Immune Competent Cells Research Articles

GRP78 plays an integral role in tumor cell inflammation-related migration induced by M2 macrophages

Macrophages are the main immune-competent cells that infiltrate in tumors. Tumor-associated macrophages (TAMs), termed M2 macrophages, facilitate tumor progress and promote metastasis. However, M2 macrophages always display an immunosuppressive phenotype, which is not in accordance with the tumor inflammatory microenvironment and inflammation-related metastasis. In this study, we established a macrophage polarization model with human monocytes and found that the conditioned medium from M2 macrophages increased GRP78 expression in tumor cells and facilitated tumor cell migration. Mechanistically, excessive GRP78 formed a protein complex with STAT3 and JAK2 to promote STAT3 phosphorylation. Furthermore, p-STAT3 facilitated the high expression of inflammatory factors IL-1β and TNF-α in tumor cells, which was important in M2 macrophage-induced metastasis. The present data demonstrate that M2 macrophages elevate tumor cell GRP78 expression to trigger an inflammatory response, which further facilitates tumor metastasis. Therefore, our study not only uncovered a new cause of GRP78 overexpression in tumor cell, but also, explained the antinomy of TAMs immunosuppressive properties and inflammation-related tumor metastasis.

Mechanical deformation induces depolarization of neutrophils.

The transition of neutrophils from a resting state to a primed state is an essential requirement for their function as competent immune cells. This transition can be caused not only by chemical signals but also by mechanical perturbation. After cessation of either, these cells gradually revert to a quiescent state over 40 to 120 min. We use two biophysical tools, an optical stretcher and a novel microcirculation mimetic, to effect physiologically relevant mechanical deformations of single nonadherent human neutrophils. We establish quantitative morphological analysis and mechanical phenotyping as label-free markers of neutrophil priming. We show that continued mechanical deformation of primed cells can cause active depolarization, which occurs two orders of magnitude faster than by spontaneous depriming. This work provides a cellular-level mechanism that potentially explains recent clinical studies demonstrating the potential importance, and physiological role, of neutrophil depriming in vivo and the pathophysiological implications when this deactivation is impaired, especially in disorders such as acute lung injury.

Hematopoietic stem cell transplantation in autoimmune disorders: From immune-regulatory processes to clinical implications.

Autoimmune diseases are characterized by the development of autoreactive T- and B-cells targeting self-antigens, which eventually can result in chronic and persistent organ damage. The autologous hematopoietic stem cell transplantation (AHSCT) opened new avenues in the treatment of patients with severe, treatment-resistant autoimmune diseases. This paper reviews the immune-regulatory mechanisms behind AHSCT, and also summarizes the experiences of clinical practice related to the therapy in organ-specific and systemic autoimmune diseases. It seems that the intricate interplay of various immune competent cells with regulatory capacity control in a synergistic manner the repopulated immune system after AHSCT, which potentially leads to a significant clinical improvement in certain autoimmune diseases. However, the widespread use of AHSCT was intrinsically limited, due to the serious side-effects of conditioning treatment and relatively high treatment-related mortality; moreover, the development of new effective and safe therapeutic approaches and the dawn of biological agents further limited its indications in the last decade. Nevertheless, with an appropriate patient selection and increased experience of transplant centres, the risks can be minimized, and AHSCT remained still a reasonable choice in multiple sclerosis and systemic sclerosis when the conventional therapy failed and further progression of disease is inevitable.

Immunomodulation by Mesenchymal Stromal Cells and Their Clinical Applications.

Mesenchymal stromal cells (MSCs) are multipotent progenitor cells that can be isolated and expanded from various sources. MSCs modulate the function of immune cells, including T and B lymphocytes, dendritic cells, and natural killer cells. An understanding of the interaction between MSCs and the inflammatory microenvironment will provide critical information in revealing the precise in vivo mechanisms involved in MSCs-mediated therapeutic effects, and for designing more practical protocols for the clinical use of these cells. In this review we describe the current knowledge of the unique biological properties of MSCs, the immunosuppressive effects on immune-competent cells and the paracrine role of soluble factors. A summary of the participation of MSCs in preclinical and clinical studies in treating autoimmune diseases and other diseases is described. We also discuss the current challenges of their use and their potential roles in cell therapies.

