Mini-symposium for

Mucosal Immunology
and Allergy therapeutics
December 16, 2015
Inohana Alumni House（ゐのはな同窓会館）
Program
13:00-13:05 Opening Remarks
Takeshi Tokuhisa (President, Chiba University)

13:05-13:20“International Center of Excellence in Mucosal Immunology andInnovative Allergy Therapeutics at Chiba University”

Toshinori Nakayama (Dean, Graduate School of Medicine, Vice President, Chiba University)

13:20-14:40

Symposium 1 : Mucosal Immunology [Chair: Satoshi Uematsu & Kiyoshi Hirahara]

Satoshi Uematsu
Shinobu Saijo
Yoshiyuki Goto
Yosuke Kurashima“Roles of innate immunity in radiation-induced intestinal injury”“Mechanisms of anti-fungal immunity”“Group 3 innate lymphoid cells regulate intestinal epithelial fucosylation”“Elucidation and Regulation of Disease-specific Mast Cell Activations”

14:40-15:00 Coffee Break
15:00-16:20

Symposium 2 : Allergy Therapeutics [Chair: Satoshi Uematsu & Kiyoshi Hirahara]“Thy1+IL-7+ lymphatic endothelial cells in iBALT are potential therapeutic targets forallergic airway inflammation”
Mark Bix“Constraint of whipworm expulsion associated with Cryptdin repression”
Yuumi Matsuoka-Nakamura “Cooperation of IL-1 and IL-36 plays a pivotal role in epicutaneous S.aureus-induced
skin inflammation via IL-17 production”
Hiroshi Nakajima“Roles of C-type lectin receptors in HDM-induced allergic airway inflammation”

Kiyoshi Hirahara

16:20-16:40

Coffee Break

16:40-17:25

Plenary Lecture[Chair: Hiroshi Kiyono]Peter Ernst (Department of Pathology, University California San Diego)“The role of purine metabolism in mucosal immune homeostasis: Adenosine regulates Th cell plasticity andthe control of Th cell-mediated enterocolitis”

Closing Remarks

Hiroshi Kiyono

17:30-Get Together Party “See-Saw”

入場無料

17:25-17:30

Organized by Future Medicine Education and Research Organization at Chiba University
Co-Organized by Leading Graduate School at Chiba UniversityInquiries

Future Medicine Education and Research Organization
Administrative Office
TEL : 043-226-2832 / FAX : 043-226-2489 / E-mail : inohana-kaikaku@chiba-u.jp

International Center of Excellence in Mucosal Immunology andInnovative Allergy Therapeutics at Chiba University
Tosihnori Nakayama
Dean, Graduate School of Medicine,Vice President, Chiba University― 2 ―Roles of innate immunity in radiation-inducedintestinal injury
Satoshi Uematsu, Naoki Takemura

Department of Mucosal Immunology, Graduate Schoolof Medicine, Chiba University, Japan
Radiation-induced intestinal fibrosis (RIF) is a serious complication afterabdominal radiotherapy. Here we show RIF is mediated by eosinophil interactions
with -smooth muscle actin (-SMA)+ stromal cells. Abdominal irradiation induced
fibrosis of the submucosa (SM) of small intestine associated with the excessive
accumulation and degranulation of eosinophils, even in the absence of lymphocytes.
Eosinophil-deficiency markedly ameliorated RIF, suggesting their importance.
Chronic crypt necrosis post-irradiation elevated extracellular adenosine triphosphatelevels, which induced C-C motif chemokine 11 (CCL11) expression by pericryptal
-SMA+ cells to recruit eosinophils to the SM via CCR3. Unexpectedly, type 2 innate
lymphoid cells were also dispensable for eosinophil activation in RIF. For activationof eosinophils, activated -SMA+ cells expressed granulocyte macrophagecolony-stimulating factor (GM-CSF), which induced expression of transforming
growth factor-1 from intestinal eosinophils, leading to the promotion of collagen
expression by -SMA+ cells. In addition, eosinophils released cytotoxic granuleproteins upon co-stimulation with GM-CSF and CCL11. Thus, radiation-induced
crypt injury triggers mutual activation of eosinophils and -SMA+ cells, leading to a
progressive fibrogenesis. Our findings provide an immunopathological mechanism ofRIF and clues for the development of therapeutic strategies for RIF.

