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The paper proposes an improved active contour model for segmenting and tracking accurate boundaries of the single lymphocyte in phase-contrast microscopic images. Active contour models have been widely used in object segmentation and tracking. However, current external-force-inspired methods are weak at handling low-contrast edges and suffer from initialization sensitivity. In order to segment low-contrast boundaries, we combine the region information of the object, extracted by morphology gray-scale reconstruction, and the edge information, extracted by the Laplacian of Gaussian filter, to obtain an improved feature map to compute the external force field for the evolution of active contours. To alleviate initial location sensitivity, we set the initial contour close to the real boundaries by performing morphological image processing. The proposed method was tested on live lymphocyte images acquired through the phase-contrast microscope from the blood samples of mice, and comparative experimental results showed the advantages of the proposed method in terms of the accuracy and the speed. Tracking experiments showed that the proposed method can accurately segment and track lymphocyte boundaries in microscopic images over time even in the presence of low-contrast edges, which will provide a good prerequisite for the quantitative analysis of lymphocyte morphology and motility. [ABSTRACT FROM AUTHOR]

Circulating tumor cells (CTCs) are tumor cells that have detached from primary tumor site and are transported via the circulation system. The importance of CTCs as prognostic biomarker is leveraged when multiple studies found that patient with cutoff of 5 CTCs per 7.5 mL blood has poor survival rate. Despite its clinical relevance, the isolation and characterization of CTCs can be quite challenging due to their large morphological variability and the rare presence of CTCs within the blood. Numerous methods have been employed and discussed in the literature for CTCs separation. In this paper, we will focus on label free CTCs isolation methods, in which the biophysical and biomechanical properties of cells (e.g., size, deformability, and electricity) are exploited for CTCs detection. To assess the present state of various isolation methods, key performance metrics such as capture efficiency, cell viability, and throughput will be reported. Finally, we discuss the challenges and future perspectives of CTC isolation technologies. [ABSTRACT FROM AUTHOR]

Metastatic melanomas are frequently refractory to most adjuvant therapies such as chemotherapies and radiotherapies. Recently, immunotherapies have shown good results in the treatment of some metastatic melanomas. Immune cell infiltration in the tumor has been associated with successful immunotherapy. More generally, tumor infiltrating lymphocytes (TILs) in the primary tumor and in metastases of melanoma patients have been demonstrated to correlate positively with favorable clinical outcomes. Altogether, these findings suggest the importance of being able to identify, quantify and characterize immune infiltration at the tumor site for a better diagnostic and treatment choice. In this paper, we used Fourier Transform Infrared (FTIR) imaging to identify and quantify different subpopulations of T cells: the cytotoxic T cells (CD8+), the helper T cells (CD4+) and the regulatory T cells (T reg). As a proof of concept, we investigated pure populations isolated from human peripheral blood from 6 healthy donors. These subpopulations were isolated from blood samples by magnetic labeling and purities were assessed by Fluorescence Activated Cell Sorting (FACS). The results presented here show that Fourier Transform Infrared (FTIR) imaging followed by supervised Partial Least Square Discriminant Analysis (PLS-DA) allows an accurate identification of CD4+ T cells and CD8+ T cells (＞86%). We then developed a PLS regression allowing the quantification of T reg in a different mix of immune cells (e.g. Peripheral Blood Mononuclear Cells (PBMCs)). Altogether, these results demonstrate the sensitivity of infrared imaging to detect the low biological variability observed in T cell subpopulations. [ABSTRACT FROM AUTHOR]

Our aim in this paper is to describe the results of treatment of acute lymphoblastic leukaemia (ALL) in Mexican children treated from 2006 to 2010 under the protocol from the Dana-Farber Cancer Institute (DFCI) 00-01. The children were younger than 16 years of age and had a diagnosis of ALL de novo. The patients were classified as standard risk if they were 1–9.9 years old and had a leucocyte count <50 × 109/L, precursor B cell immunophenotype, no mediastinal mass, CSF free of blasts, and a good response to prednisone. The rest of the patients were defined as high risk. Of a total of 302 children, 51.7% were at high risk. The global survival rate was 63.9%, and the event-free survival rate was 52.3% after an average follow-up of 3.9 years. The percentages of patients who died were 7% on induction and 14.2% in complete remission; death was associated mainly with infection (21.5%). The relapse rate was 26.2%. The main factor associated with the occurrence of an event was a leucocyte count >100 × 109/L. The poor outcomes were associated with toxic death during induction, complete remission, and relapse. These factors remain the main obstacles to the success of this treatment in our population. [ABSTRACT FROM AUTHOR]

Telomeres are essential in maintaining chromosome integrity and in controlling cellular replication. Attrition of telomere length in peripheral blood mononuclear cells (PBMCs) with age is well documented from cross-sectional studies. But the actual in vivo changes in telomere lengths and its relationship with the contributing factors within the individuals with age have not been fully addressed. In the present paper, we report a longitudinal analysis of telomere length in the PBMCs, lymphocytes and monocytes of 216 human subjects aged from 20-90 years assessed at 0-, 5- and 12-year follow-up. For the 5- and 12-year follow-up, telomere length in the PBMCs decreased in 34% and 46%, exhibited no detectable change in 56% and 47% and increased in 10% and 7% of the subjects respectively. The rate of telomere change was distinct for T-cells, B-cells and monocytes for any given subject. Telomerase activity declined with age in the resting T-cells and B-cells and the activated T-cells. Finally, a significant portion of telomere attrition in T-cells with age was explained by a decline in the telomerase activity, decreased naıve cells and the change in physiological conditions such as elevated blood glucose and interleukin (IL)-6 levels. These findings show that changes in the telomere length of the PBMCs with age in vivo occur at different rates in different individuals and cell types and reveal that changes in the telomere length in the T-cells with age is influenced by the telomerase activity, naïve T-cell percentage and changes in health conditions. [ABSTRACT FROM AUTHOR]

In this paper we present VDJSeq-Solver, a methodology and tool to identify clonal lymphocyte populations from paired-end RNA Sequencing reads derived from the sequencing of mRNA neoplastic cells. The tool detects the main clone that characterises the tissue of interest by recognizing the most abundant V(D)J rearrangement among the existing ones in the sample under study. The exact sequence of the clone identified is capable of accounting for the modifications introduced by the enzymatic processes. The proposed tool overcomes limitations of currently available lymphocyte rearrangements recognition methods, working on a single sequence at a time, that are not applicable to high-throughput sequencing data. In this work, VDJSeq-Solver has been applied to correctly detect the main clone and identify its sequence on five Mantle Cell Lymphoma samples; then the tool has been tested on twelve Diffuse Large B-Cell Lymphoma samples. In order to comply with the privacy, ethics and intellectual property policies of the University Hospital and the University of Verona, data is available upon request to after signing a mandatory Materials Transfer Agreement. VDJSeq-Solver JAVA/Perl/Bash software implementation is free and available at . [ABSTRACT FROM AUTHOR]

