W B Ershler, W H Sun, N Binkley, S Gravenstein, M J Volk, G Kamoske, R G Klopp, E B Roecker, R A Daynes, R Weindruch
{"title":"Interleukin-6 and aging: blood levels and mononuclear cell production increase with advancing age and in vitro production is modifiable by dietary restriction.","authors":"W B Ershler, W H Sun, N Binkley, S Gravenstein, M J Volk, G Kamoske, R G Klopp, E B Roecker, R A Daynes, R Weindruch","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Interleukin-6 (IL-6) is a multifunctional cytokine that is proving to be a major contributor to the acute phase inflammatory response. IL-6 expression is normally low and serum levels are usually nondetectable in the absence of inflammation. With advancing age, however, serum levels become detectable and it is proposed that this reflects an age-associated loss in the normal regulation of gene expression for this molecule. There is also speculation that IL-6 may contribute to the pathogenesis of several diseases that are common in late-life including lymphoma, osteoporosis, and Alzheimer's disease. In this report we demonstrate that plasma levels of IL-6 rise with advancing age in well-selected healthy elderly people and comparably in old rhesus monkeys. That this change reflects a primary aging process is suggested by our findings in C57BL/6 mice in which the age-associated increase in the in vitro synthesis of IL-6 is largely prevented by life span-extending dietary restriction.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 4","pages":"225-30"},"PeriodicalIF":0.0,"publicationDate":"1993-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19208366","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The functional state of the beta cell modulates IL-1 and TNF-induced cytotoxicity.","authors":"V Mehta, W Hao, B M Brooks-Worrell, J P Palmer","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Insulin-dependent diabetes is an autoimmune disease specifically targeting the pancreatic beta cells and several observations, both experimental and clinical, suggest that the interaction of the immune system with the beta cells is in part determined by the functional state of the target cells, increased beta cell activity resulting in augmented immunologic mechanisms and vice versa for suppressed beta cell activity and decreased immune attack. In this study we investigated whether cytokine induced islet cell cytotoxicity in vitro was in part dependent on the functional state of the beta cells. Cytotoxicity of cultured rat islets was induced by IL-1 (100 pg/ml) and TNF (62.5 ng/ml) individually and in combination and beta cell activity was modulated by culturing the islets in media containing 3.3, 5.5, 11, and 20 mmol/liter glucose. Both IL-1 and TNF were cytotoxic when administered individually and the combination of IL-1 and TNF was more cytotoxic than either cytokine alone. Maximum cytotoxicity was observed at 11 mmol/liter glucose with cytotoxicity being reduced at 5.5 mmol/liter glucose and further reduced at 3.3 mmol/liter glucose. Interestingly, the degree of cytotoxicity was lower in 20 mmol/liter glucose compared to 11 mmol/liter. These results firmly establish that islet cytotoxicity of IL-1 and TNF is highly dependent on the functional state of the beta cells. This suggests that during the IDDM disease process as some beta cells are destroyed, the compensatory increased activity of the remaining beta cells may increase their susceptibility to cytokine attack. Furthermore, our observations provide rational support for the use of beta cell rest as intervention therapy for IDDM.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 4","pages":"255-9"},"PeriodicalIF":0.0,"publicationDate":"1993-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19208369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M Gatanaga, E A Grosen, R A Burger, G A Granger, T Gatanaga
{"title":"Release of soluble TNF/LT receptors from a human ovarian tumor cell line (PA-1) by stimulation with cytokines in vitro.","authors":"M Gatanaga, E A Grosen, R A Burger, G A Granger, T Gatanaga","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We have demonstrated the presence of the 55- and 75-kDa receptor for tumor necrosis factor (TNF) and lymphotoxin (LT) (TNF-R) in serum, and ascites from women with ovarian cancer. The present studies were initiated to begin to examine the possible cellular source of these receptors in women with ovarian cancer. Human ovarian tumor cells (PA-1) were cocultured for 24-48 hr with various levels of recombinant human cytokines (IL-1 beta, IL-4, IFN-gamma) and the supernatants were assayed by ELISA for the soluble forms of each receptor. PA-1 cells spontaneously release the 55-kDa TNF-R and low levels of the 75-kDa TNF-R. The release of both 55- and 75-kDa TNF-R was stimulated when PA-1 cells were cultured with IL-1 beta and IFN-gamma but unaffected by IL-4. The level of 55-kDa TNF-R was elevated slightly over spontaneous release but the level of 75-kDa TNF-R increased dramatically. IFN-gamma was the most potent stimulator of receptor release particularly of the 75-kDa TNF-R. IFN-gamma also induced increased expression of cell membrane TNF-R measured by binding of 125I-labeled TNF. Membrane TNF-Rs which were induced by IFN-gamma were the 75-kDa type, because TNF binding was blocked by anti-75-kDa TNF-R antibody. These data suggest that IFN-gamma selectively induced release and expression of 75-kDa TNF-Rs.