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Study was conducted on mouse spleen cells, cultured and incubated in-vitro with Holy basil and Thai basil, to observe their effect on proliferation. Four dilutions, namely 1:1, 1:5, 1:25, and 1:125...

This paper introduces a configurable combined laser, ultrasound, and elasticity (CLUE) imaging platform. The CLUE platform enables imaging sequences capable of simultaneously providing quantitative acoustic, optical, and mechanical contrast for comprehensive diagnosis and monitoring of complex diseases, such as cancer. The CLUE imaging platform was developed on a Verasonics ultrasound scanner integrated with a pulsed laser, and it was designed to be modular and scalable to allow researchers to create their own specific imaging sequences efficiently. The CLUE imaging platform and sequence were demonstrated in a tissue-mimicking phantom containing a stiff inclusion labeled with optically-activated nanodroplets and in an ex vivo mouse spleen. We have shown that CLUE imaging can simultaneously capture multi-functional imaging signals providing quantitative information on tissue.

Spleen is one of the important organs for studying graft-versus-host disease development using cryo-imaging modality. Up to this point, there is no algorithm developed to segment spleen tissues in the cryo-imaging data. In this study, we developed a new, automatic segmentation algorithm for spleen tissues in the cryo-images for the first time. The algorithm consisted of (1) pre-processing, (2) predicting, and (3) post-processing. The predicting model consisted of two U-Nets to separate spleen from background and white pulp from red pulp. The results generated by the algorithm were compared against the ground truth generated by cryo-imaging experts. The Dice similarity coefficients were about 86% for red pulp class and 87% for white pulp class. We also used the algorithm to measure spleen volumes and performed the T-cells proliferation assays on the results. We found that there was a significant difference between ratios of white pulp volume to spleen volume between allogeneic spleen and syngeneic spleen. The results were consistent with the previous biological reports. In conclusion, the algorithm accurately and objectively segmented the spleen tissues with similar quality to the experts. By incorporating the algorithm into the T-cell proliferation assay workflow, it should greatly increase throughput of the process as compared to manual segmentation by humans. We expect that this algorithm will help accelerate the research for further understanding of graft-versus-host disease and development of the cure.

We successfully developed a liver-like organ from a stroma-engineered mouse spleen with implanted human liver cells., Regenerating human organs remains an unmet medical challenge. Suitable transplants are scarce, while engineered tissues have a long way to go toward clinical use. Here, we demonstrate a different strategy that successfully transformed an existing, functionally dispensable organ to regenerate another functionally vital one in the body. Specifically, we injected a tumor extract into the mouse spleen to remodel its tissue structure into an immunosuppressive and proregenerative microenvironment. We implanted autologous, allogeneic, or xenogeneic liver cells (either primary or immortalized), which survived and proliferated in the remodeled spleen, without exerting adverse responses. Notably, the allografted primary liver cells exerted typical hepatic functions to rescue the host mice from severe liver damages including 90% hepatectomy. Our approach shows its competence in overcoming the key challenges in tissue regeneration, including insufficient transplants, immune rejection, and poor vascularization. It may be ready for translation into new therapies to regenerate large, complex human tissue/organs.

Mass cytometry, also called cytometry by time-of-flight (CyTOF), is an emerging powerful proteomic analysis technique that utilizes metal chelated polymer (MCP) as mass tags for interrogating high-dimensional biomarkers simultaneously on millions of individual cells. However, under the typical polymer-based mass tag system, the sensitivity and multiplexing detection ability has been highly restricted. Herein, a new structure mass tag based on a nanometal organic framework (NMOF) is reported for multiparameter and sensitive single-cell biomarker interrogating in CyTOF. A uniform-sized Zr-NMOF (33 nm) carrying 105 metal ions is synthesized under modulator/reaction time coregulation, which is monodispersed and colloidally stable in water for over one-year storage. On functionalization with an antibody, the Zr mass tag exhibits specific molecular recognition properties and minimal cross-reaction toward nontargeted cells. In addition, the Zr-mass tag is compatible with MCP mass tags in a multiparameter assay for mouse spleen cells staining, which exploits four additional channels, m/z = 90, 91, 92, 94, for single-cell immunoassays in CyTOF. Compared to the MCP mass tag, the Zr-mass tag provides an additional fivefold signal amplification. This work provides the fundamental technical capability for exploiting NMOF-based mass tags for CyTOF application, which opens up possibility of high-dimensional single-cell immune profiling, low abundant antigen detection, and development of new barcoding systems.

This study was conducted to assess the in vivo and ex vivo immunomodulatory effect of the ethanol leaves extract of Moringa peregrina in Balb/c mice. For this study, five groups of 5 Balb/c mice were given a single acute subtoxic oral dose of the ethanolic extract at 1.13, 11.30, 23.40 and 113.4 mg/kg and the immunomodulatory effect was assessed on the 6th day following the ingestion. In the (non-functional) assessment, the effect of the extract on the body weight, relative lymphoid organ weight, splenic cellularity and peripheral blood hematologic parameters were evaluated. While in the immunomodulation assessment (functional), we investigated the effect of the extract on the proliferative capacity of splenic lymphocytes and peripheral T and B lymphocytes using mitogen blastogenesis, mixed allogeneic MLR and IgM-Plaque forming cells assays. The ingestion of M. peregrina extract caused a significant increase in the body weight, weight and number of cells of spleen and lymph nodes of the treated mice. Furthermore, the count of RBCs, WBCs, platelets, hemoglobin concentration and PCV % were increased by the extract treatment in a dose-dependent manner. M. peregrina enhanced the proliferative responses of splenic lymphocytes for both T cell and B-cell mitogens. Likewise, the mixed lymphocyte reaction MLR assay has revealed a T-cell dependent proliferation enhancement in the extract treated mice. Moreover, the oral administration of M. peregrina leaves extracts significantly increased PFCs/10 6 splenocytes in a dose-dependent manner. In conclusion, subtoxic acute doses of M. peregrina extract demonstrated significant potential as an immunomodulatory agent even at the lowest dose of 1.13 mg/kg.

