Tumor growth is associated with aberrant myelopoiesis, including the accumulation of CD11b ؉ Gr-1 ؉ myeloid-derived suppressor cells (MDSCs) that have the potential to promote tumor growth. However, the identity, growth, and migration of tumor-associated MDSCs remain undefined. We demonstrate herein that MDSCs at tumor site were composed primarily of bone marrow-derived CD11b ؉ Gr-1 hi Ly-6C int neutrophils and

Background: Interleukin-31 (IL-31), a novel cytokine, is upregulated in atopic dermatitis skin lesions in humans and skin lesions in the NC ⁄ Nga mice, a murine model for atopic dermatitis.Objective: Here, we investigated the effect of a monoclonal IL-31 antibody on scratching behaviour, weight gain and dermatitis in NC ⁄ Nga mice.Methods: Mice were divided into three groups, n = 10 in each group. Mice were given monoclonal IL-31 ratanti-mouse antibody 10 mg ⁄ kg or albumin intraperitoneally every fifth day for seven weeks. In addition, the mice in one group were not exposed to any form of intervention. The dermatitis score was clinically assessed twice a week. The scratching behaviour was automatically detected and objectively evaluated.Results: Intervention with IL-31 antibody 10 mg ⁄ kg intraperitoneally every fifth day in NC ⁄ Nga mice from age 7 weeks reduced the scratching behaviour, but did not have any impact on weight gain or dermatitis.

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A single donor-specific transfusion induces alloantibodies and Treg cells

AimThymic epithelial cells (TECs) are thought to play an essential role in T cell development and have been detected mainly in mice using lectin binding and antibodies to keratins. Our aim in the present study was to create a precise map of rat TECs using antibodies to putative markers and novel monoclonal antibodies (i.e., ED 18/19/21 and anti-CD205 antibodies) and compare it with a map from mouse counterparts and that of rat thymic dendritic cells.ResultsRat TECs were subdivided on the basis of phenotype into three subsets; ED18+ED19+/−keratin 5 (K5)+K8+CD205+ class II MHC (MHCII)+ cortical TECs (cTECs), ED18+ED21−K5−K8+ Ulex europaeus lectin 1 (UEA-1)+CD205− medullary TECs (mTEC1s), and ED18+ED21+K5+K8dullUEA-1−CD205− medullary TECs (mTEC2s). Thymic nurse cells were defined in cytosmears as an ED18+ED19+/−K5+K8+ subset of cTECs. mTEC1s preferentially expressed MHCII, claudin-3, claudin-4, and autoimmune regulator (AIRE). Use of ED18 and ED21 antibodies revealed three subsets of TECs in mice as well. We also detected two distinct TEC-free areas in the subcapsular cortex and in the medulla. Rat dendritic cells in the cortex were MHCII+CD103+ but negative for TEC markers, including CD205. Those in the medulla were MHCII+CD103+ and CD205+ cells were found only in the TEC-free area.ConclusionBoth rats and mice have three TEC subsets with similar phenotypes that can be identified using known markers and new monoclonal antibodies. These findings will facilitate further analysis of TEC subsets and DCs and help to define their roles in thymic selection and in pathological states such as autoimmune disorders.

Background Lymphocyte recruitment into the portal tract is crucial not only for homeostatic immune surveillance but also for many liver diseases. However, the exact route of entry for lymphocytes into portal tract is still obscure. We investigated this question using a rat hepatic allograft rejection model. Methods A migration route was analyzed by immunohistological methods including a recently developed scanning electron microscopy method. Transmigrationassociated molecules such as selectins, integrins, and chemokines and their receptors expressed by hepatic vessels and recruited T-cells were analyzed by immunohistochemistry and flow cytometry.Results The immunoelectron microscopic analysis clearly showed CD8b ? cells passing through the portal vein (PV) endothelia. Furthermore, the migrating pathway seemed to pass through the endothelial cell body. Local vascular cell adhesion molecule-1 (VCAM-1) expression was induced in PV endothelial cells from day 2 after liver transplantation. Although intercellular adhesion molecule-1 (ICAM-1) expression was also upregulated, it was restricted to sinusoidal endothelia. Recipient T-cells in the graft perfusate were CD25 ? CD44 ? ICAM-1 ? CXCR3 ? CCR5 -and upregulated a4b1 or aLb2 integrins. Immunohistochemistry showed the expression of CXCL10 in donor MHCII high cells in the portal tract as well as endothelial walls of PV. Conclusions We show for the first time direct evidence of T-cell transmigration across PV endothelial cells during hepatic allograft rejection. Interactions between VCAM-1 on endothelia and a4b1 integrin on recipient effector T-cells putatively play critical roles in adhesion and transmigration through endothelia. A chemokine axis of CXCL10 and CXCR3 also may be involved.

