Ex vivo expansion of long-term culture initiating marrow cells by IL-10, SCF, and IL-3.

BACKGROUND: Ex vivo expansion of progentior cells may shorten hematopoietic regeneration after myeloablative chemoradiotherapy, increase target cells for gene therapy, and improve purging of progenitor cell components. STUDY DESIGN AND METHODS: Marrow cells were incubated for 1 week in suspension culture with and without IL-10, IL-3, and SCF. As long-term culture initiating cells (LTC-ICs) represent early hematopoietic progenitors in vitro, these cells were quantified at initiation and after a 1-week culture period in a limiting dilution assays. Additionally, immunophenotyping of cells before and after culture was performed. RESULTS: In six experiments, marrow cells cultured for 1 week with IL-10, IL-3, and SCF showed a significant increase (almost doubling) in LTC-ICs as compared with marrow cells before expansion. Additionally, an increased proliferative capacity of LTC-ICs was achieved with a sevenfold increase of committed colony-forming cells and a 10-fold proliferation of high proliferative potential colony-forming cells. Immunophenotyping revealed a sevenfold increase of CD34+ CD45 RA- cells in IL-10-, IL-3-, SCF-stimulated suspension cultures. In unstimulated cultures, no LTC-ICs were maintained after 1 week. CONCLUSION: Expansion of LTC-ICs by IL-10, IL-3, and SCF has not been shown so far. This in vitro model allows expansion of LTC-IC if compared with the input of progenitor cells without extensive progenitor cell manipulation. This should be an attractive model for in vitro purging, gene transfer, or expansion of progenitor cells to allow rapid engraftment after myeloablative chemotherapy.

The role of soluble CD23 in distinguishing stable and progressive forms of B-chronic lymphocytic leukemia.

Soluble CD23 (sCD23) has been recognized as an important prognostic parameter in patients with chronic lymphocytic leukemia (B-CLL) at early clinical stages. There is, however, no clear information on its prognostic significance in advanced stages and on its role as an indicator for aggressive or indolent courses of disease. Therefore, sCD23 was measured in the serum of 145 patients at diagnosis and serial determinations were carried out for 8 years in 38 patients. The results indicate that in patients with identical clinical stages at first presentation the disease could take different courses depending on initial sCD23 concentrations below or above specific threshold levels (860 and 5900U/ml). sCD23 higher than these thresholds was associated with faster progression into upper clinical stages. Furthermore, sCD23-doubling time (sCD23-DT) indicated that patients with long DT progressed slowly, while those with short DT had more aggressive disease. Particularly in patients with advanced disease stages, long sCD23-DT indicated development of smoldering disease. Since sCD23 levels reflect total tumor mass, determination of sCD23-DT has probably a better predictive value than lymphocyte doubling time. It appears that B-CLL patients can be divided into different risk categories according to initial determinations of sCD23 and that sCD23-DT is an additional important parameter in predicting disease progression.