Platelet factor 4 upregulates matrix metalloproteinase-1 production in gingival fibroblasts.

Periodontitis is a highly prevalent chronic inflammatory disease that causes tooth loss, morbidity and confers an increased risk for systemic disease. Tissue destruction during periodontitis is due in large part to collagen-degrading matrix metalloproteinases (MMPs) released by resident cells of the periodontium in response to proinflammatory cytokines. Platelets are immune-competent blood cells with a newly recognized role in chronic inflammation; however, their role in the pathogenesis of periodontitis is undefined. Consequently, the objective of this study was to assess the effect of platelet factor 4 (PF4), a major platelet-derived cytokine, on MMP-1 (collagenase) expression in human gingival fibroblasts (HGFs). HGFs were cultured in the presence or absence of recombinant PF4. Pro-MMP-1 secretion was quantified by enzyme-linked immunosorbent assay analysis of the cell culture supernatants. MMP-1 transcription was quantified by real-time polymerase chain reaction. Regulation of MMP-1 production by the p44/42 MAP kinase (MAPK) signaling pathway was examined in the presence or absence of PF4. Exposure to PF4 caused a ~ 2-3-fold increase in MMP-1 transcription and secretion from cultured HGFs. PF4 treatment also enhanced phosphorylation of p44/42 MAPK, which has been previously shown to induce MMP-1 expression in fibroblasts. Blockade of p44/42 MAPK signaling with the cell-permeant inhibitors PD98059 and PD184352 abrogated PF4-induced pro-MMP-1 transcription upregulation and release from cultured HGFs. We conclude that PF4 upregulates MMP-1 expression in HGFs in a p44/42 MAPK-dependent manner. These findings point to a previously unidentified role for platelets in the pathogenesis of periodontal diseases.

Targeting the Monocyte-Macrophage Lineage in Solid Organ Transplantation.

There is an unmet clinical need for immunotherapeutic strategies that specifically target the active immune cells participating in the process of rejection after solid organ transplantation. The monocyte–macrophage cell lineage is increasingly recognized as a major player in acute and chronic allograft immunopathology. The dominant presence of cells of this lineage in rejecting allograft tissue is associated with worse graft function and survival. Monocytes and macrophages contribute to alloimmunity via diverse pathways: antigen processing and presentation, costimulation, pro-inflammatory cytokine production, and tissue repair. Cross talk with other recipient immune competent cells and donor endothelial cells leads to amplification of inflammation and a cytolytic response in the graft. Surprisingly, little is known about therapeutic manipulation of the function of cells of the monocyte–macrophage lineage in transplantation by immunosuppressive agents. Although not primarily designed to target monocyte–macrophage lineage cells, multiple categories of currently prescribed immunosuppressive drugs, such as mycophenolate mofetil, mammalian target of rapamycin inhibitors, and calcineurin inhibitors, do have limited inhibitory effects. These effects include diminishing the degree of cytokine production, thereby blocking costimulation and inhibiting the migration of monocytes to the site of rejection. Outside the field of transplantation, some clinical studies have shown that the monoclonal antibodies canakinumab, tocilizumab, and infliximab are effective in inhibiting monocyte functions. Indirect effects have also been shown for simvastatin, a lipid lowering drug, and bromodomain and extra-terminal motif inhibitors that reduce the cytokine production by monocytes–macrophages in patients with diabetes mellitus and rheumatoid arthritis. To date, detailed knowledge concerning the origin, the developmental requirements, and functions of diverse specialized monocyte–macrophage subsets justifies research for therapeutic manipulation. Here, we will discuss the effects of currently prescribed immunosuppressive drugs on monocyte/macrophage features and the future challenges.