Non-Hodgkin lymphomas (NHLs) are a diverse group of blood cancers derived from lymphocytes that vary significantly in their severity. Surgery is not often used as a treatment because of the efficacy of chemotherapy, biological therapy, radiotherapy and hematopoietic stem cell transplantation. We reviewed the natural history and possible role of surgery for NHL. Surgery may be useful in confirming or refuting an equivocal radiological diagnosis through biopsy, removing symptomatic limited disease from an affected organ and in splenectomy for primary splenic lymphoma. Emergency abdominal surgery for acute complications of NHL provides palliation and diagnosis. There is as yet no consensus as to the optimum treatment for symptomatic limited disease affecting an organ and timing of chemotherapy perioperatively. Prospective randomized trials are required. Introduction Hodgkin lymphoma (HL, Hodgkin disease), described by Thomas Hodgkin in 1832, was the first form of lymphoma described and defined.1 Other forms were later described but because Hodgkin lymphoma was much more radiation-sensitive than other forms, its diagnosis was important for oncologists and their patients and thus research originally focused on it. The Rappaport classification, proposed in 1956 and 1966, became the first widely accepted classification of lymphomas other than Hodgkin.2 Surgery is not often used as a treatment because of the efficacy of chemotherapy, biological therapy, radiotherapy and hematopoietic stem cell transplantation.3-5 However, it may have some definitive roles in planned (elective) surgery but usually a palliative one in the acute abdomen.6,7 Despite the difficulty and paucity of prospective randomized trials this paper reviewed the possible role of surgery in non-Hodgkin’s lymphoma (NHL). [ABSTRACT FROM AUTHOR]

Lymphocyte invasion by HIV-1 is a complex, highly regulated process involving many different types of molecules that is prompted by the virus's association with viral receptors located at the cell-surface membrane that culminates in the formation of a fusion pore through which the virus enters the cell. A great deal of work has been done to identify the key actors in the process and determine the regulatory interactions; however, there have been no reports to date of attempts being made to fully understand the system dynamics through a systemic, quantitative modeling approach. In this paper, we introduce a dynamic mathematical model that integrates the available information on the molecular events involved in lymphocyte invasion. Our model shows that moesin activation is induced by virus signaling, while filamin-A is mobilized by the receptor capping. Actin disaggregation from the cap is facilitated by cofilin. Cofilin is inactivated by HIV-1 signaling in activated lymphocytes, while in resting lymphocytes another signal is required to activate cofilin in the later stages in order to accelerate the decay of the aggregated actin as a restriction factor for the viral entry. Furthermore, stopping the activation signaling of moesin is sufficient to liberate the actin filaments from the cap. The model also shows the positive effect of gelsolin on actin capping by means of the nucleation effect. These findings allow us to propose novel approaches in the search for new therapeutic strategies. In particular, gelsolin inhibition is seen as a promising target for preventing HIV-1 entry into lymphocytes, due to its role in facilitating the capping needed for the invasion. Also it is shown that HIV-1 should overcome the cortical actin barrier during early infection and predicts the different susceptibility of CD4+ T cells to be infected in terms of actin cytoskeleton dynamics driven by associated cellular factors. [ABSTRACT FROM AUTHOR]

In this paper we apply a parametric semi-Markov process to model the dynamic evolution of HIV-1 infected patients. The seriousness of the infection is rendered by the CD4+ T-lymphocyte counts. For this purpose we introduce the main features of nonhomogeneous semi-Markov models. After determining the transition probabilities and the waiting time distributions in each state of the disease, we solve the evolution equations of the process in order to estimate the interval transition probabilities. These quantities appear to be of fundamental importance for clinical predictions. We also estimate the survival probabilities for HIV infected patients and compare them with respect to certain categories, such as gender, age group or type of antiretroviral therapy. Finally we attach a reward structure to the aforementioned semi-Markov processes in order to estimate clinical costs. For this purpose we generate random trajectories from the semi-Markov processes through Monte Carlo simulation. The proposed model is then applied to a large database provided by ISS (Istituto Superiore di Sanità, Rome, Italy), and all the quantities of interest are computed. [ABSTRACT FROM AUTHOR]

Shortened telomere length (TL) and oxidative stress have been described in several vascular disorders at both the tissue and circulating level. However, to our knowledge, there are no reports about TL associated with varicose vein (VV) disease. This paper aimed to evaluate, at the tissue and circulating level, TL and oxidative stress in VV disease, compared to the corresponding counterparts from abdominal aortic aneurysm (AAA) patients and control healthy subjects. TL was measured using quantitative fluorescence in situ hybridization (Q-FISH). Oxidative stress was evaluated by measuring the malondialdehyde (MDA) concentration by thiobarbituric acid reactive substance/s (TBARS) assay. At the vascular tissue level, VV patients had shortened TL and a high MDA concentration, similarly to AAA patients. Conversely, blood lymphocytes and epidermal cells from VV patients had a TL similar to healthy controls and significantly longer than the same cells from AAA patients. Moreover, the MDA concentration in plasma from VV patients was significantly lower than from the AAA group. Linear regression analysis showed a statistically significant inverse correlation between the blood lymphocyte TL and plasma MDA level. Our results suggest that, unlike AAA, telomere attrition in VV tissue is not a systemic phenomenon but it may be attributable to tissue microenvironment conditions and possibly to high local oxidative stress. [ABSTRACT FROM PUBLISHER]

The purpose of this paper is to establish the cytogenetic analyses of human peripheral blood samples caused by simulation of partial-body exposures. Either accidental or occupational partial-body exposure to ionizing radiation poses significant health hazards that are indicated by induction of chromosome aberrations (CA). The percentages of mixtures of blood samples irradiated in vitro with 2 Gy of gamma rays were 10, 25, 50, 75 and 100.0%. Lymphocytes were cultured for 48 hr, harvested with standard procedures and then first-division metaphase cells were analyzed. It showed that the frequencies of unstable CA depend on the proportion of the irradiated blood. All frequencies of the observed CA was lower than that of predicted or calculated from 100% exposed blood, except in one case, indicating a phenomenon of "dilution" of the un-irradiated into irradiated lymphocytes that may take place a bystander effects. Our data showed that the quantification of CA in human peripheral blood lymphocytes may be an important tool of dose assessment for partial-body exposure to ionizing radiation. [ABSTRACT FROM AUTHOR]

