Spleen Lymphoid Cells Research Articles

Presence of TSHR in NK Cells and Action of TSH on NK Cells

Introduction: Thyroid-stimulating hormone receptor (TSHR) is widely expressed in human tissues and cells. TSHR is not only involved in thyroid disease but also in the neuroendocrine-immune regulatory network. However, no study has exclusively focused on the expression and function of TSHR in natural killer (NK) cells. Methods: We studied TSHR expression using reverse transcription PCR to verify TSHR mRNA transcripts in human and mouse NK cells. Human and mouse thyroid and liver tissues as well as peripheral blood mononuclear cells (PBMCs) or spleen lymphoid cells (SLCs) were used as controls. The TSHR protein levels in NK-92 cells were determined by immunofluorescence staining. The function of TSHR in NK cells was investigated by measuring the TSH-stimulated cAMP levels. Results: TSHR mRNA was detected in human and mouse NK cells as well as in NK-92 cells and had the same sequence as that of thyroid-derived, PBMC-derived, and liver-derived mRNA. The TSHR protein was also expressed in the cell membrane of NK-92 cells. Furthermore, the cAMP levels in NK-92 cells were significantly higher after adding 10² mIU/mL of bovine TSH at p < 0.05, which stimulated cAMP production in NK-92 cells. Conclusions: Our findings confirm that TSHR is present and functional in NK cells and provide key clues for the potential regulatory effects of TSH on TSHR in NK cells in the immune system.

Nanoceria distribution and effects are mouse-strain dependent

Prior studies showed nanoparticle clearance was different in C57BL/6 versus BALB/c mice, strains prone to Th1 and Th2 immune responses, respectively. Objective: Assess nanoceria (cerium oxide, CeO2 nanoparticle) uptake time course and organ distribution, cellular and oxidative stress, and bioprocessing as a function of mouse strain. Methods: C57BL/6 and BALB/c female mice were i.p. injected with 10 mg/kg nanoceria or vehicle and terminated 0.5 to 24 h later. Organs were collected for cerium analysis; light and electron microscopy with elemental mapping; and protein carbonyl, IL-1β, and caspase-1 determination. Results: Peripheral organ cerium significantly increased, generally more in C57BL/6 mice. Caspase-1 was significantly elevated in the liver at 6 h, to a greater extent in BALB/c mice, suggesting inflammasome pathway activation. Light microscopy revealed greater liver vacuolation in C57BL/6 mice and a nanoceria-induced decrease in BALB/c but not C57BL/6 mice vacuolation. Nanoceria increased spleen lymphoid white pulp cell density in BALB/c but not C57BL/6 mice. Electron microscopy showed intracellular nanoceria particles bioprocessed to form crystalline cerium phosphate nanoneedles. Ferritin accumulation was greatly increased proximal to the nanoceria, forming core-shell-like structures in C57BL/6 but even distribution in BALB/c mice. Conclusions: BALB/c mice were more responsive to nanoceria-induced effects, e.g. liver caspase-1 activation, reduced liver vacuolation, and increased spleen cell density. Nanoceria uptake, initiation of bioprocessing, and crystalline cerium phosphate nanoneedle formation were rapid. Ferritin greatly increased with a macrophage phenotype-dependent distribution. Further study will be needed to understand the mechanisms underlying the observed differences.

ВОЗРАСТНЫЕ ОСОБЕННОСТИ МОРФОГЕНЕТИЧЕСКИХ СВОЙСТВ ЛИМФОИДНЫХ КЛЕТОК СЕЛЕЗЕНКИ

Aim. The aim of the study is to determine the features of the morphogenetic properties of the spleen lymphoid cells in animals of different age groups. Materials and research methods. In the experiments, a model of adoptive transfer of lymphoid cells of the spleen of nephrectomized animals from different age groups was used. The studies were carried out on male Wistar rats of mature (8-10 months, weighing 200-250 g) and old (19-22 months, weighing 400-500 g) age. Study groups: control group — model of adoptive transfer of splenocytes from mature animals to mature ones; second — experimental group: model of adaptive transfer of lymphoid cells from old — old animals; the third group — transplantation of lymphoid cells from old animals — to mature ones; the fourth group — transplantation of lymphoid cells from mature animals — to old ones. The morphogenetic properties of the spleen cells of nephrectomized animals from different age groups were studied 19 hours after unilateral nephrectomy (donor interval). The resulting suspension was injected intravenously into recipients at 400×10³ cells per 0.2 kg of animal weight. 8 hours before slaughter, the animals received a single intraperitoneal dose of 2 mg/kg of vinblastine. Recipients were sacrificed 40, 48 and 56 hours after splenocyte transfer. Research results. Transplantation of splenocytes obtained from old donors to old recipients did not reveal any stimulation of proliferative processes in the epithelium of the kidney tubules of the latter. The transfer of the same splenocytes to mature donors is accompanied by the manifestation of their morphogenetic function. At the same time, the activity of proliferative processes in the kidneys of the recipients was not less pronounced than in the control group.

