Nerve growth factor prevents apoptosis of cord blood-derived human cultured mast cells synergistically with stem cell factor.
BACKGROUND
Stem cell factor (SCF) has been identified as a critical survival factor of human mast cells. Other cytokines which possess survival promotion activity on human mast cells are less known.


OBJECTIVE
We examined the survival promotion activity of nerve growth factor (NGF) on cord blood-derived human cultured mast cells.


METHODS
Expression and function of NGF receptors on the mast cells were examined by RT PCR, flowcytometric analysis, immunoprecipitaion and western blotting. The survival promotion activity of NGF to the mast cells was examined. To evaluate the proliferating activity of NGF on the human cultured mast cells, flow cytometric analysis with propidium iodide staining was applied. To confirm whether the human mast cell growth activity of NGF was caused by a suppression of apoptosis, the proportion of the cells containing in situ DNA fragmentation was counted.


RESULTS
The human cultured mast cells expressed the high affinity receptor p140trk but not the low affinity receptor p75LNGFR. NGF induced the phosphorylation of p140trk. NGF alone could not support the survival of the mast cells, however, the addition of NGF to the culture medium containing recombinant SCF led to a significant increase of the number of survival mast cells. No significant changes of the cell cycle from G0/G1 phase to the S/G2 + M phases were observed by NGF. In contrast, the addition of NGF to the medium with SCF showed a significant inhibitory effect on the apoptosis of the mast cells.


CONCLUSIONS
NGF may act as a key factor to promote the survival of human mast cells synergistically with SCF through the prevention of apoptosis.
Kanbe
N
N
Department of Dermatology; Second Department of Pathology, Gunma University School of Medicine, Maebashi, Japan.
Kurosawa
M
M
Miyachi
Y
Y
Kanbe
M
M
Saitoh
H
H
Matsuda
H
H
D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004357:Drug Synergism; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D008407:Mast Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D020932:Nerve Growth Factor / Q000494:pharmacology*; D017475:Receptors, Nerve Growth Factor / Q000032:analysis / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D017475:Receptors, Nerve Growth Factor; D019089:Stem Cell Factor; D020932:Nerve Growth Factor