Antigenic similarity between stimulators of immunogenesis of a polypeptide nature from the thymus and cerebral cortex

Rabbit antisera against low molecular weight polypeptides from the thymus (thymosin and thymarin), cortex (cortexin) and white matter of the brain of the calves were cross-absorbed with these polypeptides and tested in the complement fixation test with these preparations and in the complement-dependent cytotoxicity test with thymic and bone marrow cells. The results showed that thymosin, thymarin and cortexin are antigenically similar, but differ in antigenic structure from polypeptide from white matter of the brain. Biological effect of polypeptides from the thymus and brain cortex is connected with thymus-depending lymphocytes and does not depend on B-cells. Cross absorbtion revealed that antisera against polypeptides from thymus and cortex of the brain contain antibody both against common antigens and antigens specific for appropriate preparation only. Antigenic set of polypeptide from the thymus (thymarin) corresponds more closely to thymic antigen as compared to polypeptide from the brain cortex (cortexin).     

Effect of highly purified thymus factor on the cellular and humoral indices of immunity in thymectomized mice]

Effect of homogeneous polypeptide thymus factor of mol weight about 5000 (thymarin-III) on cellular and humoral immune responses of thymectomized adult CBA mice was studied. Thymectomy proved to greatly decrease the number of T-cells in the spleen. Accordingly, the capability of these mice to produce both IgM and IgG antibody-forming cells in response to the thymus-dependent antigen (sheep red blood cells) was significantly depressed. Subcutaneous injections of thymarin-III (1 microgram per g of body weight) for 7 days completely restored the T-cell spleen population and normalized the animals' immune response.     

Isolation and evaluation of the specificity of an antiserum to the thymic polypeptide factor

The specificity of antisera, obtained by the immunization of rabbits with the conjugated antigenic preparations of the thymic factor, have been evaluated by the method of immunochemical analysis. To carry out the comparative study, polypeptides isolated from the pineal body, cortex and white matter of the brain, Thy-1 antigen from the cerebral cortex and insulin have been used. The polypeptides of the thymus and the brain have been found to differ in their amino acid composition and molecular weight. The thymic factor possesses specific antigenic determinants which are absent in the tested preparations of cerebral polypeptides and insulin. The rabbit antisera obtained in this investigation are highly specific and can be used for the immunochemical determination of the thymic factor in the blood and other biological fluids.     

Effect of substances of a polypeptide nature isolated from the cerebral cortex on the immune response of mice

The influence of low molecular weight polypeptides (Mol wt below 10,000) isolated by acetic acid extraction from the cortex and white matter of the brain and from the bone marrow of cows on the primary immune response to SRBC was studied in experiments on 248 male CBA mice. The results showed that after the subcutaneous injections of the preparations from the cortex (where the theta-antigen cross-reacting with the thymocytes was localized) for 5 days prior to and 3 days post immunization, hemagglutinin titres and the quantity of direct (IgM) and indirect (IgG) antibody-forming cells increased 2.5 times in comparison with the control. The preparations from the white matter of the brain and from the bone marrow failed to demonstrate such an effect.     

Effect of antibodies to rat and human cerebral cortex and white matter on heterologous T- and B-cells]

A study was made of the cytotoxic and complement-fixation activity of the antisera to the cortex and white matter of the rat and human brain upon the mouse and rat thymus and bone marrow cells. The cytotoxicity test proved to be more sensitive and precise. Cytotoxins to rodent thymocytes were revealed on ly in the antisera against the human brain cortex; at the same time they were revealed both in the antisera against the cortex and against the white matter of the rat brain (much more was found in the former). The sera against the rat brain cortex lost their cytotoxicity after the exhaustion with the same antigen, but retained it when the exhaustion was carried out by the white matter.     

