T-lymphocyte function from peripheral blood stem-cell donors is inhibited by activated granulocytes.
BACKGROUND
PBSC transplant provides 10 times more T cells than BMT However, the incidence and severity of acute GvHD is similar among recipients of both types of transplants. Studies in mouse models suggest that the similar clinical outcome in BMT and PBSCT is due to differences in the lymphokine profiles.


METHODS
PBMC, PBMC from G-CSF mobilized donors (G-PBMC)and BM mononuclear cells (BM-MC) were analyzed by flow cytometry and ELISA to detect gamma-IFN and IL-4 production. Hematoxylin and eosin staining was used to identify morphology and annexin/propidium-iodide was used for apoptosis assays.


RESULTS
We show decreased production of gamma-interferon (85%) and IL-4 (60%) in G-PBMC when compared with either PBMC or BM-MCT cells on ex vivo assays. Surprisingly, 85% of fresh G-PBMC is composed of low-density granulocytes (LDG), which undergo apoptosis after 48 h in culture. At this same time, gamma-IFN production from G-PBMC T cell was reverted. In vitro, G-CSF converts granulocytes into LDGs, able to inhibit T-cell function by H2O2 production, and not through immune-deviation towards a Th2-type phenotype.


CONCLUSIONS
We show that the estimated numbers of Th1 and Th2 cells infused in BMT and PBSCT do not differ significantly. These findings are discussed with reference to the relatively low incidence of acute GvHD in PBSCT shown in the literature. We suggest that these results might depend on the high number of granulocytes and progenitors infused. The potential use of granulocytes as immunosupressive short-term therapy is now being investigated by our group using a mouse experimental model.
Vasconcelos
Z F M
ZF
Centro Nacional de Transplante de Medula Osea, Instituto Nacional do Câncer, Rio de Janeiro, Brazil.
Santos
B M
BM
Costa
E S
ES
Lima
M
M
Tabak
D G
DG
Bouzas
L F
LF
Azevedo
W M
WM
Barcinski
M A
MA
Bonomo
A
A
D017304:Annexin A5 / Q000032:analysis; D015703:Antigens, CD / Q000032:analysis; D017209:Apoptosis / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D016026:Bone Marrow Transplantation; D017252:CD3 Complex / Q000032:analysis; D002374:Catalase / Q000494:pharmacology; D002452:Cell Count; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006098:Granulocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007371:Interferon-gamma / Q000032:analysis; D015847:Interleukin-4 / Q000032:analysis; D015759:Ionomycin / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D051917:Leukosialin; D007959:Lymphocyte Culture Test, Mixed; D018375:Neutrophil Activation / Q000187:drug effects / Q000502:physiology; D036102:Peripheral Blood Stem Cell Transplantation; D012795:Sialoglycoproteins / Q000032:analysis; D013601:T-Lymphocytes / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000031:analogs & derivatives* / Q000494:pharmacology; D013997:Time Factors
D017304:Annexin A5; D015703:Antigens, CD; D017252:CD3 Complex; D051917:Leukosialin; C465014:SPN protein, human; D012795:Sialoglycoproteins; D016179:Granulocyte Colony-Stimulating Factor; D015847:Interleukin-4; D015759:Ionomycin; C016658:phorbolol myristate acetate; D007371:Interferon-gamma; D006861:Hydrogen Peroxide; D002374:Catalase; D013755:Tetradecanoylphorbol Acetate