Immunological Capacity Research Articles

Variation in Major Histocompatibility Complex diversity in invasive cane toad populations

Context The cane toad (Rhinella marina), a native species of central and southern America, was introduced to Australia in 1935 as a biocontrol agent after a complex history of prior introductions. The population rapidly expanded and has since spread through much of the Australian landmass, with severe impacts on the endemic wildlife, primarily via toxicity to predators. The invasion process has taken its toll on the cane toad, with changes in the immunological capacity across the Australian invasive population. Aims To investigate the immunogenetic underpinnings of these changes, we studied the diversity of the Major Histocompatiblity Complex (MHC) genes in introduced cane toad populations. Methods We studied the diversity of two MHC genes (the classical class I UA locus and a class II DAB locus) and compared these with neutral microsatellite markers in toads from the Australian site of introduction and the Australian invasion front. We also included toads from Hawai’i, the original source of the Australian toads, to infer founder effect. Key results Diversity across all markers was low across Australian and Hawai’ian samples, consistent with a reduction in genetic diversity through multiple founder effects during the course of the successive translocations. In Australia, allelic diversity at the microsatellite markers and the UA locus was reduced at the invasion front, whereas all three alleles at the DAB locus were maintained in the invasion-front toads. Conclusions Loss of allelic diversity observed at the microsatellite markers and the UA locus could be the result of drift and bottlenecking along the invasion process, however, the persistence of DAB diversity warrants further investigation to disentangle the evolutionary forces influencing this locus. Implications Through the use of different molecular markers, we provide a preliminary description of the adaptive genetic processes occurring in this invasive population. The extremely limited MHC diversity may represent low immunogenetic competence across the Australian population, which could be exploited for invasive species management.

The Laparoscopically Harvested Omental Free Flap: A Compelling Option for Craniofacial and Cranial Base Reconstruction.

Background Management of craniofacial and cranial base tumors is a challenge due to the anatomic intricacies associated with the calvarium, the pathological diversity of lesions that present, and the potential complications. Clinical outcomes in laparoscopically harvested omentum free flaps for cranial base and craniofacial reconstruction are presented in this paper, in the largest case series to date. Methods A retrospective single-center experience for over 10 years with laparoscopically harvested omentum flaps used to reconstruct craniofacial and cranial base defects. Results A total of 13 patients underwent craniofacial or cranial base reconstruction using laparoscopically harvested omentum free flaps. The mean patient age was 48 years. The anterior skull base represented the most common site of reconstruction. A total of 12 of the flaps survived (92%), with one flap failure due to infection. All patients demonstrated satisfactory aesthetic and functional outcomes. There were no perioperative or intra-abdominal complications. Conclusions The laparoscopically harvested omentum free flap is a safe and effective tool in the armamentarium of the reconstructive surgeon. It is the ideal option to treat complex, three-dimensional subcutaneous defects, such as those encountered in craniofacial and cranial base reconstruction. Its unique angiogenic and immunologic capacity makes it an excellent flap for the previously irradiated and/or infected wound bed.

Innate immune defense in the inner ear - mucines are expressed by the human endolymphatic sac.

The human endolymphatic sac has been shown recently to have immunological capacities and has thus been proposed as the main entity protecting the inner ear from pathogen invasion, equivalent to mucosa-associated lymphoid tissue (MALT). Although the sac expresses molecules of the innate immune system, the potential expression of members of the important mucin family has not been detailed. Thus, this paper explores endolymphatic sac expression of a number of mucins and mucin precursors. Twelve fresh tissue samples from the human endolymphatic sac were obtained during translabyrinthine surgery. The expression of Mucin 1, 2, 5B/AC and 16, as well as the core structure elements (mucin precursors) T-antigen, Tn-antigen and Sialyl-Tn-antigen was investigated by immunohistochemistry. The endolymphatic sac epithelium expressed MUC1 (both apically towards the endolymphatic sac (ES) lumen and basally towards the capillary network), MUC 16 and Tn-antigen. There was no labeling after incubation with antibodies against T-antigen, sialyl-Tn-antigen, MUC2 and MUC5B/AC. We conclude that the human endolymphatic sac epithelium expresses a number of mucin molecules, which supports the hypothesis of the sac as the primary immunological tissue structure of the inner ear, equivalent to MALT in other organs. The mucins may also play a role in the formation and continuous homeostasis of the inner ear fluids, as well as the pathogenesis of Meniere's disease.

