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Anatomy of vasculature of 850 spleen specimens and its application in partial splenectomy.

BACKGROUND: Anatomic knowledge of vasculature of the splenic lobe and segment is of great clinical significance in partial resections and transplantation of the spleen. METHODS: The methods of two-colored corrosion casting, roentgenography, and anatomic dissection were used to evaluate the vasculature of the splenic hilum and intraspleen. On this basis splenic lobectomy and segmentectomy were performed on 42 patients with traumatized spleens. RESULTS: In the observation of 850 spleen specimens the spleen showed a single lobar artery in 7 cases (0.8%), two lobar arteries in 730 cases (86%), three lobar arteries in 104 cases (12.2%), and multiple lobar arteries (i.e., more than three lobar arteries) in 9 cases (1%). In a subgroup of 276 specimens 17%, 53%, 24%, 4%, 1%, and 1% of spleen specimens had three, four, five, six, seven, and eight segmental arteries, respectively. The result from a subgroup of 280 specimens indicated that mean percentages of the existence of the superior and inferior polar arteries and of the coexistence of both polar arteries were 31.3%, 38.8%, and 13.3%, respectively. Relative avascular planes between segments or lobes were seen. Basic steps of splenic lobectomy or segmentectomy include mobilization of the injured spleen, ligation of vessels in the lobe or segment, transection of the splenic parenchyma, and sutures of the cut surface of the remaining spleen. The postoperative courses of all 42 patients undergoing partial splenectomy were uneventful. No postoperative bleeding and necrosis of the remaining spleen or infectious complications were registered. CONCLUSIONS: Anatomically the spleen is defined with two primary lobes (the superior lobe and inferior lobe), one accessory lobe, and three to five segments. This new classification facilitates surgeons to perform partial resections of the spleen and allotransplantation of the hemispleen from a living related donor in human beings.

Adolescent↗

Addition of allogeneic spleen cells causes rejection of intrastriatal embryonic mesencephalic allografts in the rat.

To address the importance of antigen-presenting cells for the survival of intracerebral neural allografts, allogeneic spleen cells were added to the graft tissue before transplantation. Dissociated embryonic, dopamine-rich mesencephalic and adult spleen tissues were prepared from either inbred Lewis or Sprague-Dawley rats. A mixture of neural and spleen cells was sterotaxically transplanted into the right striatum of adult Sprague-Dawley rats. Controls were neural allografts without addition of allogeneic spleen cells and syngeneic neural grafts with or without the addition of syngeneic spleen cells. Six weeks after transplantation, brain sections were processed immunocytochemically for tyrosine hydroxylase, specific for grafted dopamine neurons, and a bank of markers for various components in the immune and inflammatory responses. The neural allografts which were mixed with allogeneic spleen cells were rejected. In these rats, there were high levels of expression of major histocompatibility complex class I and II antigens, intense cellular infiltration including macrophages and activated microglial cells, and a presence of cluster of differentiation 4- and 8-immunoreactive cells in the graft sites. Moreover, there were increased levels of intercellular adhesion molecule-1, tumour necrosis factor-alpha and interleukin-6 in and around the grafts which were undergoing rejection. In contrast, syngeneic neural grafts survived well regardless of whether they were mixed with syngeneic spleen cells or not, and control neural allografts also exhibited unimpaired survival. No significant difference was observed in the number of grafted dopamine neurons among these three latter groups. The levels of expression of the different markers for inflammation and rejection were generally lower in these grafts than in implants of combined allogeneic neural and spleen cells. In summary, intrastriatal neural allografts, which normally survive well in our animal model, were rejected if allogeneic spleen cells from the same donor were added to the graft tissue. The added spleen cells caused strong host immune and inflammatory responses. The study gave support to the notion that immunological privilege of the brain does not provide absolute protection to immunogenetically histoincompatible neural grafts.

Animals↗

Transplantation of fetal liver tissue suspension into the spleens of adult syngenic rats: effects of different cytotoxins on cytochrome P450 mediated monooxygenase functions and on oxidative state.