― 3 ―Mechanisms of anti-fungal immunity
Mari Iwasawa1, Yuumi Matsuoka-Nakamura1 andShinobu Saijo212

Dermatology, Chiba University, Chiba Japan
Medical Mycology Research Center, Chiba University, Chiba Japan

Myeloid cells are key player in anti-microbial responses. Members of C-type
lectin receptors (CLRs) surpass among the specialized receptors that expressed inthese cells to orchestrate immune responses these responses. They are characterized
by the extracellular domain called CRD (carbohydrate binding domain). Although
they had been suggested to be members of pattern recognition receptors (PRRs), theirfunctions have long been remained unclear. Therefore, we aimed to clarify themolecular mechanisms of C-type lectins in the host defense against microbialinfections. Dectin-1 was first reported as a dendritic cell (DC)-specific C-type lectin
receptor, and is also highly expressed in macrophages and neutrophils. It has a CRD
in its extracellular carboxyl terminus and an immunoreceptor tyrosine-based
activation motif (ITAM) in its intracellular amino terminus, and is suggested to be thereceptor for -1, 3-linked and/or -1, 6-linked glucans (-glucans) found in the cell
walls of fungi. Dectin-2 is also expressed in DCs and macrophages, and has a CRD,
but has no known signaling motif in its intracellular domain. We have generated
Dectin-1 and Decttin-2-deficient mice to determine the roles of these molecules in thedefense against pathogenic fungi. In vitro, -glucan-induced cytokine production
from wild-type DCs and macrophages was abolished in Dectin-1-deficie cells, and-mannan-induced cytokine production was abolished in Dectin-2-deficient cells. Invivo, Dectin-1-deficient mice were more susceptible than wild-type mice toPneumocystis carinii infection, and Dectin-2-deficient mice were more susceptible toCandida albicans (C. albicans)infection. Interestingly, candida preferentially induced
Th17 cell differentiation, which was markedly reduced by Dectin-2 deficiency.
Because IL-17A-deficient mice were more susceptible to candida infection, it wassuggested that Dectin-2 signaling preferentially induces Th17 cell differentiation andTh17 cell-derived IL-17A is important for the host defense against C. albicans.
Thus, Dectin-1 and Dectin-2 play important roles for the defense against fungal
infections as a protective innate and acquired immunity. In this mini symposium, I
would like to share my recent findings.

― 4 ―Group 3 innate lymphoid cells regulate
intestinal epithelial fucosylation
Yoshiyuki Goto
Division of Molecular Immunology, Medical Mycology
Research Center, Chiba University
Intestine is a unique organ which is constitutively exposed by external antigens
including commensal and pathogenic bacteria. Intestinal epithelial cells and immune
cells create barrier system against harmful antigens. At the same time, host provide
peaceful symbiotic environment for commensal bacteria. 1, 2-fucose expressed onintestinal epithelial cells, catalyzed by fucosyltransferase 2 (Fut2), is one of majorcarbohydrate moiety which mediates host–microbiota symbiosis. Since specificsymbionts such as Bacteroides utilize epithelial 1, 2-fucose as a dietarycarbohydrate, dysfunction of Fut2 leads to dysbiosis of gut microflora andpredisposes to the development of host diseases. Indeed, human FUT2 non-sense
polymorphisms are associated with various inflammatory diseases such as Crohn’sdisease, type I diabetes and primary sclerosing cholangitis. Therefore, it is important
to identify the mechanism of the induction of epithelial Fut2 and 1, 2-fucose(fucosylation). Here, we show that segmented filamentous bacteria (SFB), one ofcommensal bacteria specifically colonized at ileal epithelium, have a potential toinduce epithelial fucosylation under physiological condition. Furthermore, group 3
innate lymphoid cells (ILC3) are also necessary for the induction of intestinal
epithelial fucosylation. Interleukin-22 and lymphotoxin produced from ILC3 arerequiredfortheinductionofepithelial

fucosylation

inacommensal

bacteria–dependent and –independent manner, respectively. Mice lacking epithelial
fucosylation are susceptible to infection by Salmonella typhimurium. Taken together,
these data uncovered the novel function of commensals-ILC3-epithelial cell networkin the induction of epithelial glycosylation, creation of gut barrier system against
pathogenic bacterial infection, and regulation of intestinal homeostasis.

