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Expression of Friend leukemia virus and spleen focus-forming virus-specific sequences in erythroid bursts and granulocyte-macrophage colonies from spleen and marrow of mice infected with Friend leukemia virus.

A large number of studies have been carried out to identify the Friend leukemia virus (FV) target cell(s). In FV-infected mice, the kinetics of "primitive" erythroid burst-forming units (P-BFU-E) is perturbed, and their proliferative rate is enhanced. These results indirectly suggest, but do not prove, that cycling P-BFU-E may serve as FV target. In vitro infection studies showed that normal erythroid colony forming units (CFU-E) and "mature" erythroid burst-forming units (M-BFU-E) are targets for FV, while the largely out-of-cycle normal P-BFU-E are not. In an attempt to shed light on these aspects, we have evaluated the expression of viral cytoplasmic RNA sequences in pools of colonies generated by P-BFU-E and granulocyte-macrophage colony forming units (CFU-GM) from spleen and marrow of polycythemic Friend virus (FVP)-infected mice, as measured by liquid hybridization with FVP- or spleen focus-forming polycythemic virus (SFFVp)-specific DNA probes. Moreover, similar assays were performed on RNAs derived from whole spleen or bone marrow from mice treated with FVP or the anemic strain of Friend virus (FVA). Control studies were performed on corresponding colonies and whole tissues from normal animals. FVP- and SFFVp-specific sequences are more abundant in RNA extracted from infected spleen as compared to marrow by a 10-fold factor. On the other hand, FVP and SFFVp-specific sequences are expressed at a comparable level in both P-BFU-E- and CFU-GM-derived colonies from spleen or marrow of FVP-treated mice. Since in vitro spread of FVP infection was excluded by control studies with addition in culture of antibody to the viral glycoprotein with a molecular weight of 70,000 (gp70) these results indicate that P-BFU-E and CFU-GM are infected in vivo by FVP.

Animals↗

[Correlation of spleen colony formation and changes in spleen weight during its recovery in irradiated mice].

In experiments on (CBA X C57Bl)F1 and BALB/c mice it was shown that with the spleen mass increasing, due to exo- and endorepopulation, during the postirradiation regeneration of the spleen, the number of splenic colonies also increases to a certain level. When the spleen mass reaches 1/3 of that lost after irradiation the number of discrete colonies decreases. When the spleen mass is restored to about 1/2 solitary colonies do not form at all. The spleen mass can considerably exceed the normal level in conditions of intensive repopulation.

Animals↗

Immunologic tolerance to HGG in mice. I. Suppression of the HGG response in normal mice with spleen cells or a spleen cell lysate from tolerant mice.

Adoptive transfer of spleen cells or spleen cell lysates from mice tolerant to human-gamma-globulin (HGG) specifically suppressed the response of normal syngeneic recipients to HGG. The suppressive activity could be transferred for over 100 days after tolerance induction. The suppression induced by both spleen cells and spleen cell lysate was found to be specific as evidenced by a normal response to a challenge with turkey-gamma-globulin or goat erythrocytes. The activity of the suppressive lysate could be removed by passing the material through an HGG immunoadsorbent column but not by passing it through an anti-HGG column or a BSA column. These results indicated that the factor had antigen specificity and was probably not antigen-antibody complexes. That this suppression was not due to a shifting of the kinetics of the antibody response has also been demonstrated. The antigen-specific suppressor factor in the tolerant spleen cell lysates was a protein with a m.w. of approximately 45,000 daltons. The kinetics of the appearance of both suppressor cells and suppressor factor were consistent with a mechanism of active suppression functioning in the maintenance of tolerance to HGG.

Animals↗

The effect of spleen volume on liver regeneration after hepatectomy--a clinical study of liver and spleen volumes by computed tomography.

BACKGROUND/AIMS: The relationship between liver regeneration and spleen volume after partial hepatectomy was studied with computed tomography (CT) in 13 patients with chronic liver disease (chronic active hepatitis: 2, pre-liver cirrhosis: 3, liver cirrhosis: 8). MATERIALS AND METHODS: Liver and spleen volumes were measured by abdominal CT before hepatectomy and 1 and 2 years after operation, and the percent volumes of the liver and spleen of abdominal capacity were compared. RESULTS: While the residual liver volume increased in all patients after 1 year, it continued to increase in 7 patients (percent recovery: 102 +/- 4.8%) but had decreased in 5 patients (90.0 +/- 4.8%) after 2 years. The mean spleen volume 2 years after hepatectomy was 2.8 +/- 0.9% in the former, and 6.0 +/- 1.2% in the latter. The percent increase in liver volume was inversely related to the spleen volume (r = -0.707). In a case of transcatheter splenic arterial embolization, the liver volume continued to be increase despite splenomegaly. CONCLUSION: Splenic volume is suggested to be related to the process of recovery of the liver.