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 4","pages":"249-53"},"PeriodicalIF":0.0,"publicationDate":"1993-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19208368","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"TR60 and TR80 tumor necrosis factor (TNF)-receptors can independently mediate cytolysis.","authors":"M Grell, P Scheurich, A Meager, K Pfizenmaier","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The human rhabdomyosarcoma cell line KYM-1 coexpresses high numbers of both tumor necrosis factor-alpha (TNF) receptors, TR60 and TR80, and is highly sensitive to TNF-induced cytotoxicity. We show here that each receptor type can mediate cytotoxicity on its own on selective stimulation. This was achieved using receptor specific antibodies, able to compete with ligand binding to the respective receptor molecules. Thus, the TR60-specific monoclonal antibody H398 was strongly agonistic, i.e., cytotoxic, in the presence of a secondary cross-linking immunoglobulin. Selective stimulation of TR80 by antibody-mediated receptor crosslinking also induced strong cytotoxicity in KYM-1 cells. Interestingly, when both receptor subsets were stimulated in parallel by limited receptor cross-linking, additive cytolytic effects were observed. In each case the respective Fab fragments showed no agonistic activity. When the natural ligand TNF was used to induce cytolysis, blocking studies with receptor specific Fab fragments indicated that the main cytotoxic effect of TNF is mediated via TR60, although these receptors represent only about 8% of the total TNF receptor number.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 3","pages":"143-8"},"PeriodicalIF":0.0,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19379114","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A Moutabarrik, I Nakanishi, M Namiki, T Hara, M Matsumoto, M Ishibashi, A Okuyama, D Zaid, T Seya
{"title":"Cytokine-mediated regulation of the surface expression of complement regulatory proteins, CD46(MCP), CD55(DAF), and CD59 on human vascular endothelial cells.","authors":"A Moutabarrik, I Nakanishi, M Namiki, T Hara, M Matsumoto, M Ishibashi, A Okuyama, D Zaid, T Seya","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Membrane cofactor protein (MCP), decay-accelerating factor (DAF), and CD59 are complement (C) regulatory proteins that protect human hematopoietic cells from damage induced by autologous complement. Using monoclonal antibodies in both western blotting studies and flow cytometry, we found that endothelial cells (EC) expressed on their surface MCP, DAF, and CD59 molecules. We tested whether stimulation of endothelial cells by known EC activators such as LPS, TNF-alpha, IL-1 beta, and IL-4 regulates the expression of these C regulatory proteins. EC activation was assessed in this study by an up-regulation of ICAM-1 expression. Treatment of EC with various concentrations of TNF-alpha, IL-1 beta, or IL-4, at different incubation times did not increase MCP expression on EC surface membrane. In contrast, the level of DAF was altered during EC activation: LPS or IL-1 beta treatment resulted in a slight increase of DAF expression; the most up-regulatory effect was obtained with IL-4. Up-regulation of surface DAF on EC required prolonged treatment of cells with these agents. The level of CD59 was far greater than that of DAF or MCP on EC, and was slightly up-regulated by TNF-alpha and down-regulated by IL-1 beta. These findings indicate that the levels of C regulatory proteins are regulated in an independent fashion on EC, MCP is not regulated by any cytokine tested, while DAF is an EC activation antigen, although it has a different augmentation profile from ICAM-1 since IL-4 up-regulates DAF but not ICAM-1.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 3","pages":"167-72"},"PeriodicalIF":0.0,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18692823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M A Walter, R Kurouglu, J B Caulfield, L O Vasconez, J A Thompson
{"title":"Enhanced peripheral nerve regeneration by acidic fibroblast growth factor.","authors":"M A Walter, R Kurouglu, J B Caulfield, L O Vasconez, J A Thompson","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Transection of peripheral nerves may cause permanent denervation with paralysis and disability in humans and represents a challenging problem in microsurgery. Physiologic repair at increasing intervals after the acute phase of injury results in progressively worse recovery, emphasizing the importance of rapid and timely reinnervation to optimize endorgan viability. Despite recent advances in microsurgical techniques, imperfect reinnervation results in partial return of neuromuscular function, even in the mildest neuropraxias. Axonal repair of mature neurons involves a complex interaction of molecular events, suggesting that the presence of specific neuronotropic factors might enhance the regeneration process. Recombinant human fibroblast growth factor (FGF-1) has been shown to induce both rapid angiogenesis and neurogenesis through a synthetic conduit across a 15-mm surgical gap in the peripheral nerve of the rat. Evidence of newly formed neural structures was confirmed postoperatively by histological examination in a temporal fashion over a 24-week interval. Functional motor recovery of regenerated nerves was evaluated by determining the amplitude and latency of compound muscle action potentials in treated animals. Electrophysiology studies demonstrated consistent return of motor function in 43 and 57% of animals harboring an FGF-1 conduit at 8- and 24-week intervals, respectively. None of the control animals exhibited restoration of motor function. Collectively, these data suggest that FGF may serve as an important mediator of controlled growth during peripheral nerve regeneration.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 3","pages":"135-41"},"PeriodicalIF":0.0,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18690528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