Differential TLR activation of murine mesenchymal stem cells generates distinct immunomodulatory effects in EAE.

BackgroundRecently, it has been observed that mesenchymal stem cells (MSCs) can modulate their immunoregulatory properties depending on the specific in-vitro activation of different Toll-like receptors (TLR), such as TLR3 and TLR4. In the present study, we evaluated the effect of polyinosinic:polycytidylic acid (poly(I:C)) and lipopolysaccharide (LPS) pretreatment on the immunological capacity of MSCs in vitro and in vivo.MethodsC57BL/6 bone marrow-derived MSCs were pretreated with poly(I:C) and LPS for 1 hour and their immunomodulatory capacity was evaluated. T-cell proliferation and their effect on Th1, Th17, and Treg differentiation/activation were measured. Next, we evaluated the therapeutic effect of MSCs in an experimental autoimmune encephalomyelitis (EAE) model, which was induced for 27 days with MOG35–55 peptide following the standard protocol. Mice were subjected to a single intraperitoneal injection (2 × 106 MSCs/100 μl) on day 4. Clinical score and body weight were monitored daily by blinded analysis. At day 27, mice were euthanized and draining lymph nodes were extracted for Th1, Th17, and Treg detection by flow cytometry.ResultsPretreatment of MSCs with poly(I:C) significantly reduced the proliferation of CD3+ T cells as well as nitric oxide secretion, an important immunosuppressive factor. Furthermore, MSCs treated with poly(I:C) reduced the differentiation/activation of proinflammatory lymphocytes, Th1 and Th17. In contrast, MSCs pretreated with LPS increased CD3+ T-cell proliferation, and induced Th1 and Th17 cells, as well as the levels of proinflammatory cytokine IL-6. Finally, we observed that intraperitoneal administration of MSCs pretreated with poly(I:C) significantly reduced the severity of EAE as well as the percentages of Th1 and Th17 proinflammatory subsets, while the pretreatment of MSCs with LPS completely reversed the therapeutic immunosuppressive effect of MSCs.ConclusionsTaken together, these data show that pretreatment of MSCs with poly(I:C) improved their immunosuppressive abilities. This may provide an opportunity to better define strategies for cell-based therapies to autoimmune diseases.

Relative maturity and the development of immunity to gastrointestinal nematodes in sheep: an overlooked paradigm?

Sheep display considerable variation in both the timing and magnitude of development of immunity to gastrointestinal nematodes (GIN). Onset of immunity is dependent on a number of factors, including antigenic stimulus, nutrition supply, age and size of the animals, the latter of which are confounded. Here, we review the factors associated with the development of immunity to GIN in sheep, particularly in the context of the role that relative maturity may have through applying the rules of genetic size scaling based on examples from published literature. Comparing animals based on their metabolic age, rather than chronological age, may provide an explanation for the timing of immune development and may reduce the variation in immune development that frequently is observed both between and within breeds. Further, this approach may help explain the phenotypic differences in animal performance between animals of varying immunological capacity to GIN through influences on mature body weight. As such, when considering factors influencing immune development to GIN, physiological age or relative maturity may be considered an overlooked paradigm. We propose it may be worthwhile to consider metabolic age when comparing the immune competence of animals to ensure the subjects are at an analogous stage of physiological development.

