Local Lymphocytic Infiltration Research Articles

Circulating Extracellular Vesicles Are Increased in Newly Diagnosed Celiac Disease Patients.

Extracellular vesicles (EVs) are a class of circulating entities that are involved in intercellular crosstalk mechanisms, participating in homeostasis maintenance, and diseases. Celiac disease is a gluten-triggered immune-mediated disorder, characterized by the inflammatory insult of the enteric mucosa following local lymphocytic infiltration, resulting in villous atrophy. The goal of this research was the assessment and characterization of circulating EVs in celiac disease patients, as well as in patients already on an adequate gluten-free regimen (GFD). For this purpose, a novel and validated technique based on polychromatic flow cytometry that allowed the identification and enumeration of different EV sub-phenotypes was applied. The analysis evidenced that the total, annexin V+, leukocyte (CD45+), and platelet (CD41a+) EV counts were significantly higher in both newly diagnosed celiac disease patients and patients under GFD compared with the healthy controls. Endothelial-derived (CD31+) and epithelial-derived (EpCAM+) EV counts were significantly lower in subjects under gluten exclusion than in celiac disease patients, although EpCAM+ EVs maintained higher counts than healthy subjects. The numbers of EpCAM+ EVs were a statistically significant predictor of intraepithelial leukocytes (IEL). These data demonstrate that EVs could represent novel and potentially powerful disease-specific biomarkers in the context of celiac disease.

Prognostic implications of preoperative systemic inflammatory markers in oral squamous cell carcinoma, and correlations with the local immune tumor microenvironment.

PurposeThe aim of this study was to investigate the prognostic significance of preoperative inflammatory markers in peripheral blood of patients with oral squamous cell carcinoma (OSCC), and to establish correlations with the infiltrate of macrophages and lymphocytes in the local immune tumor microenvironment (TME).Materials and MethodsNeutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), lymphocyte-monocyte ratio (LMR), and systemic immune-inflammation index (SII) were retrospectively evaluated in a cohort of 348 OSCC patients, and correlated with overall (OS) and disease-specific survival (DSS). Immunohistochemical analysis of tumoral and stromal infiltration of CD8+, CD4+, FOXP3+ and CD20+ lymphocytes and CD68+ and CD163+ macrophages was performed in a subset of 119 OSCC patient samples, and correlations further assessed.ResultsNLR, SII, and LMR were significantly associated with a poorer OS in univariate analysis; however, only NLR remained a significant independent predictor in the multivariate analysis (HR = 1.626, p = 0.04). NLR and SII were inversely and significantly correlated with stromal infiltration of CD8+, CD4+, and CD20+ lymphocytes. Moreover, a significant correlation between LMR was also found to significantly associate with stromal infiltration of CD8+, CD4+, and CD20+ lymphocytes, stromal CD68+ and CD163+ macrophages, and also tumoral infiltration of CD4+ and CD20+ lymphocytes.ConclusionsPreoperative NLR, SII, and LMR may serve as valuable systemic markers to predict OSCC patient survival, with NLR emerging as an independent predictor of poor OS. Moreover, strong significant correlations were exclusively observed between systemic inflammatory markers and the local stromal infiltration of lymphocytes in the TME.

Value of a Signature of Immune-Related Genes in Predicting the Prognosis of Melanoma and Its Responses to Immune Checkpoint Blocker Therapies.

Melanoma is becoming increasingly common worldwide, with high rates of transformation into malignancy compared to other skin lesions. The prognosis of patients with melanoma at an advanced stage is highly unsatisfying despite the development of immunotherapy, target therapy, or combinative therapy. The major barrier to exploiting immune checkpoint therapies and achieving the best benefits clinically is resistance that can easily develop if regimens are not selected appropriately. In this study, we investigated the possibility of using immune-related genes to predict patient survival and their responses to immune checkpoint blocker therapies with the expression profiles available at The Cancer Genome Atlas (TCGA) Program plus expression data from the Gene Expression Omnibus (GEO) for validation. A five gene signature that is highly correlated with the local infiltration of cytotoxic lymphocytes in the tumor microenvironment was identified, and a scoring model was developed with stepwise regression after multivariate Cox analyses. The score calculated strongly correlates with Breslow depth, and this model effectively predicts the prognosis of patients with melanoma, whether primary or metastasized. It also depicts the heterogenous immune-related nature of melanoma by revealing different predicted responses to immune checkpoint blocker therapies through its correlation to tumor immune dysfunction and exclusion (TIDE) score.