Effect of electric field during incubation of eggs on the immune responses of hatched chickens

The effects of electric field (EF) during incubation of eggs on the immunocompetence of chickens were investigated over a 42-day experimental period. Eggs from a meat-type breeder flock were incubated under EF of 30 kV/m, 60 Hz during the first 18 days of incubation as compared with the control incubation (C). Chickens from the two incubation treatments were fed ad libitum and their immune system were monitored. Measurements were made of body weight (BW), and lymphoid organs weight (thymus, spleen, and bursa of Fabricius (BOF)) of birds at 21 and 42 days of age. Immune systems of birds were tested for specific antibody responses to sheep red blood cell (SRBC) and Newcastle disease vaccine (NDV), in vivo T-lymphocyte proliferation responses to phytohemagglutinin (PHA), and in vitro to concanavalin A (Con-A). EF incubation of eggs did not significantly (P > 0.05) influence BW of bird, absolute weight of lymphoid organs and weight of thymus, and BOF as a percentage of BW of bird (% BW) at 21 and 42 days of age, humoral immune responses as measured by antibody responses to SRBC and NDV, and cell-mediated immune responses as measured by T-lymphocyte proliferation responses to PHA, and Con-A of birds when compared with those of the C treatment. EF incubation of eggs significantly (P < 0.05) increased spleen weight as a % BW at 21 and 42 days of age when compared with those incubated under the C treatment. Birds at 42 days of age had significantly (P < 0.01) higher BW, lymphoid organ weight, and weight of BOF as a % BW, and lower spleen weight as a % BW when compared with those of 21 days of age. It is concluded that the incubation of eggs under EF of 30 kV/m, 60 Hz increased spleen weight as a % BW, without altering cell-mediated and humoral immune responses and, consequently, immunocompetence of meat chickens during the rearing period of 42 days.     

Identification with a Monoclonal Antibody of a Phylogenetically Conserved Brain-Specific Determinant on a 130,000 Molecular Weight Glycoprotein of Human Brain

Human brain glycoproteins depleted of Thy-1 antigen were used to immunise Balb/c mice for monoclonal antibody production. The F3-87-8 antibody described in this paper interacts with a determinant present in large amounts on all human brain subregions studied (cerebral cortical grey matter, white matter, caudate, thalamus, dentate nucleus, putamen, cerebellar cortex) but absent from all other tissues examined (liver, heart, kidney, spleen, thymus, lymph node, erythrocyte, adrenal gland, and peripheral nerve). The determinant is conserved in mammalian evolution, as the brains of the rat and dog have amounts equal to that found in human brain. Balb/c mouse brain has approximately one-third as much antigen activity as these other mammalian brains, whereas brains of the frog and chicken have no detectable antigenic activity. Developmental studies showed that 16-week human foetal brain and neonatal dog brain had little or no antigen activity, indicating a dramatic increase in the amount of the determinant with brain maturation. Biochemical studies showed that the F3-87-8-bearing molecule was a major sialoglycoprotein of human brain with an apparent molecular weight of 130,000. It was shown by immunofluorescence to be particularly localised in what appeared to be fibre tracts in the thalamus and basal ganglia, and in the dentate nucleus, although all regions including grey matter were stained.     

免疫应答期间脑和淋巴器官中去甲肾上腺素含量的变化

应用高效液相色谱－电化学检测法（ＨＰＬＣ－ＥＣＤ）测定大鼠在用绵羊红细胞（ＳＲＢＣ）免疫后第２－７天期间,下丘脑、海马、脑干、胸腺和脾脏中去甲肾上腺素（ＮＡ）含量的变化。实验结果表明,下丘脑内ＮＡ含量在免疫后第４－７天明显增加,其中第７天有回降趋势。海马内ＮＡ含量在第４－５天显著增多。而胸腺和脾脏中ＮＡ水平在第４－５天均明显降低。脑干在免疫应答期间无明显改变。以上结果说明,体液免疫应答可影响脑和淋     

Some endocrine aspects of the thymus gland.

