ABSTRACT
The nervous system in the intestine controls motility, secretion, sensory perception, and immune function. Peptidergic neurones with neurotransmitters such as substance P and nerve growth factors have been the main focus of neuroimmunomodulation research in the gut. This review summarises the present knowledge concerning the role of the sympathetic nervous system (SNS) in modulating intestinal inflammation. The role of the SNS for gut inflammation is compared with its role in rheumatoid arthritis which demonstrates notable similarities. Nerve fibres of the SNS not only enter the enteric plexuses but also innervate the mucosa and gut associated lymphoid tissue (GALT). The SNS has pro- and anti-inflammatory functions. Neurotransmitters such as norepinephrine, adenosine, and others can evoke remarkably different opposing effects depending on concentration (presence of sympathetic nerve fibres and extent of neurotransmitter release), receptor affinity at different receptor subtypes, expression of adrenoceptors, availability of cotransmitters, and timing of SNS activity in relation to the inflammatory course. This review attempts to integrate the different perspectives of the pro- and anti-inflammatory effects of the SNS on inflammatory disease of the gut.
Subject(s)
Gastroenteritis/physiopathology , Intestines/innervation , Sympathetic Nervous System/physiopathology , Humans , Intestines/immunology , Neurotransmitter Agents/physiology , Sympathetic Nervous System/anatomy & histologyABSTRACT
BACKGROUND: The resident flora plays a critical role in initiation and perpetuation of intestinal inflammation, as demonstrated in experimental models of colitis where animals fail to develop disease under germ free conditions. However, the importance of exposure to commensal bacteria before the onset of colitis is unclear. Our aim was to investigate the influence of previous exposure of donor animals to bacterial antigens on colitis development using a transfer model. METHODS: Clinical course and histology were evaluated after transfer of CD4(+)CD62L(+) lymphocytes from germ free and conventionally housed donor mice into SCID recipients. Cotransfer of CD4(+)CD62L(+) cells with CD4(+)CD62L(- )lymphocytes from both groups of mice was initiated. Lymphocytes were analysed by FACS, polarisation potential of cells determined, and cytokines measured within the supernatant by enzyme linked immunosorbent assay. RESULTS: Animals that received cells from germ free donors developed an earlier onset of colitis compared with mice reconstituted with lymphocytes from conventionally housed animals. Additionally, CD4(+)CD62L(- )cells from germ free mice were not able to abrogate colitis induced by cotransfer with CD4(+)CD62L(+) lymphocytes whereas CD4(+)CD62L(- )T cells from normal mice ameliorated disease. The higher percentage of CD4(+)GITR(+) expressing lymphocytes and the production of interleukin 10 after priming by dendritic cells suggests the presence of T(reg) cells within the CD4(+)CD62L(+) lymphocyte subset derived from conventional housed mice and assumes a lack of T(reg) cells within germ free mice. CONCLUSION: The results indicate that bacterial antigens are crucial for the generation and/or expansion of T(reg) cells in a healthy individual. Therefore, bacterial colonisation is of great importance in maintaining the immunological balance.
Subject(s)
Antigens, Bacterial/immunology , CD4-Positive T-Lymphocytes/immunology , Colitis/immunology , Intestines/microbiology , T-Lymphocyte Subsets/immunology , Adoptive Transfer , Animals , CD4-Positive T-Lymphocytes/transplantation , Coculture Techniques , Colitis/microbiology , DNA-Binding Proteins/metabolism , Dendritic Cells/immunology , Female , Forkhead Transcription Factors , Germ-Free Life/immunology , Interferon-gamma/biosynthesis , Interleukin-10/biosynthesis , Intestines/immunology , L-Selectin/analysis , Mice , Mice, Inbred BALB C , Mice, SCID , T-Lymphocyte Subsets/transplantationABSTRACT
BACKGROUND: The glycoprotein (gp) 96 links the adaptive with the innate immune system. It is a chaperone with a binding domain for peptides generated by proteasomal degradation. During cellular stress, peptide loaded gp96 can be released and presented to T cells by antigen presenting cells (APCs). METHODS: mRNAs from in vitro differentiated macrophages (iv mac) and normal intestinal macrophages (IMACs) were compared by subtractive hybridisation and Affymetrix GeneChip analysis. Differentiation induced expression of gp96 was investigated in the multicellular spheroid (MCS) model. In vivo gp96 protein expression was detected by double labelling immunohistochemistry of human colon and in the CD4+ CD62L+ T cell transfer mouse model. RESULTS: Five of 76 clones obtained by subtractive hybridisation revealed >99% sequence homology to gp96. Affymetrix GeneChip analysis confirmed induction of gp96 in IMACs. Gp96 mRNA was detected in IMACs from normal and intestinal bowel disease mucosa. Induction of gp96 protein was observed after seven days in the MCS model of IMAC differentiation. Immunohistochemistry confirmed the presence of gp96 protein in IMACs in normal mucosa as well as in mucosa from patients with ulcerative colitis and diverticulitis. In mucosa from Crohn's disease (CD) patients, gp96 protein was not detectable. In the CD4+ CD62L+ T cell transfer mouse model, gp96 was verifiable in non-activated IMACs. CONCLUSION: Gp96 is induced during differentiation of normal IMACs but is not detected in IMACs in CD mucosa. As gp96 has been described as having a role in tolerance induction, this may be relevant for loss of tolerance against luminal bacteria found in CD patients.