Syngenic fetal liver tissue suspensions were transplanted into the spleens of adult male Fisher 344 inbred rats. Four months after surgery, transplant recipients and age matched control rats were treated with different cytotoxins (allyl alcohol [AAL], bromobenzene [BBZ], carbon tetrachloride [CCl4], or thioacetamide [TAA]) or the respective solvents 24 or 48 hours before sacrifice. Effects of the cytotoxins on P450 mediated monooxygenase functions in liver and spleen 9,000 g supernatants were assessed by measuring the model reactions ethoxyresorufin O-deethylation (EROD), ethoxycoumarin O-deethylation (ECOD), pentoxyresorufin O-depentylation (PROD), and ethylmorphine N-demethylation (EMND). Additionally, the influence on the oxidative state was investigated by assessing the liver and spleen tissue content of lipid peroxidation (LPO) products and of reduced and oxidized glutathione (GSH;GSSG). The livers of both solvent treated transplant recipients and control rats displayed regular EROD, ECOD, PROD and EMND activities. After AAL treatment EROD and EMND activities within the livers were not affected, but ECOD and PROD activities were increased. BBZ administration caused a decrease in EROD and EMND activities, ECOD activity remained unaffected, and PROD activity was even increased. CCl4 and TAA administration caused a strong reduction in the activity of all four model reactions. Spleens of control rats displayed almost no P450 mediated monooxygenase functions, independent whether the rats had been treated with the cytotoxins or not. In the transplant containing spleens, however, significant EROD and ECOD, but hardly any PROD or EMND activities were seen. After AAL administration EROD activity was not affected in the transplant containing spleens, but ECOD activity was increased. BBZ treatment led to a decrease in EROD and an elevation in ECOD activity. CCl4 and TAA strongly reduced the activity of both of these model reactions. The tissue content of LPO products within livers and transplant containing spleens was significantly increased after BBZ and CCl4 treatment. An elevation in LPO products was also seen in the spleens of the control rats due to CCl4 administration. Tissue GSH and GSSG content in both livers and transplant containing spleens were strongly reduced after BBZ treatment. After CCl4 administration only a significant decrease in liver GSSG contents was seen. TAA treatment caused a reduction in the GSH and GSSG content in the spleens of both transplant recipients and control rats, but not in the livers. From these results it can be concluded, that the effects of cytotoxins like AAL, BBZ, CCl4 or TAA on P450 dependent monooxygenase functions and on oxidative state are exerted in the ectopic intrasplenic liver cell transplants in a similar way as in normal orthotopic liver.

7-Alkoxycoumarin O-Dealkylase↗

Transplantation of fetal liver tissue suspension into the spleens of adult syngenic rats: inducibility of cytochrome P450 dependent monooxygenase functions by beta-naphthoflavone, phenobarbital and dexamethasone.

In the present study the effects of beta-naphthoflavone (BNF), phenobarbital (PB) and dexamethasone (DEX) on cytochrome P450 (P450) dependent monooxygenase functions were investigated in intrasplenic liver cell explants in comparison to adult liver. Fetal liver tissue suspensions were transplanted into the spleens of 60-90 days old adult male syngenic Fisher 344 inbred rats. 2, 4 or 6 months after surgery, transplant recipients and age matched controls were orally treated with BNF (1x50 mg/kg body weight (b.wt.)), PB (1x50 mg/kg b.wt.), DEX (for 3 days 4 mg/kg b.wt. per day), or the respective solvents (dimethylsulfoxide or 0.9% NaCl). The animals were sacrificed 24 (BNF, DEX) or 48 (PB) hours after the last treatment. P450 mediated monooxygenase functions were measured in spleen and liver 9000 g supernatants by three model reactions for different P450 subtypes: ethoxyresorufin O-deethylation (EROD; 1A), ethoxycoumarin O-deethylation (ECOD; 1A, 2A, 2B), and ethylmorphine N-demethylation (END; 3A). Spleen weights were significantly higher in transplanted rats, compared to controls, at all three time points after surgery. Induction with PB or DEX, and in some cases also with BNF, lead to a significant increase in liver weights of transplant recipients and control rats independent of the time after transplantation. In contrast, there was no influence on spleen weights due to BNF or PB. At all time points after surgery, with DEX a marked decrease in body weights, weights of adrenal glands and of lymphatic organs like thymus glands and spleens was observed, with the weights of the transplant containing spleens being still higher in comparison to control organs. Spleens of control animals displayed nearly no P450 mediated monooxygenase functions neither without nor with induction. After transplantation, however, significant EROD and ECOD, but hardly any END activities were seen in the host organs at all three time points after surgery. In transplant containing spleens EROD and ECOD were significantly increased after BNF or PB treatment at all three time points after surgery, and ECOD after DEX administration, but at 4 and 6 months after transplantation only. END was only induced after DEX treatment at 6 months after transplantation. With the livers of both transplant recipients and control rats EROD and ECOD were increased after BNF induction and EROD, ECOD, and END after PB treatment at all three time points after transplantation. After DEX administration END was significantly enhanced only at 2 and 4 months after transplantation, ECOD was decreased at 2 and 4 months, and EROD was diminished at all three time points after surgery. Transplantation of fetal liver tissue suspensions into the spleens did not influence monooxygenase functions and their inducibility within the respective livers of the animals. These results demonstrate that transplanted liver cells originating from syngenic fetal liver tissue suspensions display P450 dependent monooxygenase functions which are, simi lar to normal adult liver, inducible by BNF, PB and DEX. Both monooxygenase functions and their inducibility within the transplant containing spleens display quantitative and qualitative developmental changes.