Immune deficiency augments the prevalence of p53 loss of heterozygosity in spontaneous tumors but not bi-directional loss of heterozygosity in bone marrow progenitors.

p53 loss of heterozygosity (LOH) is a frequent event in tumors of somatic and Li-Fraumeni syndrome patients harboring p53 mutation. Here, we focused on resolving a possible crosstalk between the immune-system and p53 LOH. Previously, we reported that p53 heterozygous bone-marrow mesenchymal progenitor cells undergo p53 LOH in-vivo. Surprisingly, the loss of either the wild-type p53 allele or mutant p53 allele was detected with a three-to-one ratio in favor of losing the mutant allele. In this study, we examined whether the immune-system can affect the LOH directionality in bone marrow progenitors. We found that mesenchymal progenitor cells derived from immune-deficient mice exhibited the same preference of losing the mutant p53 allele as immune-competent matched cells, nevertheless, these animals showed a significantly shorter tumor-free survival, indicating the possible involvement of immune surveillance in this model. Surprisingly, spontaneous tumors of p53 heterozygous immune-deficient mice exhibited a significantly higher incidence of p53 LOH compared to that observed in tumors derived of p53 heterozygous immune-competent mice. These findings indicate that the immune-system may affect the p53 LOH prevalence in spontaneous tumors. Thus suggesting that the immune-system may recognize and clear cells that underwent p53 LOH, whereas in immune-compromised mice, those cells will form tumors with shorter latency. In individuals with a competent immune-system, p53 LOH independent pathways may induce malignant transformation which requires a longer tumor latency. Moreover, this data may imply that the current immunotherapy treatment aimed at abrogating the inhibition of cellular immune checkpoints may be beneficial for LFS patients.

Abstract A21: 3D tumor models in bioreactors recapitulate microenvironment and disease progression

