Maintaining hyporesponsiveness and polarization potential of T cells after in vitro mixture of G-CSF mobilized peripheral blood grafts and G-CSF primed bone marrow grafts in different proportions.
OBJECTIVE
Granulocyte colony-stimulating factor (G-CSF) primed bone marrow grafts (G-BM) plus G-CSF mobilized peripheral blood grafts (G-PB) were used successfully in haploidentical transplantation and the incidence of graft-versus-host disease (GVHD) was not higher compared with that in patients with HLA-matched donors. The immunological characteristics of T cells in G-BM and G-PB mixed in vitro in different proportions were investigated.


METHODS
Lymphocyte proliferation ability, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) secreted by T cells were determined using a monotetrazolium (MTT) assay and sandwich enzyme-linked immunosorbent assay (ELISA), respectively. T cell subgroups, dendritic cells (DC) subsets, and monocytes were analyzed using flow cytometry in steady-state bone marrow (SS-BM), G-PB, and G-BM and calculated in mixture grafts.


RESULTS
The quantities of IFN-gamma and IL-4 secreted by lymphocytes per microliter in the three mixture grafts were 2- to 4-fold lower than in G-PB and 1- to 3-fold higher than in SS-BM and G-BM, while the IL-4/IFN-gamma ratio was higher than SS-BM and G-PB and lower than G-BM, although no significant difference was confirmed. Lymphocyte proliferation ability in the three mixture grafts was comparable to G-BM and significantly lower than SS-BM and G-PB. Lymphocytes, monocytes, T cell subsets, and DC subsets were 2- to 8-fold lower than in G-PB and 2- to 22-fold higher than in SS-BM and G-BM. The DC1/DC2 ratio was significantly higher in SS-BM than G-PB, G-BM, and the three mixture grafts (P<0.05).


CONCLUSIONS
Our results suggest that T cell hyporesponsiveness and polarization of T cell from Th1 to Th2 could be maintained after in vitro mixture of G-PB and G-BM in different proportions.
Huang
Xiao-Jun
XJ
Peking University Institute of Hematology, People's Hospital, 42 Bei-Li-Shi-Lu Beijing, 100044, China. xjhrm@medmail.com.cn
Chang
Ying Jun
YJ
Zhao
Xiang Yu
XY
D016026:Bone Marrow Transplantation / Q000276:immunology*; D049109:Cell Proliferation; D003713:Dendritic Cells / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000096:biosynthesis; D015847:Interleukin-4 / Q000096:biosynthesis; D008213:Lymphocyte Activation / Q000276:immunology; D009000:Monocytes; D036102:Peripheral Blood Stem Cell Transplantation; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology*; D018417:Th1 Cells / Q000187:drug effects / Q000276:immunology; D018418:Th2 Cells / Q000187:drug effects / Q000276:immunology
D016179:Granulocyte Colony-Stimulating Factor; D015847:Interleukin-4; D007371:Interferon-gamma