Experience of menopause in women with inflammatory bowel disease: pilot study.

OBJECTIVE: Inflammatory bowel diseases (IBD) are debilitating chronic intestinal diseases requiring extensive medical intervention. Little is known how IBD symptoms and treatments affect menopause experience and quality of life. The study's goal was to investigate the relationship between IBD and menopause. METHOD: Women with IBD, between the ages of 30 and 65 years, were recruited from an outpatient IBD clinic. They completed surveys on obstetric, medical, and IBD history and clinical disease activity. Quality of life was assessed using the validated menopause-specific quality of life (MENQOL) questionnaire. RESULTS: Seventy-one women (47 Crohn's disease, 22 ulcerative colitis, and two indeterminate colitis, median age 45 years) enrolled into the study. Younger age of IBD diagnosis was correlated with younger age of last menstrual period (r = 0.697). IBD severity affected menopause-related quality of life in three MENQOL domains (psychosocial, physical, and sexual); the fourth domain (vasomotor) did not appear to be affected by the severity of IBD clinical disease. CONCLUSION: Women with IBD may experience additional challenges when going through the menopause transition. Our findings support the need for further studies to better inform patients and clinicians on the relationship between IBD and menopause to optimize patient care.

Experience of menopause in aboriginal women: a systematic review.

Every woman experiences the menopause transition period in a very individual way. Menopause symptoms and management are greatly influenced by socioeconomic status in addition to genetic background and medical history. Because of their very unique cultural heritage and often holistic view of health and well-being, menopause symptoms and management might differ greatly in aboriginals compared to non-aboriginals. Our aim was to investigate the extent and scope of the current literature in describing the menopause experience of aboriginal women. Our systematic literature review included nine health-related databases using the keywords 'menopause' and 'climacteric symptoms' in combination with various keywords describing aboriginal populations. Data were collected from selected articles and descriptive analysis was applied. Twenty-eight relevant articles were included in our analysis. These articles represent data from 12 countries and aboriginal groups from at least eight distinctive geographical regions. Knowledge of menopause and symptom experience vary greatly among study groups. The average age of menopause onset appears earlier in most aboriginal groups, often attributed to malnutrition and a harsher lifestyle. This literature review highlights a need for further research of the menopause transition period among aboriginal women to fully explore understanding and treatment of menopause symptoms and ultimately advance an important dialogue about women's health care.

An imbalance in mucosal cytokine profile causes transient intestinal inflammation following an animal's first exposure to faecal bacteria and antigens.

Intestinal microflora play a critical role in the initiation and perpetuation of chronic inflammatory bowel diseases. In genetically susceptible hosts, bacterial colonization results in rapid-onset chronic intestinal inflammation. Nevertheless, the intestinal and systemic immune response to faecal bacteria and antigen exposure into a sterile intestinal lumen of a post-weaned animal with a mature immune system are not understood clearly. This study examined the effects of faecal bacteria and antigen exposure on the intestinal mucosal and systemic immune system in healthy axenic mice. Axenic wild-type mice were inoculated orally with a crude faecal slurry solution derived from conventionally raised mice and were analysed prior to and then at days 3, 7, 14 and 28 post-treatment. Ingestion of faecal slurry resulted in a transient, early onset of proinflammatory interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and interleukin (IL)-17 response that was maximal at day 3. In contrast, the transient release of the anti-inflammatory cytokines IL-10 and IL-4 occurred later and was maximal at day 7. Both responses subsided by day 14. This early cytokine imbalance was associated with a brief rise in colonic and caecal histopathological injury score at day 7. The bacterial antigen-specific systemic response was found to follow the intestinal immune response with a maximal release of both pro- and anti-inflammatory cytokines at day 7. Thus, first exposure of healthy axenic wild-type mice to normal faecal flora and antigens results in an early proinflammatory cytokine response and transient colonic inflammation that then resolves in conjunction with a subsequent anti-inflammatory cytokine profile.

VSL#3 probiotic upregulates intestinal mucosal alkaline sphingomyelinase and reduces inflammation.

