Genome-wide association study of individual differences of human lymphocyte profiles using large-scale cytometry data.

Human immune systems are very complex, and the basis for individual differences in immune phenotypes is largely unclear. One reason is that the phenotype of the immune system is so complex that it is very difficult to describe its features and quantify differences between samples. To identify the genetic factors that cause individual differences in whole lymphocyte profiles and their changes after vaccination without having to rely on biological assumptions, we performed a genome-wide association study (GWAS), using cytometry data. Here, we applied computational analysis to the cytometry data of 301 people before receiving an influenza vaccine, and 1, 7, and 90 days after the vaccination to extract the feature statistics of the lymphocyte profiles in a nonparametric and data-driven manner. We analyzed two types of cytometry data: measurements of six markers for B cell classification and seven markers for T cell classification. The coordinate values calculated by this method can be treated as feature statistics of the lymphocyte profile. Next, we examined the genetic basis of individual differences in human immune phenotypes with a GWAS for the feature statistics, and we newly identified seven significant and 36 suggestive single-nucleotide polymorphisms associated with the individual differences in lymphocyte profiles and their change after vaccination. This study provides a new workflow for performing combined analyses of cytometry data and other types of genomics data.

Array tomography: characterizing FAC-sorted populations of zebrafish immune cells by their 3D ultrastructure.

For 3D reconstructions of whole immune cells from zebrafish, isolated from adult animals by FAC-sorting we employed array tomography on hundreds of serial sections deposited on silicon wafers. Image stacks were either recorded manually or automatically with the newly released ZEISS Atlas 5 Array Tomography platform on a Zeiss FEGSEM. To characterize different populations of immune cells, organelle inventories were created by segmenting individual cells. In addition, arrays were used for quantification of cell populations with respect to the various cell types they contained. The detection of immunological synapses in cocultures of cell populations from thymus or WKM with cancer cells helped to identify the cytotoxic nature of these cells. Our results demonstrate the practicality and benefit of AT for high-throughput ultrastructural imaging of substantial volumes.

Centriole polarisation to the immunological synapse directs secretion from cytolytic cells of both the innate and adaptive immune systems.

BACKGROUND: Cytolytic cells of the immune system destroy pathogen-infected cells by polarised exocytosis of secretory lysosomes containing the pore-forming protein perforin. Precise delivery of this lethal hit is essential to ensuring that only the target cell is destroyed. In cytotoxic T lymphocytes (CTLs), this is accomplished by an unusual movement of the centrosome to contact the plasma membrane at the centre of the immunological synapse formed between killer and target cells. Secretory lysosomes are directed towards the centrosome along microtubules and delivered precisely to the point of target cell recognition within the immunological synapse, identified by the centrosome. We asked whether this mechanism of directing secretory lysosome release is unique to CTL or whether natural killer (NK) and invariant NKT (iNKT) cytolytic cells of the innate immune system use a similar mechanism to focus perforin-bearing lysosome release. RESULTS: NK cells were conjugated with B-cell targets lacking major histocompatibility complex class I 721.221 cells, and iNKT cells were conjugated with glycolipid-pulsed CD1-bearing targets, then prepared for thin-section electron microscopy. High-resolution electron micrographs of the immunological synapse formed between NK and iNKT cytolytic cells with their targets revealed that in both NK and iNKT cells, the centrioles could be found associated (or 'docked') with the plasma membrane within the immunological synapse. Secretory clefts were visible within the synapses formed by both NK and iNKT cells, and secretory lysosomes were polarised along microtubules leading towards the docked centrosome. The Golgi apparatus and recycling endosomes were also polarised towards the centrosome at the plasma membrane within the synapse. CONCLUSIONS: These results reveal that, like CTLs of the adaptive immune system, the centrosomes of NK and iNKT cells (cytolytic cells of the innate immune system) direct secretory lysosomes to the immunological synapse. Morphologically, the overall structure of the immunological synapses formed by NK and iNKT cells are very similar to those formed by CTLs, with both exocytic and endocytic organelles polarised towards the centrosome at the plasma membrane, which forms a focal point for exocytosis and endocytosis within the immunological synapse. We conclude that centrosomal polarisation provides a rapid, responsive and precise mechanism for secretory lysosome delivery to the immunological synapse in CTLs, NK cells and iNKT cells.