Abstract The high attrition rates observed in cancer drug discovery (up to 95% of failure of drugs tested in phase I trials) have raised the awareness of the scientific and industrial communities towards the need for more predictive pre-clinical models. These models should be more representative of the disease and consequently help to eliminate at pre-clinical stages drug candidates that lack efficacy or safety. Tumor microenvironment is composed by a network of fibroblasts, endothelial cells, immune-competent cells within the extracellular matrix (ECM). Interactions between these components are critical for tumor initiation, proliferation, migration and metastasis. The design of in vitro cancer cell models that recapitulate the tumor microenvironment and 3D architecture provides higher physiological relevance as they more closely resemble the in vivo cellular context. We have established methodologies for scalable generation of 3D cancer cell models in stirred-tank culture systems, and applied these to a large panel of tumor cell lines from different pathologies, including breast, colon, hepatic and lung tumor cell lines. Large numbers of spheroids were obtained per culture (typically 1000-1500 spheroids/mL) with representative characteristics of native tumors, such as morphology, proliferation and hypoxia gradients, in a cell-line dependent mode. With the aim of increasing the relevance of spheroids as tumor cell models, several aspects of tumor microenvironment were incorporated, such as the presence of stromal cells (fibroblasts and monocytes) and specific physical parameters, namely by embedding cells in a polymeric matrix. Heterotypic 3D breast and Non-Small Cell Lung Carcinoma (NSCLC) cancer models, based on co-cultures of tumor cells with stromal cells were established by using an alginate matrix to provide physical support to cells. Tumor spheroids were microencapsulated alone or with fibroblasts and monocytes, thus allowing the establishment of an epithelial tumor compartment and a stromal compartment of increasing complexity. Cultures were performed in stirred-tank vessels for 15 days with continuous monitoring. In both breast and lung tumor models, the presence of fibroblasts was associated with secretion of pro-inflammatory cytokines and accumulation of collagen in the microcapsules. Long-term culture (up to 15 days) resulted in phenotypic alterations in co-cultured breast tumor spheroids, such as loss of cell polarity, reduced cell-cell adhesions, collective cell migration and increased angiogenic potential. In contrast, the effects of fibroblasts were not as significant in NSCLC co-cultures using H1650, H1437 and H157 cell lines suggesting that the effect of tumor-stroma cross-talk is cell line dependent. Moreover, these models have also been shown as feasible tools for drug screening by assessing the effect of chemotherapeutic and specific inhibitors compounds on mono- and co-cultures. In conclusion, we have developed scalable, robust and versatile methodologies for the generation and culture of 3D cancer models, enabling long-term in vitro recapitulation of tumor-stroma crosstalk, via reconstruction of key aspects of the tumor microenvironment, allowing continuous monitoring of disease progression events in vitro. In addition, it is easily transferable to industry for feeding high-throughput systems or miniaturized bioreactors used in drug development, target validation and target identification. Acknowledgments: We acknowledge support from the Innovative Medicines Initiative Joint Undertaking (IMI grant agreement n° 115188), resources composed of financial contribution from EU - FP7 and EFPIA companies in kind contribution. iNOVA4Health Research Unit (LISBOA-01-0145-FEDER-007344), which is cofunded by Fundação para a Ciência e Tecnologia / Ministério da Ciência e do Ensino Superior, through national funds, and by FEDER under the PT2020 Partnership Agreement, is acknowledged. This research has also received support from Fundação para a Ciência e Tecnologia, Portugal—PTDC/BBB-BIO/1240/2012. MFE, SA and CP are the recipients of PhD fellowships from FCT (SFRH/BD/52208/2013, PD/BD/105768/2014 and SFRH/BD/52202/2013, respectively). Citation Format: Vitor E Santo, Marta F Estrada, Sofia P Rebelo, Sofia Abreu, Catarina Pinto, Elizabeth Anderson, Wolfgang Sommergruber, Paula Alves, Erwin Boghaert, Catarina Brito. 3D tumor models in bioreactors recapitulate microenvironment and disease progression. [abstract]. In: Proceedings of the AACR Special Conference on Engineering and Physical Sciences in Oncology; 2016 Jun 25-28; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2017;77(2 Suppl):Abstract nr A21.

Methods of Assessing STING Activation and Trafficking.

The signaling adapter protein STING is crucial for the host immune response to cytosolic DNA and cyclic dinucleotides. Under basal conditions, STING resides on the endoplasmic reticulum (ER ) , but upon activation, it traffics through secretory pathway to cytoplasmic vesicles, where STING activates downstream immune signaling. Classical STING activation and trafficking are triggered by binding of cyclic dinucleotide ligands. STING signaling can also be activated by gain-of-function mutations that lead to constitutive trafficking of STING. These gain-of-function mutations are associated with several human diseases such as STING-associated vasculopathy with onset in infancy (SAVI), systemic lupus erythematosus (SLE), or familial chilblain lupus (FCL). This dynamic activation pathway presents a challenge to study. We describe methods here for measuring ligand-dependent and ligand-independent activation of STING signaling in HEK293T cells. We also describe a retroviral-based reconstitution assay to study STING protein trafficking and activation in immune competent cells such as mouse embryonic fibroblasts (MEF), which avoids the use of plasmid DNA. These methods will expedite research regarding STING trafficking and signaling dynamics in the settings of infection and autoimmune diseases.

