Phenotype of exogenous microparticle-containing pigment cells of the human Peyer’s patch in inflamed and normal ileum

OBJECTIVES: Pigmented cells, that contain inert, submicron-sized dietary particles, are a consistent feature of the base of human Peyer's patches (PP). We aimed (i) to phenotype these intestinal pigment cells (PC) in archival tissue specimens and (ii) to establish whether PC phenotype is altered in inflammatory conditions, especially Crohn's disease (CD). METHODS: PCs contained within PP were identified by routine haematoxylin and eosin (H&E) staining and dark field microscopy of archival ileal sections for: adenocarcinoma (n = 16), colonic CD (n = 23), non-CD colitis (n = 10). Paraffin-embedded serial sections were graded for microscopic inflammation and then investigated immunohistochemically with antibodies against CD68, MAC387, CD14, CD11b, CD15, CD1a, S100, HLA-DR, CD86 and Cathepsin D. Analyses were by light and confocal microscopies. RESULTS: The majority of PCs were CD68 positive (circa 80%) with a minority (circa 20%) staining for MAC387. Microparticles were mainly identified within cathepsin D negative lysosomal compartments. Histological inflammatory grade and disease type had no influence on cell phenotype. CONCLUSIONS: The microparticle-containing PCs of the PP base are mainly mature macrophages (CD68) of low metabolic and immunological activity. There is no evidence of differential PC phenotype or activation in differing disease states, including CD.     

Immunohistochemical localization of prolactin receptor (PRLR) to Hodgkin’s and Reed-Sternberg cells of Hodgkin’s lymphoma

Prolactin receptor (PRLR), a type-1 cytokine receptor, is overexpressed in a number of cancer types. It has attracted much attention for putative pro-oncogenic roles, which however, remains controversial in some malignancies. In this study, we reported the localization of PRLR to the Hodgkin’s and Reed-Sternberg (HRS) cells of Hodgkin’s lymphoma (HL), a neoplasm of predominantly B cell origin. Immunohistochemistry performed on 5-μm thick FFPE sections revealed expression of PRLR in HRS cells. Cellular immunofluorescence experiments showed that the HL-derived cell lines, Hs445, KMH2 and L428 overexpressed PRLR. The PRLR immunofluorescent signal was depleted after treating the cell lines with 10 μM of siRNA for 48 h. We also tested whether PRLR is involved in the growth of HL, in vitro. One-way analysis of variance (ANOVA) on cell growth data obtain from WST-1 cell proliferation assay and trypan blue exclusion assay and hemocytometry showed that siRNA-depletion of PRLR expression resulted in decreased growth in all three cell lines. These results offered only a short insight into the involvement of PRLR in HL. As a result, further investigation is required to decipher the precise role(s) of PRLR in the pathogenesis of HL.     

Age-Related Features of Expression of Cell Renewal Factors in the Intestinal Peyer’s Patches

Immunohistochemical study with markers of proapoptotic protein P53 and proliferation protein Ki-67 revealed an age-related decrease in proliferative activity of lymphocyte in the intestinal Peyer’s patches and an increase in the ratio of apoptotic cells in humans. These changes aggravate atrophy and involution of the intestinal epithelium during aging.     

β2-Microglobulin-free HLA-G activates natural killer cells by increasing cytotoxicity and proinflammatory cytokine production