The system influence of allogeneic adipose tissue derived mesenchymal stem cells on the functional state of immune organs

The studies were conducted on 2–3-months-old males of mice weighing 20–24 g. Оur work was to study the functional state of the organs of the immune system of C57Bl/6 mice after introduction of allogeneic MSCs of adipose tissue origin. Obtaining and cultivating of MSCs were carried out in a sterile laminar box with compliance of conditions of asepsis and antiseptics. C57Bl/6 mice adipose tissue cultured in a CO2 incubator at 37 °C and 5% CO2 in DMEM with 10–15% of fetal bovine serum, 1% of antibiotic-antimycotic solution (Sigma-Aldrich, USA). The following groups of animals were formed: 1 group – intact (control group); 2 group – animals, to whom 0.5 ml of 0.9% NaCl solution (placebo) were injected into the caudal vein; 3 group – animals, to whom were injected 104 of allogeneic MSCs from adipose tissue in 0.5 ml of phosphate buffer solution into the caudal vein. The weight index, content of lymphoid cells of thymus and spleen in C57Bl/6 mice investigated after the introduction of MSCs on 7, 18 and 25 days. To assess the content of lymphocytes in lymphoid organs, the latter were weighed. Whole thymus and 50 mg of spleen were triturated and filtered through the kapron tissue. After that, the tissue homogenate was applied to the gradient of ficoll-urografin (density 1.077) in a ratio of 3:2. The test tubes were centrifuged at a rate of 1500 rpm for 30-40 minutes. After centrifugation the layer of lymphocytes which was above the gradient was collected by a Pasteur pipette and washed twice with an arbitrary amount of Hanks' solution by centrifugation at a rate of 1500 for 10 minutes. 1 ml of Hanks's solution was added to lymphocytes after washing. Lymphocytes were counted in the Goryaev chamber. Calculation of the cells of lymphoid organs was performed on 1 mg of tissue. The administration of allogenic adipose derived mesenchymal stem cells affects on the central and peripheral organs of the immune system. Administration of allogenic adipose derived mesenchymal stem cells cause a significant increase in the content of lymphoid cells in the thymus at 7, 18 and 25 days by 71, 57 and 53% respectively (P &lt; 0.05) compared to the control. Weight index of the thymus directly correlates with the content of lymphoid cells and its value was r = 0.57, P &lt; 0.01 on 7 day and 18 day r = 0.50, P &lt; 0.05. Quantity lymphoid cells in the spleen significant increase at the 7 and 18 days of the immune response by 33 and 19%, respectively (P &lt; 0.01, P &lt; 0.05) compared to the control under administration of allogenic adipose derived mesenchymal stem cells. On the 25th day of experience, the content of lymphoid cells in spleen and spleen index values return to normal. Weight index of the spleen directly correlates with the content of lymphoid cells – r = 0.91–0.94 (P &lt; 0.001).

Effect of age and inoculation route on the infection of duck Tembusu virus in Goslings