Evolution and development of immunological structures in the lamprey.

Comparative immunology has been revitalized by the integration of genomics approaches, which allow a foothold into addressing problems that previously had been difficult to study. One such problem had been the enigmatic finding of overt immune anatomical structures in the lamprey, yet its apparent lack of bona fide immunoglobulin or T cell receptor molecules. The genomic characterization of a novel extended locus that undergoes rearrangements to generate receptor diversity and the subsequent implementation of this diversity in the immune system of lampreys have generated considerable interest as well as new avenues for investigation. Here, we review the anatomical structures of the lamprey that exhibit lympho-hematopoietic characteristics, with the ultimate goal of reconciling these data with contemporary molecular findings. By integrating these datasets we seek to better understand how an alternative adaptive immune system could have evolved.

Two-photon imaging of the immune system: a custom technology platform for high-speed, multicolor tissue imaging of immune responses.

Modern imaging approaches are proving important for addressing contemporary issues in the immune system. These approaches are especially useful for characterizing the complex orchestration of immune responses in vivo. Multicolor, two-photon imaging has been proven to be especially enabling for such studies because of its superior tissue penetration, reduced image degradation by light scattering leading to better resolution, and its high image quality deep inside tissues. Here, we examine the functional requirements of two-photon imaging instruments necessary for such immune studies. These requirements include frame rate, spatial resolution and the number of emission channels. We use this discussion as a starting point to compare commercial systems and to introduce a custom technology platform that meets those requirements. This platform is noteworthy because it is very cost-effective, flexible and experimentally useful. Representative data collected with this instrument is used to demonstrate the utility of this platform. Finally, as the field is rapidly evolving, consideration is given to some of the cutting-edge developments in multiphoton microscopy that will likely improve signal strength, depth penetration and/or the experimental usefulness of this approach.

[Changes in liver and in some organs of immune system of animals exposed to twenty-four-hour illumination].

The aim of this research was to study the morpho-functional organization of liver and some organs of immune system in animals exposed to continous twenty-four-hour illumination (CI) for 14 days. Using the methods of light and electron microscopy and morphometric analysis of histological preparations, liver and regional lymph nodes (hepatic in rats and iliac in mice) were studied. It was found that CI resulted in a state of dyschronosis, which was structurally manifested by the changes of diurnal variations of morphometric parameters of liver and its draining lymph nodes, disturbances of hemo- and lymphocirculation, destruction of cellular membranes. Response of central and peripheral organs of immune system to CI included the changes of diurnal variations of blast forms and mature lymphocytes, that was also indicative of dyschronosis development.

Luteinizing hormone-releasing hormone (LHRH) receptors in the neuroendocrine-immune network. Biochemical bases and implications for reproductive physiopathology.