In studies of the mouse thymus, lymphocyte mitoses are seen to be most frequent in the thymus cortex. There is evidence from thymic grafts that a hypothetical factor, thymopoietin, may stimulate mitosis of thymic lymphocytes. It is a factor which is postulated to act in conjunction with the PAS-positive mesenchymal reticular cells and epithelial reticular cells of the cortex. The thymus medulla is necessary for the integrity of thymic grafts, and may also elaborate a secretion for maintaining the cellular functions of the gland. Thymectomy has been used as a gauge for judging normal thymic function and results, in the mouse, in lymphopenia, degeneration of spleen and lymph nodes, delayed rejection of skin allografts, reduced ability of spleen cells to mount the graft versus host reaction, and reduced primary immune response to certain antigens. Correction of these deficiencies offers a means of evaluating various thymic extracts and grafts. Lymphocytosis-stimulating hormone (LSH) is known to maintain the peripheral lymphoid organs and cause lymphocytosis in the thymectomized animal. Diffusion chamber studies of thymic grafts also show restored lymphoid tissue by a cell-free factor (CIF). These two factors may be the same and probably represent the basis of the highly purified lymphocyte-stimulating proteins, LSHr and LSHh, which restore the L/P ratio in thymectomized animals and may stimulate lymphopoiesis in spleen and lymph nodes. LSHr, unlike LSHh, increases the total lymphocyte count. LSHr has been found to increase the humoral antibody response in neonatal mice both by the PFC technique and by direct hemolysis of sheep erythrocytes. Homeostatic thymic hormone (HTH) is a thymic extract of small molecular weight and contains nucleic acid. In the thymectomized guinea pig it has been found to maintain normal levels of lymphocytes in the blood, spleen and lymph nodes, to restore antibody titers to typhoid H antigen and to restore the toxic allergic reaction. Thymic humoral factor (THF) is of smaller molecular weight (less than 1,000) and probably is not a protein. It also enhances lymphoid proliferation in neonatally thymectomized mice. There is evidence that THF participates in humoral antibody formation because it stimulates PFC formation from neonatally thymectomized mice after inoculation with sheep erythrocytes. Its effects on cell-mediated immunity are seen from findings that injection of THF restores the ability of thymectomized mice to reject skin allografts. THF enables spleen cells from thymectomized or neonatal animals to mount the graft versus host reaction, and causes maturation of bone marrow cells and spleen or lymph node cells so that they can participate in the graft versus host reaction. It has been reported to stimulate lymphocytes to kill isogeneic tumor cells in vitro. Thymosin is protein extracted from the thymus. It has been found to alleviate leukopenia slightly and provide some improvement in lymphoid histology in thymectomized mice...     

Histology and histochemical enzyme‐staining patterns of major immune organs in Epinephelus malabaricus

The histological architecture of major immune organs in the Malabar grouper Epinephelus malabaricus was investigated. The novel characteristics such as melanomacrophage centres (MMCs) appeared in the thymus and lymphopoietic tissue formed as foci in the head kidney. Leukocyte distribution in organs was identified by enzyme histochemistry. β‐glucuronidase (BG) reactive cells in the cortex region of the thymus were botryoidally aggregated. Both acid phosphatase (AcP) and BG reactive cells concentrated within the specialized lymphopoietic foci in the head kidney, suggesting that the foci might be functional. Primitive histological characters of immunity were observed in the spleen. Although leukocyte aggregation was demonstrated in the spleen, additional enzyme histochemistry indicated that the aggregate might not be the equivalent of white pulp found in other vertebrates. The histological evidence did not support intestinal involvement in the immune system: there was no demonstrable gut associated lymphoid tissue. The limited distribution in the cortex and medulla boundary and the condensed format of the lymphocytes suggests a functional role for the MMC in the thymus of E. malabaricus . The MMC appearance in the thymus of a teleost was unusual.     

Neonatal exposure to propylthiouracil induces a shift in lymphoid cell sub-populations in the developing postnatal male rat spleen and thymus