Postoperative atrial fibrillation (AF) is a common complication of coronary artery bypass grafting (CABG). The mean platelet volume (MPV) is an important marker of platelet activity and is associated with cardiovascular risk factors. We investigated whether the MPV is associated with the development of AF after CABG. This study included 208 patients undergoing elective isolated CABG. We evaluated the standard preoperative 12-lead electrocardiograph (ECG) recorded at a paper speed of 25 mm/s obtained for each patient from our hospital records before surgery. All study patients underwent standard CABG requiring cardiopulmonary bypass without concurrent valvular surgery. Forty-three patients were excluded. After CABG, all patients were monitored by telemetry and 12-lead ECGs. AF was defined using the established Society of Thoracic Surgeons definition. Postoperative AF occurred in 38 (22%) patients. The hemoglobin and platelet and leukocyte counts were similar in the groups with and without AF. However, the MPV and neutrophil/lymphocyte ratio were significantly higher in the AF group (8.9 [1.4] vs. 7.9 [1.2], p < 0.001 and 3.2 ± 1.9 vs. 2.6 ± 1.2, p = 0.005, respectively). In addition, the C-reactive protein (CRP) levels were significantly higher in the AF group (8.9 [19.6] vs. 5.3 [8.7], p = 0.025). Multivariate logistic regression analysis showed that MPV and CRP were independent predictors of postoperative AF (odds ratio [OR] 2.564, 95% confidence interval [CI] 1.326-4.958, p = 0.005; OR 1.055, 95% CI 1.000-1.114, p = 0.050, respectively). Our results show that increased platelet activity is associated with the development of AF after CABG. [ABSTRACT FROM AUTHOR]

Many studies have suggested a link between the spatial organization of genomes and fundamental biological processes such as genome reprogramming, gene expression, and differentiation. Multicolor fluorescence in situ hybridization on three-dimensionally preserved nuclei (3D-FISH), in combination with confocal microscopy, has become an effective technique for analyzing 3D genome structure and spatial patterns of defined nucleus targets including entire chromosome territories and single gene loci. This technique usually requires the simultaneous visualization of numerous targets labeled with different colored fluorochromes. Thus, the number of channels and lasers must be sufficient for the commonly used labeling scheme of 3D-FISH, 'one probe-one target'. However, these channels and lasers are usually restricted by a given microscope system. This paper presents a method for simultaneously delineating multiple targets in 3D-FISH using limited channels, lasers, and fluorochromes. In contrast to other labeling schemes, this method is convenient and simple for multicolor 3D-FISH studies, which may result in widespread adoption of the technique. Lastly, as an application of the method, the nucleus locations of chromosome territory 18/21 and centromere 18/21/13 in normal human lymphocytes were analyzed, which might present evidence of a radial higher order chromatin arrangement. [ABSTRACT FROM AUTHOR]

Interest in cell heterogeneity and differentiation has recently led to increased use of time-lapse microscopy. Previous studies have shown that cell fate may be determined well in advance of the event. We used a mixture of automation and manual review of time-lapse live cell imaging to track the positions, contours, divisions, deaths and lineage of 44 B-lymphocyte founders and their 631 progeny in vitro over a period of 108 hours. Using this data to train a Support Vector Machine classifier, we were retrospectively able to predict the fates of individual lymphocytes with more than 90% accuracy, using only time-lapse imaging captured prior to mitosis or death of 90% of all cells. The motivation for this paper is to explore the impact of labour-efficient assistive software tools that allow larger and more ambitious live-cell time-lapse microscopy studies. After training on this data, we show that machine learning methods can be used for realtime prediction of individual cell fates. These techniques could lead to realtime cell culture segregation for purposes such as phenotype screening. We were able to produce a large volume of data with less effort than previously reported, due to the image processing, computer vision, tracking and human-computer interaction tools used. We describe the workflow of the software-assisted experiments and the graphical interfaces that were needed. To validate our results we used our methods to reproduce a variety of published data about lymphocyte populations and behaviour. We also make all our data publicly available, including a large quantity of lymphocyte spatio-temporal dynamics and related lineage information. [ABSTRACT FROM AUTHOR]

Abstract: Cytogenetic analysis of peripheral blood lymphocytes is the most sensitive and reliable method currently available for in vivo assessment of the biological effects of exposure to radiation and provides the most informative measurement of radiation induced health risks. Data indicates that space missions of a few months or more can induce measureable increases in the yield of chromosome damage in the blood lymphocytes of astronauts that can be used to estimate an organ dose equivalent, and biodosimetry estimates lie within the range expected from physical dosimetry. Space biodosimetry poses some unique challenges compared to terrestrial biological assessments of radiation exposures, but data provides a direct measurement of space radiation damage, which takes into account individual radiosensitivity in the presence of confounding factors such as microgravity and other stress conditions. Moreover if chromosome damage persists in the blood for many years, results can be used for retrospective dose reconstruction. In contrast to physical measurements, which are external to body and require multiple devices to detect all radiation types all of which have poor sensitivity to neutrons, biodosimetry is internal and includes the effects of shielding provided by the body itself plus chromosome damage shows excellent sensitivity to protons, heavy ions, and neutrons. In addition, chromosome damage is reflective of cancer risk and biodosimetry values can therefore be used to validate and develop risk assessment models that can be used to characterize health risk incurred by crewmembers. The current paper presents a review of astronaut biodosimetry data, along with recently derived data on the relative cancer risk estimated using the quantitative approach derived from the European Study Group on Cytogenetic Biomarkers and Health database. [Copyright &y& Elsevier]

Background: Atopic asthma is characterized by intermittent exacerbations triggered by exposure to allergen. Exacerbations are characterized by an acute inflammatory reaction in the airways, with recruitment of both innate and adaptive immune cells. These cell populations as well as soluble factors are critical for initiating and controlling the inflammatory processes in allergic asthma. Detailed data on the numbers and types of cells recruited following allergen challenge is lacking. In this paper we present an extensive phenotypic analysis of the inflammatory cell infiltrate present in the bronchoalveolar lavage (BAL) fluid following bronchoscopically directed allergen challenge in mild atopic asthmatics. Methods: A re-analysis of pooled data obtained prior to intervention in our randomized, placebo controlled, double blinded study (costimulation inhibition in asthma trial [CIA]) was performed. Twenty-four subjects underwent bronchoscopically directed segmental allergen challenge followed by BAL collection 48 hours later. The BAL fluid was analyzed by multi-color flow cytometry for immune cell populations and multi-plex ELISA for cytokine detection. Results: Allergen instillation induced pro-inflammatory cytokines (IL-6) and immune modulating cytokines (IL-2, IFN- γ, and IL-10) along with an increase in lymphocytes and suppressor cells (Tregs and MDSC). Interestingly, membrane expression of CD30 was identified on lymphocytes, especially Tregs, but not eosinophils. Soluble CD30 was also detected in the BAL fluid after allergen challenge in adult atopic asthmatics. Conclusions: After segmental allergen challenge of adult atopic asthmatics, cell types associated with a proinflammatory as well as an anti-inflammatory response are detected within the BAL fluid of the lung. [ABSTRACT FROM AUTHOR]

