共查询到20条相似文献,搜索用时 15 毫秒
1.
Stimulation of synthesis of immunoglobulin (Ig) in vitro by Con A and anti-Ig in cultures of rabbit lymphoid cells has been analyzed qualitatively using an assay that measures the incorporation of [3H]leucine into newly synthesized proteins, followed by the specific absorption of tritiated immunoglobulin by staphylococcal protein A. Whereas Con A stimulates Ig production by spleen cells only if T lymphocytes are present, anti-immunoglobulin serum enhances Ig synthesis in the absence of T lymphocytes. In contrast, neither Con A nor anti-immunoglobulin serum stimulates peripheral blood lymphocytes to produce enhanced levels of Ig. It is concluded that both Con A and anti-immunoglobulin serum do not activate resting B cells but drive differentiation of B cells which are already synthesizing Ig. Anti-Ig acts directly whereas stimulation of B-cell Ig synthesis by Con A occurs indirectly through stimulation of T cells. 相似文献
2.
THE division of lymphocytes into thymus-derived (T) cells and bursa-equivalent-derived (B) cells is well established (reviewed in refs. 1–3). The result of antigenic stimulation in the B line of lymphocytes is a differentiation process, involving clonal expansion and ultrastructural changes, to give a specialized population of cells which synthesize and secrete immunoglobulin. In the study of these processes a major problem is the small number of cells involved in response to antigen, usually less than 1% of the total lymphocyte population. Clearly a system for activating large numbers of lymphocytes into immunoglobulin synthesis would offer considerable advantages. This seems to occur when mouse B lymphocytes are stimulated by pokeweed mitogen (PWM). In our experimental conditions, however, IgM is the only immunoglobulin class to be synthesized. The rational basis for our experiments rests on three previous observations: (1) PWM-stimulated lymphocytes develop rough endoplasmic reticulum4,5 and might therefore be expected to be secreting cells; (2) a small proportion of enlarged (“blast”) lymphoid cells in PWM-treated human blood lymphocyte cultures contain immunoglobulin demonstrated by immunofluorescence6 and (3) the recent demonstration that mouse B lymphocytes are activated by PWM7. 相似文献
3.
D H Boldt R P MacDermott S F Speckart G S Nash 《Journal of immunology (Baltimore, Md. : 1950)》1977,118(4):1495-1498
A large proportion of DNA synthesized in vitro by human lymphocytes stimulated with plant mitogens or specific antigens is selectively excreted from the cells. To determine if DNA excretion differs among various types of lymphocytes, we examined purified human lymphocyte subpopulations for DNA synthesis and excretion in response to stimulation by L-PHA. The relative proportion of newly synthesized DNA that is excreted by unseparated mononuclear cells, macrophage-depleted cells, T, and B lymphocytes is identical despite great differences in the magnitude of their responses. Low levels of both DNA synthesis and excretion by macrophage-depleted cells and B cells can be increased by reconstitution with macrophages and T cells, respectively. These data indicate that DNA exretion is a general property of lymphocytes stimulated to undergo DNA synthesis by plant mitogens. 相似文献
4.
We have measured the concentration of mRNAs coding for immunoglobulins, k and lambda type light chains and gamma 1 type heavy chain, in mouse spleen cells activated by bacterial lipopolysaccharide or sheep red blood cells. These mRNAs were quantitated by hybridization to radioactive DNA complementary to highly purified immunoglobulin mRNAs from mouse myelomas. In the lipopolysaccharide-stimulated spleen cells, only light chain mRNA accumulated, whereas gamma 1 type heavy chain mRNA remained unvaried. The light chain mRNA concentration also increased in purified bone-marrow-derived lymphocytes. The lipopolysaccharide-induced light chain mRNA was similar to light chain mRNAs purified from myelomas. The accumulation and disappearance of light chain mRNA in bone-marrow-derived lymphocytes coincide with the kinetics of synthesis of immunoglobulin M which is the major species induced by lipopolysaccharide. In sheep red blood cell stimulated spleen, the specific accumulation of k type light chain and gamma 1 type heavy chain mRNAs parallels immunoglobulin G synthesis. These results seem to indicate that the increment of immunoglobulin mRNA concentration in bone-marrow-derived lymphocytes is important for induction of immunoglobulin synthesis. 相似文献
5.