Subject(s)
Antigens, Neoplasm/metabolism , Crohn Disease/metabolism , Intestinal Mucosa/metabolism , Macrophages/metabolism , Animals , Antigens, Neoplasm/genetics , Antigens, Neoplasm/physiology , Cell Differentiation/physiology , Cells, Cultured , Colitis/metabolism , Disease Models, Animal , Gene Expression , Humans , Immunoenzyme Techniques , Mice , Mice, Inbred BALB C , Mice, SCID , Oligonucleotide Array Sequence Analysis/methods , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Spheroids, CellularABSTRACT
BACKGROUND AND METHODS: Cytosin-guanosin dinucleotide (CpG) motifs of bacterial DNA are known to be potent activators of innate immunity. We have shown previously that administration of CpG containing oligodeoxynucleotide (CpG-ODN) to mice before the onset of dextran sodium sulphate induced colitis ameliorated colitis and inhibited induction of proinflammatory cytokines. To investigate the possible involvement of CD4(+) T cells in the prophylactic CpG-ODN effects, we used the SCID transfer model of colitis. RESULTS: CD4(+)CD62L(+) T cells from CpG-ODN treated donors did not induce significant intestinal inflammation in SCID recipients, in contrast with control cells. Additionally, cotransfer of these cells with CD4(+)CD62L(+) cells from normal mice protected recipient animals from colitis, indicating regulatory activity. Also, CD4(+)CD62L(+) cells from toll-like receptor 9 deficient animals induced a significantly more severe colitis in SCID recipients than cells from wild-type littermate controls, suggesting a similar protective role of "endogenous" bacterial DNA leading to a less "aggressive" phenotype of these cells. There was no detectable difference in regulatory T cell surface markers between aggressive and attenuated cell pools but attenuated cell pools showed reduced proliferation in vitro and in vivo and produced less interferon gamma, interleukin (IL)-5, and IL-6 after anti-CD3 stimulation. CONCLUSIONS: Collectively, our data support the concept that both endogenous bacterial DNA and exogenously supplied CpG motifs of bacterial DNA induce regulatory properties in CD4(+) T cells. Therefore, bacterial DNA derived from the normal gut flora may contribute essentially to the homeostasis between effector and regulatory immune mechanisms in healthy individuals to protect them from chronic intestinal inflammation.