This paper describes the immunophenotype of cellular inflammatory infiltrates in chronic cholangitis in six common dolphins (Delphinus delphis), four striped dolphins (Stenella coeruleoalba), three Atlantic spotted dolphins (Stenella frontalis) and one pygmy sperm whale (Kogia Breviceps) found stranded along the coasts of the Canary Islands (Spain). A panel of 5 antibodies previously tested in dolphins (anti-CD3, -IgG, -MHC class II, -S100 protein and -lysozyme) were used. The present work also reports cross reactivity with dolphin antigens of two antibodies not used to date in dolphins (anti-mouse iNOS and anti-mouse Foxp3). The most common type of cholangitis found was chronic granulomatous cholangitis, associated with the presence of the parasite Campula spp., or its eggs in bile ducts. The cellular composition of the hepatic inflammatory infiltrate associated to chronic parasitic cholangitis was closely similar to that found in the cortex of control lymph nodes, including the presence of S100+ and MHC class II+ dendritic- like cells in lymphoid follicles and interfollicular areas. Only occasional macrophages expressed iNOS, whereas Foxp3+ lymphocytes were not found in any of the lesions described in the different types of cholangitis. [ABSTRACT FROM AUTHOR]

An amendment to this paper has been published and can be accessed via a link at the top of the paper. [ABSTRACT FROM AUTHOR]

An amendment to this paper has been published and can be accessed via a link at the top of the paper. [ABSTRACT FROM AUTHOR]

A conference paper on the correlation between in vivo accumulation and in vitro adhesion of liver associated lymphocytes in and around liver adenocarcinoma metastases is presented. Results reveal that more peripheral and sinusoidal blood cells in vitro adhered to the sites of CC531 than to the liver tissue. There were more peripheral blood mononuclear cells (PBMs) than liver sinusoidal washout cells (LSWCs) adhering to both liver and tumor.

Simple Summary: Lymphoma is a heterogeneous group of neoplasms including over 70 different subtypes. Its biological characteristic of deriving from lymphoid tissues makes it ideal for immunotherapy. In this paper, we provide insights into lymphoma-specific clinical trials based on cytokine-induced killer (CIK) cell therapy. We also reviewed pre-clinical lymphoma models where CIK cells have been used along with other synergetic tumor-targeting immune modules to improve their therapeutic potential. From a broader perspective, we will highlight that CIK cell therapy has potential, and in this rapidly evolving landscape of cancer therapies its optimization (as a personalized therapeutic approach) will be beneficial in lymphomas. Cancer is a complex disease where resistance to therapies and relapses often pose a serious clinical challenge. The scenario is even more complicated when the cancer type itself is heterogeneous in nature, e.g., lymphoma, a cancer of the lymphocytes which constitutes more than 70 different subtypes. Indeed, the treatment options continue to expand in lymphomas. Herein, we provide insights into lymphoma-specific clinical trials based on cytokine-induced killer (CIK) cell therapy and other pre-clinical lymphoma models where CIK cells have been used along with other synergetic tumor-targeting immune modules to improve their therapeutic potential. From a broader perspective, we will highlight that CIK cell therapy has potential, and in this rapidly evolving landscape of cancer therapies its optimization (as a personalized therapeutic approach) will be beneficial in lymphomas. [ABSTRACT FROM AUTHOR]

Abstract: The characterization of circulating tumor cells (CTCs) could substantially improve the management of cancer patients. However, their study is still a matter of debate, often due to lymphocyte contamination. In the present paper, an investigation of CTCs was carried out for the first time using DEPArray, a dielectrophoresis-based platform able to detect and sort pure CTCs. Analyses were conducted on peripheral blood (PB) samples from patients with metastatic colon cancer. After 100% pure cell recovery and whole genome amplification, KRAS gene mutation of CTCs was screened and compared to gene status in the primary tumor tissue. CTCs were found in 21 colon cancer patients (52.5%), with more than three tumor cells per 7.5ml. KRAS gene mutation analysis, showed a mutational concordance between CTCs and primary tumor in 50% of matched cases. The present study demonstrates for the first time the feasibility of analyzing at the molecular level pure CTCs avoiding lymphocyte contamination using an innovative instrumentation, and a KRAS discordance between CTCs and primary tissue. Our results present dielectrophoresis-based procedures as a new standard in single cell analysis and recovery and invite careful reflection on the value of CTCs characterization. [Copyright &y& Elsevier]

Human African trypanosomiasis (HAT) is caused by trypanosomes of the species Trypanosoma brucei and belongs to the neglected tropical diseases. Presently, WHO has listed 36 countries as being endemic for sleeping sickness. No vaccine is available, and disease treatment is difficult and has life-threatening side effects. Therefore, there is a crucial need to search for new therapeutic targets against the parasite. Trypanosome excreted-secreted proteins could be promising targets, as the total secretome was shown to inhibit, in vitro, host dendritic cell maturation and their ability to induce lymphocytic allogenic responses. The secretome was found surprisingly rich in various proteins and unexpectedly rich in diverse peptidases, covering more than ten peptidase families or subfamilies. Given their abundance, one may speculate that they would play a genuine role not only in classical 'housekeeping' tasks but also in pathogenesis. The paper reviews the deleterious role of proteases from trypanosomes, owing to their capacity to degrade host circulating or structural proteins, as well as proteic hormones, causing severe damage and preventing host immune response. In addition, proteases account for a number of drug targets, such drugs being used to treat severe diseases such AIDS. This review underlines the importance of secreted proteins and especially of secreted proteases as potential targets in HAT-fighting strategies. It points out the need to conduct further investigations on the specific role of each of these various proteases in order to identify those playing a central role in sleeping sickness and would be suitable for drug targeting. © 2013 BioFactors, 39(4):407-414, 2013 [ABSTRACT FROM AUTHOR]

This paper describes the optical mobilities of blood cell components. Blood cells are heterogeneous, and their optical behaviors depend on size, morphology, and other optical properties. In a step toward the label-free separation of blood cells, the optical mobility resulting from the optical scattering and cell properties was derived and evaluated for each cell component. The optical mobilities of red blood cells, lymphocytes, granulocytes, and monocytes were measured under various flow conditions of a cross-type optical particle separator. [ABSTRACT FROM AUTHOR]

The localization of cervical lymph nodes is extremely important in practice for the positive and differential diagnosis as well as the staging of cervical lymphadenopathies. Ultrasonography represents the first line imaging method in the diagnosis of cervical lymphadenopathies due to its excellent resolution and high diagnosis accuracy. The present paper aims to illustrate the ultrasonographic identification of the anatomical landmarks used for the definition of cervical lymphatic spaces. The application of standardized views allows a delineation of clear anatomical landmarks and an accurate localization of the cervical lymph nodes. [ABSTRACT FROM AUTHOR]