It seems apparent that the brain-pituitary-reproductive axis and the brain-thymus-lymphoid axis are linked by an array of internal mechanisms of communication that use similar signals (neurotransmitters, peptides, growth factors, hormones) acting on similar recognition targets. Moreover, such communication networks form the basis and control of each step and every level of reproductive physiology. This work has focused on the LHRH system, a primary central and peripheral clock of both neuroendocrine and immune functions. From the initiation of a sexually organized response, the detection of sexual odors, and the induction of mating behavior, extrahypothalamic and hypothalamic LHRH orchestrates the neuroendocrine modulation of gonadotropin secretion, while its expression within the ovary directly controls specific events such as follicular atresia. The presence of LHRH receptors in oocytes clearly anticipates a potential action of the decapeptide during the process of fertilization and/or implantation. Within the thymus and other peripheral immune organs, LHRH plays a unique role of immunomodulator, contributing to the sex-dependent changes in immune responsiveness during the estrous-menstrual cycle as well as pregnancy. The reciprocity of the neuroendocrine-immune signaling systems is further supported by the ability of sex steroids to modulate thymus-dependent immune functions via direct effects on specific target genes involved in the development of sex dimorphism and sex-dimorphic immune responses, including the downregulation of immune response observed during pregnancy. Such cyclic changes in immune responsiveness could have a physiological implication, such as the decrease or suppression in cell-mediated immunity observed in the postovulatory phase of the cycle and in pregnancy, respectively, and might play a role during the implantation process and the establishment of pregnancy. In this context, the ability of corticosterone to directly inhibit both GR transcript levels as well as a cell-mediated immune response within the thymus, and the modulation of such an inhibitory effect by the sex steroid hormone milieu, may offer an explanation and a molecular mechanism whereby stress may be deleterious for reproduction, also via immunomodulation. On the other hand, hormonally mediated alterations in immunity might also have a pathological implication in sexually related immune diseases. For example, in mouse and humans, lupus erythematosus is more prevalent in females and estrogen accelerates the disease process, while menstruation is known to exacerbate idiopathic thrombocytopenia purpura. Sex steroid hormone milieu might also have a role in controlling the stress response through immunomodulation. Within the placenta, an intricate network of signaling systems controls a delicate interplay between the neuroendocrine hormones, growth factors, and cytokines that are susceptible to play a major local role in the processes of implantation and the establishment and completion of pregnancy. The neuroendocrine and immunomodulatory role of LHRH continues well after parturition because the presence of LHRH-like material within the mammary gland and milk participates in the physiological modulation of hypophyseal, gonadal, and immune functions of the pups. Such a significant role played by the hypothalamic peptide in the modulation of immune responsiveness would indicate LHRH as the signal conveying information to both neuroendocrine and immune cells, with the role of informing and then transducing the messages into appropriate biological responses.(ABSTRACT TRUNCATED)

Making sense of molecular signatures in the immune system.

The development of Functional Genomics technologies has opened new avenues to investigate the complexity of the immune system. Microarray technology has been particularly successful because of its relatively low cost and high genome coverage. Consequently to our ability to monitor the expression of a significant proportion of an organism genome, our understanding of the molecular dynamics behind cell differentiation and cell response has greatly improved. Molecular signatures associated to immune cells have provided important tools to investigate the molecular basis of diseases and have been often associated to diagnostic and prognostic markers. The availability of such large collection of data has stimulated the application of complex machine learning techniques in the attempt to link molecular signatures and cell physiology. Here we review the most recent developments in the analysis of molecular signatures in the immune system.

Cellular elements of the immune system in the larynx cancer, SEM study.

Observations that certain primary tumors (solid ones) are infiltrated by cells which participate in immune responses tend us to examine this problem in spontaneously growing larynx carcinoma (ca. planoepitheliale). Our observations in SEM revealed within but also in surrounding of cancer infiltrations the presence of cellular elements belonging to the immune system like: lymphocytes, monocytes and macrophages. The most numerous of them, lymphocytes, are brought to the surrounding and tumor's territory by very rich vascular network (angiogenesis phenomenon). Bidirectional transmigration of the lymphocytes, observed mainly within the postcapillary venules, takes place through the cytoplasm of endothelial cells. It is most probably that they are lymphocytes of T-type which are involved in a cell mediated mechanisms of the immune response against a tumor. The lymphocytes are responsible also for the presence of monocytes and especially macrophages within a tumor's territory. These facts suggest that the human organism, in certain degree, is able to fight against malignant tissue by similar mechanisms which are involved in the rejection of the graft.

Morphological alterations in the lymphoreticular system in acquired immunodeficiency syndrome.

Acquired immunodeficiency syndrome (AIDS) manifests a profound deficiency in cellular and humoral immunity causing opportunistic infections with high mortality. Intensive searching for accurate diagnosis, effective treatment, and reliable preventions are in progress. Diagnostic findings include lymphocytopenia, decreased T-helper/T-suppressor ratio and antibodies against human T-lymphotropic retrovirus-III. Specific morphological markers for the diagnosis of AIDS are not yet available at this time. Consistent findings in the lymphoreticular system include a reactive hyperplasia in the onset to lymphocyte depletion in it's advance stage. The frequently mentioned ultrastructural changes in lymphoreticular cells are tubulo-reticular structures, test tube and ring-shaped forms, multivesicular and virus-like particles. These are, however, nonspecific for the diagnosis of AIDS.