Subject(s)
CD4-Positive T-Lymphocytes/immunology , Colitis/immunology , GC Rich Sequence/immunology , Oligonucleotides/immunology , Animals , Cells, Cultured , Colitis/metabolism , Disease Models, Animal , Interferon-gamma/immunology , Interleukins/immunology , Intestinal Mucosa/immunology , L-Selectin/immunology , Lymph Nodes/immunology , Mesentery/immunology , Mice , Mice, Inbred BALB C , Mice, SCID , Rectum/immunology , Weight Loss/physiologyABSTRACT
Intestinal inflammation in inflammatory bowel disease (IBD) and experimental models of colitis is characterized by a dysregulated intestinal immune response with elevated levels of Th1 cytokines. The luminal flora has been implicated as a major factor contributing to the initiation and perpetuation of inflammation in experimental colitis by mechanisms not known. Bacterial DNA contains unmethylated cytosin-guanosin dinucleotides (CpG) which strongly activate Th1-mediated immune responses. To test whether these CpG-motifs modulate intestinal inflammation we treated mice with dextran sulphate sodium (DSS)-induced colitis with CpG-containing oligodeoxynucleotides (CpG-ODN). CpG-ODN given after the onset of DSS colitis aggravated the disease, as indicated by a significantly increased loss of body weight and a 30% increase of the histological score. Further, we found a severe increase of proinflammatory cytokines (interleukin (IL)-6: 40-fold; interferon (IFN)-gamma: 11-fold). In a pretreatment setting CpG-ODN reduced weight loss significantly and reduced intestinal inflammation by 45%. Colonic IFN-gamma and IL-6 mRNA levels were reduced by 75%, and IL-10 was elevated by 400% compared to controls. The prophylactic CpG-effect was not imitated by IL-12 because IL-12 pretreatment was not protective. In time-course experiments, CpG-ODN pretreatment over 5 days resulted in a tolerance effect concerning its IFN-gamma-inducing quality, and during the following days of colitis induction IL-10 secretion from mesenterial lymph node cells was elevated compared to controls. Therefore, the prophylactic effect of CpG-ODN might be explained by its tolerizing effect and/or the increased ability for IL-10 production during the consecutive intestinal inflammation.
Subject(s)
Colitis/immunology , Oligodeoxyribonucleotides/immunology , Acute Disease , Adjuvants, Immunologic , Animals , Body Weight , Colitis/pathology , Colitis/prevention & control , Colon/immunology , CpG Islands/immunology , Dextran Sulfate , Disease Models, Animal , Female , Immune Tolerance , Interferon-gamma/biosynthesis , Interleukin-12/immunology , Interleukin-6/biosynthesis , Lymph Nodes/immunology , Mesentery , Mice , Mice, Inbred BALB C , Oligodeoxyribonucleotides/therapeutic use , Weight LossABSTRACT
Integrins are important for T cell interactions with endothelial cells. Because the integrin alpha(E)beta(7) is expressed on some circulating gut-homing T cells and as T cell numbers are reduced in the intestinal lamina propria of alpha(E)-deficient mice, we evaluated whether alpha(E)beta(7) mediates binding to intestinal endothelial cells. We found that anti-alpha(E)beta(7) mAbs partially blocked the binding of cultured intraepithelial T cells to human intestinal microvascular endothelial cells (HIMEC). Furthermore, alpha(E)beta(7)-transfected K562 cells bound more efficiently than vector-transfected K562 cells to HIMEC. Finally, HIMEC bound directly to an alpha(E)beta(7)-Fc fusion protein. These interactions were partially blocked by anti-alpha(E)beta(7) mAbs, and endothelial cell binding to the alpha(E)beta(7)-Fc was dependent upon the metal ion-dependent adhesion site within the alpha(E) A domain. Of note, the HIMEC lacked expression of E-cadherin, the only known alpha(E)beta(7) counterreceptor as assessed by functional studies, flow cytometry, and RT-PCR. Thus, HIMEC/alpha(E)beta(7) binding was independent of E-cadherin. In addition, this interaction appeared to be tissue selective, as HIMEC bound to the alpha(E)beta(7)-Fc, whereas microvascular endothelial cells from the skin did not. Finally, there was evidence for an alpha(E)beta(7) ligand on intestinal endothelial cells in vivo, as alpha(E)beta(7) expression enhanced lymphocyte binding around vessels in the lamina propria in tissue sections. Thus, we have defined a novel interaction for alpha(E)beta(7) at a nonepithelial location. These studies suggest a role for alpha(E)beta(7) in interactions with the intestinal endothelium that may have implications for intestinal T cell homing or functional responses.