Duck Tembusu virus (TMUV) is an emerging flavivirus that has caused variable levels of outbreaks in poultry in recent years. In order to study the effect of age and inoculation routes on the TMUV infection, one hundred healthy domestic 5-day-old and 20-day-old goslings were equally divided into five groups and four experimental groups of goslings were infected with the TMUV-SDSG strain by intravenous and intranasal routes, respectively. Severe clinical signs were observed in goslings infected at 5 days of age, including listlessness, growth retardation, severe neurological dysfunction and even death. However, goslings infected at 20 days of age showed mild symptoms and no mortality. The severity of gross lesions gradually reduced as goslings matured. The severe histopathological changes were observed in 5-day-old infected goslings, including cerebral edema, viral encephalitis, myocardial necrosis, hepatic steatosis, spleen lymphoid cell depletion, pancreatic epithelial cell shedding and interstitial hemorrhage. However, 20-day-old infected goslings showed mild histopathological changes. Viral loads in different tissues were detected by the SYBR Green I real-time PCR assay. The level of viral loads in most of tissues 5-day-old infected goslings was higher than that of 20-day-old infected goslings, correlating with the severity of clinical symptoms and lesions in these tissues. 20-day-old infected goslings developed significantly higher serum neutralizing antibody titers than 5-day-old infected goslings. Furthermore, goslings infected with TMUV intravenously demonstrated more severe clinical signs, lesions and higher viral loads in tissues than those of goslings infected with TMUV intranasally. Therefore, age and inoculation routes can affect the pathogenicity of TMUV in geese and younger geese are more susceptible to the virus. Age and inoculation route factors should be considered in study of the pathogenicity, pathogenesis, folumation of prevention and therapy strategies of TMUV infection in geese.

ID: 129: Role of the IL-36 pathway in inflammatory responses in mouse neonates

Background: IL-36 synergizes with IL-12 to induce T helper (Th)1 differentiation. To investigate IL-36’s TH1-biased immunostimulatory potential, neonates (1wk-old) were immunized with aluminum (Alum)-adsorbed tetanus toxoid (TT/Alum) and recombinant IL-36 β . Unexpectedly, the combination of IL-36 β (0.1 μ g/g) with TT/Alum resulted in rapid mortality, which was not observed in 1 wk old IL-36R deficient or in adult wild-type mice. Objective: Understand the mechanisms of IL-36-induced toxicity in early life. Methods: 1 wk-, 3 wk- old (infants) and adult (6–8 wk) mice were immunized with TT/Alum, different doses of IL-36 β or a combination thereof. Cytokine levels were measured in serum and organ total RNA. Liver and spleen lymphoid and myeloid cells were sorted. Results: Upon IL-36 injection, mRNA levels of TNF- α , IFN- γ , and of several other cytokines were significantly and specifically increased in the liver of neonates, as compared to 3 wkold and adult mice. Liver IL-36R mRNA levels were elevated in neonates and rapidly decreased with age, which correlated with a higher number of leukocytes, especially B cells, monocytes and neutrophils in neonatal liver. IL-36R mRNA was expressed in dendritic cells,and to a lesser extent in B cells, T cells, NK/NKT cells, monocytes and neutrophils. TNF- α and IFN- γ were mainly produced by myeloid (monocytes and neutrophils) and lymphoid (NK1.1+ and T cells) cells respectively. Importantly, mice injected with a monoclonal anti-TNF- α antibody were fully protected from IL-36-induced lethality. Conclusion: IL-36-induced toxicity in neonates is associated with a higher number of liver IL-36R expressing myeloid cells, leading to excessive TNF- α production.

Clinicopathological studies on alpha-cypermethrin induced toxicity in wistar rats (Rattus norvegicus)

The clinicopathological changes were studied in alpha-cypermethrin induced toxicity in wistar rats. Forty adult wistar rats were divided randomly into four equal groups Viz., group I, II, III and IV. Group I rats received only Dimethyl sulphoxide (DMSO) as vehicle and served as control. Group II, III and IV rats were orally administered daily with alpha-cypermethrin at the dose rate of 4.83, 7.25 and 14.50 mg/kg, respectively after dissolving it in DMSO for a period of 28 days. After 14 day of study onwards, alpha-cypermethrin dosed rats showed behavioral changes within 15 minutes to 2 hrs after dosing. Group III rats received 7.25 mg/kg showed tendency to push head to the ground, whereas elevated startle response and abnormal gait with spayed hind limb were observed in group IV rats received 14.50 mg/kg of alphacypermethrin. The significant (P<0.05) decrease in body weights was observed in rats of group III and IV from day 14 onwards when compared with the control. All the rats exposed to alpha-cypermethrin at three different dose levels revealed dose dependent pathological changes. The changes comprised of varying degrees of congestion, hemorrhages, degeneration and necrosis in various visceral organs and lysis of lymphoid cells in spleen. The severity and distribution of such lesions were found higher in rats of group III and IV as compared to group II and control, group I rats.

Effect of isopropyl methylphosphonofluoridate (IMPF) poisoning on selected immunological parameters of angiogenesis.