Human leukocyte antigen-G (HLA-G) is a nonclassical HLA class-I molecule and plays a role in tissue specific immunoregulation. Many studies have addressed functional aspects of β2-microglobulin (β2m)-associated HLA-G1. β2m-free HLA-G has been found in human placental cytotrophoblasts and pancreatic β cells although its function remains unclear. In the present study, we investigated the function of β2m-free HLA-G by transfecting HLA-G1 and -G3 into human β2m deficient rat pancreatic β cell carcinoma (BRIN-BD11) cells. RT-PCR and western blots studies confirmed high expression of HLA-G1 and -G3 in -G1 and -G3 transfectants, respectively. HLA-G1 and -G3 were detected mainly in intracellular compartments of BRIN-BD11 transductants by confocal fluorescent microscopy and flow cytometry. Functional analysis revealed that β2m-free HLA-G promoted xenogeneic cytotoxic lysis of BRIN-BD11 cells by natural killer (NK) cells and increased production of IL-1β, TNF-α, and IFN-γ. Stimulation of cytotoxic lysis was impaired by blocking the MAPK and DNA-PKcs pathways in NK cells. Importantly, treatment with 33mAb, a KLR2DL4 receptor agonist, induced NK-mediated cytotoxic lysis of BRIN-BD11 cells transfected with a mock vector. Our data suggest that β2m-free HLA-G activates NK cells via engagement of KLR2DL4 receptors.     

The differential effects of IL-1 and TNF-α on proinflammatory cytokine and matrix metalloproteinase expression in human chondrosarcoma cells

Objective and design:Interleukin-1 (IL-1), tumor necrosis factor- (TNF-), and matrix metalloproteinases (MMPs) play important roles in the pathogenesis of osteoarthritis (OA). In the present study, using Affymetrix oligonucleotide array technology and real-time quantitative RT-PCR we have investigated the molecular mechanisms underlying the differential effect of IL-1 and TNF- on gene expression in the human chondrosarcoma cell line, SW1353. Materials and methods:SW1353 cells were stimulated singularly with IL-1, TNF-, Phorbol 12-myristate 13-acetate (PMA), or treated with the combination of cytokine and PMA. Total RNA was collected at multiple time points over a 24-h period followed by biotinylated cRNA target preparation and hybridization onto the Affymetrix HG-U95Av2 array. The differential expression patterns of several cytokine and MMP genes were further confirmed by real time quantitative RT-PCR, Western blot, and ELISA. Results:Our microarray experiments have broadly confirmed previously published data on chondrocyte gene expression regulated by IL-1 and TNF-. The expression pattern of proIL-1, MMP-1, and MMP-13 in chondrocytes is differentially regulated when stimulated with proinflammatory cytokines. IL-1, but not TNF-, can induce IL-6, bone morphogenic protein 2 (BMP-2), and cyclooxygenase (COX-2) expression in SW1353 cells. Additionally, our Western blot results provide the first evidence that IL-1 is produced in the proform in IL-1-activated chondrosarcoma cells and that additional signals are required for its posttranslational processing/activation. Conclusions:IL-1 and TNF- each activate a distinct set of genes in chondrosarcoma cells, and gene expression in these cells is regulated by groups of genes related in part by their function. Chondrocyte IL-1 appears to serve an important role in the pathogenesis OA contributing to joint inflammation and cartilage destruction.Received 15 September 2003; returned for revision 16 October 2003; accepted by J. S. Skotnicki 11 March 2004     

Proliferation and apoptosis of Peyer’s patches and its lymphocytes in experimental terminal ileitis

This study will provide guide for the terminal ileitis in clinical diagnosis and treatment. The animals were been done terminal ileum-cecum side to side anastomosis, terminal ileum operation line and only anesthesia treatment, respectively. The model group presented acute inflammation after surgery for 2 weeks and the inflammation was limited to the mucosal layer. Animals presented chronic inflammation to 8 weeks, mucosal membrane was given priority to with lymphocytic infiltrates. In 2 weeks and 4 weeks, the number of Peyer’s patches (PP knot) and PP knot lymphocytes increased significantly in the model group (P < 0.05, P < 0.01). At 8 weeks, the suture group and the model group presented a large number of lymphocytic apoptosis (P < 0.01). Rat ileal PP knot lymphocyte small molecule DNA showed typical “trapezoid” bands. We observed apparent morphology of apoptosis and crescent-shaped nucleus. Continuous immune response in terminal ileitis plays a considerable role in the process of the disease.     