― 5 ―Elucidation and Regulation of Disease-specific Mast
Cell Activations
Yosuke Kurashima
Division of Mucosal Immunology,Department of Microbiologyand Immunology, The Institute of Medical Science,
The University of Tokyo, TokyoSince the discovery of mast cells in 1878, and IgE 50 years ago, the important rolesof mast cells in both allergic diseases and parasitic infections have been extensivelyrecognized. A few years after the discovery of IgE, it was reported that histamine isreleased from mast cells upon the stimulation of extracellular ATP; however, thediseases related to the extracellular ATP-mediated mast cell activations are issued
relatively recently. In this symposium, we would like to introduce our recent findings,indicating the ATP-mast cell pathway as a new drug target against inflammatory bowel
diseases. In addition, a unique skin-barrier homeostatic network operating throughretinoic acid-metabolism-mediated inhibition of ATP-dependent MC activation byfibroblasts. At the end of the presentation, I would like to share the results obtained
recently indicating the important roles of mast cells in both development and inhibitionof food allergy.

― 6 ―Thy1+IL-7+ lymphatic endothelial cells in iBALT
are potetial therapeutic targets for allergic airway
inflammation
Kiyoshi Hirahara1, Kenta Shinoda2, Toshinori Nakayama2
1. Department of Advanced Allergology of the Airway,Graduate School of Medicine, Chiba University, Chiba,Japan.
2. Department of Immunology, Graduate School of Medicine, Chiba University,Chiba, Japan.
Memory CD4+ T helper (Th) cells are crucial to long-term protection against
pathogens.

But they can also be pathogenic and drive chronic inflammatory

disorders including allergic airway inflammation such as asthma.

In 2011, we

identified that IL-5-producing memory Th2 cell subsets are critical for the pathologyof allergic inflammation, and function as “memory type pathogenic Th2 (Tpath2)
cells (Endo et al. Immunity, 2011).

Very recently, we found that Interleukin-33(IL-33)-ST2 signals as a key pathway for the induction of memory-type Tpath2 cells
during the eosinophilic airway inflammation in both mice and human systems (Endoet al. Immunity, 2015).
But It has been still unclear how memory type pathogenic Th2 (Tpath2) aremaintained, particularly at the site of local inflammation. We found that ectopic
lymphoid-like structures called inducible bronchus-associated lymphoid tissue(iBALT) are formed during chronic allergic inflammation in the lung, and thatmemory-type

Tpath2cells

aremaintained

withintheiBALT structures.

Maintenance of the Tpath2 cells within iBALT is supported by novel
Thy1+IL-7-producing lymphatic endothelial cells (LECs) within the lung.
Moreover, ectopic lymphoid structures consisting of memory CD4+ T cells were
found in nasal polyps of eosinophilic chronic rhinosinusitis patients, indicating thatthe persistence of inflammation is controlled by these structures.

Therefore, the cellcomponents that organize and contribute to iBALT formation and the functional
molecules that maintain pathogenic inflammatory memory Th2 cells are likely to be
excellent targets for the treatment of chronic allergic airway inflammation.
― 7 ―Constraint of whipworm expulsion associatedwith Cryptdin repression
Mark Bix
Department of Innovative Therapeutics, School of Medicine,
Chiba University, Japan
Cryptdins (also known as alpha-defensins) are a major class of anti-microbial
peptides secreted by Paneth cells located at the base of intestinal Crypts ofLieberkuhn. Cryptdins exhibit potent anti-microbial activity toward viruses, bacteria,fungi and protozoa. However, activity toward helminths has not been described. I will
present evidence that under inflammatory conditions the JmjC-family epigenetic
regulatory protein Mina represses Cryptdin expression. Further, during intestinal
whipworm infection of Mina KO mice, where Cryptdin expression is de-repressed,
helminth expulsion is accelerated, raising the possibility that Cryptdins may possess
anti-helminthic activity.

