Immunolabeling Of Cells Research Articles

Preliminary results of prognostic significance of proliferating cell nuclear antigen expression in advanced primary larynx carcinomas and lymph node metastases

IntroductionThe aim of this study was to investigate the prognostic significance of proliferating cell nuclear antigen (PCNA) expression in laryngeal carcinoma in relation to clinicopathological features. Special emphasis was placed on examining the relationship of PCNA expression in the primary tumour and PCNA expression in corresponding lymph node metastases obtained from the same patients.Material and methodsThe study included 60 patients with advanced larynx carcinoma who had received treatment and follow-up for at least 5 years. Sixty laryngeal carcinoma specimens and metastatic lymph nodes from 24 patients were examined for immunohistochemical PCNA expression.ResultsThe percentages of PCNA positive cells were significantly higher in the primary tumours which developed lymph node metastases than in those without metastases. The fraction of PCNA immunolabelled cells in metastatic lymph nodes increased significantly when compared with the PCNA positive cell score in their corresponding primary tumours obtained from the same patient. There was a significant difference in PCNA index score in primary tumours between the group of patients who survived a 5-year period and those who died within 5 years after treatment.ConclusionsOur data demonstrate that a high proliferation index in primary larynx tumours is retained and increased in corresponding lymph node metastases. Measurement of the fraction of cancer cells stained for PCNA in primary larynx carcinomas can be helpful in selecting tumours with high aggressiveness potential that are more likely to develop neck metastases and thereby in identifying patients who need elective lymph node dissection or additional treatment.

Observation of immuno-labeled cells at high resolution using soft X-ray microscope at Ritsumeikan University SR Center

Mouse fibroblast cell line NIH3T3 cells were labeled with the heavy metal (silver and gold) and observed intracellular structure under an X-ray microscope. Microtubules, Golgi apparatus and early endosomes of NIH3T3 cells were stained with immuno-gold nanoparticles, and immuno-staining was intensified by silver or gold enhancement procedure. Using a transmission soft X-ray microscope beamline (BL12) at Ritsumeikan University SR center, we observed immuno-stained NIH3T3 cells with several wavelengths just below and above oxygen edge (λ = 2.32 nm). Using this method, cytoskeleton (microtubules) and organelles (Golgi apparatus and early endosomes) were successfully imaged with high resolution. Thus, immuno-gold silver and gold enhancement technique is useful for specific labeling of intracellular structure under an X-ray microscope.

Labeling of BSA and imaging of mouse T‐lymphocyte as well as mouse spleen tissue by l‐glutathione capped CdTe quantum dots

L-glutathione capped highly fluorescent CdTe quantum dots (QDs) were prepared by an aqueous approach and used as fluorescent labels to link albumin bovine serum (BSA) and rat anti-mouse CD4, which was expressed on mouse T-lymphocyte and mouse spleen tissue. The sharp and narrow emission peaks showed that the as-prepared QDs have desirable dispersibility, uniformity and good fluorescence properties. Both CdTe-BSA and CdTe-CD4 conjugates showed an enhancement of fluorescence intensity over that of bare CdTe QDs. The experimental result of gel electrophoresis confirmed the successful conjugation of CdTe-BSA and CdTe-CD4. The fluorescent microscopic images of CdTe-CD4 labeled mouse T-lymphocyte cells and mouse spleen tissue were compared with that obtained from fluorescein isothiocyanate labeling. It was demonstrated that the CdTe QDs-based probe exhibited much better photostability and fluorescence intensity than fluorescein isothiocyanate, showing a good application potential in the immuno-labeling of cells and tissues.