CD49b is a member of the integrin family, expressed on basophils, natural killer (NK) cells and a subset CD4(+) T cells in the spleen. This protocol describes the adoptive transfer of basophil-enriched CD49b(+) cells obtained from mouse spleens by magnetic enrichment. This protocol can be used to assess the contribution of basophils or basophil-derived mediators to a certain immune response.

Ultrasound-switchable fluorescence (USF) imaging was recently developed to overcome the limitation of the poor spatial resolution of the conventional fluorescence imaging in centimeter-deep tissue. However, in vivo USF imaging has not been achieved so far because of the lack of stable near-infrared contrast agents in a biological environment and the lack of data about their biodistributions. In this study, for the first time, we achieved in vivo USF imaging successfully in mice with high resolution. USF imaging in porcine heart tissue and mouse breast tumor via local injections were studied and demonstrated. In vivo and ex vivo USF imaging of the mouse spleen via intravenous injections was also successfully achieved. The results showed that the USF contrast agent adopted in this study was very stable in a biological environment, and it was mainly accumulated into the spleen of the mice. All the results of USF imaging were validated by a commercial micro-CT., Comment: 14 pages, 6 figures

Background and aim Sodium nitrite (NaNO2) is an inorganic salt with numerous applications in a variety of industries, as well as in medicine. Nevertheless, exposure to high levels of NaNO2 is toxic for animals and humans. Sodium nitrite intoxication is shown to decrease the activity of major antioxidant defence enzymes which is dependent on the maintenance of specific ion equilibrium. The aim of the present study was to investigate the effect of acute NaNO2 intoxication on the content of the essential metals iron (Fe), calcium (Ca) and zinc (Zn) in mouse spleen. Methods Mature male ICR mice were divided into four groups and subjected to acute NaNO2 exposure by a single intraperitoneal injection of 120 mg/kg body weight. Animals in each group were sacrificed at certain time interval after treatment (1 h, 5 h, 1 day and 2 days). Spleens were excised and processed for atomic absorption spectrometry analysis of Fe, Ca and Zn content. Results At the first hour after treatment, a decrease in Fe and Ca levels was observed. One day following NaNO2 administration, Zn concentration reached its lowest value and Ca levels remained lower, compared to the untreated controls. In contrast, Fe concentration increased on the first and second day after treatment. Conclusion The results of the present study demonstrate that acute NaNO2 intoxication provokes changes in the endogenous levels of Fe, Ca and Zn in mouse spleen. These findings suggest disruption of the ionic balance and impact on the activity of antioxidant defence enzymes.

The spleen is an important organ of the immune system responsible for filtering blood and initiating immune responses to blood-borne antigens. Various blood, lymphoid, and hematopoietic cell types may be isolated from spleen samples for further study of the immune system. Preparing a true single cell suspension of the primary tissue sample will optimize cell separation by avoiding additional cell loss and enabling maximum labeling of the target cells. This protocol describes how to harvest cells from a spleen sample, and prepare a single cell suspension prior to performing cell isolation.

Acanthopanax senticosus is an herb that has been used as a traditional remedy and medicine source. Its anti-inflammatory and, anti-oxidative effects have been reported in previous studies. This study aimed to investigate the effect of Acanthopanax senticosus water extracts on mouse macrophage cell in vitro. Mouse splenocyte proliferation increased after application of Acanthopanax senticosus water extract supplement of 5, 10, 50, 100, 250, 500, 1,000 μg/mL after 48 h pre-treatment with a mitogen (ConA or LPS). The production of cytokines secreted by LPS and non LPS stimulated macrophages was detected by ELISA assay using a cytokine kit. Cytokine production (IL-2, IFN-γ, and TNF-α) increased after water extract supplementation. The result of this in vitro study, showed that splenocyte proliferation and cytokine production by activated peritoneal macrophages were increased after Acanthopanax senticosus water extract in the range of 500~1,000 μL/mL. Thus, it is suggested that supplementation with Acanthopanax senticosus water extracts may enhance immune function by regulating splenocyte proliferation and enhancing cytokine production by activated macrophage.Key words: splenocyte proliferation, Acanthopanax senticosus, macrophage, cytokine, immune

Our screening project, namely, search for new immunomodulatory constituents from Ascomycetous fungi, was guided by the effects on mitogen-induced proliferations of mouse spleen lymphocytes. On the project, the defatted crude extracts from Gelasinospora multiforis, G. heterospora, G. longispora, G. kobi, Diplogelasinospora grovesii, Emericella aurantio-brunnea, Eupenicillium crustaceum, etc., submitted to the solvent partition followed by fractionation with repeated chromatography monitored by immunomodulatory activity to afford many active constituents, of which molecular structures including absolute configurations and immunomodulatory activities were elucidated. All of these immunomodulatory constituents isolated on the project were practically not immunostimulants but immunosuppressants.