CD11c+ B Cells Participate in the Pathogenesis of Graves' Disease by Secreting Thyroid Autoantibodies and Cytokines.

Graves’ disease (GD) is a common autoimmune disorder with an elevation in pathogenic autoantibodies, specifically anti-thyrotropin receptor antibodies (TRAbs), which are secreted by autoreactive B cells. To date, there has been little research on self-reactive B cells in GD. In the current study, we reported that a unique B-cell subset, CD11c+ B cells, was expanded in the peripheral blood (PB) of GD patients, as detected by flow cytometry. The frequency of CD11c+ B cells was positively correlated with serum TRAb levels. The flow cytometry data showed that CD11c expression was higher in a variety of B-cell subsets and that CD11c+ B cells presented a distinct immunophenotype compared to paired CD11c- B cells. Immunohistochemical and immunofluorescence staining indicated the presence of CD11c+CD19+ B cells in lymphocyte infiltration areas of the GD thyroid. Flow cytometric analysis of PB and fine-needle aspiration (FNA) samples showed that compared to PB CD11c+ B cells, CD11c+ B cells in the thyroid accumulated and further differentiated. We found that CD11c+ B cells from the PB of GD patients were induced to differentiate into autoreactive antibody-secreting cells (ASCs) capable of secreting TRAbs in vitro. Luminex liquid suspension chip detection data showed that CD11c+ B cells also secreted a variety of cytokines, including proinflammatory cytokines, anti-inflammatory cytokines, and chemokines, which might play roles in regulating the local inflammatory response and infiltration of lymphocytes in the thyroid. In addition, we performed a chemotaxis assay in a Transwell chamber to verify that CD11c+ B cells were recruited by thyroid follicular cells (TFCs) via the CXCR3-CXCL10 axis. In conclusion, our study determined that CD11c+ B cells were involved in the pathogenesis of GD in multiple ways and might represent a promising immunotherapeutic target in the future.

NOTCH1 Intracellular Domain and the Tumor Microenvironment as Prognostic Markers in HNSCC.

Simple SummaryIn the head and neck, a large proportion of squamous cell carcinoma demonstrate a mutation of the NOTCH1 gene. The aim of this project was to investigate the role of NOTCH1 and immunological characteristics and highlight a potential rationale for therapy. We found that a high expression of NOTCH1 intracellular domain in these patients is associated with reduced overall survival. In vitro experiments additionally showed a reduction of migration and proliferation of cancer cells when NOTCH1 was knocked down. NOTCH1 is, therefore, most likely involved in migration and proliferation of head and neck squamous cell carcinoma and is a prognostic marker in these patients.(1) Background: NOTCH1 is the second most common mutated gene in whole-exome sequencing of HNSCC. The aim of this project was to gain further insight into the relevance of NOTCH1 in HNSCC, potentially establishing NOTCH1 as a prognostic marker or therapeutic target; (2) Methods: NOTCH1 was silenced via RNA interference in six HNSCC cell lines and the impact was evaluated in migration and proliferation assays. Subsequently, the protein expression of NOTCH1 intracellular domain (NICD) and NOTCH1 mRNA expression were examined in 70 oropharyngeal squamous cell cancer tissue samples. Lastly, the NICD expression was compared with the local infiltration of lymphocytes, measured with the immunoscore; (3) Results: Knockdown of NOTCH1 decreased migration and proliferation. A high NICD expression was associated with lower OS. A high immunoscore resulted in significantly better OS. NICD expression was independent of the immunoscore and as a whole differentiated three distinct prognostic groups; (4) Conclusions: These data suggest that NOTCH1 is involved in migration and proliferation of HNSCC cell lines. In vivo, NICD expression was associated with overall survival and could, therefore, be used as a prognostic marker. NICD expression differs from NOTCH1 mRNA levels, potentially explaining the previously suggested bimodal role as an oncogene and tumor suppressor in HNSCC.