Immunolabeling and NIR-Excited Fluorescent Imaging of HeLa Cells by Using NaYF4:Yb,Er Upconversion Nanoparticles

Upconversion fluorescent nanoparticles can convert a longer wavelength radiation (e.g., near-infrared light) into a shorter wavelength fluorescence (e.g., visible light) and thus have emerged as a new class of fluorescent probes for biomedical imaging. Rare-earth doped beta-NaYF(4):Yb,Er upconversion nanoparticles (UCNPs) with strong UC fluorescence were synthesized in this work by using a solvothermal approach. The UCNPs were coated with a thin layer of SiO(2) to form core-shell nanoparticles via a typical Stober method, which were further modified with amino groups. After surface functionalization, the rabbit anti-CEA8 antibodies were covalently linked to the UCNPs to form the antibody-UCNP conjugates. The antibody-UCNP conjugates were used as fluorescent biolabels for the detection of carcinoembryonic antigen (CEA), a cancer biomarker expressed on the surface of HeLa cells. The successful conjugation of antibody to the UCNPs was found to lead to the specific attachment of the UCNPs onto the surface of the HeLa cells, which further resulted in the bright green UC fluorescence from the UCNP-labeled cells under 980 nm near-infrared (NIR) excitation and enabled the fluorescent imaging and detection of the HeLa cells. These results indicate that the amino-functionalized UCNPs can be used as fluorescent probes in cell immunolabeling and imaging. Because the UCNPs can be excited with a NIR light to exhibit strong visible fluorescence and the NIR light is safe to the body and can penetrate tissue as deep as several inches, our work suggests that, with proper cell-targeting or tumor-homing peptides or proteins conjugated, the NaYF(4):Yb,Er UCNPs can find potential applications in the in vivo imaging, detection, and diagnosis of cancers.

Adhesion of pancreatic beta cells to biopolymer films

Dramatic reversal of Type 1 diabetes in patients receiving pancreatic islet transplants continues to prompt vigorous research concerning the basic mechanisms underlying patient turnaround. At the most fundamental level, transplanted islets must maintain viability and function in vitro and in vivo and should be protected from host immune rejection. Our previous reports showed enhancement of islet viability and insulin secretion per tissue mass for small islets (<125 mum) as compared with large islets (>125 mum), thus, demonstrating the effect of enhancing the mass transport of islets (i.e. increasing tissue surface area to volume ratio). Here, we report the facile dispersion of rat islets into individual cells that are layered onto the surface of a biopolymer film towards the ultimate goal of improving mass transport in islet tissue. The tightly packed structure of intact islets was disrupted by incubating in calcium-free media resulting in fragmented islets, which were further dispersed into individual or small groups of cells by using a low concentration of papain. The dispersed cells were screened for adhesion to a range of biopolymers and the nature of cell adhesion was characterized for selected groups by quantifying adherent cells, measuring the surface area coverage of the cells, and immunolabeling cells for adhesion proteins interacting with selected biopolymers. Finally, beta cells in suspension were centrifuged to form controlled numbers of cell layers on films for future work determining the mass transport limitations in the adhered tissue constructs. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 676-685, 2009.This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com.

Single-dose and chronic corticosterone treatment alters c-Fos or FosB immunoreactivity in the rat cerebral cortex