Evidence of the connections between the immune system and the thyroid axis is increasingly strong; however, much of the data are focused on immune effects of altered thyroid status in adults or rodents with congenital defects of the pituitary/thyroid axis. The object of the present study was to determine the effects of PTU-induced hypothyroidism on the developing immune system of the rat by focussing on both the spleen and thymus gland. Male Sprague-Dawley rat pups were exposed to PTU through maternal milk by giving the mothers 0.02% PTU in their drinking water starting on the pups' day of birth until day 24 (d24), shortly before weaning on d28. Animals were sampled on days 14, 22, 30, and 91. The mean body weight was decreased in the PTU-treated animals on days 14, 22, and 30. The mean spleen and thymic weights and cellularity were all decreased in the PTU-treated animals on d22 and d30. PTU exposure increased the proportion of NK cells in the spleen on days 14, 22, and 30. The proportion of T-cells was increased on days 22 and 30 with a particular increase in the CD4+ T-cells, resulting in an increase in the ratio of helper T-cells to suppressor/cytotoxic T-cells at d22. PTU also decreased the proportion of splenic B-cells at days 14, 22, and 30 which could explain the increased proportion of both NK and T-cells during these sampling periods. PTU treatment decreased the lytic ability of NK cells at d22, but no functional differences were observed at days 14, 30, 91, despite the increased proportion of NK cells in PTU-exposed animals at days 14, 22, and 30. PTU exposure also increased the proportion of CD4+CD8- cells in the thymus on d22 and caused an increase in both the CD4+CD8- and CD4-CD8+ populations on d30. These data suggest that the effects of temporary, PTU-induced hypothyroidism on the cell populations in the spleen partially result from transient changes in thymic T-cell development, including a shift towards increased CD4+CD8- cells. The data also suggest that temporary hypothyroidism early in development decreases B-cell development in a transient fashion. Temporary hypothyroidism induced from birth to the latter stages of the weaning period induced transitory effects on the spleen, thymus, and immune cell sub-populations--all of which recovered to normal values when the animals matured.     

Selective adherence of lymphocytes to myelinated areas of rat brain.

An in vitro system developed for studying lymphocyte binding to high endothelial venules (HEV) of lymph nodes was used to determine if there are similar binding sites in other organs of the rat. Thoracic duct lymphocytes (TDL) adhered selectively and uniformly to white matter when overlaid onto glutaraldehyde-fixed tissue sections of cerebellum and cerebrum. The pattern of TDL adherence to cerebellar sections showed that binding to nonmyelinated areas was negligible. Comparison of TDL-white matter to TDL-HEV binding demonstrated that the density of adherence to each site was quantitatively similar. In contrast, lymphocytes exhibited little tendency to bind to tissue sections of liver, spleen, heart, thymus, and salivary glands. TDL adherence to cerebellar white matter occurred rapidly, was cell dose dependent and optimal at 7 degrees C. White matter binding was also a property of spleen lymphocytes but the thymus was deficient in cells with this capability. The affinity of TDL and spleen lymphocytes for myelinated areas of the brain suggests the presence of myelin binding receptors on these cells.     

Prothymosin alpha and parathymosin: mRNA and polypeptide levels in rodent tissues

Blot hybridization analyses have established the presence of mRNAs for prothymosin alpha (ProT alpha) and for parathymosin (ParaT) in rat and mouse lung, liver, kidney, and brain, confirming the biosynthesis of these peptides in nonlymphoid tissues. In these tissues the levels of mRNAs paralleled the content of the polypeptides, determined with specific radioimmunoassays. The mRNA levels also confirmed the reciprocal relation between the two polypeptides; ProT alpha and its mRNA were found in highest concentrations in spleen and thymus, followed by lung, kidney, and brain, with lowest concentrations in liver. On the other hand, liver contained highest concentrations of ParaT and the mRNA for ParaT, with lowest levels present in spleen and thymus. In comparison to tissues from young (6-8 week) mice, older (18 month) mice contained lower concentrations (20-40%) of both polypeptides, with qualitatively similar decreases in mRNA content.     

Complex Regional Pain Syndrome Type I Affects Brain Structure in Prefrontal and Motor Cortex

The complex regional pain syndrome (CRPS) is a rare but debilitating pain disorder that mostly occurs after injuries to the upper limb. A number of studies indicated altered brain function in CRPS, whereas possible influences on brain structure remain poorly investigated.We acquired structural magnetic resonance imaging data from CRPS type I patients and applied voxel-by-voxel statistics to compare white and gray matter brain segments of CRPS patients with matched controls. Patients and controls were statistically compared in two different ways: First, we applied a 2-sample ttest to compare whole brain white and gray matter structure between patients and controls. Second, we aimed to assess structural alterations specifically of the primary somatosensory (S1) and motor cortex (M1) contralateral to the CRPS affected side. To this end, MRI scans of patients with left-sided CRPS (and matched controls) were horizontally flipped before preprocessing and region-of-interest-based group comparison. The unpaired ttest of the “non-flipped” data revealed that CRPS patients presented increased gray matter density in the dorsomedial prefrontal cortex. The same test applied to the “flipped” data showed further increases in gray matter density, not in the S1, but in the M1 contralateral to the CRPS-affected limb which were inversely related to decreased white matter density of the internal capsule within the ipsilateral brain hemisphere. The gray-white matter interaction between motor cortex and internal capsule suggests compensatory mechanisms within the central motor system possibly due to motor dysfunction. Altered gray matter structure in dorsomedial prefrontal cortex may occur in response to emotional processes such as pain-related suffering or elevated analgesic top-down control.     