A case of early autoimmune gastritis with characteristic endoscopic findings

Significant atrophic gastritis in the fundic gland region is a well-known endoscopic finding observed in autoimmune gastritis (AIG). The endoscopic features of early AIG have not been reported. Iron deficiency, vitamin B12 deficiency, anemia, or neurological symptoms may not be observed in the early stages of AIG, and it may thus be difficult to diagnose early AIG based on clinical findings. We treated a 50-year-old Japanese female whose condition was suspected to be early AIG. The endoscopic findings showed normal gastric pyloric gland mucosa, and diffuse reddened and edematous gastric fundic gland mucosa. Pathologically, local infiltration of lymphocytes and decrease of parietal cells was present in a deep part of the gastric fundic gland mucosa. Blood tests showed that the titer of parietal cell antibody (PCA) was 1:320 and the gastrin level was 820 pg/ml. We determined that the patient had AIG because she also had Hashimoto’s disease, the PCA titer was high, the serum gastrin level was slightly increased, and inflammation was observed only in the gastric body on the endoscopic images. To the best of our knowledge, this is the first case report of endoscopic findings that suggest early AIG, before atrophic changes were observed.

Calcium signaling and cell volume regulation are altered in Sjögren's Syndrome

Objective. Sjögren's Syndrome (SS) is a chronic autoimmune disease, leading to deficient secretion from salivary and lacrimal glands. Saliva production is normally increased by cholinergic innervation, giving rise to intracellular calcium signaling and water transport through water channels (aquaporins, AQPs). The aim of this study was to investigate possible pathophysiological changes in cell volume regulation, AQP expression and localization, and intracellular calcium signaling in glandular cells from SS patients compared to controls. Materials and methods. A total of 35 SS patients and 41 non-SS controls were included. Real time qPCR was combined with immunohistochemistry to analyze the mRNA expression and cellular distribution of AQP1, 3 and 5. Cell volume regulation and intracellular calcium signaling were examined in fresh acinar cells. Results. We show for the first time a reduced mRNA expression of AQP1 and 5 in SS compared to controls, accompanied by a decrease in staining intensity of AQP1, 3 and 5 in areas adjacent to local lymphocytic infiltration. Furthermore, we observed that the SS cells' capacity for volume regulation was abnormal. Similarly, the calcium response after parasympathetic agonist (carbachol) stimulation was markedly decreased in SS cells. Conclusions. It is concluded that mRNA expression of AQP1 and 5, protein distribution of AQP1, 3 and 5, glandular cell volume regulation and intracellular calcium signaling are all altered in SS, pointing to possible pathophysiological mechanisms in SS.

Clinical significance of microsatellite instability in colorectal cancer

Colorectal cancer is a heterogeneous tumor type with regard to molecular pathogenesis and genetic instability. The majority of colorectal cancers display chromosomal instability and follow the classical adenoma-carcinoma sequence of tumor progression. A subset of about 15% of colorectal cancers, however, displays DNA mismatch repair (MMR) deficiency and the high-level microsatellite instability (MSI-H) phenotype. MSI-H colorectal cancers can occur as sporadic tumors or in the context of hereditary non-polyposis colorectal cancer (HNPCC) or Lynch syndrome. The MSI-H phenotype is a hallmark of Lynch syndrome-associated cancers, which is of diagnostic relevance for the identification of Lynch syndrome mutation carriers. MSI-H colorectal cancers are characterized by a distinct clinical behavior, which results from their particular molecular pathogenesis and gives microsatellite instability testing its clinical significance. The MSI-H phenotype shows association with proximal tumor localization, a dense local lymphocyte infiltration, and a low frequency of distant organ metastasis. Moreover, MSI-H colorectal cancers have a better prognosis than their microsatellite-stable counterparts. A distinct responsiveness of MSI-H colorectal cancer patients towards chemotherapy has been shown in several studies. The clinical characteristics of MSI-H cancers are closely linked to their molecular pathogenesis, and research on the molecular alteration characteristic of MSI-H cancers may provide the basis for novel diagnostic or therapeutic approaches.

