2014 Fiscal Year Annual Research Report
Project/Area Number |
14J03239
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Research Institution | Kyoto University |
Principal Investigator |
趙 艶春 京都大学, 医学研究科, 特別研究員(DC2)
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Project Period (FY) |
2014-04-25 – 2016-03-31
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Keywords | Blood-thymus barrier / Endothelial cells / Claudin-5 |
Outline of Annual Research Achievements |
(1) Examine the T-cell number and population type in Cld5 KO mice Since Cld5 KO mice die quickly after birth, I examined the newborn Cld5 KO and control wild type fetuses. I expected to find an obvious change in thymocytes populations after BTB disruption in the Cld5 KO mice, however the T-cell number and T cell population in Cld5 KO mice thymus are similar to those in control mice. This result suggested that at least in normal physiological status, Cld5 KO fetuses might in relatively stable condition even without BTB. (2) Examine the affects of BTB disruption on T-cell selection using OT-II-TCR Cld5 KO mice I generated OT-II-TCR Cld5 KO fetuses by mating OT-II-TCR Cld5+/- mice. Ova-peptide (323-339) was intravenously injected into pregnant mice. However, the result showed that ova-peptide (323-339) did not reach to the fetus thymus. So I changed to use ova-peptide (257-264), which has smaller size, and OT-I-TCR Cld5 KO. I found that ova-peptide (257-264) could reach to OT-I-TCR Cld5 KO fetuses after intravenous injection to OT-I-TCR Cld5+/- pregnant mice. Since OT-I-TCR Cld5 KO fetuses are rare, I am still collecting the data at present. (3) Study about thymic vascular endothelial cells by FACS analysis I established the method to examine Cld5 expression by FACS analysis after intracellular staining. By this method, I can study about thymic vascular endothelial cells and compare Cld5+ and Cld5- vascular endothelial cells in more detail in the following research.
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Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
There are two issues to clarify in this research: (A) the molecular basis of BTB; (B) the immunological function of BTB. For the issue (A), the results up to now suggested that Cld5 is the molecular basis of BTB. This is the basic issue for this research. It makes it possible to use Cld5 KO mice to study about the immunological function of BTB, the issue (B). However, Cld5 KO mice die quickly after birth, which makes the research more difficult. And the examination of Cld5 KO fetuses showed that their T cell development (CD4 CD8 proportion) were similar to that in wild type fetuses. Although the immunological functions of BTB remain unclear, and some of the planned experiments about issue (B) did not show the expected results, those challenges will be helpful for finding out a better research direction and improving the research methods in the following year.
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Strategy for Future Research Activity |
(1) Although Cld5 KO fetuses in normal condition do not show obvious difference on T cell development, whether they are more sensitive to some abnormal conditions, for example inflammation, remain unclear. I am planning to study about it in the following year. (2) I will continue to collect the data about OT-I-TCR Cld5 KO fetuses after ova-peptide injection into OT-I-TCR Cld5+/- pregnant mice. (3) Because the system for studying about BTB function remain a lot of problems, I turned to think that it might be better to collect more basic information about the thymic vascular endothelial cells. I will study about various markers on thymic vascular endothelial cells and compare Cld5+ and Cld5- endothelial cell in more detail by the FACS analysis. Including the Cld5 expression and thymic microenvironment changes during aging.
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Research Products
(1 results)