BLOOD, METABOLIC AND ENDOCRINE BIOMARKERS IN DONKEYS (Equus africanus asinus) SUPPLEMENTED WITH DIFFERENT ENERGY SOURCES

Blood and endocrine biomarkers have rarely been described in donkeys. This study aimed to analyze the effects of isoenergetic supplementation with three energy sources for 8wk on blood and endocrine biomarkers in donkeys. Fifteen donkeys were used and divided into 3 groups (extruded feed (ExF), extruded feed plus corn (C+ExF), and corn (C) and were supplemented isocalorically. Blood samples were collected at three times (pre-test, 4 and 8 weeks after supplementation). After 8wk, there were not significant difference between the treatment but detected between the phase, where results showed decreased values of red blood cells, hemoglobin, hematocrit, other cells, total plasma protein, albumin, creatinine, uric acid, GGT, AF and NEFA (P 0.05), however, differences were significant for [insulin] (P < 0.05). ExF group exhibiting the highest insulin concentration (P < 0.05). As for the experimental phases, there were differences for glucose (P < 0.05), with the highest mean concentration observed at the end of blood collection (+ 4 h). It was concluded that supplementation with three different combination of energy source in adult donkeys improve homeostasis of the energetic biomarker, reducing NEFA concentration without changes in post-prandial glucose but modifying post-prandial insulin, and increase immunological capacity associated with an increase in the lymphocyte number.

Investigation of three lignin complexes with antioxidant and immunological capacities from Inonotus obliquus

Mushroom Inonotus obliquus (I. obliquus), a folk medicine, has been widely used to treat several human malicious tumors since 16th century. In this study, three homogenous biomolecules (designated IOA1, IOA2 and IOA3) were prepared from the alkali extract of I. obliquus. Their molecular weights were measured to be 6.1×104, 2.9×104 and 3.5×104g/mol respectively and all of them were characterized as lignin–carbohydrate complexes mainly comprised lignin as well as −25% carbohydrates. Antioxidant assays indicated that all of them exhibited pronounced reductive power and strong scavenging activities on DPPH and hydroxyl radicals in vitro. Immunological tests showed that they could also significantly stimulate nitric oxide production and phagocytic activity in RAW 264.7 macrophages. These results implied that the lignin–carbohydrate complexes extracted from I. obliquus might be used as novel natural antioxidants or immunostimulants in functional foods or pharmaceutical candidates.

The immunological capacity in the larvae of Pacific oyster Crassostrea gigas.

As the immune system has not fully developed during early developmental stages, bivalve larvae are more susceptible for pathogens, which frequently leads to the significant mortality in hatcheries. In the present study, the development of immune system and its response against bacteria challenge were investigated in order to characterize the repertoire of immunological capacity of Pacific oyster Crassostrea gigas during the ontogenesis. The phagocytosis was firstly observed in the early D-veliger larvae (17hpf), especially in their velum site, which indicated the appearance of functional hemocytes during early D-veliger larvae stage. The whole-mount immunofluorescence assay of three pattern recognition receptors (integrin β-1, caspase-3 and C-type lectin 3) and one immune effector gene (IL17-5) was performed in blastula, early D-veliger and umbo larvae, suggested that velum and digestive gland were the potential sites of immune system in the larvae. The lowest activities of antioxidant enzymes (superoxide dismutase and catalase) and hydrolytic enzyme (lysozyme), as well as descended expression levels of 12 immune genes at the transition between embryogenesis and planktonic, indicated that the larvae at hatching (9hpf) were in hypo-immunity. While the ascending activities of enzymes and expression levels of seven immune genes during the trochophore stage (15hpf) suggested the initiation of immune system. The steadily increasing trend of all the 12 candidate genes at the early umbo larvae (120h) hinted that the immune system was well developed at this stage. After bacterial challenge, some immune recognition (TLR4) and immune effector (IL17-5 and defh2) genes were activated in blastula stage (4hpf), and other immune genes were up regulated in D-veliger larvae, indicating that the zygotic immune system could respond earlier against the bacterial challenge during its development. These results indicated that the cellular and humoral immune components appeared at trochophore stage, and the cellular immune system was activated with its occurrence, while the humoral immune system executed until the early umbo larval stage. The immune system emerged earlier to aid larvae in defending bacterial challenge during the early stages of oyster development.