Chrysin suppresses LPS-stimulated proinflammatory responses by blocking NF-κB and JNK activations in microglia cells

Neuroinflammation mediated by microglia has been implicated in neurodegenerative diseases. Suppression of microglial activation may therefore contribute to neuronal cell survival. Chrysin is present in honey and propolis and in low concentrations in fruits, vegetables, and certain beverages. It has been reported that chrysin has potent anti-inflammation, anti-cancer, and anti-oxidation properties. In the present study, we investigated the effects of chrysin on the production of proinflammatory mediators in lipopolysaccharide (LPS)-stimulated microglia. Chrysin significantly inhibited the release of nitric oxide (NO) and proinflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). The expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were also significantly inhibited by chrysin. Furthermore, chrysin inhibited the activations of c-Jun N-terminal kinase (JNK) and nuclear factor-κB (NF-κB), which are signaling molecules involved in neuroinflammation. These results suggest that chrysin may act as a potential therapeutic agent for various neurodegenerative diseases involving neuroinflammation.     

Patients’ understanding of and responses to multiplex genetic susceptibility test results

PurposeExamination of patients’ responses to direct-to-consumer genetic susceptibility tests is needed to inform clinical practice. This study examined patients’ recall and interpretation of, and responses to, genetic susceptibility test results provided directly by mail.MethodsThis observational study had three prospective assessments (before testing, 10 days after receiving results, and 3 months later). Participants were 199 patients aged 25–40 years who received free genetic susceptibility testing for eight common health conditions.ResultsMore than 80% of the patients correctly recalled their results for the eight health conditions. Patients were unlikely to interpret genetic results as deterministic of health outcomes (mean = 6.0, s.d. = 0.8 on a scale of 1–7, 1 indicating strongly deterministic). In multivariate analysis, patients with the least deterministic interpretations were white (P = 0.0098), more educated (P = 0.0093), and least confused by results (P = 0.001). Only 1% talked about their results with a provider.ConclusionFindings suggest that most patients will correctly recall their results and will not interpret genetics as the sole cause of diseases. The subset of those confused by results could benefit from consultation with a health-care provider, which could emphasize that health habits currently are the best predictors of risk. Providers could leverage patients’ interest in genetic tests to encourage behavior changes to reduce disease risk.Genet Med advance online publication 5 April 2012     

Systemic cytokine response in humans with chiclero’s ulcers

In this work, we report the existence of enhanced concentrations of IL-2 (Th1 cytokine), TNF-α (mainly a Th1 cytokine), IL-12p40, a strong delayed type hypersensitivity response, and the absence of a serum-specific antibody response (IgG or IgE) in patients with active chiclero’s ulcers. There was a low serum concentration of IL-6 (a Th2 cytokine) that was further reduced once patients healed following antimonial therapy (i.e., 4 months after treatment started). There was an absence of serum IL-4 (a Th2 cytokine) in patients before and after healing. The serum content of nitric oxide, transforming growth factor-β1, and IL-12p70 in patients with progressive chiclero’s ulcers was not different from that of controls. Patients had an alteration in blood-cell counts and showed a poor T cell proliferation after in vitro stimulation with Leishmania mexicana gp63. We concluded that patients with progressive chiclero’s ulcers develop a Th1 type of response at this stage of the disease which correlates with increases in circulating eosinophils and B cells and a reduction of circulating CD8⁺ T lymphocytes.     

Post-mortem Peyer’s patches: Their potential application in forensic medicine

Pro-inflammatory mediators hold important functions in human body in response to infection, trauma and vascular disease. However, their action is down regulated by the release of anti-inflammatory cytokines, thus restoring a balance which reflects the immune status of a given individual.

Recent studies have stressed out the importance of circulating levels of cytokines for forensic purposes even if there is a lack of studies regarding the role of post-mortem mucosa-associated lymphoid tissue. In this respect, Peyer’s patches (PP), represent one of the most important immunological site of the body and the major component of the gut -associated lymphoid tissue.