Animals↗

The distribution of iron and iron binding proteins in spleen with reference to Hodgkin's disease.

The distribution of iron and iron binding proteins (IBP) have been compared with control spleen tissue in an attempt to establish a pattern of staining restricted to Hodgkin's disease (HD). All but one of the HD spleens examined stained for ferritin, which was largely present in red pulp dendritic macrophages (DM). In spleens histologically involved with HD heavy deposits of ferritin were seen around tumour nodules. Staining for ferritin increased with involvement of the spleen in HD but DM still represented the bulk of positive cells. However, ferritin positive DM were frequently seen in control spleens, and often in large numbers. Staining of ferric iron by Perls technique was less prominent than ferritin but this observation was also true of the non-HD spleens studied. Patterns of staining with transferrin were equivalent in both groups of spleens with DM being the most frequently positive cell type. Polymorphous macrophages showing erythrophagocytosis were present in the red pulp sinuses of all groups of spleens and although these cells have been considered as precursors of the Reed-Sternberg cell their presence seemed related to total splenic ferritin regardless of the disease process. These cells marked as macrophages and their presence was not restricted to HD. The results show that there is no particular appearance of iron or IBP distribution which is restricted to HD spleens. However, staining for ferritin and iron increased in HD spleens with tumour involvement and could contribute to circulatory abnormalities in this disease.

Ferritins↗

Analysis of loss of heterozygosity and X chromosome inactivation in spleens with myeloproliferative disorders and acute myeloid leukemia.

Neoplastic myeloid proliferations are seen in the spleens of some patients with acute and chronic myeloproliferative disorders. Both acute myeloid leukemia (AML) and chronic myeloproliferative disorders have a variety of underlying cytogenetic defects that can be evaluated by loss of heterozygosity (LOH) studies. LOH studies have advantages over conventional cytogenetics by allowing the use of archival tissues. We evaluated the spleens in AML and chronic myeloproliferative disorders with neoplastic myeloid proliferations for the presence of LOH at several chromosome loci, and X-chromosome inactivation. A total of 17 spleens were evaluated (chronic myelogenous leukemia = 6; chronic idiopathic myelofibrosis = 6; essential thrombocythemia = 1; AML arising from previous chronic myeloproliferative disorders = 4). We examined LOH loci 7q (D7S2554), 8q (D8S263), 9p (D9S157, D9S161), 13q (D13S319), common sites of genetic abnormality in chronic myeloproliferative disorders, and TP53. In six cases, spleen LOH findings were compared to those of concurrent or preceding bone marrow biopsies. Five spleens of female patients were evaluated for the presence of clonality using X-chromosome inactivation. Of the 16 cases analyzed, 14 (88%) had at least one abnormal LOH locus, with 6/16 with two abnormal loci. The abnormalities were distributed as follows: D9S161-7/15 (47%), TP53-6/16 (38%), D7S2554-5/16 (31%), D9S157-5/15 (33%), D8S263-3/14 (21%), and D13S319-2/14 (14%). Of the six bone marrows, 4/6 showed concordance in bone marrow and spleen specimens, with additional LOH abnormalities being identified in the spleen specimens of all four cases. X-chromosome inactivation studies were showed nonrandom (clonal) patterns in two cases. Our results show that allelic losses were common in the neoplastic extramedullary hematopoiesis found in spleens of chronic myeloproliferative disorders and AML. Comparison of spleen and bone marrow specimens by LOH demonstrated additional abnormalities in the spleen compared to the marrow.