Fipronil Induced Hepatotoxicity, Genotoxicity, Oxidative Stress and the Possible Ameliorative Effect of Ginseng

Fipronil (FPN) is a wide-spectrum insecticide and has been extensively used in agriculture since 1990s. The current study was deliberated to inspect the retrograde effects of exposure to the FPN on the hepatic tissues of male rats at concentration 10 mg/L in drinking water for 30 days. Serum aspartate aminotransferases (AST), alanine aminotransferases (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activity and tumor necrosis factor –α (TNF-α) were significantly increased in FPN-treated rats. Oxidative stress biomarkers such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reduced (GSH) were significantly decreased, while malondialdehyde level (MDA) was significantly increased in treating rats in a concentration dependent manner. From the obtained results, it can be concluded that FPN elicited lipid peroxidation, oxidative stress and liver injury in rats and ginseng has protective effect against lipid peroxidation induced by FPN as ginseng lowers the level of MDA and increased levels of SOD and CAT and also ginseng improved the immunological capacities by diminishing the level of TNF-α. These pathophysiological changes in liver tissues could be due to the toxic effect of FPN that related to a production of free radicals.

Ontogenic development of kidney, thymus and spleen and phenotypic expression of CD3 and CD4 receptors on the lymphocytes of cobia (Rachycentroncanadum).

In the present study was evaluated the ontogenic of immunocompetent organs of cobia up to 53 days after hatching (dah) through histology and immunohistochemistry techniques. The kidney was the first lymphohematopoietic organ to appear, at 1 dah, followed by the spleen at 5 dah and the thymus at 7 dah. The first CD3 receptors on the lymphocytes were observed in 27% of the thymic tissue at 7 dah and in 99% at 53 dah. The phenotypic expression of CD3 receptors was registered in 10% of the kidney at 8 dah and in 32% at 53 dah. CD4 receptors were observed in 5% and 63% of the thymic area at 7 and 53 dah, respectively. In the kidney, T4 lymphocytes were first observed at 13 dah in 9% of the organ and in 28% at 53 dah, defining the functional development of the specific system associated with immunological memory capacity.

Expression of histamine receptors in the human endolymphatic sac: the molecular rationale for betahistine use in Menieres disease.

The human endolymphatic sac (ES) is situated in a duplicature of the dura in the posterior cranial fossa and constitutes a part of the inner ear. The sac possesses immunological capacities and is responsible for a major part of the trans-epithelial ion transport occurring within the inner ear, via molecular mechanisms similar to that of the kidney collecting duct epithelia. Dysfunction of the trans-epithelial ion transport has been hypothesized as the reason for the endolymphatic hydrops occurring in Menieres diseases. Thus, candidate drug selection for medical treatment of Menieres disease has been based on a potential capability of improving trans-epithelial ion transport. However, recent human studies seems to rule out diuretic therapy and The Committee for Medicinal Products for Human Use redrew the recommendation for trimetazidine (Vastarel) treatment in the management of Meniere disease in 2012. This leaves betahistine (Betaserc) as the only drug for potential prevention of the incapacitating attacks of dizziness, tinnitus and hearing loss. However, the histamine receptors targeted by betahistine have never been demonstrated in the human ES. Accordingly, this study aims to investigate the expression of histamine receptors of the human ES epithelium and sub-epithelial stroma. Following sampling of human endolymphatic sac tissue during translabyrinthine surgery, the expression of histamine receptor genes was determined by cDNA microarray analysis. Results were subsequently verified by immuno-histochemistry. The combined results of microarrays and immuno-histochemistry showed expression of the histamine receptor HRH1 in the epithelial lining of the ES, whereas HRH3 was expressed exclusively in the sub-epithelial capillary network. Receptors HRH2 and -4 were not expressed. The present data provide the first direct evidence of a molecular rationale for betahistine treatment in Menieres disease. A potential betahistine effect in Menieres disease may primarily be through the H3-receptor antagonism, leading to inhibition of vestibular neuro-transmission and central vaso-dilation. The H1-receptor localization in the ES epithelium suggests an immuno-regulatory effect.