BACKGROUND: Alkaline sphingomyelinase, an enzyme found exclusively in bile and the intestinal brush border, hydrolyzes sphingomyelin into ceramide, sphingosine and sphingosine-1-phosphate, thereby inducing epithelial apoptosis. Reduced levels of alkaline sphingomyelinase have been found in premalignant and malignant intestinal epithelia and in ulcerative colitis tissue. Probiotic bacteria can be a source of sphingomyelinase. OBJECTIVE: To determine the effect of VSL#3 probiotic therapy on mucosal levels of alkaline sphingomyelinase, both in a mouse model of colitis and in patients with ulcerative colitis. METHODS: Interleukin-10 gene-deficient (IL10KO) and wild type control mice were treated with VSL#3 (10(9) colony-forming units per day) for three weeks, after which alkaline sphingomyelinase activity was measured in ileal and colonic tissue. As well, 15 patients with ulcerative colitis were treated with VSL#3 (900 billion bacteria two times per day for five weeks). Alkaline sphingomyelinase activity was measured through biopsies and comparison of ulcerative colitis disease activity index scores obtained before and after treatment. RESULTS: Lowered alkaline sphingomyelinase levels were seen in the colon (P=0.02) and ileum (P=0.04) of IL10KO mice, as compared with controls. Treatment of these mice with VSL#3 resulted in upregulation of mucosal alkaline sphingomyelinase activity in both the colon (P=0.04) and the ileum (P=0.01). VSL#3 treatment of human patients who had ulcerative colitis decreased mean (+/- SEM) ulcerative colitis disease activity index scores from 5.3+/-1.8946 to 0.70+/-0.34 (P=0.02) and increased mucosal alkaline sphingomyelinase activity. CONCLUSION: Mucosal alkaline sphingomyelinase activity is reduced in the intestine of IL10KO mice with colitis and in humans with ulcerative colitis. VSL#3 probiotic therapy upregulates mucosal alkaline sphingomyelinase activity.

beta7 Integrin expression is not required for the localization of T cells to the intestine and colitis pathogenesis.

beta7 Integrins have been shown to have an important role in the localization of T cells to the intestine. Utilizing two different experimental mouse models of inflammatory bowel disease (IBD), this study was undertaken to determine if beta7 integrin expression is critical for T cell localization to the intestine and colitis pathogenesis. Transfer of CD4+ CD45RBhigh cells into immunodeficient mice results in colitis. To examine the role of beta7 integrins, donor cells were obtained from beta7 integrin gene-deficient animals and disease induction was examined following transfer into severe combined immunodeficiency (SCID) mice. Additionally, beta7 integrin gene-deficient animals were crossed to IL-2-deficient mice and the onset of spontaneous colitis that normally occurs in IL-2-deficient animals was examined. No differences in the onset or severity of spontaneous colitis was noted in animals that were deficient in both beta7 integrin and IL-2. In contrast, the onset of colitis in recipients of T cells from beta7 integrin-deficient donors was delayed significantly. In mice receiving beta7 integrin negative cells, the initial lack of colitis appeared to correlate with fewer numbers of CD3+beta7 integrin -/- donor lymphocytes present in the host colon. The eventual development of disease, however, was associated with increased numbers of donor beta7 integrin -/- lymphocytes. These results show that beta7 integrin expression is not absolutely required for T cell localization to the intestine and colitis pathogenesis.

Systemic activation and antigen-driven oligoclonal expansion of T cells in a mouse model of colitis.

Transfer of CD4+CD45RBhigh T cells into immunodeficient mice results in both the expansion of the transferred T cells and colitis. Here we show that colitis pathogenesis requires expression of MHC class II molecules by the immune-deficient host. Analysis of the TCRbeta repertoire of the cells found in the large intestine of diseased mice revealed a population with restricted TCR diversity. Furthermore, nucleotide sequence analysis demonstrated the selection for particular CDR3beta amino acid sequence motifs. Collectively, these data indicate that the expansion of T cells in the intestine and colitis pathogenesis are likely to require the activation of Ag-specific T cells, as opposed to nonspecific or superantigen-mediated events. There is relatively little overlap, however, when the TCR repertoires of different individuals are compared, suggesting that a number of Ags can contribute to T cell expansion and the generation of a T cell population in the intestine. Surprisingly, many of the expanded clones found in the large intestine also were found in the spleen and elsewhere, although inflammation is localized to the colon. Additionally, donor-derived T cells appear to be activated in both the intestine and the spleen at early time points after cell transfer. Together, these results strongly suggest that disease induction in this model involves either the early and systemic activation of antigen-specific T cells or the rapid dispersal of T cells activated at a particular site.