Fermented Ethnic Medicine (<i>Pueraria mirifica</i>) Designed by Lactobacillus Regulated Leukocyte Subsets via Activation of Complement Components

A plant fermentation was carried out by Lactobaccilli against the Rhizome from Pueraria milifica (f-PMF). This material was evidenced by safe in animal toxic study. The main aim of this study was to revise the traditional way of hot water extraction to fermentation so as to use up the original material and finding new activity. We tried to show the new activity through phytoestrogen and immune-competent cells from the host that administrated either of original remedy and the new fermented sample, plus activated water SRE. In mice, compromised host was prepared by cancer chemotherapeutic agent (Mitomycin-C). After administration of f-PMF to immno-suppressed animals, the effects by both samples were augmented by lymphocyte in number and functions, macrophage activities, anti-oxidative activity. However, the intense of effect was much more by fermented one but not by conventional one. The anti-oxidative assay was also carried out ex-vivo system by peritoneal macrophage that we proposed as suitable system for evaluating anti-oxidative assay. In our clinical study by 20 healthy volunteers, granulocyte and lymphocyte ratio was regulated as neutral in peripheral white blood cells, increasing one, two and three weeks after the administration of f-PMF. We have found the significant regulation of blood chemical factors that were important makers for the lifestyle-related diseases. The mechanism of augmentation by probing directory with immuno-electrophoretic method, generating new complement component, especially found by alternative pathway of complement. So we discussed the process concerning designed f-PMF molecule for activation of complement component and bound for the biological activity of each physical component. In a limited condition, fewer numbers of volunteers, the breast size was tending to increase along with the administration time. Including these evidences, we discussed the possibility of this traditional ethnic medicine, originally found and spread in the highland area in Thailand and Myanmar.

Research progress of active polypeptides ofCordyceps militaris

It is recognized at home and abroad that Cordyceps militaris is a fungus that can be edible and used for medicinal application, and also a common cordyceps taishanensis that is widely distributed and with very high medicinal value in China. Active Cordyceps militaris polypeptide has shown several highlights in its absorption mechanism, such as directly absorbed without the need for digestion, and absorbed quickly and completely, and more and more international researchers are attaching great importance to it. Physiological and pharmacological activities of Cordyceps militaris polypeptide are mainly to activate related enzyme systems, promote intermediary metabolisms or control the DNA transcription and translation, simultaneously activate the reticuloendothelial system and macrophage, promote the transformation of lymphocytes, and as a nonspecific immune enhancer and regulator, activate the immune competent cells, especially lymphocytes, lymphokines, mononuclear macrophage system and NK cells, thereby attacking the target cells to play an antitumor role, and also exerting its anti-aging, anticoagulant, hypolipidemic to enhance the immunity, improve the liver function and delay the aging. In this paper, the immunity improvement, anticancer, antioxidation and antimicrobial activities of active polypeptides from Cordyceps militaris are reviewed, in order to provide a solid theoretical help for the further development and application of active polypeptides of Cordyceps militaris .

Parameters of innate and adaptive immune response in children with virus-induced and allergen-induced bronchial asthma phenotypes