The idiotypic network theory (INT) was conceived by the Danish immunologist Niels Kaj Jerne in 1973/1974. It proposes an overall view of the immune system as a network of lymphocytes and antibodies. The paper tries to offer a reconstruction of the genesis of the theory, now generally discarded and of mostly historical interest, first of all, by taking into account the context in which Jerne's theoretical proposal was advanced. It is argued the theory challenged, in a sense, the supremacy of the clonal selection theory (CST), this being regarded as the predominant paradigm in the immunological scenario. As CST found shortcomings in explaining certain phenomena, anomalies, one could view INT as a competing paradigm claiming to be able to make sense of such phenomena in its own conceptual framework. After a summary outline of the historical background and some relevant terminological elucidations, a narrative of the various phases of elaboration of the theory is proposed, up to its official public presentation. [ABSTRACT FROM AUTHOR]

Estaquier et al. provide commentary on our paper that elucidated the mechanism by which HIV-1 causes cell death in activated CD4 T lymphocytes. We showed that proviral DNA integration triggers DNA-PK dependent death signaling, leading to p53 phosphorylation and cell demise (Cooper A et al. Nature 498:376-379, 2013). They have raised several hypothetical points that we further clarify here. [ABSTRACT FROM AUTHOR]

Posttransplant lymphoproliferative disease (PTLD) after lung transplantation occurs due to immunosuppressant therapy which limits antiviral host immunity and permits Epstein-Barr viral (EBV) replication and transformation of B cells. Mechanistically, EBV survives due to latency, escape from cytotoxic T cell responses, and down regulation of host immunity to EBV. Clinical presentation of EBV may occur within the lung allograft early posttransplantation or later onset which is more likely to be disseminated. Improvements in monitoring through EBV viral load have provided a means of earlier detection; yet, sensitivity and specificity of EBV load monitoring after lung transplantation may require further optimization. Once PTLD develops, staging and tissue diagnosis are essential to appropriate histopathological classification, prognosis, and guidance for therapy. The overall paradigm to treat PTLD has evolved over the past several years and depends upon assessment of risk such as EBV-naïve status, clinical presentation, and stage and sites of disease. In general, clinical practice involves reduction in immunosuppression, anti-CD20 biologic therapy, and/or use of plasma cell inhibition, followed by chemotherapy for refractory PTLD. This paper focuses upon the immunobiology of EBV and PTLD, as well as the clinical presentation, diagnosis, prognosis, and emerging treatments for PTLD after lung transplantation. [ABSTRACT FROM AUTHOR]

Background: Supervised machine learning approaches have been recently adopted in the inference of transcriptional targets from high throughput trascriptomic and proteomic data showing major improvements from with respect to the state of the art of reverse gene regulatory network methods. Beside traditional unsupervised techniques, a supervised classifier learns, from known examples, a function that is able to recognize new relationships for new data. In the context of gene regulatory inference a supervised classifier is coerced to learn from positive and unlabeled examples, as the counter negative examples are unavailable or hard to collect. Such a condition could limit the performance of the classifier especially when the amount of training examples is low. Results: In this paper we improve the supervised identification of transcriptional targets by selecting reliable counter negative examples from the unlabeled set. We introduce an heuristic based on the known topology of transcriptional networks that in fact restores the conventional positive/negative training condition and shows a significant improvement of the classification performance. We empirically evaluate the proposed heuristic with the experimental datasets of Escherichia coli and show an example of application in the prediction of BCL6 direct core targets in normal germinal center human B cells obtaining a precision of 60%. Conclusions: The availability of only positive examples in learning transcriptional relationships negatively affects the performance of supervised classifiers. We show that the selection of reliable negative examples, a practice adopted in text mining approaches, improves the performance of such classifiers opening new perspectives in the identification of new transcriptional targets. [ABSTRACT FROM AUTHOR]

The use of optical dielectrophoresis (ODEP) to manipulate microparticles and biological cells has become increasingly popular due to its tremendous flexibility in providing reconfigurable electrode patterns and flow channels. ODEP enables the parallel and free manipulation of small particles on a photoconductive surface on which light is projected, thus eliminating the need for complex electrode design and fabrication processes. In this paper, we demonstrate that mouse cells comprising melan-a cells, RAW 267.4 macrophage cells, peripheral white blood cells and lymphocytes, can be manipulated in an opto-electrokinetics (OEK) device with appropriate DEP parameters. Our OEK device generates a non-rotating electric field and exerts a localized DEP force on optical electrodes. Hitherto, we are the first group to report that among all the cells investigated, melan-a cells, lymphocytes and white blood cells were found to undergo self-rotation in the device in the presence of a DEP force. The rotational speed of the cells depended on the voltage and frequency applied and the cells' distance from the optical center. We discuss a possible mechanism for explaining this new observation of induced self-rotation based on the physical properties of cells. We believe that this rotation phenomenon can be used to identify cell type and to elucidate the dielectric and physical properties of cells. [ABSTRACT FROM AUTHOR]

The study of magnetic effects on biological system was started in response to Dr. Nose's wish for the development of extracorporeal immunomodulation therapy. In the extracorporeal immunomodulation system, monocytes or lymphocytes in blood are stimulated by the interactions with immune-active materials. The concept of magnetic field application was introduced in order to enhance the cell activation. Therefore, the applicability of magnetic field was studied. This paper was concerned with in vitro study of magnetic field effects on macrophage/monocytes and lymphocytes activations. Time-varying magnetic field could enhance macrophages/monocytes and lymphocytes activations. [ABSTRACT FROM AUTHOR]

Human SLC25A13 gene encodes citrin, the liver-type aspartate–glutamate carrier isoform 2, and SLC25A13 mutations lead to citrin deficiency (CD). The definitive diagnosis of CD relies on SLC25A13 analysis, but conventional DNA analysis could not identify all SLC25A13 mutations. We investigated transcriptional features of SLC25A13 gene in peripheral blood lymphocytes (PBLs) from CD patients and healthy volunteers. SLC25A13 mutations were explored by PCR/LA-PCR, PCR-RFLP and direct sequencing. SLC25A13 cDNA was amplified by RT-PCR, cloned and then sequenced. All diagnoses of the CD patients were confirmed, including a heterozygote of g.2T>C and an unknown mutation yielding an aberrant transcript r.16_212dup. Twenty-eight alternative splice variants (ASVs) were identified from normal SLC25A13 alleles. Among them, r.213_328del took account for 53.7%, the normal transcript r.=, 16.6%, and the remaining 26 novel ASVs, collectively 29.3%, of all cDNA clones. Moreover, similar ASVs, all reflecting corresponsive mutations, were detected from the mutated alleles. These results indicated that the normal SLC25A13 transcript could be cloned, and the abundance of the ASV r.213_328del predicted the existence of a constructively novel protein isoform for this gene in human PBLs. And, the 26 novel ASVs, along with the novel aberrant transcript r.16_212dup and the SNP g.2T>C, enriched the transcript/variation spectrum of SLC25A13 gene in human beings. The findings in this paper, for the first time, uncovered the marked transcript diversity of SLC25A13 gene in human PBLs, and suggested that cDNA cloning analysis of this gene in human PBLs might be a feasible tool for CD molecular diagnosis. [ABSTRACT FROM AUTHOR]