[The function of the immune, reproductive and endocrine body systems during irradiation under exposure to stress factors of a non-radiation nature]. / Sostoianie immunnoi, reproduktivnoi i éndokrinnoi sistem organizma pri obluchenii v usloviiakh vozdeistviia stress-faktorov neradiatsionnoi prirody.

Diverse indexes of the state of immune, reproductive and endocrine systems were studied by morphological, biomechanical and biological methods under the effect of non-radiation stress factors, accompanying irradiation of the animals and their changes under effect of the ionizing irradiation 7 and 360 sGy in doze. It was demonstrated that short-time extreme effect lead to hormonal homeostasis destabilization, compensatory-adaptive transformations in immune system and also influence the indexes of copulation efficiency and intrauterine losses. Ionizing irradiation modifies the response to stress-factors effect with modification depending on the irradiation doze.

[An electron microscopic study of the effector link in immune reactions in stomach cancer]. / Elektronno-mikroskopicheskoe izuchenie éffektornogo zvena immunnykh reaktsii pri rake zheludka.

21 surgically removed gastric carcinomas of different histologic structure and stage of development were studied ultrastructurally. A variety of morphological manifestations of local immune reactions and nonspecific antitumour resistance were observed. These manifestations were characterized by the contacts of lymphocytes, macrophages, polymorphonuclear leukocytes, plasma cells with each other and with tumour cells and by the presence of variable changes in tumour cells. The causes of such a variety are discussed and they may be due to different types of cytotoxicity, stage of the interaction between the effector and the target and so on.

[Effect of the neurosensitization of female rats on the functional indices of the DNA-, RNA- and protein-synthesizing neuronal apparatus of the cerebral cortex and of the immune system in their progeny]. / Vliianie neirosensibilizatsii samok krys na nekotorye pokazateli sostoianiia DNK-RNK-beloksinteziruiushchego apparata neironov kory golovnogo mozga i immunnoi sistemy u potomstva.

Neurosensitization effects on the state of the ribonucleoprotein particles and chromatin in nuclei of the Vth layer cortical neurons were studied in female rats. Anti-brain antibodies were investigated in their offspring on the 30th and 60th days. On the 60th day the electron-cytochemical changes were found in the nuclei suggesting the deterioration of nucleolar DNA processing and inhibition of nucleolar functional activity. Anti-brain antibodies were also found in the blood. Direct antibodies interference in the function of cellular DNA-RNA-protein-synthetizing system seems plausible.

[Research of ultra-structural pathological changes of nervous, endocrine and immune system in heroin addicts].

OBJECTIVE: To investigate ultrastructural pathological changes of Heroin-Addicts. METHODS: Heroin-Addicts' central nervous system, endocrine system, immune system and reproductive system in 4 cases are observed by using transmission electron microscope(TEM). RESULTS: The changes of central nervous system are mitochondrion swelling, crista fragmentation and disappear. Endoplasmic reticulum dilation, nervous fibres and cell organelles reduction; mitochondrion swelling, Partial crista fragmentation and endoplasmic reticulum dilation are also found in endocrine system; Lymphocytes reduction, cytoplasm ingredient reduction and dead lymphocytes increase in immune system; in reproductive system, spermatogenic cells and cell organelles are reduced in the male and follicle disappeared in the female. CONCLUSION: Ultra-structural pathological changes of heroin-addicts are presented acute, chronic oxygen deficiency degeneration and necrosis.

[The effect of immunomudulin on the structural-functional status of the organs of the immune and hematopoietic systems in chronic toxic hepatitis]. / Vliianie immunomodulina na strukturno-funktsional'noe sostoianie organov immunnoi i krovetvornoi sistem v usloviiakh khronicheskogo toksicheskogo gepatita.