An opposite pattern of selection of a single T cell antigen receptor in the thymus and among intraepithelial lymphocytes.

The differentiation of intestinal intraepithelial lymphocytes (IEL) remains controversial, which may be due in part to the phenotypic complexity of these T cells. We have investigated here the development of IEL in mice on the recombination activating gene (RAG)-2(-/-) background which express a T cell antigen receptor (TCR) transgene specific for an H-Y peptide presented by Db (H-Y/Db x RAG-2(-) mice). In contrast to the thymus, the small intestine in female H-Y/Db x RAG-2(-) mice is severely deficient in the number of IEL; TCR transgene+ CD8alphaalpha and CD8alphabeta are virtually absent. This is similar to the number and phenotype of IEL in transgenic mice that do not express the Db class I molecule, and which therefore fail positive selection. Paradoxically, in male mice, the small intestine contains large numbers of TCR+ IEL that express high levels of CD8alphaalpha homodimers. The IEL isolated from male mice are functional, as they respond upon TCR cross-linking, although they are not autoreactive to stimulator cells from male mice. We hypothesize that the H-Y/Db TCR fails to undergo selection in IEL of female mice due to the reduced avidity of the TCR for major histocompatibility complex peptide in conjunction with the CD8alphaalpha homodimers expressed by many cells in this lineage. By contrast, this reduced TCR/CD8alphaalpha avidity may permit positive rather than negative selection of this TCR in male mice. Therefore, the data presented provide conclusive evidence that a TCR which is positively selected in the thymus will not necessarily be selected in IEL, and furthermore, that the expression of a distinct CD8 isoform by IEL may be a critical determinant of the differential pattern of selection of these T cells.

Generation of intestinal mucosal lymphocytes in SCID mice reconstituted with mature, thymus-derived T cells.

Transfer of peripheral lymph node lymphocytes to SCID mice leads to the long term establishment of mucosal T lymphocytes within the epithelium and lamina propria of the small and large intestines. Analysis of engrafted intraepithelial lymphocytes (IEL) showed that they had acquired a surface phenotype that in several respects is typical of IEL. In addition, the functional profile of engrafted IEL derived from lymph node T cells was similar to that of normal IEL; as the donor-derived T cells exhibited a strong cytolytic activity, a poor proliferative response to mitogenic stimuli, and a tendency to home and expand specifically in the intestine upon transfer to secondary SCID recipients. Optimal engraftment of intestinal T cells required bacterial flora, as the number of lymphocytes was greatly reduced in SCID recipients with a reduced flora. These results demonstrate that mature, thymus-derived T cells can migrate to the intestine and become functionally specialized to the intestinal milieu. The acquisition of phenotypic markers characteristic of the intestinal microenvironment by engrafted cells suggests that T cell migration of lymphocytes to the SCID intestine is not aberrant, but it may reflect processes that are ongoing in immunocompetent mice. Furthermore, these data suggest that the homing and/or expansion of typical, thymus-derived T cells in the intestine may be driven by luminal Ags such as those derived from bacterial flora.

Mouse CD1 is mainly expressed on hemopoietic-derived cells.