Aim. To analyze the immune mechanisms of activation of virus-induced and allergen-induced bronchial asthma phenotypes in children. Materials and м ethods. We have performed an integrated assessment of parameters of innate and acquired immunity in 98 children with bronchial asthma (BA) at the age of 3–11 years in groups with virus-induced (n = 49) and allergen-induced (n = 49) phenotypes. Bronchial asthma phenotypes were verified in accordance with PRACTALL international consensus report (2008). Study exclusion criteria were: severe course of bronchial asthma and application of immunocorrecting drugs during preceding six months. To analyze immune competent cells we used flow cytofluorometer COULTER EPICS XL by Beckman Coulter Inc. Cytokine levels were identified using immunoenzyme method and reagents by R & D Diagnostics Inc. (USA); IgE – using reagents by Alkor Bio company (Saint-Petersburg); production of cytokines – using reagents by VektorBest (Novosibirsk city). Statistical processing of data was performed by Statistica. 10 program with critical significance level p < 0,05, correlations were analyzed using Spearman rank correlation coefficient. Results. Children with bronchial asthma showed enhanced expression of T-lymphocyte activation marker HLA–DR+(р < 0,05) with reduced level of CD3+CD95+- (р < 0,01), high level of serum IgE (р < 0,01), the inverse correlation between serum levels of IFNγ and IgE (r = –0,16), IFNγ and IL-13 (r = –0,25), direct correlation between IL-17A and IgE (r = 0,48), IgE and IL-13 (r = 0,56). The children with allergeninduced BA phenotype showed low expression of CD95+ on T-lymphocyte cells, predominance of Тh-2 profile cytokines, high mitogen-induced production of IL-4 and low induction of IFNγ production. Levels of cytotoxic T-lymphocytes, natural killers, expression of activation markers and CD95+ on T-lymphocyte cells and mitogen-induced synthesis of IFNγ by cells in children with virus-induced BA phenotype were lower than the same indices in healthy children (at р < 0,05; р < 0,01; р < 0,05; р < 0,01; р < 0,01). Levels of leucocytes, neutrocytes, absolute number of T-lymphocytes, T-helpers, B-lymphocytes (р < 0,001) and Th-2 – Th-17 cytokine profile in blood serum in this group were higher than the same indices in children with allergen-induced phenotype. Conclusion. Children with virus-induced BA phenotype showed presence of immune-mediated factors predisposing them to persistent viral infections and initiation of body sensibilization with development of eosinophilic inflammation, such as impairment of cellular cytotoxicity, predominance of Th-2 – Th-17 profile cytokines, inhibition of induction of IFNγ production by blood cells. The changes of thr adaptive response system in children with activation of allergen-induced BA phenotype are characterized by enhanced proliferation, inhibition of negative regulation processes, activation of synthesis of Th2 profile cytokines, and enhanced synthesis of IgE.

Fermented Medicinal Plant Designed by Lactobacillus Hits Alternative Complement Pathway and Controlled hA1c Level

Background: A Type II Diabetes mellitus showed many symptoms that brought serious for the patients. One of the problem was suppressed activity of the phagocyte for the foreign microorganism. Objective: The objective of this study was to find excellent method to recover the suppressed phagocytic cell activity both in animal and human. Methods: A wild medicinal herbs were prepared by Yeast and Lactobaccilli against 80 sorts of wild plants including medicinal remedies (FWP80). These products were exhibited by safe in animal safety test. The trial was made to investigate a recovery of immune-competent cells both in mice and human that exhibited phagocytic depression. Results: Our results in animal model showed that FWP80 augmented the macrophage activity, both chemotaxis and phagocytosis. In clinical study with 20 volunteers diagnosed type II DM, the granulocyte and lymphocyte ratio was produced to as modulate as neutral, 30 days after the introduction by FWP80. Conclusion: In mice, depressed phagocytic cell by Streptozotocin, the macrophage activities were recovered by FWP80. We proposed an idea that FWP80 exhibited remedy effect via controlling complement component. The mechanism of activation was proved by activating alternative pathway of complement by the technic, immuneelectrophoresis. We tried to evaluate more to the anti-oxidative activities of FWP80, directory employing peritoneal macrophage in vivo.

&amp;lt;i&amp;gt;Lactobacilli&amp;lt;/i&amp;gt; Enjoyed Fermented Herbs on to the Last Fragment and Regulated Leucocyte Subsets and Anti-Oxidative Activity