The aim of this study was to evaluate post-mortem PP immune response in 40 serial autopsy cases of people who died from natural and traumatic death. The study examined spontaneous release of the following cytokines by fresh isolated PP cells: interleukin (IL)-12, tumor necrosis factor (TNF)-α, IL-10, IL-6, IL-1β, and IL-8. Results will show that higher levels of TNF-α, IL-6, IL-1β, and IL-8 are statistically correlated with the traumatic death group. From a forensic point of view these data demonstrate that fundamental lymphoid organs, such as PP, may have a potential in diagnosing the cause of death.     

What’s self got to do with it: Sources of heterogeneity among naive T cells

There is a long-standing assumption that naive CD4⁺ and CD8⁺ T cells are largely homogeneous populations despite the extraordinary diversity of their T cell receptors (TCR). The self-immunopeptidome plays a key role in the selection of the naive T cell repertoire in the thymus, and self-peptides are also an important driver of differences between individual naive T cells with regard to their subsequent functional contributions to an immune response. Accumulating evidence suggests that as early as the β-selection stage of T cell development, when only one of the recombined chains of the mature TCR is expressed, signaling thresholds may be established for positive selection of immature thymocytes. Stochastic encounters subsequently made with self-ligands during positive selection in the thymus imprint functional biases that a T cell will carry with it throughout its lifetime, although ongoing interactions with self in the periphery ensure a level of plasticity in the gene expression wiring of naive T cells. Identifying the sources of heterogeneity in the naive T cell population and which functional attributes of T cells can be modulated through post-thymic interventions versus those that are fixed during T cell development, could enable us to better select or generate T cells with particular traits to improve the efficacy of T cell therapies.     

Microglial cells and Parkinson’s disease

Chronic inflammation mediated by microglial cells is the fundamental process contributing to the death of dopamine (DA)-producing neurons in the brain. Production of inflammatory products by these microglial cells characterizes the slow destructive process in Parkinson’s disease (PD). The activation of microglial cells and the generation of pro-inflammatory cytokines that characterize PD are mediated by several different signaling pathways, with the activation of the respiratory burst by microglial cells being a critical event in the ultimate toxicity of DA-neurons. The work on our lab is concerned with understanding the mechanisms of activation, response, and therapeutic targets of microglial cells, with the aim to provide more effective treatments for PD and other inflammatory diseases of the CNS.     

The effects of bisphenol A on some plasma cytokine levels and distribution of CD8+ and CD4+ T lymphocytes in spleen,ileal Peyer’s patch and bronchus associated lymphoid tissue in rats

The effects of bisphenol A on the some plasma cytokine levels and distribution of CD8⁺ and CD4⁺ T lymphocytes in spleen, ilealPeyer’s patch and bronchus-associated lymphoid tissue in rats were investigated. A total of fourty male Wistar Albino rats were divided into five groups including 8 rats in each one: control, vehicle, BPA 5, BPA 50 and BPA 500 groups. Doses of 5, 50 and 500?μg/kg BPA were dissolved in ethanol, then mixed with corn oil. The control group received no treatment. The vehicle group was given the ethanol-corn oil mixture. BPA 5, BPA 50 and BPA 500 groups were given, respectively, 5, 50, and 500?μg/kg/day orally. In blood samples, IL-4, IL-6, IL-10 and TNF-α plasma levels were determined with ELISA. Tissue samples (spleen, ileal Peyer’s patches and lung) were processed by means of routine histological techniques. CD4 and CD8 were stained immunohistochemically. Data obtained from this study showed that, BPA causes the alteration on immune parameters including cytokine profile, distribution of CD8⁺ and CD4⁺ T lymhpocytes in spleen and ileal Peyer’s patches. Present study indicated that BPA may affect immune systems even at lower doses.Disruption of immun system cells and cytokine levels can result in harmful outcomes triggering autoimmune diseases and immunodeficiencies.     