Carcinoma, Hepatocellular↗

THE REGENERATION OF F1 HOST CELL SPLEEN AND THYMUS AT ECTOPIC SITES IN F1 ANIMALS INDUCED BY IMPLANTATION OF PARENTAL SPLEEN AND THYMUS.

The ability of parental to F(1) spleen grafts to regenerate was equal to that of parental to parental or to F(1) or F(1) spleen grafts. Cytotoxicity analysis of parental to F(1) spleen and thymic grafts indicated that these "regenerated grafts" were composed of F(1) lymphocytes. Regenerated parental to F(1) spleen grafts were incapable of producing runt disease in newborn F(1) animals.

Animals↗

Cells with marrow and spleen repopulating ability and forming spleen colonies on day 16, 12, and 8 are sequentially ordered on the basis of increasing rhodamine 123 retention.

Mouse bone marrow cells (BMC) were subjected to countercurrent centrifugal elutriation and subsequently separated on the basis of light scatter and fluorescence intensity after being labeled with the supravital dye Rhodamine 123 (Rh-123). The sorted cells were then assayed for their in vivo spleen colony-forming ability (day -8, -12, and -16 CFU-S) and their ability to repopulate the bone marrow or spleen over a 13-day period with CFU-S-12, CFU-GM, or nucleated cells. Cells with marrow repopulating ability (MRA), as measured by the ability of the sorted cells to repopulate the marrow with secondary CFU-S-12 or CFU-GM, had low affinity for Rh-123. These cells showed minimal spleen colony-forming ability, and the ratio of MRA to CFU-S-12 in this preparation was 309. Cells with spleen repopulating ability (SRA), CFU-S-16, CFU-S-12, and CFU-S-8 retained increasing amounts of Rh-123, respectively, and CFU-S-8 were almost exclusively found among cells with high Rh-123 affinity. These cells also included about half of all day-12 CFU-S, and the ratio of MRA to day-12 CFU-S was 0. The results show that MRA cells, SRA cells, CFU-S-16, CFU-S-12, and CFU-S-8 can be sequentially ordered on the basis of increasing mitochondrial activity. The data also demonstrate for the first time, and without the application of negative selection by the use of cytostatic agents, that MRA cells are a separate class of primitive hemopoietic stem cells that fully meet the criteria of pre-CFU-S.

Animals↗

TrkB mRNA and protein in mouse spleen: structure of the spleen of functionally deficient TrkB mice.

Whereas it is nowadays clear that neurotrophins are involved in the regulation of various aspects of the functioning of immune system, knowledge of their actual immunomodulatory roles is still fragmentary and incomplete. In this respect, knock-out mouse models remain particularly unexplored. In the present study, the expression of the TrkB neurotrophin receptor in murine spleen was addressed at the mRNA (reverse transcription/polymerase chain reaction) and protein (Western blot) levels. Once the presence of TrkB at both levels was demonstrated, the age-dependent changes in the pattern of expression of the receptor were analyzed and quantified, and TrkB-positive cells were identified by immunohistochemistry. TrkB-immunoreactive cells, identified as red pulp macrophages, were detected in the spleen throughout postnatal development and adult life; their numbers peaked at the age of 15 days. The absence of functional TrkB did not appear to result in morphological changes as assessed by light and electron microscopy of spleens from 15-day-old mice knockout for the trkB gene. The present results support the idea that, in the murine spleen, TrkB and its ligands are involved in macrophage physiology in a developmentally regulated fashion, but they do not seem to be essential for macrophage survival.

Animals↗

Role of spleen or spleen products in the deficiency in morphine-induced analgesia in diabetic mice.

We examined the possibility that the spleen or factor(s) derived from spleen mononuclear cells are involved in the deficient mu-opioid receptor-mediated analgesia encountered in diabetic mice. Splenectomized diabetic mice had a significantly higher sensitivity to morphine analgesia than untreated or sham-operated diabetic mice. Naive recipient mice injected with mononuclear spleen cells from diabetic mice exhibited a lower sensitivity to morphine analgesia than vehicle-treated naive mice. These results suggest that some factor(s) derived from spleen mononuclear cells may play an important, direct or indirect role in the selective reduction in mu-agonist-mediated analgesia in diabetic mice.

Analgesia↗

Purification and characterization of cytoplasmic 5'(3')-nucleotidase from rabbit spleen: characteristic differences of the enzyme from the rat spleen nucleotidase.