Acetylcholinesterase (AChE) and cholinergic receptors play an important role in the immune system, including lymphocyte-induced angiogenesis. However, their exact role is not fully understood. The presented work tests the influence of isopropyl methylphosphonofluoridate (IMPF), an irreversible inhibitor of AChE, on selected immune parameters associated with angiogenesis in mice. The levels of VEGF, bFGF, TNF-α, and IFN-γ production were measured, together with the ability of lymphoid spleen cells to induce local GvH reaction after a single dose of IMPF. Experiments were performed in male BALB/c mice. Acetylcholinesterase activity in erythrocytes was determined by the Ellman`s procedure. Levels of cytokines were measured in serum using standard commercial ELISA kits. Influence of IMPF intoxication upon angiogenesis was examined by the LIA test, according to the Sidky and Auerbach procedure. The results showed a 6- and 8-fold increase in VEGF at days 1 and 7 of the experiment, respectively, as well as a decrease (at days 14 and 21 after administration), followed by a significant increase (day 1) in bFGF levels. A statistically significant decrease in the concentration of IFN-γ was observed throughout all experiments. The maximum decrease in the level of TNF-α was found at days 1 and 7 after administration of IMPF. Additionally, a significant decrease was found in the ability to form new blood vessels following IMPF administration. This study revealed that IMPF has a significant effect on the regulation of lymphocyte-induced angiogenesis, which is related with the modulation of angiogenic and pro-inflammatory cytokines secretion. The observed differences suggest a possible derangement of certain elements of the neuronal and/or non-neuronal cholinergic system.

Immunohistochemical localization of arginase‐1 expression in B6 and BALB/c mice

Arginase is the final enzyme of the urea cycle hydrolyzing L‐arginine to ornithine and urea. The isoform arginase‐1 has been suggested to play a role in cardiovascular disease, cystic fibrosis, asthma and cancer. The goal of this study was to develop and validate immunohistochemical detection of arginase‐1 in formalin‐fixed paraffin embedded mouse tissues from B6 and BALB/c strains. Immunohistochemistry was performed using a primary Ab (polyclonal rabbit anti‐arginase‐1) with optimization/validation using the liver, where zone 1–2 hepatocytes express arginase‐1, but zone 3 lacks cellular expression. Tissues with prominent arginase‐1 cellular expression included liver, endocrine pancreas, lymphoid tissues, esophagus, skin, lung, thyroid, parathyroid, bone marrow, intestine, spleen, salivary glands and clitoral glands. Various disease states were also examined. Normal lymphoid cells in spleen had moderate diffuse arginase‐1 expression, but neoplastic lymphocytes lacked arginase‐1 expression. Hepatocytes, in a liver infiltrated by lymphoma, had enhanced arginase‐1 expression. Lungs with eosinophilic crystalline pneumonia commonly had alveolar macrophages with strong immunostaining. Endothelial cells of vessels had common immunostaining, while erythrocytes and platelets were negative. Smooth muscle of various tissues had multifocal uncommon immunostaining. There was no evidence of strain‐dependent differences in arginase‐1 expression. The diversity of tissues and cells that expressed arginase‐1 emphasizes the potential importance of arginase‐1 in disease models.

Influence of Stimforte on the immunologic characteristics of mice against induced immunosuppression