Eighty sorts of herbs were fermented by Lactobaccilli (f-ESH). This material was proved by as safe in animal safety experiment. This work was aimed to revise the traditional way of hot water extraction to the fermentation, in order to use up the original material and searching new activity as well. We tried to prove the new activity by fermentation for immune-competent cells in the host that administrated both of original remedy and the new fermented sample. In rodents, immune-compromised host was prepared with cancer chemotherapeutic agent. Our observations showed that the antigen stimulated animal increased the antibody producing cell for heterologous erythrocyte by compromised host by f-ESH. Moreover, by administration of f-ESH to immno-suppressed hosts, these samples regulated lymphocyte in number and functions, macrophage activities, and regulating anti-oxidative activity by phagocytic cells. The anti-oxidative assay was carried out ex-vivo system by peritoneal excaudate cell that we propose as suitable system for evaluating antioxidative assay. With these evidences, the original COF only augmented the level of lymphocytes in number, while f-ESH regulated the ratio of granulocytes and lymphocyte. In clinical study with 20 healthy volunteers, granulocyte and lymphocyte ratio was also resulted as neutral in peripheral white blood cells, 1, 2 and 3 weeks after the administration of f-ESH. We discussed the significance and mechanism of cleving complement components Fb by f-ESH in this text. Fb. The new cleavage of complement was directly evident by immune-electrophoretic method. In conclusion, this modification of food materials introduced new style of food intake resulted in the significant regulation such as antibody producing cells and anti-oxidative activity for phagocytic cells comparing traditional processing by hot water extraction.

The monitoring of laboratory indices of immune status of healthy children residing in conditions of intensive immunotropic zone of industrial town.

The complex evaluation was implemented concerning to quantitative and functional indices of immune-competent cells of peripheral blood, cytokine and immunoglobulin profile of blood serum with detection of confidence ranges of immunologic parameters of healthy children aged 3-11 years (n=98) residing in intensive immunotropic zone with the highest ecological risk. The multi-parametric flow cytofluorimeter COULTER EPICS XL (Beckman Coulter Inc.) with selection of of panels of monoclonal antibodies with multi-color combination of fluorichromуes was applied to analyze leukocytes, sub-population composition of lymphocites and processes of activation of cells of peripheral blood. The levels of cytokines were detected using immunoenzyme technique applying reagents by «R&D Diagnostics Inc.» (USA). The reagents' kit of «Vektor-Best» (Novosibirsk) was applied to detect spontaneous and mitogen-induced production of cytokines by cells of total blood The immuneturbidimetry technique was applied to detect content of immunoglobulins in blood serum. The total and specific IgE was detected using solid-phase immune enzyme analysis with reagents' kits by «Alkor Bio Company» (St. Petersburg). The statistical data processing was implemented using software Statistica 10 with data verification 95%-99%. In junior school children as compared with children aged 3-6 years, statistically reliable greater number of leukocytes were detected with preservation of high ratio of neutrophils, prevalence of helper sub-population of T-lymphocytes, expressed activation of B-lymphocytes, higher levels of serum IgA, IgG, common and specific IgE (latent sensibilization by domestic allergens), prevalence of intensity of production of interleukins IL-13 over IL-4 and significantly higher provision with natural killers.

Exposure of Eisenia andrei (Oligochaeta; Lumbricidea) to Cadmium Polluted Soil Inhibits Earthworm Maturation and Reproduction but not Restoration of Experimentally Depleted Coelomocytes or Regeneration of Amputated Segments.

Lumbricid earthworms are often exposed to polluted soil. They are also commonly subjected to various stimuli and attacks by predators that induce extrusion of coelomocyte-containing coelomic fluid and/or the loss of body segments followed by the renewal of immune-competent cells and regeneration of tissues/organs. The aim of our investigations was to test the effects of exposure of the earthworm Eisenia andrei to cadmium-polluted soil, combined with electrostimulation-induced depletion of coelomocytes (i.e. amoebocytes and chloragocyte-derived eleocytes) or the surgical amputation of posterior segments, on earthworm maturation, reproductive output, and regenerative processes. Experimental worms were maintained up to 7 weeks either in unpolluted soil or in soil spiked with cadmium chloride (500 mg/kg air-dried soil). In juvenile worms, sexual maturation (measured by clitellum formation) was delayed and cocoon production was inhibited in Cd-exposed worms. Coelomocytes were significantly depleted by electrostimulation and the kinetics of their recovery was similar in worms kept in clean and cadmium polluted soils, in both exposure conditions amoebocyte recovery was faster than recovery of riboflavin-storing eleocytes. In adult worms, soil cadmium exposure inhibited reproduction but, at macroanatomical level, had a negligible effect on regeneration of amputated posterior segments, visible only on histological cross-sections.