― 8 ―Cooperation of IL-1 and IL-36 plays a pivotal
role in epicutaneous S.aureus-induced skininflammation via IL-17 production
Seitaro Nakagawa1, Yuumi Matsuoka-Nakamura1,Gabriel Nunez2, Hiroyuki Matsue1
12

Dermatology, Chiba University, Chiba, Japan
Pathology and Comprehensive Cancer Center, University of Michigan, MI, USAAlthough S. aureus is known to colonize the skin of atopic dermatitis patients, little is
known about the mechanisms of S.aureus-induced eczematous inflammation. MyD88
is known to be an adaptor in TLR- and IL-1 receptor (R)-mediated signaling
pathways for innate immunity to microorganisms. Myd88-/- mice reportedly exhibited
more severe skin inflammation than WT mice due to high bacteria burden if S.aureus
was injected subcutaneously. Contrary to the subcutaneous injection, we found thatMyd88-/- mice showed no skin inflammation, whereas WT mice showed severe
eczematous lesions with the similar epicutaneous colonization of S. aureus, indicatingthat MyD88 signaling promoted skin inflammation but not cleared S. aureus in our
epicutaneous disease model. When applied on keratinocyte (KC)-specific Myd88
knock-down, K14-CreMyd88-/- mice, the skin inflammation was also dramaticallyreduced, indicating the involvement of MyD88 in KC. To explore the upstream
signals, we next applied S. aureus on Tlr2/4-/- and Il1r-/- mice. Tlr2/4-/- mice showed
severe inflammation as WT mice, whereas Il1r-/- mice showed moderate
inflammation, indicating that the inflammation was partially IL-1-mediated. Whenapplied on Il1r-/- mice with IL-36R-neutralizing antibody (IL-36R Ab), theinflammation was significantly reduced, showing additive IL-36 involvement. Upon
the infection, little mRNAs and proteins of IL-17 were detected in K14-CreMyd88-/-,
Il1r-/- with or without IL-36RAb mice. In accordance with these, Il17a-/--Il17f--/- mice
showed significantly less skin inflammation upon the infection. In conclusion, KC iscentral for the IL-1-mediated inflammation via IL-17 production and otherwise IL-36
involvement in the skin inflammation of this epicutaneous disease model.

― 9 ―Roles of C-type lectin receptors in HDM-induced
allergic airway inflammation
Hiroshi Nakajima, Takashi Ito, Koichi Hirose
Department of Allergy and Clinical Immunology, Graduate
School of Medicine, Chiba University, Japan
Background: The fact that sensitization against fungi is closely related to the severityof asthma suggests that immune systems recognizing fungi are involved in thepathogenesis of severe asthma. Recently, C-type lectin receptors, Dectin-1 (genesymbol Clec7a) and Dectin-2 (Clec4n), have been shown to function as not only
major pattern recognition receptors for fungi but also receptors for some components
of house dust mite (HDM) extract, a major allergen for allergic asthma. However, theroles of Dectin-1 and Dectin-2 in the induction of HDM-induced allergic airway
inflammation remain largely unknown.
Objectives: To determine the roles of Dectin-1 and Dectin-2 in HDM-induced allergic
airway inflammation.
Methods: We examined the roles of Dectin-1 and Dectin-2 in the induction ofHDM-induced Th2 and Th17 cell differentiation and subsequent allergic airway
inflammation by using Clec7a-deficient (Clec7a-/-) mice and Clec4n-deficient(Clec4n-/-) mice. We also investigated Dectin-1- or Dectin-2-expressing cells in thelung and their roles in HDM-induced allergic airway inflammation.
Results: Both Clec7a-/- mice and Clec4n-/- mice showed significantly attenuatedHDM-induced allergic airway inflammation and decreased Th2 and Th17 celldifferentiation as compared with wild-type (WT) mice. Both Dectin-1 and Dectin-2
were expressed on CD11b+ dendritic cells (DCs). CD11b+ DCs isolated from
Clec7a-/- mice or Clec4n-/- mice expressed lower levels of proinflammatory cytokinesand co-stimulatory molecules which could induce Th2 and Th17 cell differentiation
than those from WT mice.
Conclusion: Dectin-1 and Dectin-2 were expressed on CD11b+ DCs and promote
HDM-induced allergic airway inflammation.

― 10 ―MEMOMEMO