The mouse CD1 (mCD1) is a class I-like molecule that is encoded outside the MHC. Recent studies demonstrate that mCD1 presents hydrophobic peptides to CD8+ T cells and also that it is recognized by a population of NK1.1+ T cells that are thought to play an immunoregulatory role because of their ability to secrete IL-4. It has previously been reported that mCD1 is expressed predominantly by intestinal epithelial cells, although most NK1.1+ T cells are located elsewhere. We, therefore, have generated new mAbs to mCD1 to investigate its tissue distribution. The principal site of mCD1 expression in normal mice is on cells in the hemopoietic series, including constitutive expression on nearly all T and B cells, on macrophages, and on dendritic cells. Other than bone marrow-derived cells, mCD1 is not widely expressed and is not detectable on great majority of intestinal epithelial cells. The B cells, but not the T cells, from beta2m-deficient mice can be recognized by two mCD1 autoreactive T hybridomas. Therefore, although we could not detect a beta2m-independent form of mCD1 using these mAbs, mCD1 in a different conformation or a mCD1-related molecule is likely to be expressed in the absence of beta2m on some cell types. The pattern of expression of mCD1 correlates with the distribution of NK1 T cells and is consistent with an important Ag-presenting function for this molecule.

Analysis of intestinal lymphocytes in mouse colitis mediated by transfer of CD4+, CD45RBhigh T cells to SCID recipients.

Transfer of specific T lymphocyte subsets isolated from the spleens of healthy donor mice into immunodeficient SCID mice leads to chronic intestinal inflammation with characteristics similar to those of human inflammatory bowel disease (IBD). CD4+, CD45RBhigh cells cause disease, whereas CD4+, CD45RBlow and CD8+, CD45RBhigh cells do not. Despite this difference, we demonstrate that all three T cell populations reconstitute the intraepithelial and lamina propria compartments of both small and large intestines of SCID recipients. Therefore, infiltration of lymphocytes alone is not sufficient for disease development. CD4+ lymphocytes that have trafficked to the SCID intestine exhibit a phenotype characteristic of normal mucosal lymphocytes. This includes high expression of alpha E integrin and CD69, expression of CD8 alpha alpha homodimers in some of the intraepithelial lymphocytes, as well as low expression of CD62L and CD45RB. The phenotype of the infiltrating mucosal cells is indistinguishable, with respect to the cell surface markers tested, regardless of whether the starting donor population is CD45RBhigh or CD45RBlow. Severe inflammation is restricted primarily to the colon despite lymphocyte infiltration throughout the length of the intestine. This suggests that some property of the colon microenvironment contributes to inflammation. Consistent with this, transfer of CD4+, CD45RBhigh cells to SCID mice that have significantly reduced numbers of enteric flora results in attenuation of the wasting and colitis. Fewer numbers of donor lymphocytes are recovered from the intraepithelial and lamina propria compartments of reduced flora SCID mice. We hypothesize that the ability of pathogenic cells to traffic to the intestine and mediate colitis may be driven by T cell reactivity to bacteria or bacterial products.

TAP-independent selection of CD8+ intestinal intraepithelial lymphocytes.

Intestinal intraepithelial lymphocytes (IEL) are mostly CD8 single positive T cells. IEL with a TCR-alpha(beta) that are CD8 single positive are absent from beta(2)-microglobulin (beta(2)m)-deficient mice, consistent with the idea that these IEL, like other TCR-alpha(beta)+, CD8+ T cells, require class I molecules for positive selection. In contrast, here we show that substantial numbers of TCR-alpha(beta)+, CD8 single positive IEL are present in mice deficient for the transporter associated with Ag processing 1 (TAP 1) gene, although T cells with this phenotype are absent from thymus, spleen, and lymph nodes of these same mice. The majority of TCR-alpha(beta)+, CD8 single positive IEL in TAP-deficient mice expresses CD8 molecules composed of alpha(alpha) homodimers and they express a diverse set of V(beta) gene segments. In addition, the number of TCR-alpha(beta)+, CD4/CD8 double positive IEL is decreased in beta(2)m-deficient mice but not in TAP-deficient mice. The dependence of the two TCR-alpha(beta)+ IEL populations that express CD8alpha(alpha) homodimers on beta(2)m as opposed to TAP molecules is striking. It suggests that TAP-independent but beta(2)m-requiring nonclassical class I molecules expressed by cells in the intestine, such as the thymus leukemia Ag and CD1, could play a pivotal role in the development and/or the accumulation of major subpopulations of TCR-alpha(beta)+ IEL.