Event Abstract Back to Event Influence of Stimforte on the immunologic characteristics of mice against induced immunosuppression Olga V. Lebedinskaya1*, Elena A. Lebedinskaya1, Evgenia V. Fadeeva1, Nelly K. Akhmatova2, Dmitriy G. Maldov2 and Mikhail V. Kiselevsky2 1 Acad. E.A. Wagner Perm State Medical Academy, Russia 2 N.N. Blokhin Russian Cancer Research Center RAMS, Russia The aim of study was the analysis of Stimforte effect on immunological parameters in mice against induced immunosuppression. Investigation was carried out with 80 BALB/c mice there were injected either with cyclophosphane (CP) and Stimforte in different combinations or with physiological solution (control group). The immunophenotype of mononuclear leukocytes (ML) isolated from animal spleen and killer activity against YAK-1 tumor line were defined. Morphological investigations of Romanosvky-Giemsa-stained smears were administrated. CP administered (100 mg/kg) within 3 days evoked immunosuppression that was manifested in nearly 7-fold lowering of ML number in mice spleen as compared with control group. ML population in CP-received animal group was represented by mature lymphocytes, whereas control group demonstrated considerable amount of young lymphoid cells in spleen. Animals against CP introduction exhibited significant decrease in natural killer (NK) and pronounced reduction in their NK-activity. Stimforte injection against CP-induced immunosuppression increased more than by 2-fold the ML content in mice spleen, when immature forms such as pro-lymphocytes were detected. In addition, Stimforte favored the normalization of NK content in spleen ML and their NK activity increment nearly to the initial values. Peripheral blood of animals under study also demonstrated the increase in granulocyte number. Thus, CP resulted in marked immunosuppression manifesting in lowering in total lymphocyte amount in mice peripheral blood and spleen, as well as declining in NK content in lymphocyte population as compared with intact animals. Stimforte administration against cytostatic-induced immunosuppression resulted in significant increase in lymphocyte number and normalization of their immunophenotype and function characteristics. Keywords: Stimforte, induced immunosuppression, cyclophosphane, immunophenotype, mononuclear leukocytes Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Immune-mediated disease pathogenesis Citation: Lebedinskaya OV, Lebedinskaya EA, Fadeeva EV, Akhmatova NK, Maldov DG and Kiselevsky MV (2013). Influence of Stimforte on the immunologic characteristics of mice against induced immunosuppression. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00845 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 20 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Ms. Olga V Lebedinskaya, Acad. E.A. Wagner Perm State Medical Academy, Perm, Russia, lebedinska@mail.ru Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Olga V Lebedinskaya Elena A Lebedinskaya Evgenia V Fadeeva Nelly K Akhmatova Dmitriy G Maldov Mikhail V Kiselevsky Google Olga V Lebedinskaya Elena A Lebedinskaya Evgenia V Fadeeva Nelly K Akhmatova Dmitriy G Maldov Mikhail V Kiselevsky Google Scholar Olga V Lebedinskaya Elena A Lebedinskaya Evgenia V Fadeeva Nelly K Akhmatova Dmitriy G Maldov Mikhail V Kiselevsky PubMed Olga V Lebedinskaya Elena A Lebedinskaya Evgenia V Fadeeva Nelly K Akhmatova Dmitriy G Maldov Mikhail V Kiselevsky Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Critical Role of cRel Subunit of NF-κB in Sepsis Survival

NF-κB is a critical regulator of gene expression during severe infections. NF-κB comprises homo- and heterodimers of proteins from the Rel family. Among them, p50 and p65 have been clearly implicated in the pathophysiology of sepsis. In contrast, the role of cRel in sepsis is still controversial and has been poorly studied in single-pathogen infections. We aimed to investigate the consequences of cRel deficiency in a cecal ligation and puncture (CLP) model of sepsis. We have approached the underlying mechanisms of host defense by analyzing bacterial clearance, systemic inflammation, and the distribution of spleen dendritic cell subsets. Moreover, by using a genome-wide technology, we have also analyzed the CLP-induced modifications in gene expression profiles both in wild-type (wt) and in rel(-/-) mice. The absence of cRel enhances mortality due to polymicrobial sepsis. Despite normal pathogen clearance, cRel deficiency leads to an altered systemic inflammatory response associated with a sustained loss of the spleen lymphoid dendritic cells. Furthermore, a whole-blood microarray study reveals that the differential outcome between wt and rel(-/-) mice during sepsis is preceded by remarkable changes in the expression of hundreds of genes involved in aspects of host-pathogen interaction, such as host survival and lipid metabolism. In conclusion, cRel is a key NF-κB member required for host antimicrobial defenses and a regulatory transcription subunit that controls the inflammatory and immune responses in severe infection.

High dose interleukin-12 exacerbates Bordetella pertussis infection and is associated with suppression of cell-mediated immunity in a murine aerosol challenge model.