Sarcomatoid Lung Carcinomas Show High Levels of Programmed Death Ligand-1 (PD-L1)

Programmed death-1 (PD-1) is a coinhibitory inducible receptor present on T-cells and macrophages. Tumor cells with increased programmed death ligand-1 (PD-L1) are believed to escape immunity through activation of PD-1/PD-L1 pathway and suppression of effector-immune responses. Recent strategies targeting the PD-1/PD-L1 axis have shown promising results in patients with several tumors types, including lung carcinomas. Preliminary data suggest that PD-L1 protein expression might have predictive response to such therapies. Sarcomatoid carcinomas (SCs) of the lung include rare subtypes of poorly differentiated non-small-cell lung carcinomas of high grade and aggressive behavior. The biology of these neoplasms is poorly understood and they frequently show increased local inflammatory and lymphocytic infiltration. Here, we report the expression of PD-L1 in 13 SCs from two large retrospective lung cancer cohorts. Using automated quantitative immunofluoresence and a mouse monoclonal antibody directed against the extracellular domain of PD-L1, we show that 9 of 13 patients (69.2%) with SCs are positive for PD-L1 and their levels are higher than in conventional non-small-cell lung carcinoma. These results provide rationale for the potential use of targeted immunotherapy in lung SCs.

Transplantation of human amniotic mesenchymal stem cells in the treatment of focal cerebral ischemia

Cerebrovascular injury is one of the three major causes of death and is the leading cause of adult disability. Despite the increasing progress in emergency treatment and early rehabilitation in patients with cerebrovascular injury, treatment options for neurological dysfunction that presents at a later stage are lacking. This study examined the potential of human amniotic mesenchymal stem cell (hAMSC) transplantation in the repair of neurological deficits in an experimental focal cerebral ischemia model. Following the isolation of hAMSCs, growth characteristics and surface antigen expression were observed. Butylated hydroxyanisole (BHA) was used to induce the cultured cells into neuron-like cells, which were identified by immunocytochemistry. The suture model was used to induce focal cerebral ischemia in rats, which were subsequently randomly divided into experimental and control groups for treatment with BrdU-labeled hAMSCs or PBS, respectively. Neurological deficits were assessed following transplantation using the neurological severity score, beam balance test and elevated body swing test. Eight weeks later, rat brain tissue was analyzed with H&E staining and BrdU immunohistochemistry, and the survival and spatial distribution of the transplanted hAMSCs were determined. The hAMSCs proliferated invitro, and it was found that neuron-specific enolase (NSE) was expressed in neurons, whereas glial fibrillary acidic protein (GFAP) was expressed in astrocytes. The focal ischemia model caused varying degrees of left limb hemiplegia accompanied by right sided Horner's Syndrome. When examined 1, 3, 6 and 8weeks later, significant recovery in neurological behavior was detected in the rats treated with hAMSC transplantation compared with the control (P<0.01). BrdU-labeled hAMSCs were concentrated near the graft site and surrounding areas, in certain cases migrating towards the ischemic lesion. Local gliosis and lymphocytic infiltration were not detected. hAMSCs exhibit great potential for proliferation and are induced to differentiate into NSE-expressing neuron-like cells following treatment with BHA. Moreover, hAMSC transplantation may improve neurological symptoms following focal cerebral ischemia.

Local infiltration of lymphocytes in large bowel tissue cancer and chronic colitis.

e21101 Background: comparative analysis of concentrations of lymphocyte subpopulations in tissue of various intestinal zones in cancer of large bowel and chronic atrophic colitis. Methods: subpopulation composition of T- (CD3, CD4, CD8) and B- (CD19) lymphocytes, natural killers NK (CD16CD56), and T-lymphocytes with antigen-recognizing receptors TCRαβ and TCRγδ were examined in bowel adenocarcinoma tissue, peritumoural zone (1-3 cm away from tumour) and in resection line (10 cm away from tumour edge) and in chronic colitis tissue. Flow cytofluorimetry was used for immunophenotyping of the lymphocytes Results: 52 cases of adenocarcinoma and 24 cases of chronic colitis were examined. CD3 level in tumour tissue (64.2±3.6%) didn’t differ from this in colitis samples (65.6±3.5%); CD3 level was significantly lower in peritumoural zone and resection line (54.9±4.0% and 51.9±3.6%, correspondingly, p&lt;0.05) than in colitis samples. The quantaty of lymphocytes with TCRαβ receptor in all tissue samples with cancer was significantly higher in comparison with one in chronic colitis samples. The level of lymphocytes with TCRαβ receptor in all tissue samples with cancer was 49.8±7.25, in peritumoural zone – 39.3±5.9%, in resection line 46.5±6.0%, compared to chronic colitis – 20.7±5.7%, p&lt;0.05. Concentration of lymphocytes with TCRγδ receptor in cancer samples was 3-5 times less (tumour – 14.2±4.8%, peritumoural zone – 9.5±1.5%, resection line – 11.3+2.1%) than in samples colitis ones (42.9±9.9%) p&lt;0.05. It was found that tumour tissue, in contrast to peritumoural tissue, accumulated T-lymphocytes (64.2±3.6% and 51.9±3.6%, correspondingly) due to CD4 cells (35.5±2.3% and 24.3±2.1%, correspondingly, p&lt;0.05). Lower level of B-cells was found in tumour (18.0±2.8% and 30.1±3.7%, correspondingly, p&lt;0.05). Conclusions: abnormalities of local cellular immunity at diseases of large bowel are probably related not only to imbalance of lymphocyte subpopulations, but also to change of local correlation of levels of αβ-T-lymphocytes capable of recirculation, and tissue γδ-T-lymphocytes. This may cause inadequacy of immune protection at cancer of large bowel.

