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Competent dendritic cells derived from CD34 + progenitors express CMRF‐44 antigen early in the differentiation pathway
Author(s) -
Monji Tatsue,
Petersons Jerilyn Tynes,
Saund Narinder K,
Vuckovic Slavica,
Hart Derek Nigel John,
AuditoreHargreaves Karen,
Risdon Grant
Publication year - 2002
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1046/j.1440-1711.2002.01075.x
Subject(s) - cd34 , cd40 , leukapheresis , progenitor cell , antigen , haematopoiesis , microbiology and biotechnology , cd14 , stem cell , immunology , flow cytometry , biology , chemistry , in vitro , cytotoxic t cell , biochemistry
Differentiation of CD34 + haematopoietic stem cells into functional dendritic cells (DC) was investigated using the mAb CMRF‐44 and other mAb against DC‐associated markers. GM‐CSF mobilized peripheral blood stem cells were obtained from healthy donors by leukapheresis. CD34 + cells were purified using CD34 + ‐positive selection, and subsequent immunomagnetic depletion of CD14 and CD2 cells. CD34 + cells were cultured in medium supplemented with one or more of GM‐CSF, TNF‐α, IL‐4 or IL‐6. CMRF‐44 Ag expression was monitored by flow cytometry, and DC function by allogeneic MLR and tetanus toxoid (TT) presentation assays. CD34 + cells quickly acquired the CMRF‐44 Ag when cultured in the presence of TNF‐α. By day 3, more than 50% of the cells were double‐positive for CD34 and CMRF‐44. CD34 expression was gradually lost, so that by day 9, the majority of the cells were CD34 ‐ /CMRF‐44 + . GM‐CSF and TNF‐α also induced CD40 expression, and up‐regulation of CD54 and MHC class II on CD34 + cells; their expression was correlated to the CMRF‐44 Ag. Day 3 CD34 + /CMRF‐44 + cells, but not CD34 + /CMRF‐44 ‐ cells, become potent APC when cultured further with GM‐CSF plus TNF‐α. These CMRF‐44 + cells were potent inducers of Th1‐type immune response in the primary allogeneic MLR and present TT to autologous CD4 + T cells. TNF‐α alone is sufficient to induce CMRF‐44 expression on CD34 + cells, but in combination with GM‐CSF expands the CMRF‐44 + population. CMRF‐44 expression correlates with DC function and may be a useful early marker for commitment of CD34 + cells to the DC differentiation pathway.