The aim of this study was to examine the effects of single-dose and chronic corticosterone treatment on the inducible transcription factor c-Fos and FosB, and thereby to estimate the effects of high-doses of corticosterone on calcium-dependent neuronal responses in the rat cerebral cortex. At the same time we investigated the distribution of interneurons containing calretinin (CR), vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY) in chronically treated animals in order to collect data on the involvement of inhibitory neurons in this process. Adult male rats were injected subcutaneously with 10mg corticosterone, whereas controls received the vehicle (sesame oil). The animals were fixed by transcardial perfusion 12 and 24h following single corticosterone injection, and the brains were processed for c-Fos and FosB immunohistochemistry. To investigate the effects of repeated corticosterone administration, rats were daily treated with the same amount of corticosterone (10mg/animal, subcutaneously) for 21 days. Controls were injected with vehicle. At the end of the experiment, the rats were perfused and immunohistochemistry was used to detect the presence of the FosB protein, CR, VIP and NPY. Quantitative evaluation of immunolabelled cells was performed in the neocortex and the hippocampus. The number of immunoreactive nuclei per unit area was used as a quantitative measure of the effects of corticosterone. It was found that a single-dose administration of corticosterone resulted in a significant, time-dependent increase of c-Fos protein immunoreactivity in the granule cell layer of the dentate gyrus, as well as in regions CA1 and CA3 of the hippocampus 12 and 24h post-injection with respect to control animals. Significant enhancement of c-Fos immunoreactivity was also observed in the neocortex at 12 and 24h post-injection. Single-dose treatment did not significantly alter FosB immunolabelling. Repeated administration of corticosterone produced a complex pattern of changes in FosB immunolabelling: significant increase in FosB immunoreactivity was detected in the granule cell layer of the dentate gyrus, with no significant changes in the CA1 and CA3 layers of the hippocampus and in the neocortex. However, a significant decrease of FosB induction in the neocortex was observed in chronically treated rats in comparison to single-dose injected animals (12h before immunohistochemistry). Analysis of immunohistochemical detection of interneuronal markers revealed a significant reduction of the CR immunolabelling in the CA3 area of the hippocampus. No changes in VIP or NPY immunoreactivity were found in the Ammon's horn 3 weeks following daily corticosterone treatment. NPY immunoreactivity was significantly attenuated in the neocortex. The present data suggest that single-dose corticosterone treatment increases immunoreactivity of c-Fos protein in a time-dependent manner, 12 and 24h post-injection in the rat hippocampus and the neocortex, whereas chronic corticosterone treatment influences FosB immunoreactivity, primarily in the dentate gyrus. Chronic corticosterone administration seems to affect CR levels in the CA3 area of the hippocampus.

Increase of gingival matured dendritic cells number in elderly patients with chronic periodontitis

Objective The purpose of this study was to evaluate the inflammatory cell subset proportions in the upper gingival connective tissue, including mature dendritic cells (DC) in elderly and younger patients with generalized chronic periodontitis in order to further understand the effect of aging on gingival inflammatory phenomenon. Methods Gingival tissue specimens presenting chronic periodontitis from 8 elderly patients aged >75 (test group, group T) and from 8 younger patients aged 50–60 (considered as controls, group C) were analysed by immunohistochemistry using monoclonal antibodies against CD45RB, CD4, CD8, CD19, CD68, DC-SIGN, DC-LAMP molecules. The number of each immunolabelled cells subset was counted using image analysis. Results The difference in the number of CD45RB + leucocytes in the upper gingival connective tissue between groups was not significant permitting to use it as reference. As compared to group C, the lymphocyte subsets/CD45RB + leucocytes ratios tended to decrease in group T but the decrease was significant only for CD4 + T lymphocytes/CD45RB + cells ratio ( p < 0.03). On the opposite, the ratios of antigen-presenting cells DC-SIGN + cells/CD45RB + cells and DC-LAMP + cells/CD45RB + cells were significantly increased ( p < 0.03 and <0.0001, respectively) in group T. Moreover, in group T the DC-LAMP + cells/DC-SIGN + cells ratio was significantly increased ( p < 0.05) showing an increased number of matured dendritic cells. Conclusion During chronic periodontitis in elderly patients, our results show a decrease in the ratio of gingival CD4 + lymphocyte subset associated with an increase in the ratios of antigen-presenting cells subsets and more particularly maturated DC-LAMP + dendritic cells.

Role of hepatic macrophages during the viral haemorrhagic fever induced by African Swine Fever Virus.