Transplantation of Human Amniotic Mesenchymal in the Treatment of Focal Cerebral Ischemia

This study examined the potential of human amniotic mesenchymal (hAMSC) transplantation in repairing neurological deficits in an experimental focal cerebral ischemia model. Following isolation of hAMSC, growth characteristics and surface antigen expression was observed. Butylated hydroxyanisole (BHA) was used to induce the cultured cells into neuron-like cells, identified by immunocytochemistry. The suture model was used to induce focal cerebral ischemia in the rats, which were subsequently randomly divided into experimental and control groups for treatment with BrdU-labeled hAMSCs or PBS, respectively. Neurological deficits were assessed after transplantation using the Neurological Severity Scores, Beam Balance Test, and Elevated Body Swing Test. Eight weeks later rat brain tissue was processed for hematoxylin-eosin staining and BrdU immunohistochemistry, and survival and spatial distribution of transplanted hAMSCs. hAMSCs proliferated in vitro, and neuron specific enolase expressed in neurons and glial fibrillary acidic protein in astrocytes. The focal ischemia model resulted in varying degrees of left limb hemiplegia accompanied by the right side of Horner’s sign. When examined 1, 3, 6 and 8 weeks later, significant recovery in the neurological behavior was detected in the rats treated with the hAMSC transplantation compared to the control (P<0.01). BrdU-labeled hAMSCs were concentrated near the graft site and surrounding areas, in some cases migrating towards the ischemic lesion. Local gliosis and lymphocytic infiltration were not detected. hAMSC exhibit great potential for proliferation, and can be induced to differentiate into NSE-expressing neuron-like cells following treatment with BHA. Moreover, hAMSC transplantation may improve neurological symptoms after focal cerebral ischemia.

Postmortem Study of Femoral Osteolysis Associated with Metal-on-Metal Articulation in Total Hip Replacement

Improved metal-on-metal articulations were reintroduced in total hip replacement to avoid the osteolysis sometimes seen with conventional ultra-high molecular weight polyethylene bearings. Osteolysis and local lymphocytic infiltration have been reported at revision of some metal-on-metal devices. We report similar and additional results in a study of second-generation metal-on-metal hip implants retrieved post mortem. Components and surrounding tissues were collected post mortem from seven patients with nine total hip replacements (Zweymüller SL stem with an Alloclassic cup) with Metasul metal-on-metal articulations. All available patient information was recorded. Radiographs of the hips were evaluated for osteolysis. Sections of joint capsule as well as of the femoral implant with surrounding bone were reviewed, and energy-dispersive x-ray analysis was used to evaluate the composition of wear products. The amount of wear was measured for each component (nine femoral heads and eight cup inserts), when possible, by a coordinate measurement machine with use of the dimensional method. The patients died between three and ten years after arthroplasty, and six of the seven were asymptomatic at the time of death. One patient, with the highest rate of total wear (i.e., wear of femoral head and acetabular cup; 7.6 microm/yr), had increasing hip pain for one year, and histological analysis confirmed the radiographic findings of osteolysis. For two other patients, histological analysis confirmed the radiographic findings of asymptomatic osteolysis. For three patients, histological analysis revealed osteolysis that had escaped conventional radiographic analysis. Joint capsule tissue showed evidence of metallosis in all hips and local lymphocytic infiltration in eight hips. Energy-dispersive x-ray analysis revealed elements attributable to CoCrMo alloy in all hips and traces of corrosion products in three hips. The postmortem findings of osteolysis and/or lymphocytic infiltration associated with eight clinically well-functioning, low wear devices (a total wear rate of <4 microm/yr) suggest there may be frequent, unappreciated femoral bone loss and local immunological response in patients with second-generation metal-on-metal hip implants. Compared with previous postmortem studies, our findings showed the extent of osteolysis was similar to that with metal-on-polyethylene articulations.