Background and aims Skin injury is the second most common clinical manifestation in patients with systemic lupus erythematosus (SLE). Oestrogen may affect the onset and development of SLE. This study was undertaken to elucidate the role of oestrogen in the development of SLE skin injury. Methods We investigated the role of oestrogen and its membrane receptor GPER1 in SLE-related skin injury in mice treated with SLE serum in vivo, and monocytes from mouse spleen in vitro. Results We found that skin injury induced by SLE serum was more severe in female mice and required monocytes. E2 promoted these effects through the membrane receptor GPER1 located in lipid rafts and that inhibition of lipid rafts and GPER1 suppressed SLE serum-induced skin inflammation and expression of inflammatory molecules. Conclusions We conclude that oestrogen promotes the development of skin injury induced by SLE serum through the membrane receptor GPER1 and that lipid rafts play an important role in the regulatory effect of GPER1 in SLE skin inflammation.

In this study, we revealed that a Mekabu (Udaria pinnantifida) extract enhanced immunoglobulin (Ig) production of mouse spleen lymphocytes. Furthermore, it was suggested that water-soluble and high molecular weight ingredients in the Mekabu extract have significant enhancing effect on Ig production. Therefore, fucoidan was estimated as the active component.

In the experiment, a systematic study on the genotoxicity of organic extracts of the water in a reservoir at wet season and dry season. And the study was done by using comet assay. The study was to appraise the waters qualities after every treatment step from the view of the DNA damages on the mouse spleen lymphocytes. The results of the experiment can provide reliable basis for adjusting the treatment reasonably to purify raw water of different qualities.

We observed degradation of β-actin mRNA and 18S rRNA in mouse spleen cells under constant temperature conditions in the different temperature group during postmortem intervals (PMIs) of 0–72 h. Thirty-nine mice were sacrificed by cervical dislocation and kept at constant temperatures of 10°C, 15°C, 20°C, 25°C, and 30°C. From 0 to 72 h after death, total RNA in spleen cells was extracted every 6 h. The cycle threshold (Ct) values of β-actin mRNA and 18S rRNA were obtained by real-time-quantitative polymerase chain reaction. The results showed that, under the conditions of different and constant temperatures after mouse death at 72 h, the Ct values of β-actin and 18S, Ct ratios of β-actin to 18S, and relative ratios of β-actin to 18S were significantly correlated with PMI. In addition, the relative degradation rates of β-actin and 18S appeared to change from fast to slow with the increase of temperature. By interpolation and fitting analysis of the data, we obtained a ternary quintic equation of the relationship between the change in the relative ratios and PMI, which can be used to infer PMI within a certain temperature range (10°C–30°C).

A part of his ‘theory of haematopoiesis’, Alexander Maksimov first proposed the existence of stem cells circa 1908. The importance and role of stem cells remained shrouded until the early 1960s when Becker and colleagues demonstrated self-renewing haematopoietic cells in mouse spleen [1], and Joseph Altman first examined neurogenesis in the mammalian adult brain [2]. In the 1980s, two groups simultaneously reported the ability to i i h t t t a w a a v d r o c d t l i i t s t m m c a u

The present study showed that a hot water extract of the fruiting body of the edible mushroom Boletopsis leucomelas, known as ‘Kurokawa’ Japanese, strongly stimulated IgA-production in mouse spleen cells in our screening experiment. The in vivo study was also conducted with the objective of enhancing adaptive immune response by oral administration of the hot water extract of B. leucomelas (BLE) in lipopolysaccharide (LPS)-challenged mice. The mice were fed a standard diet with or without 0.16% BLE. The mice were also orally administered sodium phosphate buffer or LPS weekly at days 7, 14 and 21. Results indicated that LPS-specific serum IgG, IgM and IgA were increased in the BLE diet group compared to the standard diet group. Interestingly, intestinal total IgA and LPS-specific IgA were significantly increased in the BLE diet group. Moreover, the

Thymosin, composed of many peptides, is distributed in animal and has immunomodulating effects. However, many factors could affect its bioactivity. Chitosan microspheres containing pig thymosin (CMPT) were prepared through W/O emulsion cross-linking method. In this study, thymosin activity between pig-thymosin and CMPT was investigated through E-rosette test and mouse spleen lymphocyte proliferation (MPLP) experiment in vitro. Results showed that CMPT has the same immunomodulatory activity as pig thymosin. When pig-thymosin was 1.5 μg / ml, E-rosette binding rate was 63%, and E-rosette binding rate of CMPT reached 60%。MPLP rate was 73.3% and 68.5%, respectively. CMPT have the capacity to keep the E-rosette binding and MPLP activity of pig thymosin. This may help to improve traditional formulation and bioavailability of thymosin, and expand its scope of application in clinic.