Abstract: This paper investigates the in vitro effect of dexamethasone on bovine CD25highCD4+, CD25lowCD4+ and CD25−CD4+ T cells. Only a small percentage of bovine CD25highCD4+ (2–4%) and CD25lowCD4+ (1–2%) cells expressed Foxp3. Dexamethasone caused considerable loss of CD25−CD4+ cells, but it increased the relative and absolute numbers of CD25highCD4+ and CD25lowCD4+ lymphocytes, while at the same time reducing the percentage of Foxp3+ cells within the latter subpopulations. Considering all these, as well as the intrinsically poor Foxp3 expression in bovine CD25+CD4+, it can be concluded that the drug most probably increased the number of activated non-regulatory CD4+ lymphocytes. It has been found that changes in cell number were at least partly caused by proapoptotic effect of the drug on CD25−CD4+ cells and antiapoptotic effect on CD25highCD4+ and CD25lowCD4+ cells. The results obtained from this study indicate that the involvement of CD4+ lymphocytes in producing the anti-inflammatory and immunosuppressive effect of dexamethasone in cattle results from the fact that the drug had a depressive effect on the production of IFN-γ by CD25−CD4+ cells. Secretion of TGF-β and IL-10 by CD4+ lymphocytes was not involved in producing these pharmacological effects, because the drug did not affect production of TGF-β and, paradoxically, it reduced the percentage of IL-10+CD4+ cells. [Copyright &y& Elsevier]

Abstract: Objective: To evaluate whether vitamin D levels are related to the risk of developing autoimmune diseases and whether supplementation with vitamin D can modify the course of the diseases. Methods: We reviewed the most relevant papers published from January 1973 to October 2011, using Medline and EMBASE and the search terms “vitamin D”; “autoimmune disease”; “autoimmunity”; “rheumatoid arthritis”; “systemic lupus erythematosus”; “scleroderma”; “systemic sclerosis”; “type 1 diabetes”; “multiple sclerosis”; and “undifferentiated connective tissue disease”. We selected studies on the environmental, genetic and epidemiologic association of vitamin D with autoimmune diseases. Using the strategy described, we identified 1268 articles. 331 articles were eliminated on the basis of the title and another 703 on the basis of the abstract, since they were considered irrelevant for the purposes of the study. Full-text examination was performed on the remaining 234 studies, and a further 15 studies were excluded from the review, since the results had been confirmed or superseded by more recent research. Finally, a systematic review was conducted on 219 articles concerning cross-sectional data on: vitamin D levels and autoimmune diseases; interventional data on vitamin D supplementation in autoimmune diseases; prospective data linking vitamin D level or intake to autoimmune disease risk. Results: Physiopathology studies confirm that hypovitaminosis D, in genetically predisposed subjects, can impair self tolerance by compromising the regulation of dendritic cells, of regulatory T-lymphocytes and of Th1 cells. Cross-sectional studies show that levels of vitamin D <30ng/mL are present in a significant percentage, not only in patients with autoimmune disease, but also in healthy subjects (30–77%), and link profound deficiency (<10ng/mL) with aggravation of symptomatology, while genetic studies associate polymorphism of vitamin D receptors to various autoimmune diseases. Among experimental studies on humans, only those on type-1 diabetes prove that the risks are significantly reduced in infants treated with vitamin D after the 7th month (OR 0.71, 95% CI, 0.60 to 0.84) and that a dose–response effect exists. Conclusions: Basic, genetic, and epidemiological studies indicate a potential role of vitamin D in the prevention of autoimmune diseases, but randomized and controlled trials are necessary to establish the clinical efficacy of vitamin D supplementation in ill or at-risk subjects. [Copyright &y& Elsevier]

Since it was first identified, thymosin alpha 1 (Tα1) has been characterized to have pleiotropic effects on several pathological conditions, in particular as a modulator of immune response and inflammation. Several properties exerted by Tα1 may be attributable to a direct action on lymphoid cells. Tα1 has been shown to exert an immune modulatory activity on both T cell and natural killer cell maturation and to have an effect on functions of mature lymphocytes, including stimulating cytokine production and cytotoxic T lymphocyte-mediated cytotoxic responses. In previous studies we have shown that Tα1 increases the expression of major histocompatibility complex class I surface molecules in murine and human tumor cell lines and in primary cultures of human macrophages. In the present paper, we describe preliminary data indicating that Tα1 is also capable of increasing the expression of tumor antigens in both experimental and human tumor cell lines. This effect, which is exerted at the level of the target tumor cells, represents an additional factor increasing the antitumor activity of Tα1. [ABSTRACT FROM AUTHOR]

Abstract: It has been reported that treatment with tricin (4′,5,7-trihydroxy-3′,5′-dimethoxyflavone), a derivative of Sasa albo-marginata, after human cytomegalovirus (HCMV) infection significantly suppressed both infectious virus production and HCMV replication in the human embryonic fibroblast cell line MRC-5. In this paper, we examined the mechanisms for the anti-HCMV effects of tricin in MRC-5 cells. Exposure of fibroblasts to tricin inhibited infectious HCMV production, with concomitant decreases in levels of transcripts of the CXC chemokine IFN-inducible T cell alpha chemoattractant (I-TAC or CXCL11) gene. We also found that the transcripts of the HCMV immediate early (IE) gene and replication of HCMV were lower in CXCL11 gene-knockdown cells. These results suggest that tricin is a novel compound with potential anti-HCMV activity and that CXCL11 is one of the chemokines involved in HCMV replication. In addition, it is possible that CXCL11 is the one of the targets of tricin. [Copyright &y& Elsevier]

Abstract: There are a lot of questions about genetic effects of ionizing radiation, the main one is does ionizing radiation induce mutations in humans? There is no direct evidence that exposure of parents to radiation leads to excess heritable disease in offspring. What is the difference between human and other species in which radiation induced mutations are easily registered? During evolution germ cell selection ex vivo has been changed to a selection in vivo and we cannot observe such selection of radiation damaged cells in human. Low radiation doses – are they harmful or beneficial? The “hormesis” phenomenon as well as radioadaptive response proves positive effects of low radiation dose. Can analysis of chromosomal aberration rate in lymphocytes be used for dosimetry? Many uncontrolled factors may be responsible for significant mistakes of this method. Why did evolution preserve the bystander effect? This paper is discussion one and its goal is to pay attention on some effects of ionizing radiation. [Copyright &y& Elsevier]