Does donor age matter? Biological and immunogenic differences between infant and aged mesenchymal stromal cells

Over the last decades, mesenchymal stromal cells MSC) have emerged as a promising therapeutic tool for cell-based therapies. However, their biological characteristics and beneficial potential may decline with patients’ age. Because most patients amenable for regenerative medicine have increased age, this could dramatically limit the autologous use of MSCs. We therefore sought to characterize the biological, immunological, and angiogenetic capacities of infant (iMSCs) versus aged MSCs (aMSCs). Both iMSCs and aMSCs showed typical spindle-shaped morphology, plastic adherence, and the same potential to differentiate into osteogenic, chondrogenic and adipogenic lineages. iMSCs and aMSCs expressed MSC-typical surface markers such as CD90 and CD105 and were negative for CD45, CD34, CD31 and CD117. iMSCs showed faster in vitro proliferation (p<0.001), migration (p<0.001), and stronger endothelial adhesion (p=0.018) than aMSC. There were no differences in HLA class I, β2 microglobulin, HLA class II, and co-stimulatory molecule expression between iMSCs and aMSCs. Both immunogenic HLA class II and CD54 and tolerogenic HLA-E expression were upregulated by IFNγ. iMSCs showed significantly higher CD29 signals and stronger endothelial adhesion than aMSCs. Interestingly, iMSCs outperformed aMSCs in different angiogenesis parameters. If not used in an autologous setting, iMSCs and aMSCs similarly provoked TH1 and TH2 responses in unidirectional ELISPOT assays and donor-specific IgM antibody production. Our results thus suggest that donor age does not affect immunogenicity and immunomodulatory properties, but markedly reduces biological properties of proliferation, migration, adhesion, and limits their angiogenic effects.

Effect of Knockdown of Vascular Cell Adhesion Molecule-1 on the Immunologic Regulation Capacity of Murine Mesenchymal Stem Cells

To establish the stably lower expression of vascular cell adhesion molecule-1 (VCAM-1) in MSC cell line (C3H10T1/2) by siRNA technology, and explore the effect of knockdown of VCAM-1 on the immunologic regulation capacity of murine MSC. The mouse GV118-VCAM-1-RNAi retrovirus vector was constructed by gene recombination technology. The recombinant plasmid was identified by restriction analysis and sequencing, and then the recombinant plasmid GV118-VCAM-1-RNAi was transfected into 293 cells by Lipofectamine, and the supernatant was collected to transfect C3H10T1/2. Moreover, the VCAM-1 lower expression on MSC was evaluated by flow cytometry and fluorescent microscopy. The knockdown VCAM-1 MSC was sorted by flow cytometry. Furthermore, the inhibitory effect of the knockdown VCAM-1 MSC on lymphocyte proliferation was tested by lymphoblast transformation assay (LTT) and mixed lymphocyte reaction assay(MLR). The recombinant retroviral vector of knockdown VCAM-1 (GV118-VCAM-1-RNAi) was successfully constructed and transfected into mouse MSC cell line C3H10T1/2. The knockdown VCAM-1/MSC was obtained by flow cytometric sorting. The LTT and MLR assay showed that the immunosuppressive effect of MSC lower-expressing VCAM-1 dramatically decreased (P<0.05). Knockdown VCAM-1 in MSC can significantly down-regulate the inhibitory capability of MSC on the proliferation of T-cells. The data of this study laid an experimental foundation for studying effect of VCAM-1 transfecting into MSC on immune function.

Ameliorative Roles of Silymarin and Nigella sativa on Hematological Parameters and Immunological Capacities of Male Mice Affected by Paracetamol

Ameliorative Roles of Silymarin and Nigella sativa on Hematological Parameters and Immunological Capacities of Male Mice Affected by Paracetamol

Low prevalence of blood parasites in a long-distance migratory raptor: the importance of host habitat.