Subject(s)
Antigens, CD/physiology , Cadherins/physiology , Endothelium, Vascular/immunology , Integrin alpha Chains , Integrins/physiology , Intestinal Mucosa/blood supply , Intestinal Mucosa/immunology , Antibodies, Blocking/metabolism , Antibodies, Blocking/pharmacology , Antibodies, Monoclonal/metabolism , Antibodies, Monoclonal/pharmacology , Antigens, CD/biosynthesis , Antigens, CD/genetics , Antigens, CD/immunology , Binding Sites, Antibody , Binding, Competitive/immunology , Cell Adhesion/genetics , Cell Adhesion/immunology , Cell Communication/immunology , Cell Line , Cell Line, Transformed , Cells, Cultured , Endothelium, Vascular/cytology , Endothelium, Vascular/metabolism , Genetic Vectors/biosynthesis , Genetic Vectors/metabolism , Humans , Immunoglobulin Fc Fragments/genetics , Immunoglobulin Fc Fragments/metabolism , Integrins/biosynthesis , Integrins/genetics , Integrins/immunology , Intestinal Mucosa/cytology , Intestinal Mucosa/metabolism , K562 Cells , Metals/metabolism , Microcirculation/cytology , Microcirculation/immunology , Microcirculation/metabolism , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/metabolism , Skin/blood supply , Skin/cytology , Skin/immunology , Skin/metabolism , Solubility , Tumor Cells, CulturedABSTRACT
Mice chimeric for integrins alpha(5), alpha(V), or alpha(4) were used to dissect the in vivo roles of these adhesion receptors during leukocyte development and traffic. No major defects were observed in the development of lymphocytes, monocytes, or granulocytes or in the traffic of lymphocytes to different lymphoid organs in the absence of alpha(5) or alpha(V) integrins. However, in agreement with previous reports, the absence of alpha(4) integrins produced major defects in development of lymphoid and myeloid lineages and a specific defect in homing of lymphocytes to Peyer's patches. In contrast, the alpha(4) integrin subunit is not essential for localization of T lymphocytes into intraepithelial and lamina propria compartments in the gut, whereas one of the partners of alpha(4), the beta(7) chain, has been shown to be essential. However, alpha(4)-deficient T lymphocytes cannot migrate properly during the inflammatory response induced by thioglycolate injection into the peritoneum. Finally, in vitro proliferation and activation of lymphocytes deficient for alpha(5), alpha(V), or alpha(4) integrins upon stimulation with different stimuli were similar to those seen in controls. These results show that integrins play distinct roles during in vivo leukocyte development and traffic.
Subject(s)
Cell Movement/immunology , Chimera/immunology , Integrins/physiology , Leukocytes/cytology , Leukocytes/immunology , Animals , Antigens, CD/genetics , Antigens, CD/physiology , Cell Division/genetics , Cell Division/immunology , Cell Movement/genetics , Integrin alpha4 , Integrin alpha5 , Integrin alphaV , Integrins/genetics , Lymphocyte Activation/genetics , Lymphocytes/cytology , Lymphocytes/immunology , Mice , Mice, Inbred C57BL , Mice, Knockout , Myeloid Cells/immunology , Peritonitis/chemically induced , Peritonitis/genetics , Peritonitis/immunology , Receptors, Lymphocyte Homing/physiology , Thioglycolates/administration & dosageABSTRACT
The mucosal lymphocyte integrin alpha E(CD103)beta 7 is thought to be important for intraepithelial lymphocyte (IEL) localization or function. We cloned the murine integrin gene encoding alpha E, localized it to chromosome 11, and generated integrin alpha E-deficient mice. In alpha E-/- mice, intestinal and vaginal IEL numbers were reduced, consistent with the known binding of alpha E beta 7 to E-cadherin expressed on epithelial cells. However, it was surprising that lamina propria T lymphocyte numbers were diminished, as E-cadherin is not expressed in the lamina propria. In contrast, peribronchial, intrapulmonary, Peyer's patch, and splenic T lymphocyte numbers were not reduced in alpha E-deficient mice. Thus, alpha E beta 7 was important for generating or maintaining the gut and vaginal T lymphocytes located diffusely within the epithelium or lamina propria but not for generating the gut-associated organized lymphoid tissues. Finally, the impact of alpha E deficiency upon intestinal IEL numbers was greater at 3-4 wk of life than in younger animals, and affected the TCR alpha beta+ CD8+ T cells more than the gamma delta T cells or the TCR alpha beta+ CD4+CD8- population. These findings suggest that alpha E beta 7 is involved in the expansion/recruitment of TCR alpha beta+ CD8+ IEL following microbial colonization. Integrin alpha E-deficient mice will provide an important tool for studying the role of alpha E beta 7 and of alpha E beta 7-expressing mucosal T lymphocytes in vivo.