In this paper, we introduce the stochasticity into an HIV-1 infection model with cytotoxic T lymphocytes (CTLs) immune response via the technique of parameter perturbation. We show that there is a positive solution as desired in any population dynamics. Then we analyze the long time behavior of this model. We obtain a sufficient condition for the stochastic asymptotic stability in the large of the infection-free equilibrium and give the conditions for the solution fluctuating around the two infection equilibria (one without CTLs being activated and the other with). Finally, we make simulations to conform to our analytical results. [ABSTRACT FROM AUTHOR]

The comet assay or single cell gel electrophoresis has proven to be a versatile and sensitive method of measuring the induction and repair of DNA damage in individual cells. However, one of the drawbacks of the assay is the bias caused by changes in the ability of cells to repair DNA damage in different cell cycle phases. Whereas the bias seems less important when G0 peripheral blood lymphocytes are studied, it might cause problems when proliferating cells are investigated. In this paper, we validate the assumption that the total comet fluorescence intensity corresponds to the position of the cell in the cell cycle and can be used to assign single cells to specific cell cycle phases. To validate the approach, we used a very homogenous blood mononuclear CD34+ cell population in G0 phase (unstimulated) or stimulated to enter the cell cycle. An analysis of the cell cycle distribution revealed that the 15 comet intensity classes and the 100 comets usually analyzed in a typical comet experiment are sufficient to obtain a reliable cell cycle distribution comparable with the results obtained by the flow cytometry for the same cell population. The effect of the cell cycle position on the results obtained by the comet assay for proliferating and non-proliferating cell populations irradiated with 3 Gy of X-radiation is also discussed. [ABSTRACT FROM AUTHOR]

Eosinophils have long been associated with allergy and parasitic infections. Today, they are considered as multifunctional leukocytes, which participate both in innate and adaptive immune response though the expression of various receptors and mediators. Although the tumor-associated eosinophilia is observed for a long time in many hematological and solid malignancies, with a generally good prognosis value, there is a lack of knowledge on the different mechanisms involved in this phenomenon. Moreover, the recent discovery in human eosinophils of different receptors and mediators, shared with lymphocytes and involved in anti-tumor defense, suggests that eosinophils can play a role in anti-tumoral immunity. We review in the present paper the current knowledge on epidemiology, recruitment, and mechanisms involved in the response of eosinophils toward tumors. [ABSTRACT FROM AUTHOR]

In this paper we present a mathematical model formulated within the framework of the kinetic theory for active particles. The model is a bilinear system of integro-differential equations of Boltzmann type and it describes the interactions between virus population and the adaptive immune system. The population of cytotoxic T lymphocytes is additionally divided into precursor and effector cells. The effects of the parameters describing the proliferation and differentiation of cytotoxic T lymphocytes are studied numerically. [ABSTRACT FROM AUTHOR]

Abstract: Gossypol is a polyphenolic aldehyde that is produced in the cotton plant. Since long it has been reported to possess antiproliferative activity against a variety of cancer cell lines as well as tumor regression in animal models. However, the toxicity of gossypol does not permit it to be an effective antitumor agent. One of the derivatives of gossypol to show promising results is apogossypolone. For example, it has been shown to specifically target tumor growth in hepatocellular carcinoma xenograft in nude mice without causing any damage to normal tissue. Using human peripheral lymphocytes, in this paper we show that both gossypol and its semi-synthetic derivative apogossypolone cause oxidative DNA breakage in these cells through the mobilization of endogenous copper ions. Such cellular DNA breakage is inhibited by copper specific chelator but nor by iron or zinc chelating agents. Similar results are obtained with isolated nuclei indicating that chromatin bound copper is mobilized in this reaction. Further, apogossypolone showed enhanced DNA breakage and increased oxidative stress in whole lymphocytes as compared with gossypol indicating that this is possibly the result of greater permeability of apogossypolone. It is well established that tissue, cellular and serum copper levels are considerably elevated in various malignancies. Therefore, cancer cells may be subject to greater electron transfer between copper ions and gossypol/apogossypolone to generate reactive oxygen species responsible for DNA cleavage. This may account for the preferential cytotoxicity of apogossypolone towards tumor cells. [Copyright &y& Elsevier]

Abstract: The bursa of Fabricius, the acknowledged central humoral immune organ, is vital to B cell differentiation. However, the regulatory function of the bursal-derived peptide on avian B cell proliferation has not been reported. BSP-II is a recently reported bursal-derived bioactive peptide. In this paper, 75 days-old chicks were twice subcutaneously immunized with BSP-II and inactivated avian influenza virus (AIV, H9N2 strain). It was proved that BSP-II induced a strongly AIV-specific HI antibody production in the immunized chicks. Also, BSP-II could enhance avian pre-B lymphocyte DT40 cell viability. To investigate the global patterns of gene expression in DT40 cells after BSP-II treatment, gene microarray was carried out. It was identified that the differentially expressed genes were involved in various pathways, of which six pathways were associated with signaling transductions, including ErbB signaling, MAPK signaling, Toll-like receptor signaling, Notch signaling, mTOR signaling, and Wnt signaling. Finally, RT-qPCR was used to confirm the microarray expression data. These results indicated the molecular basis of pre-B lymphocyte viability with BSP-II treatment, which provided a potential mechanism of the bursa of Fabricius on pre-B lymphocyte viability, differentiation, and development. These results are valid for the mechanism of the bursa of Fabricius on B lymphocytes development. [Copyright &y& Elsevier]

Abstract: In this paper we evaluate the role of human γδ T cells in control of Francisella tularensis infection. Using an in vitro model of infection, a reduction in bacterial numbers was detected in the presence of human γδ T cells for both attenuated LVS and virulent SCHU S4 strains of F. tularensis. Antibody neutralisation of IFN-γ caused an increase in survival of F. tularensis LVS suggesting that γδ T cell-mediated control of F. tularensis infection is partially mediated by IFN-γ. [Copyright &y& Elsevier]

Active contours and active shape models (ASM) have been widely employed in image segmentation. A major limitation of active contours, however, is in their 1) inability to resolve boundaries of intersecting objects and to 2) handle occlusion. Multiple overlapping objects are typically segmented out as a single object. On the other hand, ASMs are limited by point correspondence issues since object landmarks need to be identified across multiple objects for initial object alignment. ASMs are also are constrained in that they can usually only segment a single object in an image. In this paper, we present a novel synergistic boundary and region-based active contour model that incorporates shape priors in a level set formulation with automated initialization based on watershed. We demonstrate an application of these synergistic active contour models using multiple level sets to segment nuclear and glandular structures on digitized histopathology images of breast and prostate biopsy specimens. Unlike previous related approaches, our model is able to resolve object overlap and separate occluded boundaries of multiple objects simultaneously. The energy functional of the active contour is comprised of three terms. The first term is the prior shape term, modeled on the object of interest, thereby constraining the deformation achievable by the active contour. The second term, a boundary-based term detects object boundaries from image gradients. The third term drives the shape prior and the contour towards the object boundary based on region statistics. The results of qualitative and quantitative evaluation on 100 prostate and 14 breast cancer histology images for the task of detecting and segmenting nuclei and lymphocytes reveals that the model easily outperforms two state of the art segmentation schemes (geodesic active contour and Rousson shape-based model) and on average is able to resolve up to 91% of overlapping/occluded structures in the images. [ABSTRACT FROM AUTHOR]