BackgroundThe low prevalence of blood parasites in some bird species may be related to the habitats they frequent, the inexistence of the right host-parasite assemblage or the immunological capacity of the host. Here, we assess the parasite load of breeding populations of Eleonora’s falcon (Falco eleonorae), a medium-sized long-distance migratory raptor that breeds on small isolated islets throughout the Mediterranean basin and overwinters in inland Madagascar.MethodsWe examined the prevalence and genetic diversity of the blood parasites belonging to the genera Plasmodium, Haemoproteus and Leucocytozoon in Eleonora’s falcon nestlings from five colonies and in adults from two colonies from nesting sites distributed throughout most of the species’ breeding range.ResultsNone of the 282 nestlings analysed were infected by blood parasites; on the other hand, the lineages of Plasmodium, Haemoproteus and Leucocytozoon were all found to infect adults. Our results support the idea of no local transmission of vector-borne parasites in marine habitats. Adult Eleonora’s falcons thus may be infected by parasites when on migration or in their wintering areas.ConclusionThe characteristics of marine environments with a lack of appropriate vectors may thus be the key factor determining the absence of local transmission of blood parasites. By comparing the parasite lineages isolated in this species with those previously found in other birds we were able to infer the most likely areas for the transmission of the various parasite lineages.

Gene expression demonstrates an immunological capacity of the human endolymphatic sac.

The purpose of the present study is to explore, demonstrate, and describe the expression of genes related to the innate immune system in the human endolymphatic sac. It is hypothesized that the endolymphatic sac has a significant immunological function in the human inner ear. DNA microarrays and immunohistochemistry were used for analyses of fresh human endolymphatic-sac tissue samples. Twelve tissue samples from the human endolymphatic sac were obtained during translabyrinthine surgery for vestibular schwannoma. Microarray technology was used to investigate tissue sample gene expression using adjacent dura mater as control. The expression of genes specific for the innate immune system was determined and results for selected key molecules verified by immunohistochemistry. A comprehensive overview of expressed genes of the innate immune system was obtained. Multiple key elements of both the cellular and humoral innate immune system were expressed, including Toll-like receptors 4 and 7, as well as beta-defensin and lactoferrin. The present data provides the first direct evidence of an immunological capacity of the human endolymphatic sac. At the molecular level, the endolymphatic sac is capable of antigen recognition and processing for initiation of an immune response. In addition, potent molecules directly toxic to invading pathogens are expressed by the sac epithelium. This evidence strongly supports the endolymphatic sac as a significant immunological entity of the inner ear. N/A.

Management of HIV in Children Using a Bovine Colostrum-Based Food Product— An Observational Field Study

Aim: Malnutrition increases the severity and mortality of HIV infection. Therefore, this field study was started and aimed to provide a major cohort of HIV-infected children with the colostrum- based food product ColoPlus (ColoPlus AB, Malmo, Sweden) and to investigate the effects of Co- loPlus on the nutritional status and immunological capacity of children including tolerability and safety of the product. Methods: In this major field program comprising 850 malnourished HIV positive children, 50 grams of ColoPlus was administrated for 4 weeks as the first meal in selected health facilities in Northern Uganda. Forty-eight of these children (8 months - 14 years of age) were recruited into a descriptive prospective study and were followed for 12 weeks. At the start (week 0) and at weeks 4, 8 and 12, CD4+ cell counts, serum albumin and hemoglobin were ana- lyzed. The remaining 802 patients were observed to register safety and tolerability of ColoPlus. Results: There was a general improvement of wellbeing of the children with increased body weight and decreased fatigue. In the descriptive study of the 48 children, there was a significant rise of the CD4+ values at week 4 (+15.4% ± 2.8%, p = 0.0001) compared to week 0, and at week 8 (+39.1% ± 3.9%, p < 0.0001), but a return towards the 0-values at week 12 (+2.1%, ±2.8%, NS). He- moglobin and serum albumin showed an almost similar trend. ColoPlus was well tolerated by all the 850 children and no side effects or adverse events were seen. Conclusion: These results show that addition of a colostrum-based food product to the daily diet is beneficial in HIV-positive malnou- rished children. An improvement in nutritional status as well as in immune capacity was seen. These effects were prolonged and remained at least 4 weeks after cessation of ColoPlus administration. * Corresponding author.