The in-vivo clearance of Bordetella pertussis infections in murine models in naive mice and animals vaccinated with whole-cell vaccine is considered to be via a Th-1-dependent mechanism in which interleukin-12 (IL)-12 may play a prominent role. It has also been demonstrated clearly that the treatment of animals with macrophage-derived IL-12 administered with an acellular vaccine can increase the efficacy of this vaccine preparation to levels seen with the whole-cell vaccine. However, the effects of exogenously added IL-12 on immune responses in non-vaccinated B. pertussis-challenged mice remain unclear, with two studies giving contradictory findings. In this study we have treated mice with escalating doses of mIL-12 (0.1-10 microg/mouse) prior to challenge with B. pertussis (using an aerosol challenge model of infection). The ability of mice to clear infection was assessed in IL-12 treated and in phosphate buffered saline (PBS) control animals at days 6 and 13 post-challenge. Lymphoid cells were isolated from spleen and cell-mediated immune responses assessed at days 1, 6 and 13 post-challenge. In addition, the direct effects of high-dose IL-12 on challenged mice was assessed by checking natural killer (NK) activity from isolated lung and spleen lymphoid cells as well as interferon-gamma (IFN-gamma) generation from isolated cells and serum at day 1 post-challenge. The results from this study show that bacterial colonization of the lungs is actually enhanced following treatment with high-dose IL-12. This is associated with impaired cellular immune responses. The mechanisms associated with the immunosuppressive effects of IL-12 are discussed.

An Interaction Between Dietary Silicon and Arginine Affects Immune Function Indicated by Con-A-Induced DNA Synthesis of Rat Splenic T-Lymphocytes

Sporadic reports have appeared that suggest silicon plays a functional role in immune function by affecting lymphocyte proliferation. In addition, there is also considerable interest in supplemental arginine as a modulator of immune function. Therefore, the purpose of this animal experiment was to determine the effect of supplemental compared to adequate arginine on immune function as measured by splenic T-lymphocyte proliferation in the presence of adequate or inadequate dietary silicon. The independent variables were, per gram of fresh diet, silicon supplements of 0 or 35 microg and arginine supplements of 0 or 5 mg. The basal diet contained 2.3 microg silicon/g and 7.82 mg L-arginine/g. After feeding the male rats (nine per treatment group) for 8 wk, spleen lymphoid cells were isolated and cultured with methyl-3[H]thymidine. Supplemental arginine significantly decreased Con-A-induced DNA synthesis of splenic T-lymphocytes, but the response to arginine was influenced by dietary silicon. The decreased DNA synthesis was more marked when rats were fed adequate silicon than when fed inadequate silicon. Also, when arginine was not supplemented, DNA synthesis was higher in lymphocytes from rats fed an adequate silicon diet than rats fed the inadequate silicon diet. These findings support the huypothesis that an interaction between silicon and arginine affects immune function and that inadequate dietary silicon impairs splenic lymphocyte proliferation in response to an immune challenge.

The beta-galactosidase gene as a marker for hematopoietic reconstitution: individual cell analysis in a murine bone marrow transplantation model.

We have used a simple, single-gene retrovirus carrying the Escherichia coli beta-galactosidase reporter gene (lacZ), termed LlacZ. This virus was found to infect immortalized myeloid and lymphoid precursor/leukemic cell lines efficiently as well as primary murine bone marrow clonogenic progenitors, without apparent modulation of growth or phenotype. Following infection of bone marrow cells, a significant proportion of progenitors--36% of lineage-negative cells with low levels of c-kit expression (lin-/c-kit(lo)) known to be enriched with pluripotent hemopoietic stem cells, and 19% of Sca1-positive cells known to be enriched with transplantable cells with lymphomyeloid-reconstituting ability--were shown to express lacZ. Use of an LlacZ-infected population of post 5-fluorouracil bone marrow cells to reconstitute lethally irradiated mice demonstrated the presence of lacZ-expressing cells in the spleen at day 12 post-transplantation with provirus detected in individual spleen colonies (CFU-S). In the long term (3-6 months following transplantation), lacZ expression was detected in hematopoietic tissues of all recipient mice. The use of two-color in situ and flow cytometry analysis combined with lineage-specific antibodies showed lacZ expression in both myeloid and lymphoid cells in spleen and bone marrow. In addition, lacZ-expressing cells were detected in secondary recipient mice injected with bone marrow cells derived from primary LlacZ recipients. Overall, these data show the efficacy of a single gene vector for stem cell transduction, the utility of beta-galactosidase as a single cell marker for stem cell transduction and reconstitution ability, and the need for protocol optimization to see high-level multilineage gene expression.