To ascertain the role played by the various liver monocyte-macrophage populations in the course of a viral hemorrhagic fever, fifteen pigs were inoculated intramuscularly with the highly virulent isolate of African Swine Fever Virus (ASFV) España-70 and slaughtered at 1-7 days post-inoculation (dpi). Samples of liver were fixed in different solutions and routinely processed for morphological, immunohistochemical and ultrastructural studies. Viral antigen (vp73) was detected from 3 dpi onward, mainly in circulating monocytes of sinusoid and Kupffer's cells (KC), as well as in portal macrophages and hepatocytes from 5 dpi. Anti-SWC3 immunolabelled cells were increased from 1 dpi, peaking between 3 and 5 dpi, thereafter declining until the end of the experiment. The significant increase in the number of sinusoidal circulating monocytes and KC expressing IL-1alpha, TNFalpha and IL-6 from 1 dpi, confirmed the secretory activation of these cells. The results show that in the course of an ASFV-induced hemorrhagic syndrome, hepatic macrophage populations undergo major quantitative and biosynthetic changes prior to virus detection, suggesting the existence of a mechanism by which the virus concentrates infectable cells, which subsequently spread the virus around the body.

MMP-13 (collagenase 3) localization in human temporomandibular joint discs with internal derangement

The normal articular disc of the human temporomandibular joint (TMJ) consists of regularly arranged collagen fibers and fibroblast-like cells. Disc tissue is subjected to various types of loading. Adaptive remodelling entails microscopic changes in which different types of collagen and non-collagen proteins, as well as matrix metalloproteinases (MMPs), are involved. Collagenase-3 (MMP-13) seems to play a key role. The aim of the study was to evaluate its involvement in TMJ disc histopathology. We studied 10 discs from patients with arthropathy of the TMJ. Paraffin wax-embedded sections were processed for histological and immunohistochemical analysis. A well-characterized mouse monoclonal anti-MMP-13 antibody (anti-collagenase 3) was used. The results showed increased MMP-13 immunoreactivity in TMJ diseased disc tissue was correlated with the severity of the histopathological changes. The more severe the pathological changes, the greater the number of immunolabelled cells detected. Nearly all chondrocyte-like cells were immunopositive, whereas fibroblast-like cells and fibrochondrocytes were more rarely labelled. MMP-13 upregulation in disc tissue from patients with TMJ internal derangement suggests that MMP-13 is involved in disc histopathology, and that disc cells actively participate in the synthesis of extracellular matrix-degrading proteinases.

Expression of transforming growth factor-beta 1 (TGF-β1) in different types of granulomatous lesions in bovine and ovine paratuberculosis

Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (Map), is manifested by a broad spectrum of clinical and lesional presentations. We have evaluated the expression of transforming growth factor-beta1 (TGF-beta1), a cytokine known to have immunosuppressor effects, by immunohistochemistry, in different paratuberculosis lesions in the intestine and lymph nodes from 20 sheep and 25 cattle. Peripheral immune responses were assessed by interferon-gamma (IFN-gamma) test and the presence of antibodies. Expression of TGF-beta1, observed in macrophages and giant cells forming the lesions, was closely related to the amount of Map. In focal and multifocal forms, usually positive to IFN-gamma test, bacilli were difficult to detect and TGF-beta1 expression was low or absent. Diffuse multibacillary lesions, negative to IFN-gamma, show large numbers of Map and the highest percentage of immunolabelled cells. Diffuse paucibacillary forms, positive to IFN-gamma, have low numbers of AFB and scant or no cells positive to TGF-beta1. The high expression of TGF-beta1 would be related to the inability of macrophages to limit the multiplication of Map.

PCNA indexing as a preclinical immunohistochemical biomarker for testicular toxicity

It is well known that toxicants such as cyclophosphamide and ethanol can have deleterious effects on normal spermatogenesis. End points such as testis weight and sperm counts have been used widely to assess gross structural and functional changes in testes resulting from toxicant exposure. Histopathological assessments are more sensitive measures of testicular health, but generally they are neither quantitative nor sensitive enough to detect early toxicity. Recently, immunolabeling cells with proliferating cell nuclear antigen (PCNA) has been used to identify proliferating spermatogonia; however, there have been no systematic attempts to quantify these changes. We have developed a sensitive, reliable and quantitative assay using immunohistochemistry on formalin fixed, paraffin embedded rat testes to assess the degree of proliferation-related toxicity. An indexing scheme was derived based on the determination of radially positioned PCNA-positive cells within similarly staged seminiferous tubules presenting a single layer of PCNA-positive cells along the basement membrane of the basal tubular compartment. An average of 60 tubules in the testes were counted per animal. Our results show significant decreases in the PCNA index in rats treated with an experimental compound that has been shown to produce testicular histopathology. The analysis provides a quick, reliable, sensitive, and quantitative means for assessing early testicular toxicity. The assay has potential utility as an in vivo biomarker for detecting early testicular toxicity of experimental compounds in preclinical development as well as for refining follow-up compounds with reduced testicular toxicity.