Intestinal intraepithelial lymphocytes respond to systemic lymphocytic choriomeningitis virus infection.

Intestinal intraepithelial lymphocytes (IEL) are a population of cells consisting mostly of CD8+ T lymphocytes. Although their function is unknown, because of their location within the epithelium it has been postulated that IEL may be involved in defense against infection of the gut mucosa by pathogens including viruses. To address this issue, we have examined IEL populations from BALB/c mice systemically infected with lymphocytic choriomeningitis virus (LCMV). Viral infection induced a virus-specific cytotoxic response by IEL at 8 days postinfection. This virus-specific cytotoxic T lymphocyte (CTL) response was MHC class I restricted, and as is true for splenic T cells, recognition of viral antigen occurred predominantly in the context of the Ld molecule. The effector cells could be depleted by treatment with anti-CD8 antibody plus complement. In vivo treatment of mice with anti-alpha beta T cell receptor (TCR) antibody during the course of viral infection abrogated the response, suggesting that the virus-specific CTL were cells that express the alpha beta rather than gamma delta TCR. Consistent with this, no virus-specific IEL response could be detected in athymic mice, which have TCR gamma delta+ but not TCR alpha beta+ IEL. LCMV antigen could not be detected in the epithelium of the intestine, suggesting that viral antigen may have been encountered elsewhere. These data demonstrate for the first time a specific response by IEL to virus given by a non-oral route, and they suggest that thymus-derived alpha beta T cells can migrate to the intestinal epithelium following activation, where they may play a role in the response to virus and perhaps other infections.

T-cell receptor gamma delta diversity and specificity of intestinal intraepithelial lymphocytes: analysis of IEL-derived hybridomas.

The phenotype, T-cell antigen receptor (TCR) usage and specificity of intestinal intraepithelial lymphocytes (IEL), and hybridomas derived from IEL have been examined. In young conventionally reared mice, approximately 50% of the IEL express TCR alpha beta and 50% express TCR gamma delta, although there is considerable variation between individuals. Here we demonstrate that T-cell hybridomas can be prepared from freshly isolated BALB/c mouse IEL. The expressed TCR of these hybridomas reflects the TCR isotype distribution of the IEL population. Analysis of the gamma delta TCR expressed by the hybridomas demonstrates that IEL express a greater number of TCR V gene segments than has been reported for gamma delta T cells in several other epithelial sites. At least five types of gamma delta TCRs are expressed by a panel of BALB/c IEL hybridomas, although use of the gamma delta TCR V gene repertoire clearly is not random. Some TCR gamma delta cells and gamma delta hybridomas have been reported to recognize purified protein derivative (PPD); however, none of the IEL hybridomas secreted IL-2 in response to PPD. These data suggest that most TCR gamma delta IEL are not likely to be PPD reactive.

Intestinal intraepithelial lymphocytes preferentially repopulate the intestinal epithelium.

We have used C.B-17 severe combined immune deficiency (SCID) mice to study the repopulation of intestinal intraepithelial cells in these mice. We have found that intestinal intraepithelial lymphocytes (IELs) injected into SCID mice preferentially repopulate the intestinal epithelium. About 5 weeks after injection we can detect significant numbers of IEL in repopulated SCID mice. Repopulation occurs in approximately 70% of the injected mice and the amount of recovered cells per mouse is variable. The recovered cells are of donor-type origin and exhibit a typical IEL phenotype. The donor-type T lymphocytes that can sometimes be found in other organs of IEL-repopulated SCID mice are generally of low number. They are not stained with antibodies against IEL-specific markers and their phenotypes appear to be more typical for T cells normally found in these sites. In contrast, the intestinal epithelium of SCID mice cannot be efficiently repopulated with lymphocytes using cells of other organs including thymocytes, Peyer's patch lymphocytes, and bone marrow cells. From our data we conclude that intestinal IELs are confined to the intestinal epithelium and possibly contain a precursor-type cell that preferentially regenerates cells of its own population.

