motta, MR, Rizzi, S, Terragna, C, Leopardi, G, Benni, M, Ronconi, S, Cantori, I, Rondelli, D, Mangianti, S, Leoni, P, Montanari, M, Cavo, M & Tura, S 1999, 'Selection and transplantation of autologous CD34+ B-lineage negative cells in advanced-phase multiple myeloma patients: A pilot study', British Journal of Haematology, vol. 107, no. 2, pp. 419-428. https://doi.org/10.1046/j.1365-2141.1999.01691.x
Lemoli, Roberto M. ; Martinelli, Giovanni ; Olivieri, Attilio ; Motta, Maria Rosa ; Rizzi, Simonetta ; Terragna, Carolina ; Leopardi, Giuliana ; Benni, Monica ; Ronconi, Sonia ; Cantori, Isabella ; Rondelli, Damiano ; Mangianti, Serena ; Leoni, Pietro ; Montanari, Mauro ; Cavo, Michele ; Tura, Sante. / Selection and transplantation of autologous CD34+ B-lineage negative cells in advanced-phase multiple myeloma patients : A pilot study. In: British Journal of Haematology. 1999 ; Vol. 107, No. 2. pp. 419-428.
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AU - Olivieri, Attilio

AU - Motta, Maria Rosa

AU - Rizzi, Simonetta

AU - Terragna, Carolina

AU - Leopardi, Giuliana

AU - Benni, Monica

AU - Ronconi, Sonia

AU - Cantori, Isabella

AU - Rondelli, Damiano

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AU - Leoni, Pietro

AU - Montanari, Mauro

AU - Cavo, Michele

AU - Tura, Sante

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N2 - The feasibility of sequential positive and negative selection of mobilized CD34+ B-lineage negative cells to achieve tumour-free autografts in multiple myeloma (MM) patients was evaluated. Peripheral blood stem cells (PBSC) of 14 patients with advanced disease were mobilized. CD34+ cells were enriched in 12 of the patients by the avidin-biotin immunoabsorption technique. Subsequently, CD10+, CD19+, CD20+ and CD56+ cells (B-lin cells) were removed by immunomagnetic depletion. Minimal residual disease (MRD) was detected by flow cytometry and PCR-based molecular analysis of the patient specific IgH complementary-determining region III (CDRIII). Positive selection of stem cells produced a median recovery of 54.7% of the initial content of CD34+ cells (median purity 71.9%). Negative depletion of B- lineage cells reduced the number of CD34+ cells to 33.3% of the baseline value (median purity 72.7%). However, long-term culture assays showed the recovery of >60% of primitive haemopoietic progenitor cells after depletion of the B-lineage-positive cells. All evaluable patients had detectable disease in PBSC collections. The first step of positive selection of CD34+ cells resulted in >2 logs of tumour cell purging. However, molecular assessment showed the persistence of the disease in 6/7 cases. Immunofluorescence analysis demonstrated 1 additional log of B-cell purging by negative depletion. More importantly, molecular evaluation of IgH CDRIII region showed the disappearance of myeloma cells in 6/7 patients. 12 patients received a median of 3.9 x 106 CD34+ B-lin- cells/kg after conditioning with high-dose melphalan and showed a rapid reconstitution of haemopoiesis. These results were similar to two similar cohorts of patients who received either unmanipulated PBSC or positively selected CD34+ cells after the same conditioning regimen. Severe extrahaematological toxicity was limited to mucositis; no late infections were observed. We concluded that autotransplantation of purified CD34+ B-lin- cells was associated with a rapid and sustained recovery of haemopoiesis and low peritransplant morbidity. Sequential positive and negative enrichment of stem cells reduced tumour cell contamination in B-cell malignancies below the lower limit of detection of molecular analysis.

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