Subject(s)
Antigens, CD/genetics , Integrin alpha Chains , Intestinal Mucosa/immunology , Intestinal Mucosa/pathology , Lymphopenia/genetics , Lymphopenia/immunology , T-Lymphocyte Subsets/immunology , Animals , CD4-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/pathology , Chromosome Mapping , Cloning, Molecular , Crosses, Genetic , Epithelial Cells/immunology , Epithelial Cells/pathology , Female , Hyaluronan Receptors/biosynthesis , Intestinal Mucosa/metabolism , Lymphocyte Count , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Molecular Sequence Data , Muridae , Peyer's Patches/cytology , Peyer's Patches/immunology , Receptors, Antigen, T-Cell, alpha-beta/metabolism , Spleen/cytology , Spleen/pathology , T-Lymphocyte Subsets/metabolism , T-Lymphocyte Subsets/pathology , Vagina/immunology , Vagina/pathologyABSTRACT
Integrin receptors are important for regulating lymphocyte recirculation and recruitment to sites of inflammation. Transfectants of the B cell lymphoma 38C13 were generated that differ exclusively in the expression of integrin beta 1 or beta 7 subunits allowing for a functional comparison of lymphocyte Peyer's patch HEV adhesion molecule 1 (LPAM-1) (alpha 4 beta 7) and very late antigen 4 (VLA-4) (alpha 4 beta 1) in an identical cellular environment. Whereas 38-beta 7 transfectants bound to purified and cellular mucosal addressin cell adhesion molecule (MAdCAM-1), unstimulated 38-beta 1 cells failed to bind MAdCAM-1. Treatment of 38-beta 1 cells with Mn2+ but not with PMA induced low level binding to MAdCAM-1. MAdCAM-1 adhesion of 38-beta 7 cells was constitutive and not enhanced by Mn2+ treatment. Similarly, MAdCAM-1-dependent adhesion to mucosal high endothelial venules was shown for 38-beta 7 but not for 38-beta 1 cells. The results therefore establish the LPAM-1-MAdCAM-1 interaction as the functionally dominant adhesion pathway for regulating lymphocyte homing to mucosal sites. Nonetheless, the activated VLA-4 on some lymphocytes may be involved in MAdCAM-1 recognition or promote binding to MAdCAM-1 in other tissues. By contrast, 38-beta 7 and 38-beta 1 transfectants did not differ in their binding capacity for vascular cell adhesion molecule 1 (VCAM-1) or fibronectin and LPAM-1 did not display any preference for interacting with either MAdCAM-1 or VCAM-1. LPAM-1 may therefore contribute significantly to cellular functions previously attributed to VLA-4. Interestingly, functional analysis of the intraepithelial lymphocyte integrin alpha IEL beta 7 which is structurally related to LPAM-1 did not reveal detectable binding activity for MAdCAM-1, VCAM-1, or fibronectin.
Subject(s)
Cell Adhesion Molecules/metabolism , Integrin beta Chains , Integrins/metabolism , Animals , Base Sequence , Blotting, Western , CHO Cells , Cell Adhesion , Cricetinae , Fibronectins/metabolism , Immunoglobulins/metabolism , Mice , Molecular Sequence Data , Mucoproteins/metabolism , Precipitin Tests , Receptors, Very Late Antigen/metabolism , Tumor Cells, Cultured , Vascular Cell Adhesion Molecule-1ABSTRACT
The L-selectin adhesion receptor plays a central role in regulating leukocyte adhesion to endothelial cells. The data presented in this report demonstrate that triggering of L-selectin results in a rapid and vigorous homotypic adhesion among normal lymphocytes as well as lymphoblastoid cells, thereby providing evidence for a novel cell-cell adhesion function of L-selectin. The cellular adhesion event induced by mAb MEL-14 was dependent on metabolic energy, an intact cytoskeleton, and the activation of intracellular protein kinases. Cell clustering did not require cross-linking of L-selectin molecules and occurred in the complete absence of divalent cations. Analysis of adhesion receptor expression and antibody inhibition experiments indicated that cluster formation did not involve LFA-1, alpha 4 integrins, beta 1 integrins, beta 7 integrins, or CD44.