To mount an immune response, lymphocytes must recirculate between the blood and lymph nodes, recognize antigens upon contact with specialized presenting cells, proliferate to expand a small number of clonally relevant tymphocytes, differentiate to antibody-producing plasma cells or effector T cells, exit from lymph nodes, migrate to tissues, and engage in host-protective activities. All of these processes involve motility and cellular interactions--events that were hidden from view until recently. Introduced to immunology by three papers in this journal in 2002, in vivo live-cell imaging studies are revealing the behavior of cells mediating adaptive and innate immunity in diverse tissue environments, providing quantitative measurement of cellular motility, interactions, and response dynamics. Here, we review themes emerging from such studies and speculate on the future of immunoimaging. [ABSTRACT FROM AUTHOR]

Ag receptor diversity involves the introduction of DNA double-stranded breaks during lymphocyte development. To ensure fidelity, cleavage is confined to the G0-G1 phase of the cell cycle. One established mechanism of regulation is through periodic degradation of the RAG2 recombinase protein. However, there are additional levels of protection. In this paper, we show that cyclical changes in the IL-7R signaling pathway functionally segregate pro-B cells according to cell cycle status. In consequence, the level of a downstream effector of IL-7 signaling, phospho-STAT5, is inversely correlated with cell cycle expression of Rag, a key gene involved in recombination. Higher levels of phopho-STAT5 in S-G2 correlate with decreased Rag expression and Rag relocalization to pericentromeric heterochromatin. These cyclical changes in transcription and locus repositioning are ablated upon transformation with v-Abl, which renders STAT5 constitutively active across the cell cycle. We propose that this activity of the IL-7R/STAT5 pathway plays a critical protective role in development, complementing regulation of RAG2 at the protein level, to ensure that recombination does not occur during replication. Our data, suggesting that pro-B cells are not a single homogeneous population, explain inconsistencies in the role of IL-7 signaling in regulating Igh recombination. [ABSTRACT FROM AUTHOR]

Obesity is a chronic inflammatory condition characterized by activation and infiltration of proinflammatory immune cells and a dysregulated production of proinflammatory cytokines. While known as a key regulator of immune natural killer (NK) cell function and development, we have recently demonstrated that reduced expression of the cytokine Interleukin-15 (IL- 15) is closely linked with increased body weight and adiposity in mice and humans. Previously, we and others have shown that obese individuals have lower circulating levels of IL-15 and NK cells. Lean IL-15 overexpressing (IL-15 tg) mice had an accumulation in adipose NK cells compared to wildtype and NK cell deficient obese IL-15-/- mice. Since IL-15 induces weight loss in IL-15-/- and diet induced obese mice and has effects on various lymphocytes, the aim of this paper was to determine if lymphocytes, particularly NK cells, play a role in IL-15 mediated weight loss. Acute IL-15 treatment resulted in an increased accumulation of NK, NKT, and CD3+ T cells in adipose tissue of B6 mice. Mice depleted of NK and NKT cells had similar weight loss comparable to controls treated with IL-15. Finally, IL-15 treatment induces significant weight loss in lymphocyte deficient RAG2-/-γc-/- mice independent of food intake. Fat pad cross-sections show decreased pad size with cytokine treatment is due to adipocyte shrinkage. These results clearly suggest that IL-15 mediates weight loss independent of lymphocytes. [ABSTRACT FROM AUTHOR]

Background: Non-obese diabetic (NOD) mice develop Sjögren's-like disease (SS-like) with loss of saliva flow and increased lymphocytic infiltrates in salivary glands (SGs). There are recent reports using multipotent mesenchymal stromal cells (MSCs) as a therapeutic strategy for autoimmune diseases due to their anti-inflammatory and immunomodulatory capabilities. This paper proposed a combined immuno- and cell-based therapy consisting of: A) an injection of complete Freund's adjuvant (CFA) to eradicate autoreactive T lymphocytes, and B) transplantations of MSCs to reselect lymphocytes. The objective of this was to test the effectiveness of CD45-/TER119- cells (MSCs) in re-establishing salivary function and in reducing the number of lymphocytic infiltrates (foci) in SGs. The second objective was to study if the mechanisms underlying a decrease in inflammation (focus score) was due to CFA, MSCs, or CFA+MSCs combined. Methodology/Principal Findings: Donor MSCs were isolated from bones of male transgenic eGFP mice. Eight week-old female NOD mice received one of the following treatments: insulin, CFA, MSC, or CFA+MSC (combined therapy). Mice were followed for 14 weeks post-therapy. CD45-/TER119- cells demonstrated characteristics of MSCs as they were positive for Sca-1, CD106, CD105, CD73, CD29, CD44, negative for CD45, TER119, CD11b, had high number of CFU-F, and differentiated into osteocytes, chondrocytes and adipocytes. Both MSC and MSC+CFA groups prevented loss of saliva flow and reduced lymphocytic infiltrations in SGs. Moreover, the influx of T and B cells decreased in all foci in MSC and MSC+CFA groups, while the frequency of Foxp3+ (Treg) cell was increased. MSC-therapy alone reduced inflammation (TNF-a, TGF-b), but the combination of MSC+CFA reduced inflammation and increased the regenerative potential of SGs (FGF-2, EGF). Conclusions/Significance: The combined use of MSC+CFA was effective in both preventing saliva secretion loss and reducing lymphocytic influx in salivary glands. [ABSTRACT FROM AUTHOR]

Abstract: Fetal cattle B-cell development proceeds via a pre-B cell stage that is characterized by the expression of surrogate light chain and recombination activation genes. In this paper, we identify a new member of bovine pre-B lymphocyte genes, VPREB2. Using RT-qPCR, we assess the expression of VPREB2 and three other surrogate light chain genes as well as RAG1 and RAG2 in fetal and adult cattle tissues. The absence of VPREB1, IGLL1, RAG1 and RAG2 expression in adult tissues and the lack of B-lymphoid differentiation in adult bone marrow – OP9 stromal cell co-culture, suggest a decline of B lymphopoiesis in adult cattle. The marked differences in the expression profiles of VPREB2 and VPREB3 in comparison to those of VPREB1, IGLL1 and RAGs suggest that the biological roles of VPREB2 and VPREB3 are unrelated to the pre-B cells. [Copyright &y& Elsevier]