The distribution of triterpene glycoside cucumarioside A₂-2, the main compound of medical lead Cumaside in immunodeficiency diseases, in mouse spleen was determined. For this purpose the stability and dynamics of glycoside content changes over time in Balb/c mouse spleen tissue homogenate as well as the study of the cucumarioside A2-2 spatial distribution in tissue sections were investigated using radiospectroscopy, MALDI-MS and MALDI Imaging Mass Spectrometry (IMS), correspondingly. Cucumarioside A₂-2 is reliably detected by MALDI-MS in the mouse spleen tissue after single intraperitoneal (i.p.) injection at a dosage of 5 mg/kg. The glycoside is stable in the spleen and does not undergo metabolic transformation in either tissue homogenates or in the intact organ within 24 h after i.p. injection. The cucumarioside A₂-2 was absorbed fairly rapidly: the glycoside maximum concentration (Cmax) in tissue homogenate was observed in the first 30 min after injection; the minimum values were registered in 3 h. These results are in agreement with those obtained in the pharmacokinetic study of (3)H-cucumarioside A₂-2. It was established by MALDI-IMS that glycoside was mainly located in the tunica serosa part of the spleen and only a small amount was detected within the red and white pulp of the organ. MALDI MS images obtained 15-30 min post dosage clearly reflect high drug concentrations in the regions surrounding the organ followed by its decline in the surface part and a very slight redistribution to the internal part of the spleen.

생리활성물질을 다량함유하고 있는 마늘의 발효산물인 흑마늘의 면역활성을 검증하기 위하여 C57BL6 마우스 비장세포를 이용하여 흑마늘이 비장세포의 활성화에 미치는 영향을 확인하였다. 흑마늘 추출물은 시판되는 남해 흑마늘 액기스를 농축하여 사용하였다. 그 결과 IL-2에서 흑마늘 추출물만 처리한 군에서 생성이 증가하였으며, LPS와 흑마늘 추출물을 함께 처리하였을 때 IL-2와 TNF- ${\alpha}$ , IFN- ${\gamma}$ 의 생성이 LPS만 처리한 군보다 증가하여 대식세포나 T림프구의 발현에 의해 일어나는 세포성 매개 면역을 활성화를 유도하는 Th1 세포의 발현을 활성화 하였다. 그리고 IL-6는 흑마늘 추출물만 처리하였을 때 후기생성이 증가하였으며, LPS와 흑마늘 추출물을 함께 처리한 경우 LPS만 처리한 군보다 IL-4와 IL-6의 생성이 증가하였다. IL-10은 LPS와 흑마늘 추출물을 함께 처리하였을 때 후기 생성이 감소하였는데, 이는 B 림프구의 활성화에 따른 항체생성 면역을 활성화하며 Th1 세포로부터 유도되는 세포성 면역반응을 억제함으로서 항체유도 체액성 면역반응으로 전환을 효과적으로 조절하는 것을 확인하였다. 따라서 흑마늘 추출물은 마우스 비장세포에서 T 림프구의 활성화에 따른 Th1 세포와 Th2 세포가 활성화되어 면역계의 세포성 면역과 체액성 면역반응을 활성화하여 면역조절에 효과를 나타내는 것으로 사료된다. 【The effect of black garlic extract on the activation of spleen cells from a C57BL6 mouse was investigated to examine immune activities of of fermented black garlic containing a variety of bioactive substances. xtract obtained from the concentration of commercial Namhae black garlic was used for the analysis of immune activities. Treatment with the extract increased the expression of interleukin-2 (IL-2) cytokine. The simultaneous administration of the extract plus lipopolysaccharide (LPS) increased the expression of IL-2, tumor necrosis factor (TNF)- ${\alpha}$ , and interferon (IFN)- ${\gamma}$ compared with that of a control group. This result suggests that cellular immunity can be induced by macrophages, resulting in the expression of T lymphocytes and T helper type 1 (Th1) cells. In addition, treatment with the extract increased the late response of IL-6 cytokines, and the extract plus LPS augmented the expression of IL-4 and IL-6 compared with that of an LPS-treated group. Meanwhile, the extract plus LPS decreased the late response of IL-10, suggesting that humoral immunity can be activated by stimulating B lymphocytes, suppressing cellular immunity, and effectively modulating the conversion into humoral immune responses. These findings demonstrate that the black garlic extract activates Th1 and Th2 cells by stimulating T lymphocytes in mouse spleen cells and leads to immunomodulation by activating cellular and humoral immune responses of the immune system.】

This study was conducted to assess the in vivo and ex vivo immunomodulatory effect of the ethanol leaves extract of Moringa peregrina in Balb/c mice. For this study, five groups of 5 Balb/c mice were given a single acute subtoxic oral dose of the ethanolic extract at 1.13, 11.30, 23.40 and 113.4 mg/kg and the immunomodulatory effect was assessed on the 6th day following the ingestion. In the (non-functional) assessment, the effect of the extract on the body weight, relative lymphoid organ weight, splenic cellularity and peripheral blood hematologic parameters were evaluated. While in the immunomodulation assessment (functional), we investigated the effect of the extract on the proliferative capacity of splenic lymphocytes and peripheral T and B lymphocytes using mitogen blastogenesis, mixed allogeneic MLR and IgM-Plaque forming cells assays. The ingestion of M. peregrina extract caused a significant increase in the body weight, weight and number of cells of spleen and lymph nodes of the treated mice. Furthermore, the count of RBCs, WBCs, platelets, hemoglobin concentration and PCV % were increased by the extract treatment in a dose-dependent manner. M. peregrina enhanced the proliferative responses of splenic lymphocytes for both T cell and B-cell mitogens. Likewise, the mixed lymphocyte reaction MLR assay has revealed a T-cell dependent proliferation enhancement in the extract treated mice. Moreover, the oral administration of M. peregrina leaves extracts significantly increased PFCs/106 splenocytes in a dose-dependent manner. In conclusion, subtoxic acute doses of M. peregrina extract demonstrated significant potential as an immunomodulatory agent even at the lowest dose of 1.13 mg/kg.