The effects of temperature and body size on immunological development and responsiveness in juvenile shortnose sturgeon (Acipenser brevirostrum)

Sturgeon are an important evolutionary taxa of which little is known regarding their responses to environmental factors. Water temperature strongly influences growth in fish; however, its effect on sturgeon immune responses is unknown. The objective of this study was to assess how 2 different temperatures affect immune responses in shortnose sturgeon (Acipenser brevirostrum) relevant immune organs such as the meningeal myeloid tissue, spleen, thymus and skin. These responses were studied in 2 different sizes of same age juvenile sturgeon kept at either 11 °C or 20 °C (4 treatment groups), before and after exposure to an ectoparasitic copepod (Dichelesthium oblongum). Based on a differential cell count, temperature was found to strongly influence immune cell production in the meningeal myeloid tissue, regardless of the fish sizes considered. Morphometric analysis of splenic white pulp showed a transient response to temperature. There were no differences between the groups in the morphometric analysis of thymus size. Splenic IRF-1 and IRF-2 had similar expression profiles, significantly higher in fish kept at 20 °C for the first 6 weeks of the study but not by 14 weeks. In the skin, IRF-1 was significantly higher in the fish kept at 11 °C over the first 6 weeks of the study. IRF-2 had a similar profile but there were no differences between the groups by the end of the trial. In conclusion, higher water temperatures (up to 20 °C) may have beneficial effects in maximizing growth and improving immunological capacity, regardless of the fish sizes considered in this study.

B cells and ectopic follicular structures: novel players in anti-tumor programming with prognostic power for patients with metastatic colorectal cancer.

Remarkably limited information is available about biological mechanisms that determine the disease entity of metastatic colorectal cancer in the liver (CRCLM) with no good clinical parameters to estimate prognosis. For the last few years, understanding the relationship between tumor characteristics and local immune response has gained increasing attention. Given the multifaceted roles of B-cell-driven responses, we aimed to elucidate the immunological imprint of B lymphocytes at the metastatic site, the interrelation with macrophages, and their prognostic relevance. Here we present novel algorithm allowing to assess a link between the local patient-specific immunological capacity and clinical outcome. The microscopy-based imaging platform was used for automated scanning of large-scale tissue sections and subsequent qualitative and quantitative analyses of immune cell subtypes using lineage markers and single-cell recognition strategy. Results indicate massive infiltration of CD45-positive leukocytes confined to the metastatic border. We report for the first time the accumulation of CD20-positive B lymphocytes at the tumor – liver interface comprising the major population within the large CD45-positive aggregates. Strikingly, functionally active, activation-induced cytidine deaminase (AID)-positive ectopic lymphoid structures were found to be assembled within the metastatic margin. Furthermore, the CD20-based data set revealed a strong prognostic power: patients with high CD20 content and/or ectopic follicles had significantly lower risk for disease recurrence as revealed by univariate analysis (p<0.001 for both) and in models adjusted for clinicopathological variables (p<0.001 and p = 0.01, respectively), and showed prolonged overall survival. In contrast, CD68 staining-derived data set did not show an association with clinical outcome. Taken together, we nominate the magnitude of B lymphocytes, including those organized in ectopic follicles, as novel prognostic marker which is superior to clinicopathological parameters. Findings emphasize anti-tumoral role of B cell-driven mechanism(s) and thus indicate a new way of thinking about potential treatment strategies for CRCLM patients.

Immunotoxic Effect of Sodium Fluoride and the Mitigating Effect of Selenium and Curcumin in Male Mice

Sodium fluoride used in the fluoridation of drinking water and it is still commonly used for that purpose to prevent dental caries. It exerts toxic effects on many soft tissues. The present study was aimed to achieving the protective effects of selenium and Curcumin extract against the toxicity and oxidative stress of sodium fluoride (NaF) that adversely affects the immune system capacity. This study evaluated the effects of Sodium fluoride on some immunological parameters and also assessed the ameliorating effects of selenium and Curcumin extract. Mature male mice (weighing 35-45 g and each group of ten animals) were given sodium fluoride (10.3 mg/Kg bw) and/or Selenium (0.5 mg/Kg) + Curcumin extract (60 mg/Kg) daily intraperitoneally (I.P) for 4 weeks. In the present study, Sodium Fluoride exposure resulted in decrease in the IgG, IgM and increasing TNF-± level with respect to the control. As a result, Sodium fluoride induced immunotoxicity which is reduced by Curcumin extract and/or selenium to great extent by restoration of the immunological capacities.