Bone Marrow Cells↗

Effects of granulocyte-macrophage colony stimulating factor (GM-CSF) in vivo on cytokine production and proliferation by spleen cells.

GM-CSF is a potent stimulator of haematopoietic cells as well as some functions of granulocytes and macrophages. GM-CSF has many clinical uses; however, little is known about the effects of GM-CSF treatment in vivo on the responses of tissue lymphocytes in terms of secretion of Th-1 and Th-2 cytokines. We investigated this issue by measuring the responses of spleen cells from mice 24 h after treatment i.p. with saline or rmGM-CSF. GM-CSF at 16.7-50.0 microg/kg significantly increased (P < 0.01) spleen cellularity 2-2.5-fold and enhanced proliferative responses of non-stimulated (no mitogen) as well as concanavalin A (Con A)-stimulated spleen cells. Secretion of IFN-gamma by Con A (2.5 microg/ml)-stimulated spleen cells was significantly (P < 0.01) increased from 1.8 microg/ml by control spleen cells to 5.2 microg/ml by GM-CSF spleen cells. IL-10 production was greater (0.25 microg/ml, P < 0.05) by Con A-stimulated spleen cells from GM-CSF-treated mice compared to control spleen cells (0.06 microg/ml). By contrast, there were no significant differences in IL-4 production by Con A-stimulated spleen cells from the different groups. These results show that GM-CSF treatment increases spleen cellularity and primes lymphocytes for enhanced responses. The enhanced production of Th-1 cytokines by primed lymphocytes may partially explain the beneficial role of in vivo administration of GM-CSF in several clinical situations.

Animals↗

Characterization of the residual adenosine deaminating activity in the spleen of a patient with combined immunodeficiency disease and adenosine deaminase deficiency.

A number of infants with an autosomal recessive form of combined immunodeficiency disease also lack adenosine deaminase (adenosine aminohydrolase; EC 3.5.4.4) activity in their erythrocytes. Other tissues from these infants contain only a few percent of the adenosine-deaminating activity present in corresponding normal tissue. The residual adenosine-deaminating activity in extracts from the spleen of a combined immunodeficient, adenosine deaminase-deficient patient was compared with adenosine deaminase from normal spleen. Affinity and immunoadsorbant column chromatography revealed distinct differences between the adenosine-deaminating activity in the patient's spleen and adenosine deaminase from normal spleen. The point of maximum activity and general configuration of the pH optimum curves were also different. erythro-9-(2-Hydroxyl-3-nonyl)adenine, a potent inhibitor of adenosine deaminase from normal spleen, had relatively little effect on the activity from the patient's spleen. In contrast, adenine was a better inhibitor of the activity in the patient's spleen than it was of the enzyme from normal tissue. An adenosine-deaminating activity with the same characteristics and specific activity as that in the patient's spleen was also isolated from normal spleen. These results suggest that the adenosine-deaminating activity in the spleen of this patient is not due to a mutant form of adenosine deaminase.

Adenine↗

Intracellular localization of enzymes in spleen. I. Reduced diphosphopyridine nucleotide cytochrome c reductase, cytochrome c oxidase, and succinic dehydrogenase in the rat and guinea pig.