Broadening cell selection criteria with micropallet arrays of adherent cells

A host of technologies exists for the separation of living, nonadherent cells, with separation decisions typically based on fluorescence or immunolabeling of cells. Methods to separate adherent cells as well as to broaden the range of possible sorting criteria would be of high value and complementary to existing strategies. Cells were cultured on arrays of releasable pallets. The arrays were screened and individual cell(s)/pallets were released and collected. Conventional fluorescence and immunolabeling of cells were compatible with the pallet arrays, as were separations based on gene expression. By varying the size of the pallet and the number of cells cultured on the array, single cells or clonal colonies of cells were isolated from a heterogeneous population. Since cells remained adherent throughout the isolation process, separations based on morphologic characteristics, for example cell shape, were feasible. Repeated measurements of each cell in an array were performed permitting the selection of cells based on their temporal behavior, e.g. growth rate. The pallet array system provides the flexibility to select and collect adherent cells based on phenotypic and temporal criteria and other characteristics not accessible by alternative methods.

Cytokine Expression in Paraffin Wax-embedded Tissues from Conventional Calves

The cross-reactivity of antibodies against human tumour necrosis factor (TNF)alpha, interleukin (IL)-1alpha, IL-1beta and porcine IL-6, and the distribution of immunolabelled cells were evaluated on paraffin wax-embedded tissues from five healthy calves. The tissues were fixed in 10% buffered formalin or Bouin's solution and processed for structural studies and immunohistochemical studies by the avidin-biotin-peroxidase technique. Bouin's solution proved to be the more suitable fixative and Tween 20 the most effective antigen unmasking technique for increasing detectable antigenicity. Constitutive expression of TNFalpha, IL-1alpha, IL-1beta and IL-6 by different cell populations, mainly macrophage-like cells, was detected. Lymphoid organs displayed a higher presence of immunolabelled cells than did lung, liver or kidney. TNFalpha and IL-1alpha appeared as the predominant cytokines, especially in the gut-associated lymphoid tissue of the ileum and in the regional mesenteric lymph nodes. The results will facilitate investigation of the role of these cytokine-producing cells in inflammatory disease processes in calves.

Determination of ghrelin immunoreactivity in the rat stomach after fasting and refeeding

Ghrelin is a recently discovered hormone secreted by cells of the stomach. The aim of this study was to investigate fasting and refeeding induced alterations on ghrelin immunolabelling of cells of the stomach. Thirty-six adult male Wistar rats were used in this study. Rats were divided into six groups. Group I: control group; Group II: rats fasted for 7 days; Group III: rats fed for 1 day after 7 days of fasting; Group IV: rats fed for 3 days after 7 days of fasting; Group V: rats fed for 5 days after 7 days of fasting; Group VI: rats fed for 7 days after 7 days of fasting. At the end of the experiment, rats were sacrificed and stomach tissues were processed for imunohistochemistry to localize ghrelin. Ghrelin-immunopositive cells were detected only in the mucosal lining of the stomach. After fasting for 7 days, the number of ghrelin-immunopositive cells increased significantly compared to the control rats. Following refeeding, the number of ghrelin-immunoreactive cells was reduced to a level comparable to the controls. Therefore, fasting and refeeding after fasting were observed to result in changes in ghrelin immunoreactivity in the cells of the stomach. We conclude that ghrelin is highly expressed in the stomach and that fasting increases the expression of ghrelin in the stomach, but this expression decreases after refeeding. Our results indicate that regulation of ghrelin is a process probably involved in the long-term control of nutritional states.

