PMID- 16836847
OWN - NLM
STAT- MEDLINE
DCOM- 20100726
LR  - 20181201
IS  - 0578-1310 (Print)
IS  - 0578-1310 (Linking)
VI  - 44
IP  - 6
DP  - 2006 Jun
TI  - [Bifidobacterium DNA upregulates Th1 type response of umbilical cord blood 
      mononuclear cell].
PG  - 415-9
AB  - OBJECTIVE: To study the effect of bifidobacterium genomic DNA on umbilical cord 
      blood mononuclear cell (CBMC), and investigate the immunoregulation of 
      bifidobacterium DNA and explore possible mechanisms by which bifidobacterium acts 
      against allergic reaction. METHODS: Bifidobacterium genomic DNA (bDNA) and human 
      DNA (hDNA) were extracted with phenol/chloroform/isoamyl alcohol and stored at 
      -20 degrees C for later use. Parts of bDNA were completely digested with DNaseI 
      (d-bDNA) at 37 degrees C. CBMCs were isolated with Ficoll from umbilical cord 
      blood and incubated at 37 degrees C in a 5% CO2 humidified incubator. These cells 
      were divided into four groups, control group: without any stimulant; bDNA group: 
      stimulated with 25 microg/ml bDNA; d-bDNA group: stimulated with 25 microg/ml 
      d-bDNA; hDNA group: stimulated with 25 microg/ml hDNA. The cells were stimulated 
      with different stimulants in vitro, at the end of incubation culture supernatant 
      and cells were collected. IL-12 and IL-10 levels in the culture supernatant were 
      measured by enzyme linked immuno sorbent assay (ELISA); cells secreting IL-4 and 
      IFN-gamma were counted by enzyme linked immunospot (ELISPOT) assay; and total RNA 
      was isolated from the cells to assay T-bet and GATA3 mRNA expression levels by 
      reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Six hours 
      after stimulation there was no significant difference in IL-12 level in 
      supernatant among the four groups; 12 hours after stimulation, IL-12 level in 
      supernatant of bDNA treated group was significantly higher than that of each of 
      the other groups, so were the results obtained at 24 hours and 48 hours after 
      stimulation (P < 0.05). No significant difference could be detected in IL-12 
      level in supernatant among the other 3 groups. On the other hand, 6 hours after 
      stimulation there was no significant difference in IL-10 level in supernatant 
      among the four groups. But 12 and 24 hours after stimulation IL-10 level in 
      supernatant of bDNA treated group was lower than that of each of the other 
      groups, but the difference was not statistically significant. The count of 
      IFN-gamma secreting cells of bDNA treated group was higher than that of the other 
      groups, while IL-4 secteting cells of bDNA treated group were lower than that of 
      the other groups. After bDNA stimulation, nuclear factor T-box expressed in T 
      cells (T-bet) mRNA expression was conspicuously enhanced as compared to the other 
      three groups (P < 0.05). GATA3 mRNA transcription in CBMC had no significant 
      change after bDNA stimulation. CONCLUSION: bDNA could promote secretion of Th1 
      type cytokine IL-12, while Th2 type cytokine IL-10 level of cell supernatant was 
      decreased. bDNA could stimulate secretion of IFN-gamma by CBMC and inhibit 
      secretion of IL-4. T-bet mRNA expression was highly enhanced after bDNA 
      stimulation. bDNA could upregulate Th1 type response, which may be one of 
      important mechanisms by which bifidobacterium inhibit allergic response.
FAU - Zhao, Hui
AU  - Zhao H
AD  - Department of Immunology, Children's Hospital, Fudan University, Shanghai 200032, 
      China.
FAU - Wang, Xiao-chuan
AU  - Wang XC
FAU - Wang, Jing-yi
AU  - Wang JY
FAU - Yu, Ye-heng
AU  - Yu YH
FAU - Wang, Chuan-qing
AU  - Wang CQ
FAU - Yang, Yi
AU  - Yang Y
LA  - chi
PT  - Comparative Study
PT  - Journal Article
PL  - China
TA  - Zhonghua Er Ke Za Zhi
JT  - Zhonghua er ke za zhi = Chinese journal of pediatrics
JID - 0417427
RN  - 0 (DNA, Bacterial)
RN  - 0 (GATA3 Transcription Factor)
RN  - 0 (RNA, Messenger)
RN  - 0 (T-Box Domain Proteins)
RN  - 0 (T-box transcription factor TBX21)
RN  - 130068-27-8 (Interleukin-10)
RN  - 187348-17-0 (Interleukin-12)
RN  - 207137-56-2 (Interleukin-4)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Bifidobacterium/cytology/*genetics
MH  - Cell Culture Techniques
MH  - DNA, Bacterial/biosynthesis/*metabolism/pharmacology
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Fetal Blood/*cytology/immunology
MH  - GATA3 Transcription Factor/genetics
MH  - Humans
MH  - Infant, Newborn
MH  - Interferon-gamma/immunology/metabolism
MH  - Interleukin-10/immunology/metabolism
MH  - Interleukin-12/immunology/metabolism
MH  - Interleukin-4/immunology/metabolism
MH  - Leukocytes, Mononuclear/*immunology/metabolism
MH  - RNA, Messenger/isolation & purification
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - T-Box Domain Proteins/genetics
MH  - Th1 Cells/drug effects/*immunology/*metabolism
EDAT- 2006/07/14 09:00
MHDA- 2010/07/27 06:00
CRDT- 2006/07/14 09:00
PHST- 2006/07/14 09:00 [pubmed]
PHST- 2010/07/27 06:00 [medline]
PHST- 2006/07/14 09:00 [entrez]
PST - ppublish
SO  - Zhonghua Er Ke Za Zhi. 2006 Jun;44(6):415-9.