1. A 5'(3')-nucleotidase, which preferably hydrolyzed 3'-dTMP and 3'-dUMP, was highly purified from rabbit spleen cytosol. 2. The enzyme also hydrolyzes 3'-UMP, 5'-dUMP and guanine nucleotides, but does not hydrolyze any adenine nucleotides or cytosine nucleotides. 3. The activity is dependent upon Mg2+, Co2+ or Mn2+; the addition of deoxyinosine stimulates the activity, and the pH optimum for the hydrolysis of 3'-dTMP is 7.0. 4. Although the catalytic properties of the enzyme are similar to the 5'(3')-nucleotidase from rat spleen, these nucleotidases differ in their molecular disposition. 5. The charge state of the rabbit enzyme is slightly basic, and the subunit M(r) is about 27 kDa, while the value of the rat enzyme is 26 kDa. 6. Immunochemical experiments with the use of antibodies against the purified nucleotidase indicate that enzymes from rabbit spleen and from rat spleen are composed of different polypeptides.

Animals↗

Purification of a 100 kDa phospholipase A2 from spleen, lung and kidney: antiserum raised to pig spleen phospholipase A2 recognizes a similar form in bovine lung, kidney and platelets, and immunoprecipitates phospholipase A2 activity.

Phospholipase A2 (PLA2) plays a key role in the production of intracellular and extracellular chemical mediators such as arachidonic acid, eicosanoids and platelet-activating factor, which modulate membrane channel activity, signal transduction, are vasoactive and chemotactic, and are implicated in many pathophysiological mechanisms of inflammation and tissue injury. We previously identified, purified and characterized an arachidonic acid-selective cytosolic 100-110 kDa PLA2 from bovine platelets and rat kidney that is activated during cell stimulation. The purification schemes previously published resulted in low yields of enzyme, insufficient for extensive biochemical characterization. We report the purification of a large-molecular-mass (100 kDa) PLA2 from pig spleen, bovine kidney and bovine lung, using a novel large-scale purification scheme. The enzyme was purified to near homogeneity from an acidified extract obtained from 4.8 kg of pig spleen by sequential use of DEAE-cellulose anionic exchange, Butyl-Toyopearl hydrophobic chromatography and DEAE-5PW h.p.l.c., and further purified by non-denaturing PAGE. This purification scheme will permit the preparation of quantities of purified native enzyme sufficient to study its properties and regulation. To generate antiserum against the PLA2 enzyme, the 100 kDa protein was excised and electroeluted from SDS/PAGE gels of the active fractions after DEAE-5PW h.p.l.c., and this was used as antigen. This polyclonal antibody against pig spleen 100 kDa PLA2 protein reacted with 100 kDa bands in preparations partially purified from bovine platelets, kidney and lung as well as pig spleen, and immunoprecipitated PLA2 activity from these sources. The antibody also immunoprecipitated a 100 kDa protein from cytosolic fractions of cultured renal mesangial cells, human erythroleukaemia cells and human monocytic U937 cells. Considerable PLA2 activity was present in the immunoprecipitates. To our knowledge this antibody is unique in its ability to permit measurement of PLA2 activity in the immunoprecipitate itself, and will be a useful tool for the study of the regulation and the activation mechanisms of the native PLA2 enzyme.

Animals↗

The function of the spleen in adults after ligation of the splenic artery of the traumatized spleen in childhood.

BACKGROUND: Ligation of the splenic artery (LSA) has been successfully used as a spleen-saving procedure in rare cases of splenic trauma in children in which management with splenorrhaphy or partial splenectomy alone was not possible. There are no data regarding the long-term effects of the procedure on the functional status of the spleen. The purpose of this study is to present and discuss our clinical and laboratory findings in adults who underwent LSA in childhood. METHODS: Our first 2 patients in whom LSA was done at ages 4 and 2 years in 1977 underwent the following examinations in the year 2000: 1, imaging of the spleen; 2, immunologic studies; and 3, peripheral blood tests. Their ages at reexamination were 27 and 25 years, respectively. RESULTS: Results were as follows: triplex ultrasound revealed normal size and echomorphology; Doppler techniques revealed normal vasculature; 99mTc-Tin colloid scanning revealed normal uptake. Immunoglobulins (IgG1 to IgG4, IgA, IgM, IgE), complement fraction (C3, C4), antibodies response to vaccinations, and peripheral blood tests all had normal results. No Howell-Jolly bodies were found. CONCLUSION: Laboratory investigations in adults with LSA during childhood disclosed undisturbed function of the spleen. LSA can be used as an adjunct to splenorrhaphy in children with rare splenic injuries involving major hilar vessels.

Adult↗

Sequential liver-spleen scanning for documentation of wandering spleen.

An additional case of wandering spleen is reported. A brief review of the etiology of this entity, as well as various findings obtained with nuclear imaging of the spleen, is presented. The authors discuss the relevance of repeated spleen scans to the clinical decision making process. Particular significance is paid to preservation of the spleen to maintain immunologic competence.

Child, Preschool↗

Naturally occurring spleen-associated suppressor activity of the newborn mouse. Requirement for two genetic restrictions in suppression of lethal graft-versus-host disease by newborn spleen cells.

