研究題目

Thymus medulla development and function during
αβT-cell development

研究組織研究代表者：Graham Anderson (Experimental Immunology, Institute for Immunology andImmunotherapy, University of Birmingham)
共同研究者：高浜洋介、大東いずみ (徳島大学先端酵素学研究所）【1】研究の概要

exciting position of being able to exploit our[1-1]本研究の目的・概要

individual and unique skill sets, to explore

The thymus is an essential primary lymphoid

key unanswered questions in thymus biology.

organ in the immune system of all vertebrates.

Additionally, these links will allow us toIt is unique in its ability to support thatdevelop new joint research projects, that will

generation of T-cells that play key roles intrain young scientists in Birmingham andimmune responses to invading pathogens andTokushima via reciprocal lab visits, andtumor

generate co-authored peer reviewed research

surveillance.

Researchin

mylaboratory has focused on the mechanisms
T-cellarticles.

In

Our overall aims are: (1) To combine

particular, we examine how cortical andfluorescent reporter mouse strains generated

medullarycells

in Tokushima (CCL21 Red mice) with those(cTEC/mTEC) are able to generate and selectgenerated in Birmingham (RANK Venus

self-tolerantmice) to generate a novel ‘dual reporter’

controlling

autoreactive

thymicdevelopment.
epithelialT-cells
T-cellsand

tomouse line, where newly defined thymic

autoimmunity. The work of my laboratory inepithelial cell subsets can be analysed andthis area has been greatly aided by successful

isolated from embryonic and adult mice. (2)collaborations with immunologists at The

To train young researchers in cellular

University of Tokushima. Most recently, by(Birmingham) and molecular (Tokushima)collaborating with Dr Izumi Ohigashi andimmunologicalProfessor Yousuke Takahama at The Institutedeveloping their own research careers andof Genome Research, we co-authored anunderpin long-term collaborations between

influential review article in Nature Reviews

next generation scientists in Birmingham andImmunology

Tokushima.

onthateliminate

thymicpredispose

epithelial

celltechniques,

toaidinbiology.
By continuing to develop our collaborative[1-2]研究の方法・経過

links with Dr Ohigashi, we are now in theOur main aim is to investigate how cTEC andmTEC lineages emerge during thymus

成果

development, and examine the functionalCurrently, following the importation ofsignificance of heterogeneity within these

CCL21Tomato reporter mice from University

lineages in the adult thymus. In the Andersonof Tokushima into University of Birmingham,lab,

we have successfully generated double

werecentlygenerated

fluorescentreporter mice in which cells expressing

reporterRANK, a key regulator of the thymus

TEC reporter mice. In initial experiments, wemedulla,green

have assessed the expression of Venus andfluorescent protein Venus. The Ohigashi lab

Tomato in both thymus sections and by flow

has also recently generated a different

cytometry in adult and fetal stages. In earlyreporter mouse in which mTEC that express

fetal stages (E15-E17) we see two clearly

the chemokine CCL21 are carry a red

separate

fluorescent label. We crossed these two

CCL21Tomato+RANKVenus-

independently generated mouse strains toCCL21Tomato-RANKVenus+.

generate a new ‘dual reporter’ mouse line

initial experiments, both populations appear

where mTEC heterogeneity can be examined

to arise at the same time, suggesting that they

through analysis of red (CCL21) and greenmay(RANK) expression. By sharing our mouse

relationship. Moreover, in the adult, while welines, and visiting and collaborating with thesee the persistence of these populations, weOhigashi lab we did: (1) Examine thealso see a double positive population of

emergence of the mTEC lineage during

mTEC that is CCL21Tomato+RANKVenus+.

embryogenesis by flow cytometric/confocalThus, the approach of generating a new TEC

analysis of the developing thymus, from

reporter mouse line has identified new TEC

embryonic day 12 through to birth. (2) Isolate

populations that need to be assessed at bothdifferent subsets of mTEC from dual reporterthe developmental and functional level.

miceareie.

labeledwith++++notpopulations

reflectaofdual

mTEC:
andIn

theseprecursor-product(green red ),In addition, by importing the CCL21Tomato

RANK+CCL21- (green+red-), RANK-CCL21+

mice from Tokushima, we used them to-

RANK CCL21 :theCCL2Tomato/RANKVenus+----(green red ), and RANK CCL21 (green red )assess the role of CCl21 in formation of theand analyse their gene expression of known

intrathymic dendritic cell (DC) pool. We

regulators of medulla development andfound that CCL21 is essential to guide themTEC function eg Aire, Relb, LTR, CD40,

migration of CCR7+ DC into the thymus.

Fezf2.

This work was recently published in Journalof【2】研究成果[2-1]本共同研究で明らかになった研究

Immunology

publication.

asacollaborative

[2-2]本共同研究による波及効果及び今[3-3]成果資料等

後の発展性
Establishing a novel dual reporter mouse line,【4】今後の課題等

in which TEC are defined by the expression

We now aim to systematically characterizeof functionally relevant molecules (RANKmTEC development in the fetal and adultand CCL21) will provide important insightstages of life in CCL21Tomato/RANKVenus

into the poorly understood field of thymus

dual reporter mice. We will define themedulla

phenotypic

developmentandfunction.

propertiesof

Tomato/Venus

Additionally, support obtained from this

subpopulations by confocal microscopy andfunding has had a significant impact on theflow cytometry using a panel of markers

training of young PhD students and postdocs(EpCAM1, Ly51, UEA1, b5t, MHCII, CD80,

in both Tokushima and Birmingham. By

CD86). We will also test the developmental

supporting reciprocal visits, scientists havepotential

met and discussed future plans, and dataisolating defined populations on the basis ofgenerated in this collaboration will beTomato/Venus expression, and incorporating

important in applying for future researchthem into reaggregate thymus organ cultures,

funding in both the UK and Japan.

where the developmental potential of purified

ofindividual

cells will be revealed.
【3】主な発表論文等[3-1]論文発表
1）Takahama Y, Ohigashi I, Baik S, Anderson G
Generation of diversity inthymic epithelial cells.
Nat. Rev. Immunol. 17:295-305 (2017).

2) Cosway EJ, Ohigashi I, Schauble K ,Parnell SM, Jenkinson WE, Luther S,Takahama Y, Anderson G. Formation of theIntrathymic Dendritic Cell Pool Requires
CCL21-mediatedrecruitment

of

CCR7+progenitors to the thymus. J. Immunol.
201:516-523 (2018).
2）[書籍][3-2]学会発表

populationsby