Intestinal intraepithelial lymphocytes are activated and cytolytic but do not proliferate as well as other T cells in response to mitogenic signals.

Compared with T lymphocytes from other organs, intestinal intraepithelial lymphocytes (IEL) proliferate weakly in response to CD3/TCR ligation, and they do not respond at all to treatment with other mitogenic stimuli. These signals also failed to induce expression of the IL-2R alpha-chain on the surface of most IEL. IEL from germ-free mice, from V gamma 1.1-transgenic mice, and from beta 2-microglobulin-deficient mice also gave a weak proliferative response. Therefore, the low proliferative response is not linked to the level of exposure to gut bacterial flora, the V gamma region expressed by the TCR-gamma delta + IEL, or the presence of class I molecules that may be recognized by CD8+ IEL. The relatively small amount of proliferation in response to TCR signaling, therefore, is not likely to be the result of induction of anergy caused by previous contact with Ag. In contrast, ligation of the CD3/TCR complex could elicit a rapid cytotoxic response and serine esterase release by IEL. The unusual functional capabilities and the activation state of IEL are independent of the TCR isotype expressed by these cells. Freshly isolated IEL have a high intracellular microtubule-associated protein kinase-2 (MAP-2K) activity level, further suggesting that these cells are activated despite their weak proliferative response. Consistent with this, MAP-2K is tyrosine-phosphorylated in both untreated and PMA-treated IEL. In contrast, MAP-2K activation and tyrosine phosphorylation occur in other T cells only when they are activated by PMA or other treatments. MAP-2K activity also is elevated in IEL from germ-free mice, demonstrating that activation does not depend on normal levels of exposure to bacterial flora. The activation of protein kinases such as MAP-2K could reflect the differentiation state of IEL or Ag receptor stimulation of some of these cells by epithelial cells in the preparation.

Depletion of mouse alpha beta T cell antigen receptor bearing lymphocytes by neonatal monoclonal antibody treatment.

Neonatal treatment with a monoclonal antibody specific for the alpha beta TCR results in mice with a long term, severe depletion in the number of alpha beta T cells in the periphery. Significant numbers of T cells reappear in the periphery about age 65 days, but these cells tend to lack expression of CD4 or CD8. Splenocytes of antibody-treated mice are less sensitive to mitogen stimulation or stimulation with MHC allogeneic cells. The level of serum IgG but not IgM was decreased by the treatment. Anti-alpha beta TCR antibody treatment decreased single-positive T lymphocytes that express high levels of the CD3/alpha beta TCR complex from the thymus, suggesting that the treatment could act in part by affecting negative selection of alpha beta TCR+ thymocytes. This treatment does not, however, detectably affect either the homing or the numbers of gamma delta T cells which are abundant in the intestinal epithelium, but which remain a minor population in the spleen and lymph nodes. This supports the hypothesis that gamma delta T cells are developmentally autonomous from alpha beta T cells. These mice provide an excellent model system for assessing the developmental and functional role of gamma delta T lymphocytes in vivo.

Characterization of a CD4-positive T-cell line derived from an athymic (nu/nu) mouse.

We have isolated a Thy-1+, CD3+, CD4+ T-cell line from the spleen of a 12-week-old nu/nu (nude) BALB/c mouse. The cell line is clonal, and it expresses an alpha beta T-cell antigen receptor. Upon activation, these cells secrete IL-2 but not IL-4, putting them in the Th1 category. The cells can be triggered to proliferate and secrete lymphokines in the presence of irradiated syngeneic or allogeneic splenic feeder cells that express a variety of MHC haplotypes. This response is MHC class II-specific, because it can be blocked by either anti-Ia or anti-CD4 antibodies. From the response pattern of this T-cell line, we conclude that it recognizes a common determinant on class II MHC antigens. This nude mouse T-lymphocyte presumably has not undergone thymic selection. Therefore its unique specificity may reflect both the bias of T-cell antigen receptor genes for encoding receptors that recognize MHC molecules and the requirement for functional thymic epithelial cells for the efficient education of a self-MHC-restricted repertoire.