1. The intracellular distribution of nitrogen, DPNH cytochrome c reductase, succinic dehydrogenase, and cytochrome c oxidase has been studied in fractions derived by differential centrifugation from rat and guinea pig spleen homogenates. 2. In the spleens of each species, the nuclear fraction accounted for 40 to 50 per cent of the total nitrogen content of the homogenate, and the mitochondrial, microsome, and supernatant fractions contained about 8, 12, and 30 per cent of the total nitrogen, respectively. 3. Per mg. of nitrogen, DPNH cytochrome c reductase was concentrated in the mitochondria and microsomes of both rat and guinea pig spleens. Seventy per cent of the total DPNH cytochrome c reductase activity was recovered in these two fractions. The reductase activity associated with the nuclear fraction was lowered markedly by isolating nuclei from rat spleens with the sucrose-CaCl(2) layering technique. The lowered activity was accompanied by the recovery of about 90 per cent of the homogenate DNA in the isolated nuclei, indicating that little, if any, of the reductase is present in spleen cell nuclei. 4. Per mg. of nitrogen, succinic dehydrogenase was concentrated about 10-fold in the mitochondria of rat spleen, and 65 per cent of the total activity was recovered in this fraction. 5. Cytochrome c oxidase was concentrated, per mg. of nitrogen, in the mitochondria of both rat and guinea pig spleens. The activity associated with the nuclear fraction was greatly diminished when this fraction was isolated from rat spleens by the sucrose-CaCl(2) layering technique. Only 50 to 70 per cent of the total cytochrome c oxidase activity of the original homogenates was recovered among the four fractions from both rat and guinea pig spleens, while the specific activities of reconstructed homogenates were only 55 to 75 per cent of those of the original whole homogenates. This was in contrast to the results with DPNH cytochrome c reductase and succinic dehydrogenase where the recovery of total enzyme activity approached 100 per cent, and the specific activities of reconstructed homogenates equalled those of the original homogenates. The recovery of cytochrome c oxidase was greatly improved when only the nuclei were separated from rat spleen homogenates. 6. Data were presented comparing the concentrations (ratio of activity per mg. of nitrogen of the fraction to activity per mg. of nitrogen of the homogenate) of DPNH cytochrome c reductase in mitochondria and microsomes derived from different organs of different animals. 7. Data were presented comparing the activities per mg. of nitrogen of DPNH cytochrome c reductase in homogenates from several organs of various animals.

Animals↗

Evidence of suppressor cell activity in spleens of mice bearing primary tumors induced by Moloney sarcoma virus.

Spleens from Moloney sarcoma virus (MSV) tumor-bearing C57BL/6N mice contained four times the normal number of mononuclear cells and displayed a markedly elevated "spontaneous" (mitogen-independent) DNA synthesis on a per cell basis. The number of macrophages were increased three-fold while there was a slight reduction in the percentage of T lymphocytes. The phytohemagglutinin (PHA) response on a per cell basis of spleens from tumor-bearing mice was decreased about 90% when compared with normal control mice. The primary in vitro immune response to sheep red blood cells was also suppressed to levels of less than 10% of normals. The PHA response could be restored by purification of MSV spleen cells by rayon adherence columns and by removal of phagocytic cells by an iron/magnet technique. The activity of suppressor cells in MSV spleens was demonstrated in mixtures with syngeneic normal spleen cells where a marked impairment of the PHA response was observed. Spleen cells from tumor-free nude mice and normal spleen cells treated by anti-theta serum plus guinea pig complement (C'), both totally unreactive to PHA, had no such effect. The inhibitor cell in MSV spleens was shown to be insensitive to inactivation by anti-theta plus C', but could be removed by the adherence columns and the iron/magnet technique. These data suggest that this suppressor cell is a cell of the monocyte/macrophage series. Suggestive evidence was also presented that the suppressor cells belong to a proliferating population in MSV spleens. Similar suppressor cells have been previously demonstrated in spleens of mice during a variety of immune responses. Our data show, that a tumor, although stimulating the immune system, nevertheless may be suppressive on certain immune functions through the activation of suppressor cells.

Animals↗

Characterization of spleen cells capable of inducing unresponsiveness in ALS-treated mice.