MCM immunocytochemistry as a first line cervical screening test in developing countries: a prospective cohort study in a regional cancer centre in India

Cervical screening is not available for the majority of women in resource-poor countries. An important factor is a lack of skilled operators necessary for high-throughput assessment of the Papanicolaou (Pap) test currently in use. We compared the efficacy of immunocytochemistry for minichromosome maintenance (MCM) proteins vs standard Pap testing at detecting disease in 455 cervical smears processed in a typical Indian screening laboratory. Conventional (non-monolayer) smears were stained manually and then examined by a cytotechnologist and a cytopathologist. The MCM test was called positive when immunolabelled cells were identified as dyskaryotic by the Pap counterstain. The MCM test was read more quickly than the Pap test (approximately 2 vs 10 min) and there was 100% inter-observer agreement compared with 85% for Pap (P<0.0001). The MCM test detected 10 biopsy-proven cancers or pre-cancers that were not detected by Pap (P=0.002; P=0.016 excluding three cases where the Pap was deemed unsatisfactory on review). The cases in question included one recurrent squamous carcinoma and one adenocarcinoma in a screening patient who would have returned to 5 year recall. There were no false positive MCM test results. We propose that MCM immunocytochemistry has considerable advantages for cervical screening in developing countries like India.

Differential distribution of transforming growth factor beta and receptors in the hyper or hypoproliferative gastric mucosa of developing and adult rats

Transforming growth factor beta (TGFbeta) isoforms are known for their antiproliferative effect on epithelial cells in vitro, but the role of each isoform in vivo is poorly understood, mainly when non-pathological conditions are considered. We correlated the presence and distribution of isoforms and receptors to physiological changes in gastric cell proliferation in developing and adult rats. We used fasting to induce either the hyper (14-day-old pups) or hypoproliferation (60-day-old rats) of the gastric epithelium. In 14-d-old pups fasting reduced only TGFbeta3 labelling in the gland. Conversely, in 60-d-old rats there was an increase of TGFbeta1 and TGFbeta3 immunolabelled cells. Receptors were detected at both ages. Therefore, the changes induced by fasting in the constitutive TGFbeta expression can be correlated to the differential epithelial proliferation in the stomach of developing and adult rats. These results suggest that one of the functional roles of TGFbeta in vivo is to locally regulate cell proliferation.

Nitric oxide synthase protein levels, not the mRNA, are downregulated in olfactory bulb interneurons of reeler mice

Homozygous mutations in the Reelin gene result in severe disruption of brain development. The histogenesis of layered regions, like the neocortex, hippocampus and the cerebellum, is most notably affected in mouse reeler mutants and similar traits are also present in mice lacking molecular components of the Reelin signalling pathway. Moreover, there is evidence for an additional role of Reelin in sustaining synaptic plasticity in adult networks. Nitric oxide is an important gaseous messenger that can modulate neuronal plasticity both in developing and mature synaptic networks and has been shown to facilitate synaptic changes in the hippocampus, cerebellum and olfactory bulb. We studied the distribution and content of neuronal nitric oxide synthase in the olfactory bulbs of reeler and wildtype mice. Immunocytochemistry reveals that Reelin and neuronal nitric oxide synthase containing interneurons are two distinct, non overlapping cell populations of the olfactory bulb. We show by in situ hybridization that both nitrergic and Reelin expressing cells represent only a subset of olfactory bulb GABAergic neurons. Immunoblots show that neuronal nitric oxide synthase protein content is decreased by two thirds in reeler mice causing a detectable loss of immunolabelled cells throughout the olfactory bulb of this strain. However, neuronal nitric oxide synthase mRNA levels, essayed by quantitative real-time RT-PCR, are unaffected in the reeler olfactory bulb. Thus, disruption of the Reelin signalling pathway may modify the turnover of neuronal nitric oxide synthase in the olfactory bulb and possibly affects nitric oxide functions in reeler mice.