Wound-induced PAL activity is suppressed by heat-shock treatments that induce the synthesis of heat-shock proteins 总被引:1,自引:0,他引:1
Wounding lettuce leaves induces the de novo synthesis of phenylalanine ammonia-lyase (PAL, EC 4.3.1.5), the accumulation of phenolic compounds, and subsequent tissue browning. A brief heat-shock at 45°C reduces the rise in wound-induced PAL, the accumulation of phenolic compounds, and tissue browning. The activity of PAL measured 24 h after wounding and the content of phenolic compounds (absorbance of methanol extract at 320 nm) measured 48 h after wounding was highly correlated (R2 > 0.90) in tissue developing the normal wound response and in tissue subjected to 0–180 s of heat-shock after wounding. The synthesis of a unique set of proteins called heat-shock proteins (hsps) is induced by these heat-shock treatments. Western-blot analyses of proteins isolated from wounded and heat-shocked Iceberg and Romaine lettuce mid-rib leaf tissue was done using antibodies against hsp 23. Only those heat-shock treatments that were effective at inducing the synthesis of hsp 23 were effective in reducing the activity of PAL induced by wounding and the subsequent accumulation of phenolic compounds. Hsps induced in non-wounded, whole leaves by exposure to 45°C for 150 s did not significantly interact with PAL previously synthesized in non-heat-shocked wounded leaves to limit its activity. The preferential synthesis of hsps over that of wound-induced PAL, rather than the presence of hsps, may be responsible for the ability of a heat-shock treatment to reduce the wound-induced increase in PAL activity. Our results support this novel concept, and the possibility that heat-shock treatments can have significant physiological effects on the response of the tissue to other stresses, not because of the specific genes they induce or repress, or the products they cause to be synthesized, but by their secondary action of influencing the synthesis of other proteins (e.g. PAL) by the suppression of non-hsps protein synthesis. 相似文献
6.
M L Bekker A T Akhmedov V S Smolina S N Naryzhny? 《Molekuliarnaia genetika, mikrobiologiia i virusologiia》1989,(5):15-19
The effect of cellular capsule elimination in Saccharomyces cerevisiae yeasts (protoplast formation) on the heat-shock protein synthesis and the synthesis of the proteins in protoplasts were studied. The methods of mono- and dimeric electrophoresis have demonstrated that (1) about 18 heat-shock proteins with the molecular masses 26-98 Kd are synthesized in cells at 41 degrees C; (2) protoplast formation per se does not induce the synthesis of heat-shock proteins, but the induction of these proteins in protoplasts at 41 degrees C is similar to the one in intact cells. The protoplast formation induces the synthesis of specific proteins different from heat-shock proteins and the synthesis is inhibited by the heat-shock. The heat-shock induces modification of 88 and 86 Kd heat-shock proteins. It inhibits the synthesis of a number of peptides (15-50 Kd) in cells and protoplasts. 相似文献
7.
2D-electrophoresis performed as described by O'-Farrel has revealed clear-cut differences in the pattern of proteins synthesized in the cells of wild-type flies and in the cells of ts-lethal. The cells of the mutant studied after heat-shock exhibit not only the lack of HSP83 and HSP35 but also the absence of a few of the heat-shock proteins belonging to the HSP70 group. Moreover, in the cells of the mutant an intensive synthesis of a protein with mol. weight 72 KDa was observed after heat-shock. This protein belongs to highly abundant heat-shock cognate proteins (HSCP) which are usually not induced by temperature elevation. 相似文献
8.
R B Lal H L Cooper J Coligan T M Chused 《Journal of immunology (Baltimore, Md. : 1950)》1987,138(11):3793-3798
New Zealand Black (NZB) mice exhibit polyclonal B cell activation and elevated immunoglobulin production, an abnormality associated with the spontaneous autoimmune disease that affects this strain. To further our understanding of this abnormality of B cell differentiation and maturation, we have employed two-dimensional polyacrylamide gel electrophoresis to analyze the proteins synthesized by lymphocytes of several strains. Two proteins were produced by lymphocytes from NZB mice but not those from normal strains. One was a 16 kd protein with a pI of 5.1, and the other was a 27.5 kd protein with a pI of 4.5. The presence of the xid gene on the NZB background suppressed production of both proteins. They were synthesized by spleen cells but not by bone marrow or lymph node cells, and production was restricted to enlarged B lymphocytes. p16 was synthesized by normal mouse strain B cells upon stimulation with LPS. The 27.5 kd protein was shown to be secreted. On the basis of partial amino acid sequence determination of proteins eluted from gels, p27.5 was identified as J chain and p16 as the C terminal fragment of mu-chain. The synthesis of two other proteins, 13 kd and 18 kd in size, was elevated in NZB spleen lymphocytes. The 18 kd protein was identified as translation initiation factor eIf-4D. The increased level of this protein may be related to the upregulation of immunoglobulin synthesis. 相似文献
9.