Viral Commensalism in Humans?

Viral Commensalism in Humans?

Transplantation of neural stem cells into the traumatized brain induces lymphocyte infiltration

Objective: This study examined the lymphocyte infiltration induced by neural stem cell grafts in the traumatized brain.Methods: Sixty Sprague-Dawley rats were assigned randomly to transplantation (n = 30) or control (n = 30) groups, and each rat was subjected to brain contusion. The neural stem cells derived from Wistar rats were transplanted into the lesion of the transplantation group, and saline was injected instead into the controls. Local lymphocyte infiltration was studied using haematoxylin and eosin staining, immunohistochemistry and flow cytometry. The immunogenicity of neural stem cells was evaluated using MHC-I expression.Results: About 6.57 ± 0.44% of the neural stem cells expressed MHC-I. In the transplantation group, histological examination and immunohistochemistry revealed significant lymphocyte infiltration in the contusion. The ratio of CD4+lymphocytes to total cells in the lesions was 13.28 ± 1.60% in the transplantation group and 0.41 ± 0.12% in the controls (p < 0.01). Likewise, the ratio of CD8+lymphocytes to total cells was 5.11 ± 1.03% in the transplantation group and 0.57 ± 0.26% in the controls (p < 0.01).Conclusions: Neural stem cells possess immunogenicity and can induce lymphocyte infiltration when transplanted into a traumatised brain. Our findings imply that immunosuppressive treatment is necessary following neural stem cell transplantation.

Transplantation of Rat Islets Transduced With Human Heme Oxygenase-1 Gene Using Adenovirus Vector

To investigate whether human heme oxygenase-1 (HO-1) gene has protective action on islets cultured in vitro, and to explore whether transduction of HO-1 gene to donor islets could enhance engrafted islets survival and suppress local lymphocytic infiltration in islet grafts. Newly isolated rat islets were isolated from the Sprague-Dawley rats and were divided into 3 groups in vitro study as follows: enhanced green fluorescent protein (EGFP) group, islets transduced with adenovirus vectors containing EGFP gene using multiplicities of infection (MOI) = 2, 5, 10, and 20 to determine the transduction efficacy; HO-1 group, islets transduced with adenovirus vectors containing human HO-1 gene using MOI = 20; and control group, mock transduced islets. Flow cytometry was used to detect apoptotic cells after induction by recombinant human tumor necrosis factor-alpha (rTNF-alpha) and cycloheximide (CHX) for 48 hours. Diabetic recipients were randomly divided into the following groups: HO-1 group (n = 9), receiving islets transduced with recombinant adenovirus-HO-1; control group (n = 9), receiving mock transduced islets; and phosphate buffer solution (PBS) group (n = 6), receiving only 0.8 mL PBS. About 1200 rat islet equivalents were transplanted into each recipient rendered by streptozotocin using the portal vein as transplant site. Allograft survival, apoptosis, and the state of lymphocytic infiltration were analyzed. After treatment with rTNF-alpha and CHX, the apoptotic ratio of islet cells was 4.22% +/- 2.38% in the HO-1 group (MOI = 20), significantly lower than 23.81% +/- 8.51% in the control group (P < 0.05), and 28.76% +/- 14.76% in the EGFP group (MOI = 20; P < 0.05). Maintenance of normoglycemia was prolonged in the HO-1 group, indicated by results that islet survival time was 10.56 +/- 4.33 days significantly longer than that of untreated islets which was 5.33 +/- 4.18 days (P < 0.05). The lymphocytic infiltration degree in engrafted islets treated with HO-1 gene was lower than that in the control group. HO-1 gene overexpression in rat islets by adenovirus transduction can protect cultured islets against rTNF-alpha and CHX-mediated cytotoxicity. HO-1 gene has cytoprotective effects on engrafted islets, which could prolong engrafted islets survival in allogenic transplantation model, and diminish the degree of lymphocytic infiltration in islet grafts. These findings suggest a potential therapeutic application for HO-1 gene in improving islet survival/function in human islet transplantation.