The aim of this study was to investigate the trafficking patterns, radiation sensitivities, and functions of conventional dendritic cell (DC) subsets in the rat liver in an allotransplantation setting. We examined DCs in the liver, hepatic lymph, and graft tissues and recipient secondary lymphoid organs after liver transplantation from rats treated or untreated by sublethal irradiation. We identified two distinct immunogenic DC subsets. One was a previously reported population that underwent blood-borne migration to the recipient's secondary lymphoid organs, inducing systemic CD8 1 T-cell responses; these DCs are a radiosensitive class II major histocompatibility complex (MHCII)1 subset. Another was a relatively radioresistant MHCII CD861 subset that steadily appeared in the hepatic lymph. After transplantation, the second subset migrated to the parathymic lymph nodes (LNs), regional peritoneal cavity nodes, or persisted in the graft. Irradiation completely eliminated the migration and immunogenicity of the first subset, but only partly suppressed the migration of the second subset and the CD8 1 T-cell response in the parathymic LNs. The grafts were acutely rejected, and intragraft CD8 1 T-cell and FoxP3 1 regulatory T-cell responses were unchanged. The radioresistant second subset up-regulated CD25 and had high allostimulating activity in the mixed leukocyte reaction, suggesting that this subset induced CD8 1 T-cell responses in the parathymic LNs and in the graft by the direct allorecognition pathway, leading to the rejection. Conclusion: Conventional rat liver DCs contain at least two distinct immunogenic passenger subsets: a radiosensitive blood-borne migrant and a relatively radioresistant lymph-borne migrant. LNs draining the peritoneal cavity should be recognized as a major site of the intrahost T-cell response by the lymph-borne migrant. This study provides key insights into liver graft rejection and highlights the clinical implications of immunogenic DC subsets. (HEPATOLOGY 2012;56:1532-1545 T he trafficking of dendritic cell (DC) subsets is important because appropriate site-specific antigen delivery is essential for immune regulation, both in the healthy state and in various immunemediated diseases.1 However, information regarding trafficking patterns of the specific DC subsets remains incomplete. [2][3][4][5] We demonstrated previously that after rat liver transplantation (LT), a small, but notable number of graft DCs systemically migrate to the recipient's secondary lymphoid organs through the bloodstream; these cells form clusters with the recipient's T cells and induce diffuse CD8 þ T-cell responses that

Immune responses are generally accompanied by antigen presentation and proliferation and differentiation of antigen-specific lymphocytes (immunoproliferation), but analysis of these events in situ on tissue sections is very difficult. We have developed a new method of simultaneous multicolor immunofluorescence staining for immunohistology and flow cytometry using a thymidine analogue, 5-ethynyl-2′-deoxyuridine (EdU). Because of the small size of azide dye using click chemistry and elimination of DNA denaturation steps, EdU staining allowed for immunofluorescence staining of at least four colors including two different markers on a single-cell surface, which is impossible with the standard 5-bromo-2′-deoxyuridine method. By using two rat models, successfully detected parameters were the cluster of differentiation antigens including phenotypic and functional markers of various immune cells, histocompatibility complex antigens, and even some nuclear transcription factors. Proliferating cells could be further sorted and used for RT-PCR analysis. This method thus enables functional in situ time-kinetic analysis of immunoproliferative responses in a distinct domain of the lymphoid organs, which are quantitatively confirmed by flow cytometry.Electronic supplementary materialThe online version of this article (doi:10.1007/s00418-015-1329-z) contains supplementary material, which is available to authorized users.