Lymphocytes obtained from pig blood by gradient centrifugation were subjected to a temperature shift (4 to 37 degrees C). The proteins released from the plasma membrane were fractionated by affinity chromatography using immunoglobulin G immobilized on fine polyamide particles. The main component liberated from the adsorbent by diethylamine buffer (pH 11.5) exhibited an apparent Mr of 18000-20000 in SDS-polyacrylamide gel electrophoresis. This crude receptor preparation possessed a substantially higher affinity to immobilized immunoglobulin G than to immobilized Fab fragment and inhibited significantly the binding of labeled immunoglobulin G to pig lymphocytes. 相似文献
10.
The mouse mesenteric lymph node cells (in the cell suspension and frozen sections) were incubated in the soluble immune complexes of peroxidase-antiperoxidase immunoglobulin G. After being washed, they were reacted with diaminobenzidine tetrahydrochloride. Light microscopically brown-colored granules were observed on the cell surface of a proportion of small lymphocytes. In frozen sections, a proportion of small lymphocytes were stained dark brown on the cell surface. Characterization and control experiments suggest that the binding of peroxidase-antiperoxidase immunoglobulin G to the cell surface is mediated by Fc receptor. Peroxidase-antiperoxidase immunoglobulin G, therefore, can be used as in indicator of Fc receptor. 相似文献
11.
The role of suppressor cells in the pathogenesis of common variable hypogammaglobulinemia and the immunodeficiency associated with myeloma. 总被引:8,自引:0,他引:8
T A Waldmann S Broder R Krakauer R P MacDermott M Durm C Goldman B Meade 《Federation proceedings》1976,35(9):2067-2072
The role of suppressor cells in the pathogenesis of immunodeficiency was analyzed using a technique that permits study of the differentiation of B lymphocytes into immunoglobulin-synthesizing plasma cells. Lymphocytes from normals synthesized 4,910 ng of IgM, 1,270 ng of IgA, and 1,625 ng of IgG per 2 X 10(6) cells when cultured for 7 days in the presence of pokeweed mitogen. In contrast the lymphocytes from patients with common variable hypogammaglobulinemia did not synthesize significant quantities of immunoglobulin. When lymphocytes from 9 of 13 patients with common variable hypogammaglobulinemia studied were cocultured with normal lymphocytes, the synthesis of immunoglobulin by the normal lymphocytes was depressed by 75-100%. A comparable suppression of immunoglobulin synthesis by normal lymphocytes was observed when they were cocultured with T cells from hypogammaglobulinemic patients. These studies suggest that in some patients the disease common variable hypogammaglobulinemia may not be due to an intrinsic defect of B cells alone but may be cuased or perpetuated by an abnormality of regulatory T cells that act to suppress B-cell maturation and antibody production. Peripheral blood lymphocytes from myeloma patients also had a drastically reduced capacity to produce polyclonal immunoglobulins. Three of 6 myeloma patients tested had circulating mononuclear cells that suppressed immunoglobulin production by cocultured normal lymphocytes. Purified T cells from myeloma patients did not mediate this suppressor effect. These observations suggest that one mechanism for the humoral immune deficiency observed in myeloma patients is a block of polyclonal B-cell maturation by suppressor cells. 相似文献
12.
Heat shock induced proteins in plant cells 总被引:1,自引:0,他引:1
Thomas Barnett Mitchell Altschuler Carl N. McDaniel Joseph P. Mascarenhas 《Genesis (New York, N.Y. : 2000)》1979,1(4):331-340
Tobacco (Nicotiana tabacum) and soybean (Glycine max) tissue culture cells were exposed to a heat shock and protein synthesis studied by SDS-polyacrylamide gel electrophoresis after labeling with radioactive amino acids. A new pattern of protein synthesis is observed in heat-shocked cells compared to that in control cells. About 12 protein bands, some newly appearing, others synthesized in greatly increased quantities in heat-shock cells, are seen. Several of the heat-shock proteins (HSPs) in both tobacco and soybean are similar in size. One of the HSPs in soybean (76K) shares peptide homology with its presumptive 25°C counterpart, indicating that the synthesis of at least some HSPs may not be due to activation of new genes. The optimum temperature for maximal induction of most HSPs is 39–40°C. Total protein synthesis decreases as heat-shock temperature is increased and is barely detectable at 45°C. The heat-shock response is maintained for a relatively short time in tobacco cells. After 3 hr at 39°C, a decrease is seen in the synthesis of the HSPs, and after 4 hr practically no HSPs are synthesized. After exposure to 39°C for 1 hr, followed by a return of tobacco cells to 26°C, recovery to the control pattern of synthesis requires greater than 6 hours. These results indicate that cells of flowering plants exhibit a heat-shock response similar to that observed in animal cells. 相似文献
13.