Spleens from newborn mice less than 6-7 days of age are known to contain naturally occurring suppressor cells, which can suppress the immune reactivity of third-party adult cells. In the present study newborn spleen cell populations are shown to possess the potential to inhibit lethal graft-versus-host (GVH) disease in sublethally gamma-irradiated hosts injected with allogeneic adult cells. However, this capacity to suppress GVH disease is controlled by at least two genetic restrictions: (1) the newborn spleen cells and the adult donor cells must be histocompatible at an H-2-linked region apparently telomeric of H-2DL, and (2) the newborn spleen cells must express a strongly stimulating non-H-2 (perhaps M1s) alloantigenic phenotype. Host animals that survive GVH remain chimeric for at least 3-4 weeks but return to the host phenotype by 8-10 weeks. Thus, it appears that in sublethally irradiated hosts the newborn cells suppress donor cell reactivity long enough for the host system to recover from the effects of irradiation.

Aging↗

Growth of Trypanosoma musculi in cultures of murine spleen cells and analysis of the requirement for supportive spleen cells.

Growth of Trypanosoma musculi in vitro has been achieved. The number of parasites increased by more than 1,500-fold in less than 8 days under the most suitable conditions. The rate and magnitude of growth was comparable to that which occurs in inoculated murine hosts. Maximum growth was displayed in cultures composed of RPMI 1640 medium supplemented with fetal calf serum, murine spleen cells, and foreign erythrocytes (sheep). No growth occurred in the absence of spleen cells. The adherent, macrophage-rich population supported parasite growth much better than did the nonadherent population. Parasite growth was excellent in the presence of irradiated spleen cells or of cells from thymectomized, irradiated, bone marrow-reconstituted mice. The important cells appeared to be macrophages. The beneficial effect of sheep erythrocytes probably resulted from preoccupation or stimulation of phagocytes. Soluble substances released by spleen cell cultures promote parasite growth, as was shown by experiments with double-compartment culture vessels. The utility of this culture system for analysis of host immune responses against the trypanosome was demonstrated.

Animals↗

Erythroid colony formation by fetal rat liver and spleen cells in vitro: inhibition by a low relative molecular mass component of fetal spleen.

Liver and spleen hematopoietic cell suspensions from 20-day-old-fetal rats were fractionated on Percoll gradients. A granulocyte-rich splenic fraction inhibited CFUe production by cultures of a CFUe-enriched liver fraction, and by cultures of unfractionated liver and spleen hematopoietic cells. Conditioned medium from the spleen cell fraction contained an inhibitor of relative molecular mass, Mr, 25-35 x 10(3). The sensitivity of spleen cells to the inhibitor varied with the age of the fetus from which they were derived (20-day-old less than 18-and 19-day-old). No such age-dependence was found for liver cells. The inhibitor affects cycling CFUe, blocks the lethal effect of AraC, does not appear to be lineage-specific and its influence can be reversed by washing.

Animals↗

[Ectopic or migrating spleen and supernumerary spleens: detection and identification with 99mTc, 51Cr and 113In radioisotopes].

Congenital malformations of the spleen are rare. We report a patient with an ectopic spleen located in the left iliac fossa, which was excised in order to prevent severe complications such as torsion of the pedicle, rupture or infection. A second patient with idiopathic thrombocytopenic purpura presented with an accessory spleen located retroperitoneally. Removal of this spleen following removal of the normotopic one resulted in cure of purpura. The clinical, radiological and radioisotopic studies used to locate and identify these malformations are described.

Adult↗

Role of the spleen in immunosuppression of gastric cancer: predominance of suppressor precursor and suppressor inducer T cells in the recirculating spleen cells.

In order to analyse the role of the spleen on immunosuppression of gastric cancer, T cell phenotypes in the spleen cells (SC) were investigated by two colour fluorescence flow cytometry, with reference to their suppressor cell activity. Suppressor T cell phenotypes of CD4+2H4+ cells (suppressor/inducer T cells) and CD8+CD11+ (suppressor T cells) were distributed predominantly in SCs from patients with gastric cancer, while they were distributed scarcely in those with liver cirrhosis. Moreover, CD4+2H4+ cells and CD8+CD11+ cells were found predominantly in SCs and splenic vein lymphocytes (SVL) respectively. Among SCs, a significantly higher proportion of CD4+2H4+ cells was found in the recirculating SCs, but fewer were found in the residual SCs. Higher activity of Concanavalin-A induced suppressor cells was found in the former and that of spontaneously activated suppressor cells was found in the latter. These results suggest the suppressor precursor and suppressor/inducer T cells might distribute predominantly in the cells recirculating from the spleen, and that suppressor cells might be matured during the migration from the spleen.

Adult↗