Effect of unilateral nephrectomy in mice on the level of the humoral immune response to T-independent antigen

The influence of unilateral nephrectomy on the degree of humoral immune response to T-independent (polyvinylpyrrolidone, PVP) and T-dependent (sheep red blood cells, SRBC) antigens was studied. The increase in the number in antibody-forming cells (AFC) and nonspecific immunoglobulin-forming cells (nIFC) was investigated by means of the adaptive transfer model. The lethally irradiated recipients were injected with the antigen and also the spleen cells of operated and intact donors. PVP did not induce significant alterations of antibody genesis in mice receiving spleen cells of unilaterally nephrectomized animals comparing with recipients of intact spleen cells. At the same time, the kidney operation induced the increase in the number of AFC and nIFC when the SRBC were used. Hence the activation of humoral immune response induced by kidney operation was related not to the direct activation of B-lymphocytes but to T-cells. The possible causes of this activation were analyzed. Spleen cells of operated animals enhance both specific and antigen-dependent nonspecific immune response.     

Expression of insulin-like growth factor II (IGF-II) and histological changes in the thymus and spleen of transgenic mice overexpressing IGF-II

 Previously, transgenic mice were constructed overexpressing human insulin-like growth factor II (IGF-II) under control of the H₂k^b promoter. The IGF-II transgene was highly expressed in thymus and spleen, and these organs showed an increase in weight. In the current study we have analyzed the sites of IGF-II mRNA expression, the distribution of IGF-II, IGF-I, and both IGF receptors, and histomorphometrical changes in thymus and spleen. With in situ mRNA hybridization, expression of the IGF-II transgene is found with high intensity in the thymic medulla and in the white pulp/marginal zone of the spleen, whereas there were scattered positive cells in the thymic cortex and in the splenic red pulp. Hybridization was restricted to non-lymphocytic cells. Immunohistochemistry revealed intense IGF-II peptide staining with the same distribution as IGF-II mRNA. There was additional intense IGF-II staining of all elements in the splenic red pulp (including trabeculae) and diffuse, low level staining in the thymic cortex. These findings were not observed in control mice. In the thymic medulla, most IGF-II producing cells co-labelled with keratin, whereas a minor population also stained for the monocyte/macrophage marker MOMA-2. In the spleen, co-labelling of IGF-II producing cells was found with MOMA-1 (marginal zone), or with the dendritic cell marker NLDC-145 (red pulp). IGF-I and both IGF receptors were found in these organs in nearly all cell types, with a similar pattern in transgenic mice and in control animals. Histomorphometric analysis revealed a marked increase of thymus cortex size and an increased trabecular size in the spleen. This suggests that IGF-II overproduction induces local effects (auto/paracrine) in the thymic cortex, but not in the thymic medulla. Trabecular growth in the spleen most likely is a distant effect (paracrine or endocrine) of IGF-II overproduction. Accepted: 5 September 1996     

Immunomodulatory effect of Moringa oleifera Lam. extract on cyclophosphamide induced toxicity in mice

Immunomodulatory effect of ethanolic extract (50%) of M. oleifera leaves (MOE) has been studied in normal and immunosuppressed mice models. Different doses of MOE i.e. 125, 250 and 500 mg/kg body weight of mice were administered orally for 15 days. Cyclophosphamide at a dose of 30 mg/kg body weight was administered orally for the next 3 days. On day 16 and 19, hematological parameters like white blood cell (WBC) count, red blood cell (RBC) count, haemoglobin level (Hb), percent neutrophils and organ weight were recorded. Effect of MOE on phagocytic activity of mice macrophages was determined by carbon clearance test. MOE showed significant dose dependent increase in WBC, percent neutrophils, weight of thymus and spleen along with phagocytic index in normal and immunosuppressed mice. The results indicate that MOE significantly reduced cyclophosphamide induced immunosuppression by stimulating both cellular and humoral immunity.     

