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Functional Differences between Subpopulations of Mobilized Peripheral Blood-Derived CD34⁺ Cells Expressing Different Levels of HLA-DR, CD33, CD38 and c-kit Antigens

^a Second Department of Internal Medicine, Shiga University of Medical Science,
^b Department of Pediatrics,
^c Second Department of Surgery,
^d Department of Hygiene, Kyoto Prefectural University of Medicine, Kyoto, Japan; Department of Biology, Faculty of Science, Niigata University, Niigata, Japan

Key Words. PBSCT • G-CSF • CD34⁺ • CD33 • CD38 • c-kit • LTC-IC

Dr. Yoshiaki Sonoda, Department of Hygiene, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyoku, Kyoto 602, Japan.

We have investigated the functional characteristics of peripheral blood-derived CD34⁺ cells mobilized by a combination of chemotherapy and G-CSF (mobilized peripheral blood-derived [MPB] CD34⁺ cells). In this study, subpopulations of MPB CD34⁺ cells have been directly compared in clonal cultures, long-term cultures with bone marrow (BM) stromal cells, and single-cell cultures. MPB CD34⁺ cells could be subdivided by expression levels of HLA-DR (DR), CD38, CD33 and c-kit antigens. The majority of MPB CD34⁺ cells expressed DR and CD38 antigens. In contrast, approximately 60% and 20% of the MPB CD34⁺ cells expressed CD33 and c-kit antigens, respectively. Interestingly, MPB CD34⁺ cells can be subdivided into three fractions which express high, low or negative levels of c-kit receptor. All types of committed progenitors were observed in populations of CD34⁺DR⁺, CD34⁺DR^–, CD34⁺CD33^–, CD34⁺CD38⁺ and CD34⁺ c-kit^low cells. Colony forming unit-granulocyte/macrophage was highly enriched in the population of CD34⁺CD33⁺ cells, whereas BFU-E was highly enriched in the population of CD34⁺ c-kit^high cells. In the population of CD34⁺CD38^– cells, however, a few myeloid progenitors were detected. In addition, limiting dilution analyses clearly showed that the long-term culture-initiating cell (LTC-IC) is enriched in the populations of CD34⁺DR^–, CD34⁺CD33^– and CD34⁺c-kit^{– or low} cells, but very few in CD34⁺ c-kit^high cells, and that CD38 antigen is not a useful marker for the enrichment of LTC-IC derived from MPB CD34⁺ cells. Moreover, single cell clone sorting experiments clearly demonstrated the functional differences between CD34⁺CD38⁺ and CD34⁺CD38^– cells as well as CD34⁺ cells expressing different levels of c-kit receptor. Our results suggest that an immunophenotype of LTC-IC is different between BM-, cord blood- and MPB-derived CD34⁺ cells and that primitive and committed progenitors existing in these sources may be functionally different.

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