Consider, for a moment, the contrasting approaches to the value of the humanities evident in these two vignettes.

Monoclonal antibodies offer high specificity and this makes it an important tool for molecular biology, biochemistry and medicine. Typically, monoclonal antibodies are generated by fusing mouse spleen cells that have been immunized with the desired antigen with myeloma cells to create immortalized hybridomas. Here, we describe the generation of monoclonal antibodies that are specific to Chikungunya virus using ClonaCell-HY system.

Dendritic cells (DCs) were discovered in 1973 by Ralph Steinman as a previously undefined cell type in the mouse spleen and are now recognized as a group of related cell populations that induce and regulate adaptive immune responses. Studies of the past decade show that, both in mice and humans, DCs are composed of subsets that differ in their localization, phenotype, and functions. These progresses in our understanding of DC biology provide a new framework for improving human health. In this review, we discuss human DC subsets in the context of their medical applications, with a particular focus on DC targeting.

To utilize ultraviolet (UV) irradiation as a means to prepare hypo-allergenic food, we investigated the molecular weight profile, secondary structure content, fluorescence spectrum, and immunoglobulin (Ig) production-regulating activity of β-lactoglobulin after UV-irradiation. UV-irradiation caused a change on the molecular size distribution, disruption of the ordered structure, and decrease of emission intensity of β-lactoglobulin. The alteration on Igs production regulating activity of β-lactoglobulin by UV-irradiation was observed in the mouse spleen lymphocytes system. These results suggest that UV-irradiation is effective for alteration of molecular properties and antigenicities of β-lactoglobulin and such treatment should be useful for the preparation of low allergenic foods.

Zusammenfassung Wir untersuchten die biologischen Eigenschaften der verschiedenen Bordetella bronchiseptica-Stamme, die aus Schweinebestanden mit unterschiedlichem Krankheitsstatus (Atemwegserkrankungen) isoliert wurden. Unser Ziel war, das Exotoxinbildungsvermogen nachzuweisen, sowie die Exotoxin-bildung als biologischer Marker fur Bordetella bronchiseptica-Stamme und daruber hinaus die Eigenschaften des Toxins naher zu bestimmen. Aus den Ergebnissen sind folgende Schlusfolgerungen zu ziehen: 1. Exotoxinbildung und Virulenzgrad sind eng miteinander verbunden. Mit zunehmender Virulenz steigt auch die Exotoxinbildung an. 2. Subletale Dosen des reinen Exotoxins und toxischer Stamme bei i. v. Applikation rufen nach 7 Tagen eine schwere Atrophie der lymphatischen Organe hervor, was sich in einer Verkleinerung der Malpighischen Korperchen der Milz ausert. Gleichzeitig wird die B-Lymphozytenzahl im Blut verringert. Demgegenuber bewirken die nicht exotoxinbildende Stamme eine auffallige Hyperplasie der lymphatischen Organe und parallel dazu einen Anstieg der B-Lymphozyten. 3. In Anbetracht des engen Zusammenhanges zwischen Toxinbildung und Virulenz ist die Reaktion der Milz zugleich ein Ausdruck fur die exotoxinbildende Eigenschaft von B. bronchiseptica, aber zugleich auch fur den Virulenzgrad der B. bronchiseptica-Stamme. Deshalb kann die im Mause-Milztest nachgewiesene Exotoxinmenge als ein biologischer Marker der B. bronchiseptica-Stamme angesehen werden, wobei zu berucksichtigen ist, das auch geringe Unterschiede in der Exotoxinbildung der Stamme mit diesem Test exakt festgestellt werden konnen. Summary Epizootiological studies on porcine atrophic rhinitis. IV. Influence of heat-labile toxin of Bordetella bronchiseptica strains on lymphatic organs of mice The biological properties of various Bordetella bronchiseptica strains isolated from pig herds of varying disease status (respiratory disease) were studied. The objective was to demonstrate ability to produce exotoxin and also to study exotoxin formation as a biological marker for B. bronchiseptica strains and also to study the properties of the toxin. The results gave the following conclusions: 1. Exotoxin formation and degree of virulence were closely related. Increased virulence was associated with increased production of toxin. 2. Sublethal doses of pure exotoxin and toxic strains given i. v. produced after 7 days a severe atrophy of the lymphatic organs as shown by smaller spleen Malpighian bodies. At the same time there was reduction of the number of B-lymphocytes in the blood. In contrast, strains that did not produce exotoxin caused an increase in size of lymphoid organs and a parallel rise in B-lymphocytes. 3. In connection with this close relationship between toxin production and virulence, the reaction of the spleen is both an expression of the exotoxin-forming character of B. bronchiseptica and also of the grade of virulence of the organism. The mouse spleen test as an indicator of exotoxin amount can thus be used as a biological marker for B. bronchiseptica strains, bearing in mind that even small differences in toxin formation of strains can be precisely determined by this test. Resume Recherches epizootologiques sur a rhinite atrophique du porc IV. Influence d'une toxine thermolabile de souches de Bordetella bronchiseptica sur des organes lymphatiques de souris On a recherche les proprietes biologiques de differentes souches de Bordetella bronchiseptica, isolees dans des exploitations porcines au statuts de sante differents (maladies de l'appareil respiratoire). Le but fut de mettre en evidence la possibilite de formation d'exotoxine et de determiner cette formation comme marqueur biologique pour des souches de Bordetella bronchiseptica et de definir en plus les proprietes de la toxine. Les conclusions suivantes ont ete titrees a partir des resultats: 1. La formation d'exotoxine et le degre de virulence sont etroitement lies. La formation d'exotoxine augmente avec la virulence. 2. Des doses subletales de l'exotoxine pure et de souches toxiques par application i/v provoquerent une grave atrophie des organes lymphatiques apres 7 jours qui s'est manifestee par une atrophie des corpuscules de Malpighi dans la rate. Le nombre des lymphocytes B a en meme temps diminue dans le sang. Les souches ne formant pas de toxine ont provoque par contre une hyperplasie manifeste des organes lymphatiques et une augmentation parallele des lymphocytes B. 3. En considerant l'etroite correlation entre la formation de toxine et la virulence, la reaction de la rate et l'expression en meme temps de la propriete de formation d'exotoxine de B. bronchiseptica et du degre de virulence des souches de B. bronchiseptica. Le test de la rate chez des souris pour la mise en evidence quantitative d'exotoxine peut donc etre considere comme un marqueur biologique des souches de B. bronchiseptica, tout en considerant que de faibles differences dans la formation d'exotoxine des souches peuvent etre mises exactement en evidence avec ce test. Resumen Estudios epizootologicos sobre la rinitis atrofica del cerdo IV. El influjo de la toxina termolabil de las estirpes Bordetella bronchiseptica sobre los organos linfaticos en los ratones Examinamos las propiedades biologicas de diversas estirpes de Bordetella bronchiseptica, aisladas de efectivos porcinos con estado desigual de salud (enfermedades de las vias respiratorias). El fin que nos proponiamos era poner en evidencia la facultad de formacion de exotoxina como marcador biologico para las estirpes Bordetella bronchiseptica y definir mas alla, en detalle, las propiedades de la toxina. De los resultados obtenidos se sacan las conclusiones siguientes: 1. La exotoxinogenia y el grado de virulencia se hallan relacionados estrechamente entre si. Con la virulencia creciente tambien aumenta la formacion de exotoxina. 2. Dosis subletales de exotoxina pura y de cepas toxicas provocan tras la aplicacion iv., al cabo de 7 dias, una atrofia grave de los organos linfaticos, lo cual se manifiesta por una achicadura de los corpusculos de Malpigio en el bazo. Al mismo tiempo disminuye el numero de linfocitos B en sangre. Por el contrario, las estirpes que no son exotoxinogenas causan una hiperplasia ***llamativa des los organos linfaticos y el aumento paralelo de linfocitos B. 3. En consideracion a la relacion estrecha entre la toxinogenia y la virulencia, la reaccion del bazo es al mismo tiempo la expresion de la propiedad exotoxinogena de B. bronchiseptica y un indicio del grado de virulencia de las estirpes de B.- bronchiseptica. Por lo tanto, la cantidad de exotoxina puesta en evidencia en la prueba del bazo de raton puedo considerarse como un marcador biologico de las cepas de B. bronchiseptica. Con esta prueba mencionada tambien se pueden poner en evidencia, con toda exactitud, diferencias incluso escasas en la exotoxinogenia de las estirpes.