Getting across the plasma membrane and beyond: intracellular uses of colloidal semiconductor nanocrystals.

Semiconductor nanocrystals (NCs) are increasingly being used as photoluminescen markers in biological imaging. Their brightness, large Stokes shift, and high photostability compared to organic fluorophores permit the exploration of biological phenomena at the single-molecule scale with superior temporal resolution and spatial precision. NCs have predominantly been used as extracellular markers for tagging and tracking membrane proteins. Successful internalization and intracellular labelling with NCs have been demonstrated for both fixed immunolabelled and live cells. However, the precise localization and subcellular compartment labelled are less clear. Generally, live cell studies are limited by the requirement of fairly invasive protocols for loading NCs and the relatively large size of NCs compared to the cellular machinery, along with the subsequent sequestration of NCs in endosomal/lysosomal compartments. For long-period observation the potential cytotoxicity of cytoplasmically loaded NCs must be evaluated. This review focuses on the challenges of intracellular uses of NCs.

Do Langerhans cells behave similarly in elderly and younger patients with chronic periodontitis?

Objective The purpose of this study was to compare the number, the distribution and the expression of markers of maturation of Langerhans cells (LC) in elderly and younger patients with chronic periodontitis in order to evidence the effect of aging on LC in inflammatory gingival tissue. Methods Gingival tissue specimens presenting chronic periodontitis from 8 elderly patients aged >75 (group E) and from 8 younger patients aged 50–60 (considered as controls, group C) were used for immunohistochemistry with monoclonal antibodies against CD45RB (leucocytes), CD1a (LC), markers of LC maturation (DC-LAMP, CD83) and number of immunolabelled cell subsets was evaluated using image analysis. Results The difference in the number of CD45RB+ leucocytes in the upper connective tissue between groups was not significant. In group E, the number of CD1a+ LC was significantly decreased ( P < 0.002) in the epithelium and significantly increased ( P < 0.0004) in the upper connective tissue. Furthermore, in group E, intraepithelial CD1a+ LC are more often observed in the upper epithelium and their dendritic processes were shorter and less numerous. Concerning the expression of markers of maturation, the numbers of intraepithelial DC-LAMP+ cells and CD83+ cells were significantly increased ( P < 0.0007 and P < 0.02, respectively) in group E. Conclusion During chronic periodontitis in elderly patients, the decrease in the number of intraepithelial LC and the alteration of dendritic processes could be balanced by a cellular distribution often observed in the upper epithelium associated with changes in cell maturation in response to bacterial elements.

Caspase 9 is decreased in psoriatic epidermis

Psoriasis is a proliferative and inflammatory disease of the skin. Caspase 9 is responsible for initiating the caspase activation cascade during apoptosis. Apoptosis is a physiological mechanism of homeostasis and development, and caspases are the executioners of apoptosis. This study reports the immunohistochemical localisation of caspase 9 in psoriatic skin and compares it with that seen in normal, healthy control skin. Skin biopsy specimens of lesions were obtained from 15 patients with plaque type psoriasis vulgaris. The specimens were labelled immunohistochemically for binding of an anti-caspase 9 primary antibody. Biopsies of healthy skin from 10 age-matched and sex-matched healthy control individuals were also analysed. The caspase 9 positive cell fraction was calculated for both epidermal and dermal cells in psoriatic lesions and healthy control skin. Counts of caspase 9 positive cells from the epidermis of psoriatic skin lesions were significantly lower than those seen in healthy skin (p<0.05). The caspase 9 immunolabelled perivascular cell counts in the dermis were not statistically significantly different in psoriatic lesions versus normal skin (p>0.05). Psoriatic epidermis contains little of the apoptotic marker, caspase 9. The results of this study are indicative of decreased apoptosis in psoriatic epidermis, and no change in the perivascular area in psoriatic lesions. These findings support the idea that decreased apoptosis is seen in psoriatic epidermal cells. Greater understanding of the nature of the disease may open new avenues for further therapeutic modalities.