C3H/He skin allografts are significantly prolonged in ALS-treated B6AF1 mice by the injection of 50 x 10(6) C3H/He spleen cells 1 week postgrafting. To identify and characterize the spleen cell active in promoting graft survival, C3H/He spleen cells were separated on a discontinuous Percoll gradient and the various fractions were assayed for the ability to prolong skin graft survival in ALS-treated B6AF1 mice. In addition, unfractionated spleen and spleen fractions were depleted of specific cell populations before injection to determine the effect of various cell populations on graft prolongation. The active cell was recovered primarily in the 52.5% Percoll fraction. Cells in the 60% fraction also had a graft-prolonging effect, but not as significant as that of the 52.5% fraction. Depletion of Thy 1, Ia and Ig-positive cells from unfractionated spleen or spleen fractions did not decrease the graft-prolonging effect. Both Fc gamma R-positive and Fc gamma R-negative cells prolonged graft survival, but the Fc gamma R- cells were the most effective. In contrast to the effect of spleen cells, lymph node cells and thymocytes are relatively ineffective in prolonging graft survival in ALS-treated mice. When lymph node lymphocytes and thymocytes were separated on a Percoll gradient, the cell population active in prolonging graft survival was recovered primarily in the 52.5% fraction. Treatment of the 52.5% fraction of lymph node lymphocytes or thymocytes with monoclonal antibody to Thy 1 before injection abrogated the graft-prolonging effect. These results indicate that the spleen cell(s) active in prolonging graft survival in ALS-treated mice is a non-T, non-B cell, as it lacks Thy 1, Ia, and Ig surface markers. Both Fc gamma R+ and Fc gamma R- spleen cells are effective in prolonging grafts, but Fc gamma R- cells are the most effective. In contrast, the active cell in lymph node and thymus is Thy 1-positive, indicating that it is a T lymphocyte.

Animals↗

Induction of transplantation tolerance in adult rats by vascularized spleen transplantation.

It was demonstrated by Bitter-Seurmann et al. that tolerance to subsequent skin allografts can be brought about, in some rat strain combinations, by heterotopic spleen allografts. We report here that in adult rats, tolerance can be also induced by vascularized spleen allografts using various strain combinations. DA (RT1av1) rats always accepted major histocompatible complex (MHC)-compatible but minor histocompatibility antigen (miH)-incompatible ACI (RT1av1) spleen grafts spontaneously, and further that those DA rats bearing ACI spleen grafts accepted subsequent ACI skin grafts indefinitely. In contrast, Fischer 344 (RT1lvl) rats always accepted not only the "weak" MHC-incompatible Lewis (RT1l) spleen grafts but also subsequent Lewis skin grafts indefinitely, when recipients were administered a short course of FK506 only at the time of primary spleen transplantation. Furthermore, in a number of MHC- and miH-incompatible strain combinations, recipients always accepted primary spleen grafts indefinitely when given a short course of FK506, but those recipients bearing donor spleens revealed split tolerance, that is, rejected subsequent donor-specific skin grafts but accepted heart grafts indefinitely. We conclude that the spleen seems to be a possible organ to induce transplantation tolerance, and that even if the MHC and miH are mismatched, unresponsiveness to at least heart grafts can be achieved by the administration of additional FK506 at the time of spleen transplantation.

Animals↗

In vitro reconstitution of anti-sheep erythrocyte antibody response of T cell-depleted spleen cells by allogeneic T cells or by factors derived from them.

Restoration of the impaired antibody response to sheep erythrocytes (SRBC) in cultures of mouse spleen cells, which were deprived of thymus-derived lymphocytes (T cells) by treatment with anti-mouse brain-associated theta (BA theta) antiserum and complement, was studied by adding a small portion of syngeneic or allogeneic normal spleen cells in vitro. Allogeneic spleen cells had a far greater effect than syngeneic spleen cells on the restoration, as far as the normal spleen cells added were able to recognize the alloantigens on the anti-BA theta serum-treated spleen cells (bone marrow-derived lymphocytes). Treatment of the allogeneic spleen cells with mitomycin C did not affect their activity in the restoration of the impaired antibody response. The possibility that the role of T cells in the antibody response to SRBC may be replaced by a nonspecific mediator derived from T cells reacting with allogeneic cells was proven by the finding that supernatant of the mixed allogeneic spleen cell cultures restored the impaired anti-SRBC antibody response of the T cell-depleted spleen cells. The effect of such culture supernatant on the restoration of the antibody response was greatest when it was added to the T cell-depleted spleen cell cultures one day after cultivation with SRBC, suggesting that the effectiveness may result from triggering of the proliferation and differentiation of antibody-forming cell precursors, which have already reacted with the antigen, to antibody-forming cells.

Animals↗

Lymphoproliferation induced in mouse spleen cells by Mycoplasma arthritidis mitogen. Reversal of the defect of nonresponder mice.