678 PUBLICATION Local lymphocyte infiltration as a major prognostic factor in rectal cancer patients

678 PUBLICATION Local lymphocyte infiltration as a major prognostic factor in rectal cancer patients

Microsatellite instability in colorectal cancer is associated with local lymphocyte infiltration and low frequency of distant metastases

Colorectal carcinomas (CRCs) with high microsatellite instability (MSI-H) share clinicopathological features distinctly different from their microsatellite stable (MSS) counterparts. Unlike MSS cancers, MSI-H CRCs occur predominantly in the right-sided colon and are often characterised by a strong lymphocyte infiltration. A poor differentiation pattern is found in most MSI-H CRCs, even though patients with MSI-H carcinomas seem to have a significantly longer survival after surgical resection. To clarify which factors contribute to the obvious paradoxon of a more favourable prognosis of MSI tumours, several clinical and histopathological features as well as the microsatellite status were evaluated in 120 colorectal cancer cases fulfilling clinical criteria (Bethesda) indicative for familial colorectal cancer. Microsatellite instablity status and lymphocyte infiltration were related to tumour stage and patients' follow-up. Statistical analysis confirmed well-known relations, such as enhanced lymphocyte infiltration accompanied by Crohn's like reaction (CLR) in MSI-H cancers (CLR+ in 27 out of 47 MSI-H vs 14 out of 71 MSS CRCs, P<0.001). However, after stratification for depth of local invasion and penetration of the primary tumour, T3 tumours displaying MSI had a significantly lower rate of distant metastases (M1 in four out of 35 MSI-H vs 20 out of 41 MSS CRCs, P<0.001). A similar tendency was observed for CLR-positive CRCs (M1 in six out of 29 CLR+ vs 17 out of 45 CLR− CRCs, P=0.13). In a logistic regression model, the MSI-H phenotype and the presence of CLR were independent predictors of a low UICC stage (P=0.006 and 0.04, respectively). These data, together with the recent definition of highly immunogenic neo-antigens expressed in MSI-H tumour cells, suggest that MSI-H CRCs elicit a protective host response that may prevent metastasis formation.

Gene transfer mediated by different viral vectors following direct cannulation of mouse submandibular salivary glands.

The salivary gland has been suggested as an accessible organ for gene transfer to express recombinant proteins locally in the saliva, as well as for secretion to the blood circulation. The aim of this study was to evaluate the efficiency of gene transfer to salivary glands using different viral vectors: adenovirus, vaccinia, herpes simplex type 1 (HSV), and two retroviral vectors (murine leukemia virus (MuLV) and lentivirus). We show, by in situ staining and beta-galactosidase reporter activity assay, that the adenoviral and vaccinia vectors were able to deliver the reporter gene efficiently to acinar and duct cells. The HSV vector was less efficient and infected only the acinar cells. The lentiviral vector infected acinar and duct cells, but at a relatively low efficiency. The MuLV vector did not infect the salivary gland unless cell proliferation was induced. Host immune responses to viral infection, inflammation, apoptosis and lymphocyte infiltration, in the transduced glands, were assessed. The DNA viral vectors induced local lymphocyte infiltration and apoptosis. In contrast, the retroviral vectors did not induce an immune response. Our results describe the outcome of salivary gland infection with each of the five different viral vectors and indicate their advantages and limitations for transferring genes to the salivary glands.

Morphological changes in mouse accessory sex glands following neonatal estrogen treatment

A single injection of beta-estradiol 17-cypionate into the mice within 5 hr after birth induced inflammation in all prostate lobes and the seminal vesicles. Neutrophils emigrated into the lumen through the basal lamina and epithelium of the seminal vesicle and the anterior prostate. Local infiltration of lymphocytes was observed in the stroma and epithelium of ventral prostates. Lymphocytes penetrated through smooth muscle cells into epithelium. This could support the hypothesis that smooth muscle cells are the target of the estrogen action of prostates in estrogenized animals.