A preliminary study towards downregulation of murine bone marrow eosinophilopoiesis mediated by small molecule inhibition of interleukin-5 receptor alpha gene in vitro.
BACKGROUND
Bone marrow eosinophilopoiesis induced by IL-5 makes a major contribution to eosinophilic airway inflammation in asthma. Bone marrow CD(34)(+) cells expressing IL-5Ralpha may be eosinophil progenitors. However, research on the effect of blocking IL-5Ralpha expression on bone marrow eosinophilopoiesis has seldom been reported.


OBJECTIVE
To explore the effect of inhibiting IL-5Ralpha expression with IL-5Ralpha short hairpin RNA-expressing vector on murine bone marrow eosinophilopoiesisin vitro.


METHODS
We constructed 4 kinds of plasmid vectors that could express small molecule inhibition, short hairpin RNA, which targeted IL-5Ralpha (P-IL-5Ralpha), and selected an effective one by transfecting B lymphoma cells in vitro. We also constructed an adenovirus vector which was inserted into an effective template sequence (Ad-IL-5Ralpha). The bone marrow cells were obtained from healthy Balb/c mice, and cultured and transfected by Ad-IL-5Ralpha in vitro. The expression of IL-5Ralpha and the count of newly produced eosinophils were detected in the cultured bone marrow cells.


RESULTS
We found that P-IL-5Ralpha-3 targeted at the sequence of CAG CTG CCT GGT TCG TCT T markedly suppressed the IL-5Ralpha expression in the B lymphoma cellsin vitro. Ad-IL-5Ralpha could suppress the IL-5Ralpha expression of murine bone marrow cellsin vitro and it could also significantly decrease the IL-5-induced eosinophilia in the cultured bone marrow cells.


CONCLUSIONS
These results indicate that the blocking of IL-5Ralpha expression by small molecule inhibition can help to effectively decrease murine bone marrow eosinophilopoiesis, and that bone marrow may be used as a critical target organ in the diseases involved in eosinophilia, such as asthma.
Mao
Hui
H
Department of Respiratory Medicine, West China Hospital of Sichuan University, Chengdu, PR China. lfy_74@vip.163.com
Wen
Fu-Qiang
FQ
Li
Su-Yun
SY
Liang
Zong-An
ZA
Liu
Chun-Tao
CT
Yin
Kai-Sheng
KS
Wang
Zeng-Li
ZL
D000818:Animals; D001249:Asthma / Q000276:immunology; D001854:Bone Marrow Cells / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D015536:Down-Regulation; D004804:Eosinophils / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D005822:Genetic Vectors; D007249:Inflammation / Q000378:metabolism / Q000628:therapy; D053666:Interleukin-5 Receptor alpha Subunit / Q000037:antagonists & inhibitors* / Q000235:genetics; D019891:Leukopoiesis / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D014162:Transfection / Q000379:methods
D053666:Interleukin-5 Receptor alpha Subunit; D012333:RNA, Messenger