Spleen cells of various mouse strains (e.g. BALB/c, C311, and CBA) reacted towards MAS (a mitogen derived from supernatants of cultured Mycoplasma arthritidis) with a marked lymphoproliferative response. This reactivity was T-cell-dependent. It was reduced by 90% after removal of macrophages by passage of the spleen cells through Sephadex G-10 columns. Addition of 2-mercaptoethanol (2-ME) to macrophage-depleted CBA spleen cells completely restored the response to MAS. Spleen cells of C57BL/6 and C57BL/10 mice were unreactive to MAS, even in the presence of macrophages, and this non-reactivity was controlled by the I-region of H-2. Other mouse strains that, similarly to C57BL/6, lack the expression of I-E on the cell surface (that is, mice of the haplotype H-2f, H-2q, and H-2s) were also non-responsive to MAS. However, the addition of 2-ME to spleen cells of non-responder mice resulted in high lymphoproliferative responses to MAS, which were as high as those of CBA spleen cells. The reaction of C57BL/6 spleen cells to MAS in the presence of 2-ME again was T-cell-dependent, as shown by data with spleen cells of homozygous nude mice and spleen cells treated by anti-thy-1 and C. A macrophage dependency of this response was also evident. When C57BL/6 spleen cells were vigorously freed of accessory cells by the use of nylon wool columns, the MAS response could no longer be restored by 2-ME.

Animals↗

Comparison of the pharmacological characteristics of 2-[125I]iodomelatonin binding sites in the lung, spleen, brain and kidney of chicken.

We have compared the pharmacological characteristics of 2-[125I]iodomelatonin binding to crude membrane preparations of the lung, spleen, brain and kidney of chicken. Saturation studies indicated significant differences (p < 0.05) in the equilibrium dissociation constant (Kd) and maximum number of binding site (Bmax) values among the four tissues studied. The descending order of affinities was lung = spleen > brain > kidney. Competition curves of 2-[125I]iodomelatonin binding to crude membrane preparations of all four chicken tissues by melatonin were studied simultaneously to reduce individual, physiological, age and interassay variations. Similar competition experiments were also performed on 2-phenylmelatonin, 2-iodomelatonin, 6-chloromelatonin, 6-hydroxymelatonin and N-acetylserotonin (NAS). Concentrations of indoles which inhibited 50% of specific 2-[125I]iodomelatonin binding (IC50) were calculated. The IC50 of 2-[125I]iodomelatonin inhibition curves by the indole compounds in different tissues showed the following descending orders of affinity: (1) melatonin: lung = spleen > brain > kidney, (2) 2-phenylmelatonin: lung = spleen = brain = kidney, (3) 2-iodomelatonin: lung = spleen = kidney > brain, (4) 6-chloromelatonin: lung = spleen = kidney > brain, (5) 6-hydroxymelatonin: kidney > lung = spleen = brain, and (6) NAS: kidney > lung = spleen > brain. The non-hydrolizable GTP analog, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), exhibited differential effects on the saturable binding of the four tissues. GTP gamma S increased the Kd of 2-[125I]iodomelatonin binding by 2- to 3-fold in the lung and spleen, 0.5-fold in the brain and 1-fold in the kidney. Based on our findings, we would like to suggest that the 2-[125I]iodomelatonin binding sites in these four tissues may belong to three different high affinity (picomolar) subtypes of melatonin receptor. We name them cML1A represented by the lung and spleen, cML1B by the brain and cML1C by the kidney.

Animals↗

Embryonic origins of spleen asymmetry.

The spleen is a vertebrate organ that has both hematopoietic and immunologic function. The embryonic origins of the spleen are obscure, with most studies describing the earliest rudiment of the spleen as a condensation of mesodermal mesenchyme on the left side of the dorsal mesogastrium. The development of spleen handedness has not been described previously, presumably because of the difficulty in assaying spleen position in the embryo and the lack of early, organ-specific molecular markers. Here we show that expression of the homeobox gene Nkx2-5 serves as a marker for spleen precursor tissue. Pre-splenic tissue is initially located in symmetric domains on both sides of the embryo but, during subsequent development, only the left side goes on to form the mature spleen. Therefore, the final location of the spleen on the left side of the body axis appears to result from preferential development of the spleen precursor cells on the left side of the embryo. Our studies indicate that the spleen and heart become asymmetric via different cellular mechanisms. Nkx2-5 may function locally as part of the laterality cascade, downstream of nodal and Pitx2, or it may direct asymmetric morphogenesis after laterality has been determined.

Animals↗

Purification and properties of a heat-stable glucocerebrosidase activating factor from control and Gaucher spleen.