Massive heat-shock polypeptide synthesis in late chicken embryos: convenient system for study of protein synthesis in highly differentiated organisms. 总被引:10,自引:6,他引:4
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In cultured eucaryotic cells, heat treatments specifically induced the rapid synthesis of the so-called heat-shock polypeptides. To ascertain the physiological importance of this phenomenon for highly differentiated organisms, we attempted to determine whether the heat-shock response occurs in a living endothermic organism at extreme temperatures, and if so, whether the response is organ specific. We developed a procedure to label proteins efficiently in 5- to 18-day-old chicken embryos. Heat-shock polypeptides of identical sizes of 85,000, 70,000, and 25,000 daltons were synthesized predominantly in chicken embryo fibroblasts and in many different organs of 18-day-old embryos at 42.5 to 44 degrees C. 相似文献
14.
T lymphocyte stress response. II. Protection of translation and DNA replication against some forms of stress by prior hyperthermic stress 总被引:1,自引:0,他引:1
We have compared the effects of a mild heat shock and febrile temperatures on heat-shock protein (hsp) synthesis and development of stress tolerance in T lymphocytes. Our previous studies demonstrated that febrile temperatures (less than or equal to 41 degrees C) induced the synthesis of hsp110, hsp90, and the constitutive or cognate form of hsp70 (hscp70; a weak induction of the strongly stress-induced hsp70 was also observed. In the studies reported herein, we demonstrate that a mild heat shock (42.5 degrees C) reverses this ratio; that is, hsp70 and not hscp70 is the predominate member of this family synthesized at this temperature. Modest heat shock also enhanced the synthesis of hsp110 and hsp90. In order to assess the relationship between hsp synthesis and the acquisition of thermotolerance, purified T cells were first incubated at 42.5 degrees C (induction temperature) and then subsequently subjected to a severe heat-shock challenge (45 degrees C, 30 min). T cells first incubated at a mild heat-shock temperature were capable of total protein synthesis at a more rapid rate following a severe heat shock than control cells (induction temperature 37 degrees C). This phenomenon, which has been previously termed translational tolerance, did not develop in cells incubated at the febrile temperature (induction temperature 41 degrees C). Protection of translation also extended to immunologically relevant proteins such as interleukin-2 and the interleukin-2 receptor. Because clonal expansion is a critical event during an immune response, the effects of hyperthermic stress on DNA replication (mitogen-induced T cell proliferation) was also evaluated in thermotolerant T cells. DNA synthesis in control cells (induction temperature 37 degrees C) was severely inhibited following heat-shock challenge at 44 degrees C or 45 degrees C; in contrast, T cells preincubated at 42.5 degrees C rapidly recovered their DNA synthetic capacity. T cells preincubated at a febrile temperature were moderately protected against hyperthermic stress. The acquisition of thermotolerance was also associated with enhanced resistance to chemical (ethanol)-induced stress but not to heavy metal toxicity (cadmium) or dexamethasone-induced immunosuppression. These studies suggest that prior hsp synthesis may protect immune function against some forms of stress (e.g., febrile episode) but would be ineffective against others such as elevated glucocorticoid levels which normally occur during an immune response. 相似文献
15.
Xenopus laevis oocytes respond to high temperature (>31°C) by the synthesis of one major (70 kilodalton) protein and by a gradual reduction in the rate of normal protein synthesis. In contrast with most other cells, the heat-shock response of Xenopus oocytes is controlled exclusively at the translational level. Enucleated or α-amanitin-injected oocytes synthesize normal levels of heat-shock protein. Thus high temperature induces the translation of preformed heat-shock mRNA. This continues for more than a day after a shift back to a normal temperature, but ceases within 2 days. Heat-shock protein synthesis can be sequentially induced and inactivated in the same oocyte over several days. We conclude that an oocyte contains 10–100 pg of heat-shock mRNA, which is synthesized during oogenesis at the normal temperature, and which is stored in an inactive state by a “masking” mechanism. 相似文献
16.