Effects of nefazodone on the immune system of mice

Mice exposed to a chronic auditory stressor and treated with nefazodone (10 mg/kg/day s.c.), showed a reduction in stress-induced suppression of thymus and spleen cellularity, and in peripheral T-Iymphocyte population. The in vitro blastogenic response of spleen lymphoid cells to mitogen concanavalin A, the in vitro and in vivo activity of phagocytosis, both measured using the zymosan and carbon clearance tests, respectively, were also assessed and nefazodone was found to partially reverse the inhibitory effect of stress on those parameters. Nefazodone did not significantly affect those parameters in unstressed mice. In conclusion, this report provides evidence on the immunoprotective effects of this novel antidepressant drug against the adverse effects of stress in mice.

A mutation in the ovine cathepsin D gene causes a congenital lysosomal storage disease with profound neurodegeneration.

The neuronal ceroid lipofuscinoses (NCLs) constitute a group of neurodegenerative storage diseases characterized by progressive psychomotor retardation, blindness and premature death. Pathologically, there is accumulation of autofluorescent material in lysosome-derived organelles in a variety of cell types, but neurons in the central nervous system appear to be selectively affected and undergo progressive death. In this report we show that a novel form of NCL, congenital ovine NCL, is caused by a deficiency in the lysosomal aspartyl proteinase cathepsin D. A single nucleotide mutation in the cathepsin D gene results in conversion of an active site aspartate to asparagine, leading to production of an enzymatically inactive but stable protein. This results in severe cerebrocortical atrophy and early death, providing strong evidence for an important role of cathepsin D in neuronal development and/or homeostasis.

Cytogenetic properties of spleen lymphoid cells in mice after bilateral nephrectomy

It is shown that mouse spleen lymphoid cells isolated 4 h after bilateral nephrectomy enhance mitotic activity of renal tubular epithelium in syngeneic recipients, but have no effect on hepatocytes and little effect on the proliferation of reticuloendothelial cells in these recipients. The cells isolated at the same times postoperation from sham-operated or unilaterally nephrectomized animals are ineffective.

Effects of fluoxetine on the immunosuppressive response to stress in mice

Mice exposed to a chronic auditory Stressor and treated with fluoxetine (5 mg/kg) showed a reduction in stress-induced suppression of thymus and spleen cellularity, and in peripheral T lymphocyte population. The blastogenic response of spleen lymphoid cells and the delayed type hypersensitivity response (DTH) to sheep red blood cells (SRBC) were also assessed and fluoxetine was found to partially reverse the inhibitory effect of stress on both parameters.

Ethanol-induced changes in lymphoid cell populations: relationship to liver pathology

Our study related the role of different types of dietary fat and pathologic liver injury to alterations in lymphoid cells in spleen, thymus and peripheral blood in chronically ethanol fed rats. Animals were fed either corn oil or saturated fats with ethanol. The most significant observation was that a profound decrease in CD4 + CD8 − , CD4 − CD8 + and macrophages was seen in animals fed corn oil and ethanol; similar changes were not seen in the saturated fatethanol group. In addition, the decline in CD4 + CD8 − , CD4 − CD8 + and B cells in the spleen correlated with severity of liver injury in the corn oil-ethanol group. These results suggest that feeding of corn oil and ethanol which leads to liver injury is associated with a significant decrease in lymphoid cells in thymus, spleen and peripheral blood; feeding saturated fat with ethanol which is not associated with liver injury prevents this decrease in lymphoid cells.

Cytometric profiles of bone marrow and spleen lymphoid cells after mercury exposure in mice

The potential immunotoxic effects of mercury chloride on murine bone marrow (bm) cell subpopulations, including analysis of maturation patterns for B-cells, were evaluated by flow cytometric analysis. CD-1 outbred mice were exposed for 28 days to relatively low doses of 25–100 ppm HgCl 2 in drinking water and the mercury-related functional cellular changes were validated in a macrophage phagocytosis assay. Lymphocyte subsets from the bone marrow population were stained with PNA lectin and a panel of monoclonal antibodies against cell surface antigens. The incidence of subset-specific staining was also monitored in spleens and thymuses. A dose-effect correlation was noted for the mercury-related activation of macrophage phagocytosis. Subchronic exposure to mercuric chloride resulted in a transient (7–14 day) decrease of the lymphoid/total bm cell ratio and affected the incidence of splenic T-cell subsets, however, without a clear dose-response correlation. The B-cell population in spleen and maturation patterns of B-cells in bm appeared to be unaffected by the mercury exposure. Overall, cytometric analysis of lymphoid cell subsets in murine bone marrow revealed transient and subset-non-specific cell fluctuations after subchronic exposure to inorganic mercury.