Objective The aim of the study was to investigate the adaptive response of low-dose radiation-induced apoptosis in mouse spleen cells and its related proteins control mechanism.

Summary Through electron microscopic observations of the normal mouse spleen, the following findings were obtained concerning denucleation mechanism of the erythroblast: 1. In mice the nuclei of the erythroblasts are denucleated by phagocytic activity of the reticular cell, located in the center of the erythroblastic islets. In a physiologic state, this mechanism is considered to be playing the main role in denucleation. 2. The protoplasmic projection of the reticular cell compresses and makes an infolding between the head and body of the orthochromatic erythroblast, and finally the head is cut off and phagocytized by this reticular cell, leaving the body to develop into a reticulocyte.

Yield of polysaccharides from Phascolosoma esulenta obtained by phosphate buffer extraction through an orthogonal experiment (L9(3)(4)) were investigated to get the best extraction conditions. The results showed that extraction temperature, ratio of phosphate buffer to raw material, extraction time, and ratio of trypsinase to raw material were the main four variables that influenced the yields of extracts. The highest yield was obtained when extraction temperature, ratio of phosphate buffer to raw material, extraction time and ratio of trypsinase to raw material were 40°C, 2, 5.5h and 1.6, respectively. The immunity-stimulating method showed that polysaccharides from P. esulenta could significantly raise liver, spleen and thymus index of mice and enhance Con A-stimulated mouse spleen cells proliferation. These results indicate that polysaccharides from P. esulenta had significantly higher immunity-stimulating activities.

Using comet assay, a statistically significant increase (p < 0.05) in the level of DNA breaks in spleen cells was revealed in male CBA/lac mice exposed to gamma-radiation (1.7 cGy/day) or 90Sr (150-250 Bq/day) for 210 days. The level of DNA breaks also increased under combined exposure to both gamma-radiation and 90Sr (p < 0.05), but to a lesser degree than under exposure to each of these factors alone. Upon additional in vitro treatment of spleen cells with hydrogen peroxide, the relative increase in the level of DNA breaks was smaller in cells of irradiated mice than in the control. The ratio of the level of DNA breaks after hydrogen peroxide treatment to that before this treatment in control mice was 4.2 +/- 0.9, compared to 1.4 +/- 0.6 in gamma-irradiated mice, 1.9 +/- 0.8 in 90Sr-irradiated mice, and 2.3 +/- 0.8 in mice exposed to both gamma- and 90Sr-irradiation.