Gaucher's disease is a lysosomal storage disease caused by a deficiency in the enzyme glucocerebrosidase. A small, heat-stable glycoprotein first obtained from Gaucher spleen (Ho, M. W., and O'Brien, J. S. (1971) Proc. Natl. Acad. Sci. U. S.A. 68, 2810-2813) has been observed to stimulate the activity of glucocerebrosidase isolated from normal tissue. It has been suggested that this material might be important in the physiological catabolism of glucocerebroside in normal individuals (Ho, M. W. (1974) in Enzyme Therapy in Lysosomal Storage Diseases (Tager, J. M., Hooghwinkel, G. J. M., and Daems, W. Th., eds) pp.239-246, North-Holland Publishing Co., Amsterdam). In order to investigate this suggestion, glucocerebrosidase activating factors were isolated and purified from control and Gaucher spleen and characterized. Although approximately the same mass of activator was isolated from both spleens, the two activators differ from one another in a number of important respects: (a) the activator from the control spleen is only 6 per cent as active (on a protein basis) as the activator from Gaucher spleen; (b) the amino acid compositions of the purified activators are significantly different; and (c) carbohydrate analysis of the purified activators indicates that the activator from Gaucher spleen is a glycoprotein, while that from control spleen is not. Comparative kinetic studies demonstrate that the anionic detergent, sodium taurocholate, and the acidic phospholipid, phosphatidylinositol, both stimulate glucocerebrosidase activity to a larger extent than the activator substance from Gaucher spleen. The activator from Gaucher spleen and human liver glucocerebrosidase both appear to contain significant hydrophobic character. We conclude that the activator is probably not physiologically important in the catabolism of glucocerebroside in normal tissues. The significance of the occurrence of this apparently unique glycoprotein activator in Gaucher spleen remains obscure; however, its presence represents another interesting aspect of Gaucher's disease that warrants further investigation.

Amino Acids↗

[Non-surgical aproach in blunt injuries to the liver and spleen].

INTRODUCTION: Non-surgical approach in blunt injuries of the liver and spleen in patients haemodynamically stable, is widely accepted as their therapeutical method of choice. Based on a retrospective analysis, the aim of this study is to assess the injured patients with blunt injuries of the liver and spleen who were treated non-surgically. SUBJECTS AND METHODOLOGY: 75 injured patients with blunt intraabdominal injuries were hospitalized from 2001 to 2003. The spleen injuries were diagnosed in 40 patients, out of which 12 of them suffered from concomittent injuries of other intraabdominal organs, the liver injuries were diagnosed in 37 cases, out of which 24 injuries were isolated. The non-surgical approach was indicated in haemodynamically stable patients with isolated injuries of the spleen and the liver of the ISt-IVth grade with haemoperitoneum up to 300 ml, diagnosed on a spiral CT examination. RESULTS: 47 (63%) patients were urgently operated for intraabdominal injuries, 7 of them (15%) exited within 30 days after the surgery. The non-surgical approach was indicated in 28 (37%) cases. In this group, 19 patients suffered from liver injuries, 7 from spleen injuries and 2 from pancreatic injuries. The severity of the injury was classified as the It degree in 7 patients, as the IInd degree in 7 patients, as the IIIrd degree in 7 patients and as the IVth degree in 5 patients. The non-surgical procedure failed in 5 (19%) cases of patients with IIIrd or IVth degree liver or spleen injuries. The failure was caused by a delayed bleeding from the injured spleen in 3 patients and by a continuous bleeding in the IVth degree liver injury and the bile collection from the biliary bladder rupture in the patient with the IIIrd degree liver injury. The spleen injury was treated using splenectomy. In the IVth degree liver injury a debridement of the dilacerated tissue of the 7th segment and a ligature of the injured branch of the right hepatic vein, were conducted. The patient with the IIIrd degree injury was treated using cholecystectomy with a suture of the liver rupture. CONCLUSION: The non-surgical approach to the treatment of the isolated blunt liver and spleen injuries is indicated for the injured patients who are haemodynamicaly stable, with Ist-IVth degree injuries, without or with minor (up to 300 ml) haemoperitoneum and with continuous intensive care. The non- surgical approach to the higher degree blunt liver and spleen injuries has higher treatment failure risk rates.

Adult↗