Long-term entry of the hepatotropic poison heliotropin induces the development of chronic active hepatitis attended with disturbance of the immune balance and hematologic disorders in it. Immunomodulin alleviates the destructive changes in the organs of the immune system, stimulates proliferation and differentiation of their cells, and promotes restoration of red blood parameters. It is suggested that immunoglobulin may be a promising measure in complex treatment of chronic liver diseases.

[The interrelationships of the coccidian Cryptosporidium parvum (Apicomplexa: Sporozoa) with the cells of the immune system in the mammalian host]. / Vzaimootnosheniia koktsidii Cryptosporidium parvum (Apicomplexa: Sporozoa) s kletkami immunnoi sistemy khoziaina-mlekopitaiushchego.

By means of an electron microscopic study of the intestine in young rats infected with Cryptosporidium parvum we observed a mass migration of immunocompetent cells of the host (eosinophils, neutrophils and macrophages) into the lumen of intestine. Some lymphocytes were also observed. Immunocompetent cells (except lymphocytes) included inside phagosomes with different endogenic states of C. parvum. Macrophages with typical extracytoplasmic parasitophorous vacuoles formed by C. parvum were also observed in the intestine lumen. Almost all stages of C. parvum could be observed on a surface of such macrophages. However, we did not find in lamina propria of intestine villi any macrophages with parasites. The place of macrophages infection is unknown. We suggest that surviving of C. parvum in macrophages is principally possible.

The newly found functions of MTOC in immunological response.

The MTOCs are present in all eukaryotic cells. In animal somatic cells, the MTOC function is played by a centrosome, which contains centrioles and PCM. The traditional view is that the MTOC is responsible for the organization of microtubular structures (the intracellular network, cilia, and flagella) in interphase cells, and the formation of the mitotic and meiotic spindle apparatus which is required for the partitioning of chromosomes in dividing cells. Recent evidence suggests that MTOC also plays a key role in the engagement of molecular motors, directional transport of granules, and polarization of subcellular structures and molecules. All of these functions are crucial for targeted cytotoxicity and the regulation of immune cells. In this review, we focus on the ultrastructural and molecular aspects of MTOCs in various aspects of immune cell functions, with specific emphasis on the formation of the IS and targeted cell killing.

Development of an immunologically tolerated combination of fluorescent proteins for in vivo two-photon imaging.

Combinations of fluorescent proteins (FPs) are routinely used for multi-parameter in vivo imaging experiments to visualize tagged proteins or cell populations of interest. Studies involving FPs are often limited by spectral overlap, toxicity, relative quantum efficiency, and the potential for immunological rejection upon transfer into a non-tolerant recipient. Here we evaluate the immunologic visibility of several commonly used FPs by the murine immune system and identify a spectrally compatible, immunologically tolerated combination of FPs well suited for in vivo two-photon imaging.

Histological and ultrastructural examinations of porcine tonsils.

The histology and ultrastructure of porcine tonsils were studied. The porcine tonsils were lymphoepithelial organs situated at the opening of both the digestive and respiratory tracts. The tonsil of the soft palate in the oropharyngeal tract and the paraepiglottic tonsil in the laryngopharynx were mainly consisted of secondary lymphoid follicles encapsulated by connective tissue. The stratified squamous epithelia covering the tonsils and their crypts were frequently heavily infiltrated by lymphoid cells. The pharyngeal and tubal tonsils (TT) were situated in the nasopharyngeal tract. The cells of the pseudostratified columnar epithelia of the pharyngeal and TT were loosely connected, with large intercellular space. They consisted of scattered lymphoid follicles, aggregations of lymphoid cells and diffuse lymphoid tissues. Many high endothelial venules, specialized for the diapedesis of lymphoid cells into the tonsillar tissue, were detected in the four porcine tonsils. Therefore, the overall structures of the tonsils (the tonsil of the soft palate, the paraepiglottic tonsil, the pharyngeal and the TT) reflect their immune functionality in the oral and intranasal immunity.