Mouse spleen lymphocytes treated with rabbit IgG anti-sheep erythrocytes (SRBC) complexed with protein A of Staphylococcus aureus (SpA) form rosettes with SRBC. The attachment of SRBC to lymphocytes was due to the binding of the SpA-IgG antibody complex to the surface of the lymphocytes and was thus considered "arming" of the cells. Normal mouse spleen cells "armed" with SpA-rabbit IgG anti-chicken erythrocytes (CRBC) kill specifically 51Cr-labeled CRBC "in vitro" in the absence of free antibodies. The killing by these "armed" cells is an effect of the cell-bound SpA-IgG antibody complex. Both the SRBC rosette formation and the cell-mediated CRBC killing was dependent on the concentration of the SpA-IgG antibody complexes used for "arming" the cells. A 100-fold increase in rosette formation or in killing of target cells was recorded for lymphocytes treated with SpA-IgG antibody complexes in comparison with cells treated with noncomplexed IgG antibodies. The specific binding of SpA-IgG antibody complexes to the Fc receptors of mouse spleen cells was demonstrated by inhibition studies. More than 60% inhibition of the rosette formation and in the killing of target cells was shown for cells treated with normal rabbit IgG or its Fc fragment before addition of the SpA-IgG antibody complex.

This work introduces minimum accumulative degeneracy, a variant of the degenerate primer design problem, which is particularly useful when a large number of sequences are to be covered by a set of restricted number of primers. A primer set, which is designed on a minimum accumulative degeneracy basis, especially helps to reduce nonspecific PCR amplification of undesired DNA fragments, as fewer primer species are present in PCR. A Boltzmann machine is designed to solve the minimum accumulative degeneracy degenerate primer design problem, called the MAD-DPD Boltzmann machine. This algorithm shows great flexibility, as it can be determined either to solve the problem with strict fidelity to covering all input sequences or to exclude some input sequences if it results in less degenerate primers. This Boltzmann machine is successfully implemented in designing a new set of primers for amplification of antibody variable fragments from mouse spleen cells, which theoretically covers more diverse antibody sequences than currently available primers. The MAD-DPD Boltzmann machine is available online at bioinf.cs.ipm.ir/download/MAD_DPD08172007.zip.

In this study, we compared with 80% ethanol extracts from peel of Poncirus trifoliata (PTP) and peel of Citrus junos (CJP) against antioxidant and immune activities. Total phenolics and flavonoids contents in PTP extracts were 60.75±1.15 and 33.75±0.15 ㎎/100 g, respectively, and those were lower than CJP extracts. Antioxidant activities of PTP were increased with the more concentration, and were similar to CJP. Antioxidant activities of PTP were increased with increasing of concentration, and were similar to those of CJP. The NO production in macrophage cell lines were increased in a dose-dependent manner, until 5 ㎎/ml of CJP and 1 ㎎/ml of PTP compared with control cells, but decreased at higher concentrations. The proliferation of mouse spleen cells were increased in a dose-dependent manner, until 1 ㎎/ml of CJP and PTP compared with control cells but decreased at higher concentrations. The NO production in macrophage cell lines treated with PTP and CJP were increased in a dose-dependent manner, compared with untreated control cells until the concentrations of 1~5 ㎎/ml (CJP) and 1 ㎎/ml (PTP) but decreased at higher concentrations than that. The proliferation of mouse spleen cells treated with PTP and CJP were increased in a dose-dependent manner, compared with untreated control cells until the concentration of 1 mg/ml but decreased at higher concentrations than that.

To determine whether interferon gamma (IFN-gamma) can be used as a biomarker of exposure to viral pathogens, 12-week-old BALB/c mice were injected intraperitoneally with coxsackievirus B3 (CVB3) or coxsackievirus B4 (CVB4) diluted in sterilized phosphate-buffered saline (PBS). Control mice were injected with PBS only. Four months after viral infection, mouse spleen cells were harvested and assayed for the release of IFN-gamma by memory T cells after in vitro stimulation with viral antigens, phytohemagglutinin (PHA), and PBS, respectively. The level of IFN-gamma was examined by an antibody-capture enzyme-linked immunosorbent assay (ELISA). A marked increase in the level of IFN-gamma was observed when memory T cells from CVB3-infected mice were incubated with CVB3 virus, but not with CVB4 or PBS. Conversely, memory T cells from mice infected by CVB4 were not stimulated to produce IFN-gamma when they were incubated with CVB3 and PBS, but did significantly produce IFN-gamma when stimulated with CVB4. T cells from mice injected with PBS did not release IFN-gamma after stimulation with CVB3 or CVB4. However, these T cells did release IFN-gamma after stimulation with PHA. Our results demonstrated that IFN-gamma produced by memory T cells is virus-specific and may have use as a biomarker in viral exposure studies. The results of this study may be extended to the study of infection by pathogens that are capable of inducing cell-mediated immune response in humans.