Generalized accumulation of neutral glycosphingolipids with G M2 ganglioside accumulation in the brain

Analyses have been made of glycosphingolipids from visceral organs and brain of a patient with an unusual lipid storage disorder diagnosed initially as classical Tay-Sachs disease. Levels of the lipids from fresh-frozen sections of gray and white matter, kidney, spleen, liver, and heart from this patient were compared with those of normal juvenile controls, and the fatty acid composition of accumulated glycosphingolipids was compared with reference compounds. This patient was found to have abnormally high concentrations of a globoside in liver, kidney, and spleen, asialo G(M2) ganglioside in brain and liver, and G(M2) ganglioside in the brain. On the basis of these findings along with the clinical manifestations of Tay-Sachs disease with visceral involvement (hepatosplenomegaly) and demonstration of total deficiency of both A and B components of beta-N-acetylhexosaminidase activity, this glycosphingolipidosis is the same as two previously reported cases of G(M2) gangliosidosis with globoside accumulation and total beta-N-acetylhexosaminidase deficiency.     

THE DISTRIBUTION OF LIPIDS IN THE HUMAN NERVOUS SYSTEM-V. GANGLIOSIDES AND ALLIED NEUTRAL GLYCOLIPIDS OF INFANT BRAIN

—Gangliosides and allied neutral glycosylceramides were isolated from human infant (2-24 months of age) cerebral cortex and white matter. The individual glycolipids were separated quantitatively by a combination of column and thin-layer chromatographic methods on silica gel, DEAE-cellulose and Sephadex G-25. In cerebral cortex G_D1a and G_M1 were the major fractions and constituted more than 70 per cent of the total gangliosides. The concentrations of neutral glycolipids, except for galactosylceramides, were very low: lactosylceramide and glucosylceramide comprised 30 and 5 nmol/g wet weight, respectively. In white matter their concentrations were 10 times higher. The ganglioside concentration was only 50 per cent of that in cerebral cortex: the difference was accounted for mainly by the much lower content of the major di- and trisialogangliosides. Stearic acid was the predominant fatty acid of all brain gangliosides. G_M3, and G_D3 had a considerable content of the very long-chain fatty acids, C₂₂-C₂₄, particularly in the white matter. Glucosylceramide and lactosylceramide had almost identical fatty acid patterns between each other in cerebral cortex and white matter. In the cerebral cortex stearic acid and in the white matter the very long-chain acids predominated. d20:1 Sphingosine comprised more than 20 per cent of total sphingosine in all the gangliosides of the G_l- and G₂-series. G_M3, and G_D3 like lactosylceramide contained significantly less of d20:1 sphingosine. The findings suggest the existence of separate compartments for the biosynthesis of the gangliosides. Glucosylceramides and lactosylceramides of white matter have the same ceramide composition as the galactosylceramides with normal fatty acids and are thus unlikely to be intermediates in the metabolism of the major brain gangliosides which have a completely different fatty acid composition.     

磁处理白术药液对小白鼠免疫器官指数影响的药效研究

目的 :从免疫学方面探讨磁处理白术药液对小白鼠免疫器官指数影响的药效作用。方法 :用不同强度的磁处理白术药液及非磁处理白术药液对小白鼠进行腹腔注射 ,连续 7d ,每天一次 ,末次给药 1 2h后处死 ,称其体重 ,取出胸腺、脾脏及肝脏 ,称重 ,计算各器官指数。结果 :与正常对照组比较 ,磁处理白术药液组对小白鼠的肝脏指数、脾脏指数均有极显著的提高 (P <0 .0 1 ) ,胸腺指数也有影响 ,但差异不显著 (P>0 .0 5 ) ;与非磁处理药液组比较 ,磁处理药液组对小白鼠的肝脏指数、脾脏指数、胸腺指数有影响 ,但差异不显著 (P >0 .0 5 )。结论 :磁处理白术药液对免疫器官指数有明显作用