R A Colbert J A Hucul K A Scorsone D A Young 《The Journal of biological chemistry》1987,262(35):16763-16766
The use of ultra high resolution giant two-dimensional gel electrophoresis has expanded the number of recognizable heat-shock proteins to 68 inductions in rat thymic lymphocytes, many of which are among the less abundant cellular proteins (Maytin, E. V., Colbert, R. A., and Young, D. A. (1985) J. Biol. Chem. 260, 2384-2392). Previous studies also show that cells receiving a prior heat shock recover more rapidly from the inhibition of protein synthesis induced by a second heat shock. In this report we use a monoclonal antibody to identify the alpha subunit of eukaryotic initiation factor-2 (eIF-2 alpha) as a heat-shock protein. Its relative rate of synthesis increases approximately 40% in the 2nd h and 5-fold in the 4th h of a continuous heat shock and is stimulated more dramatically, 15-fold, in the 3rd h of recovery from a 1-h heat shock. These results suggest that the induction of eIF-2 alpha in the heat-shock response may be important for restoring the cell's ability to initiate protein synthesis. In addition to identifying a function for one of the heat-shock proteins, our findings draw attention to the likelihood that other low-abundance heat-shock proteins may play critical roles in the heat-shock response. 相似文献
17.
A high proportion of H2 antigen-activated thymus-derived thoracic duct lymphocytes stained positively with rabbit anti-mouse immunoglobulin reagents as detected by indirect immunofluorescence. In view of the specificity of the reagents used and the fact that T cells from other sources e.g., thymus, were not stained by this technique, it was concluded that the material detected on H-2 antigen-activated thymus-derived thoracic duct lymphocytes was indeed immunoglobulin. When H-2 antigen-activated thymus-derived thoracic duct lymphocytes were cultured in vitro at 37 °C for 18 hr, with or without prior treatment with chymotrypsin, surface immunoglobulin could no longer be detected on the cells. This suggested that immunoglobulin had not been synthesized by the cells but absorbed from elsewhere. 相似文献
18.
Asymmetric segregation of heat-shock proteins upon cell division in Caulobacter crescentus 总被引:15,自引:0,他引:15
Three Caulobacter crescentus heat-shock proteins were shown to be immunologically related to the Escherichia coli heat-shock proteins GroEL, Lon and DnaK. A fourth heat-shock protein was detected with antibody to the C. crescentus RNA polymerase. This 37,000 Mr heat-shock protein might be related to the E. coli 32,000 Mr heat-shock sigma subunit. The synthesis of the major C. crescentus RNA polymerase sigma factor was not induced by heat shock. The E. coli GroEL protein and the related protein from C. crescentus were also induced by treatment with hydrogen peroxide. Like some of the proteins in the heat-shock protein families of Drosophila and yeast, the four heat-shock proteins in C. crescentus were found to be regulated developmentally under normal conditions. All four proteins were synthesized in the predivisional cell, but the progeny showed cell type-specific bias in the level of enhanced synthesis after heat shock. The 92,000 Mr Lon homolog and the 37,000 Mr RNA polymerase subunit were preferentially synthesized in the stalked cell, whereas the synthesis of the 62,000 Mr GroEL homolog was enhanced in the progeny swarmer cell. Furthermore, the four heat-shock proteins synthesized in the predivisional cell were partitioned in a specific manner upon cell division. The stalked cell, which initiates chromosome replication immediately upon division, received the Lon homolog, the DnaK homolog and the 37,000 Mr RNA polymerase subunit. The GroEL homolog, however, was distributed equally to both the stalked cell and the swarmer cell. These results provide access to the functions of C. crescentus heat-shock proteins under both normal and stress conditions. They also allow an investigation of the regulatory signals that modulate the asymmetric distribution of proteins and their subsequent cell type-specific expression in the initial stages of a developmental program. 相似文献
19.
J. Schwaber 《Experimental cell research》1975,93(2):343-354
The fusion of human lymphocytes and TEPC-15 mouse myeloma cells, which had not been adapted to culture, resulted in the establishment of in vitro hybrid cell cultures. Ten clones of this somatic cell hybrid were examined. There was preferential exclusion of human chromosomes: between two and five human chromosomes were identified in the hybrid clones by Giemsa banding. All of the clones had the mouse parental histocompatibility antigens, but only four clones also retained the human parental histocompatibility antigens. Secretion of parental immunoglobulin was determined by SDS-gel electrophoresis of species-specific immune precipitates. Synthesis of parental immunoglobulin by individual hybrid cells was determined by double label fluorescent antibody staining. Individual cells from six of the clones secreted and synthesized both human and mouse parental immunoglobulins. Three clones secreted only one parental immunoglobulin. Cells from one of these clones secreted and synthesized only human immunoglobulin. Cells from the remaining two clones secreted only one parental species of immunoglobulin but synthesized both human and mouse immunoglobulins. Finally, one clone did not secrete immunoglobulin, yet the individual cells synthesized both human and mouse parental species of immunoglobulin. 相似文献
20.