Aim: To study the relationship between primary structures of oligodeoxynucleo-tides (ODN) containing unmethylated deoxycytidyl-deoxyguanosine (CpG) dinucleotide motifs and their immunostimulatory activities in mouse spleen cells. Methods: A series of CpG ODN with different primary structures were synthesized. Their capabilities to stimulate mouse spleen cell proliferation were determined by [3H]thymidine incorporation assay. Cytokine (interleukin [IL]-6, IL-12, and IFN-α) secretion spectra induced by CpG ODN were assessed by ELISA. The ability of CpG ODN to activate natural killer cells was evaluated by standard 4 h 51Cr-release assay. Flow cytometry was utilized to examine the expressions of various lymphocyte surface molecules on diverse immunocytes. An effective CpG ODN for murine, ODN1826, was set as the template of modification and the positive control. Results: The immunostimulatory activities of CpG ODN with different sequences and compositions varied markedly, both in character and in extent. It was useless for improving the immunostimulatory activity of ODN1826 by simply increasing the functional hexameric CpG motif number, modifying the site of CpG motifs, or changing the distance between multi-CpG motifs. However, an addition of a self-complementary palindrome structure at the 3′-end, but not the 5′-end of CpG ODN, aroused marked improvement in its activity. Several designed ODN had superior comprehensive immunostimulatory properties compared to ODN1826. Conclusion: The immunostimulatory activity of a CpG ODN was relevant to its primary structure. It was useless for promoting immunostimulatory activity to simply change CpG motif number, space, or distance. The 3′-end palindrome structure of CpG ODN is associated with enhanced immunostimulatory activity.

Quantitative features of the proteome are extremely useful for studying cellular processes at a molecular level. In this study, we attempted to construct quantitative reference proteome maps of the mouse spleen and lymph node based on 2-DE followed by protein identification using MS. We analyzed more than 1000 spots on the 2-DE images and consequently were able to determine that 919 spots were derived from 328 different genes. To obtain statistically reliable information of the protein levels from these 2-DE images, we measured the volumes of the respective spots on 2-DE images obtained by four to six independent experimental runs. These measurements were used to calculate the variability of the volumes of the respective spots on 2-DE following subcellular fractionation, which enabled us to discriminate differentially produced proteins from those within the range of intrinsic variability. More importantly, while the 2-DE data have been traditionally collected in a gel image-based manner, the resultant quantitative 2-DE data could be analyzed using the same procedure as that for mRNA expression profiles. This greatly assists in bridging the gap between the analyses of transcriptomes and proteomes and enables the integration of this data on the same informational platform.

A total number of 31 types of seaweed were assessed with regard to their effects on the proliferation of mouse spleen and thymus cells in a culture, using an MTT reduction assay. Acetone:dichloromethane (1∶1) extracts of three seaweed plants:Derbesia marina, Sargassum sp., andHisikia fuziformis, exhibited significantly positive effects on the survival of mouse spleen and thymus cellsin vitro. The acetone: dichloromethane (1∶1) extracts ofSargassum sp., in particular, much more potent effects on thymus cell activation than did any of the other types of seaweed. However, the methanol extracts ofSargassum ringgoldianium andChondrus crispus exerted a stimulatory influence only on the proliferation of mouse spleen cells, whereas the methanol extracts ofGrateloupia lanceolata exhibited significant cell proliferation properties in both spleen and thymus cells.

To develop a new functional food material, edible crude saponin was isolated from soybean cake using HP-20 resin and its anticancer effect and immune-activity were investigated. The saponin significantly inhibited the growth of cancer cells such as A549, MCF-7 and SW480 at a concentration of 1,000 g/mL. Morphological changes was observed in the AS49 cells surface treated with the saponin of 1,000 g/mL concentration. The proliferation of mouse spleen cells treated with saponin was increased in a dose-dependent manner compared with untreated control cells until the concentration of 1 g/mL but decreased at higher concentrations than that. The NO production in marcrophage cell lines (RAW 264.7) treated with saponin was increased in a dosedependent manner compared with untreated control cells.

Theophylline (TH) is a methylxanthine widely used in clinical practice. The genotoxic effects of TH was investigated in mouse spleen, and spermatogonia cells. Also damage of DNA and molecular detection of apoptosis were applied. TH was administered orally as single dose of 25, 50, 75 and 120 mg kg−1 b.wt. and a multiple treatment with a daily dose of 25, 50 and 75 mg kg−1 b.wt. for 3 and 7 successive days. TH induced a significant increase in the percentage of chromosomal aberrations in somatic and germ cells which was dose and time dependent.Also, TH induced DNA damage and apoptosis in a dose and time dependent manner in mouse liver cells. However, induced DNA damage and apoptosis was maximally detected at a concentration of 120 mg TH kg−1 b.wt. after oral treatment for 1 d.

Group X secretory phospholipase A2 (sPLA2-X) and cytosolic phospholipase A2 alpha (cPLA2alpha) are involved in the release of arachidonic acid (AA) from membrane phospholipids linked to the eicosanoid production in various pathological states. Recent studies have indicated the presence of various types of cross-talk between sPLA2s and cPLA2alpha resulting in effective AA release. Here we examined the dependence of sPLA2-X-induced potent AA release on the cPLA2alpha activation by using specific cPLA2alpha or sPLA2 inhibitors as well as cPLA2alpha-deficient mice. We found that Pyrrophenone, a cPLA2alpha-specific inhibitor, did not suppress the sPLA2-X-induced potent AA release and prostaglandin E2 formation in mouse spleen cells. Furthermore, the amount of AA released by sPLA2-X from spleen cells was not significantly altered by cPLA2alpha deficiency. These results suggest that sPLA2-X induces potent AA release without activation of cPLA2a, which might be relevant to eicosanoid production in some pathological states where cPLA2a is not activated.