COMPARATIVE ACTIVITY OF INFLUENZA VACCINES: EFFECT ON LYMPHOCYTE SUBPOPULATION STRUCTURE

Study subpopulation structure of lymphocytes in healthy individuals under the effect of various influenza vaccines in an in vitro system. Evaluation of immune- phenotype features of PBMCs, activated in vitro by immune-adjuvanted and unadjuvanted vaccines against influenza in healthy individuals, was carried out by using flow cytometry method. Grippol plus vaccine caused a more pronounced stimulating effect compared with subunit and split-vaccines on NK-cells, cells with markers of early activation CD45/CD25, induced the quantity of natural regulatory cells (CD4/CD25/Foxp3), increase of the number of B-cells and reduced the amount of cell types with apoptosis marker CD45/CD95. Immune-adjuvanted vaccine Grippol plus induced formation of effectors of both innate and adaptive immunity and possessed the most powerful potential regarding activation of various types of immune-competent cells compared with unadjuvanted vaccines.

Profile features immunohistochemical endometrium in patients uterine form of infertility

The article presented data of the complex examination of 79 women aged of 24-43 years with uterine form of infertility, including the determination of immunohistochemical markers (ER, PR, Ki-67, CD34, CD56, CD68, CD138, VEGF) in endometrium. As a result of the study there was revealed the decline in the number of CD56-positive cells, decrease or no expression of CD138, the increase in expression of CD34, decreased VEGF expression. The deterioration of implantation processes in patients with uterine form of infertility, presumably is related to the low content of immune competent cells and inhibition of angiogenesis, as well as a weak proliferative activity of the endometrium, as evidenced by a decrease expression of Ki-67.

Microglia in Physiology and Disease.

As the immune-competent cells of the brain, microglia play an increasingly important role in maintaining normal brain function. They invade the brain early in development, transform into a highly ramified phenotype, and constantly screen their environment. Microglia are activated by any type of pathologic event or change in brain homeostasis. This activation process is highly diverse and depends on the context and type of the stressor or pathology. Microglia can strongly influence the pathologic outcome or response to a stressor due to the release of a plethora of substances, including cytokines, chemokines, and growth factors. They are the professional phagocytes of the brain and help orchestrate the immunological response by interacting with infiltrating immune cells. We describe here the diversity of microglia phenotypes and their responses in health, aging, and disease. We also review the current literature about the impact of lifestyle on microglia responses and discuss treatment options that modulate microglial phenotypes.

Canthaxanthin: From molecule to function.

The aim of this review is to summarize the relevant literature about the use of canthaxanthin in food science and nutrition research. Canthaxanthin is a red-orange carotenoid that belongs to the xanthophyll group. This naturally occurring pigment is present in bacteria, algae and some fungi. Canthaxanthin is also responsible for the color of flamingo feathers, koi carp skin and crustacean shells. Canthaxanthin is widely used in poultry (broiler, laying hens) as a feed additive. Canthaxanthin can be obtained by total synthesis. The canthaxanthin-mediated color of foods is an important quality criterion for consumers. Recently, the potential health-promoting effects of canthaxanthin have been discussed. Canthaxanthin enrichment of LDL has the potential to protect cholesterol from oxidation. In addition to its free radical scavenging and antioxidant properties (e.g., the induction of catalase and superoxide dismutase), canthaxanthin's immunomodulatory activity (e.g., enhancing the proliferation and function of immune competent cells) and its important role in gap junction communication (e.g., induction of the transmembrane protein connexin 43) have been reported. Many studies regarding the potential health benefits of canthaxanthin have been conducted in vitro and should be validated in appropriate in vivo models.