In vivo induction of T-cell hyporesponsiveness and alteration of immunological cells of bone marrow grafts using granulocyte colony-stimulating factor.
OBJECTIVE
Granulocyte colony-stimulating factor (G-CSF)-primed bone marrow (G-BM) transplantation is associated with a low incidence of graft-versus-host disease (GVHD) and has been used successfully in patients with human leukocyte antigen (HLA) matched/mismatched donors. This study evaluated the function of T cells and the quantities of immunological cells of G-BM.


METHODS
Bone marrow was obtained from fifteen donors by aspiration. 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 cell (DC) subsets, CD4(+)CD25(+) regulatory T cells and the expression of CD28/CD80/CD86 molecules on monocytes, B and T cells were analyzed using flow cytometry.


RESULTS
G-CSF treatment decreased the quantities of IFN-gamma secretion dramatically (p=0.007) and IL-4 moderately (p=0.027), leading to higher ratios of IL-4/IFN-gamma (p=0.004). We confirmed T-cell hyporesponsiveness and lower expression of CD28/CD80/CD86 on monocytes, B and T cells. The absolute values of lymphocytes, T cell subgroups, CD3+CD4-CD8- cells, CD8+CD28- cells and B cells in bone marrow grafts were similar before and after G-CSF treatment. The number of monocytes per microliter was increased 2.13-fold, while the numbers of CD4(+)CD25(+) regulatory T cells were unchanged. DC2 were preferentially increased.


CONCLUSIONS
Our results suggest that bone marrow T-cell hyporesponsiveness could be induced and that the increase of monocytes and DC2 and the downregulation of CD28/CD80/CD86 co-stimulatory signals were produced by in vivo use of G-CSF; this may be related to the preferential increase of monocytes and DC2 and the downregulation of CD28/CD80/CD86 co-stimulatory signals.
Jun
Huang Xiao
HX
Peking University, Institute of Hematology, People's Hospital, 42 Bei-Li-Shi-Lu Beijing 100044, PR of China. xjhrm@medmail.com.cn
Jun
Chang Ying
CY
Yu
Zhao Xiang
ZX
D000293:Adolescent; D000328:Adult; D000368:Aged; D015703:Antigens, CD / Q000032:analysis; D018122:B7-1 Antigen / Q000032:analysis; D051940:B7-2 Antigen / Q000032:analysis; D016026:Bone Marrow Transplantation / Q000009:adverse effects / Q000276:immunology*; D018106:CD28 Antigens / Q000032:analysis; D002452:Cell Count; D003713:Dendritic Cells / Q000145:classification / Q000187:drug effects; D005260:Female; D000069585:Filgrastim; D006086:Graft vs Host Disease / Q000517:prevention & control; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D006801:Humans; D007111:Immunity, Cellular / Q000187:drug effects*; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D008213:Lymphocyte Activation; D016131:Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000187:drug effects; D036102:Peripheral Blood Stem Cell Transplantation; D011994:Recombinant Proteins; D016176:T-Lymphocyte Subsets / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D018417:Th1 Cells / Q000187:drug effects / Q000276:immunology; D018418:Th2 Cells / Q000187:drug effects / Q000276:immunology; D014184:Transplantation, Homologous / Q000009:adverse effects / Q000276:immunology*
D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D018106:CD28 Antigens; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D015847:Interleukin-4; D007371:Interferon-gamma; D000069585:Filgrastim