Postmortem morphology and viability of human Peyer’s Patches in distal ileum: A technical note

The intestinal mucosa contains a highly specialized immune system which plays a central role in the induction of immune reactions. In the small bowel, Gut-Associated Lymphoid Tissue (GALT) is organized in lymphoid aggregates which are known as Peyer’s Patches (PP). Even though human PP involvement in systemic immunity has been described, little is known about their anatomy and morphology and viability. The aim of this study was to examine PP according to their macroscopic anatomy, distribution and cell viability after death. Specimens from the distal ileum were obtained from 72 serial autopsy cases: PP were identified and, parts of them were analyzed for histological examination. Moreover, viability of recovered PP cells was assessed by the trypan blue exclusion test. Most of the PP (90%) were situated on the antimesenteric border of ileum, and the greatest density of PP occurred in the most distal segment. The number of PP varied with age, with the maximum number observed in 21- to 30-years old cadavers. Histological examination showed their remarkable architectural preservation at different post-mortem intervals (PMI), while the mucosal surface underwent autolysis. In 56% of cases PP cells were still viable, especially at PMI < 24 hours after death. These data confirm that human PP are still well preserved in a remarkable percentage of cadavers also several hours after death, and their availability may be helpful in various fields of research.     

Differential proteomics analysis of mononuclear cells in cerebrospinal fluid of Parkinson’s disease

Parkinson’s disease (PD) is one common neurodegenerative disease featured with degeneration of dopaminergic neurons in substantia nigra. Multiple factors participate in the pathogenesis and progression of PD. In this study, we investigated the proteomics profiles of mononuclear cells in cerebrospinal fluids from both PD patients and normal people, in order to explore the correlation between disease factors and PD. Cerebrospinal fluid samples were collected from both PD and normal people and were separated for mononuclear cells in vitro. Proteins were then extracted and separated by 2-dimensional gel electrophoresis. Proteins with differential expressions were identified by comparison to standard proteome expression profile map, followed by software and database analysis. In PD patients, there were 8 proteins with consistent expression profile and 16 proteins with differential expressions. Those differential proteins identified include cytoskeleton proteins (actin, myosin), signal transduction proteins (adenosine cyclase binding protein 1, calcium binding protein, talin) and anti-oxidation factor (thioredoxin peroxide reductase). PD patients had differential protein expressional profiles in the mononuclear cells of cerebrospinal fluids compared to normal people, suggesting the potential involvement of cytoskeleton and signal transduction proteins in apoptosis of neuronal apoptosis and PD pathogenesis.     

Developmental study on the ileal Peyer’s patches of sheep,and cytokeratin-18 as a possible marker for M cells in follicle associated epithelium

Peyer's patches are known as the immune sensors of the intestine because of their ability to transport luminal antigens. The objective of this study was both to assess the prenatal and postnatal development of sheep ileal Peyer’s patches with respect to histomorphology, distribution of CD4⁺ and CD8⁺ cells, and localization of proliferating and apoptotic cells, and to examine the morphology of M cells and expression of CK18 in follicle associated epithelium (FAE). We also hypothesized that CK18 could be a potential marker for M cell. Peyer’s patches completed their histomorphological development in prenatal period and involuted in the postnatal period. The distribution of the CD4⁺ and CD8⁺ cells was similar in the last trimester of pregnancy (days 120–150) and the postnatal period, but differed in the early stages of foetal development (days 70–120). In the prenatal period, the follicular area displayed high levels of proliferation and apoptosis. We observed CK18 immunoreaction only in FAE. While M cells were devoid of microfolds in the early stages of the prenatal period, these cells acquired a prismatic shape and bore distinct apical microfolds in the late prenatal period and postnatal period. As a result, it was determined that, in sheep, the development of the ileal Peyer’s patches occurred in the prenatal period, independent of exogenous antigenic stimulation, and in association with high levels of lymphopoiesis and apoptosis in the follicles. We found, for the first time, that CK18 is a novel and reliable marker for FAE in sheep ileal Peyer’s patches. We suggest that CK18 positive cells in FAE are M cells.     

Approaches to the Early Diagnosis of Parkinson’s Disease

Neuroscience and Behavioral Physiology - Parkinson’s disease (PD) has a long period of a “cryptic” course of the neurodegenerative process. Some patients show nonmotor symptoms...