B B Aggarwal, K Graff, B Samal, M Higuchi, W S Liao
{"title":"Regulation of two forms of the TNF receptors by phorbol ester and dibutyryl cyclic adenosine 3',5'-monophosphate in human histiocytic lymphoma cell line U-937.","authors":"B B Aggarwal, K Graff, B Samal, M Higuchi, W S Liao","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Recently the cDNA for two different forms of TNF receptor, with gene products of molecular masses of 60 and 80 kDa, have been cloned. In the present report, we investigated the effects of phorbol ester and dibutyryl cAMP on the regulation of the transcript for each type of TNF receptor in U-937 cells. Our results indicate that exposure of these cells to either phorbol ester or dibutyryl cAMP increases the steady state mRNA levels of the 80 kDa form. This effect is dose- and time-dependent. The induction of the p80 receptor transcript by PMA and dibutyryl cAMP was additive suggesting independent mechanisms of induction. Under identical conditions, both agents failed to induce the transcript for the p60 form of the TNF receptor. As demonstrated by actinomycin D pulse-chase experiment, the mRNA for the p80 receptor was found to be highly stable with an approximate half-life of 16 h. No significant change in the half-life was observed when cells were treated with phorbol ester. The mechanisms by which phorbol ester and dibutyryl cAMP induce the upregulation of p80 receptor mRNA appear to be different. Induction of receptor transcript by cycloheximide suggests the presence of a labile repressor protein. Interestingly, the effect of cycloheximide on the induction of the p80 mRNA was found to be additive with that of dibutyryl cAMP but not with phorbol ester. 1-(5-Isoquinolinylsufonyl)-2-methylpiperazine (H7) and N[2-(methylamino) ethyl]-5-isoquinolinesulfonamide (H-8), inhibitors of protein kinase C and protein kinase A, respectively, both inhibited the phorbol ester-mediated induction of the p80-transcript but not that mediated through dibutyryl cAMP. Since dibutyryl cAMP undergoes intracellular dissociation into cAMP and butyric acid, we found that exposure of cells to sodium butyrate alone could induce p80 mRNA in a dose-dependent manner, thus suggesting the role of histone hyperacetylation. Furthermore forskolin treatment, an intracellular inducer of cAMP, increased the receptor transcript level whereas isobutylmethylxanthine, an inhibitor of phosphodiesterase, had no effect. Interestingly, while the p80 form of the TNF receptor mRNA levels was elevated by both phorbol ester and dibutyryl cAMP, only dibutyryl cAMP increased the TNF binding; phorbol ester treatment decreased the binding activity. Thus, our results demonstrate that the genes for the two forms of TNF receptors are differentially regulated. Furthermore, the mechanism of regulation by PMA differs from that by dibutyryl cAMP.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 3","pages":"149-58"},"PeriodicalIF":0.0,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18690529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The effects of thermal injury on serum interleukin 1 activity in rats.","authors":"A C Drost, B Larsen, L H Aulick","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Interleukin 1 (IL-1) is present in the blood of burned patients but its pathophysiologic role is not fully understood. Rat models would be useful research tools, if this cytokine could be identified in a complex fluid like blood. We describe a methodology, which revealed IL-1 activity from the serum of burned rats. Serum was collected from 37 rats with 30% total body surface burns and 9 unburned controls. To vary the burn response, the wounds of 17 rats were seeded with nonvirulent Pseudomonas aeruginosa at the time of injury. IL-1 activity was assessed by its capacity to induce IL-2 secretion in murine lymphoma cells (LBRM-33-1A5). Only after the serum had been fractionated, concentrated, and dialyzed, was IL-1 activity uncovered. Sera from burned rats contained five times more IL-1 activity than those from control animals (p < 0.05). There was no difference in serum IL-1 activity between burned and burn-seeded animals. The IL-1 activity was heat labile, and not produced by P. aeruginosa endotoxin, TNF-alpha, or endogenous IL-2 in rat serum. These results confirm that serum IL-1 levels are increased following thermal injury, and that there is no apparent relationship between IL-1 levels and infection. The serum preparation scheme presented in this study offers a reasonable approach to the measurement of serum IL-1 levels in rat models of disease and injury.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 3","pages":"181-5"},"PeriodicalIF":0.0,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19333612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H Ikegami, N Arai, J Moriyasu, M Taniguchi, K Tsusaki, N Honji, K Kohno, S Ando, M Kurimoto
{"title":"Analysis of the site on a TNF-alpha molecule which affects type II TNF receptor binding in human cells.","authors":"H Ikegami, N Arai, J Moriyasu, M Taniguchi, K Tsusaki, N Honji, K Kohno, S Ando, M Kurimoto","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The epitope region on the TNF-alpha molecule recognized by monoclonal antibody (mAb) 3-D-6, which neutralizes the cytotoxic activity on murine LM cells, has been determined as Gly24-Gln-Leu-Gln-Trp-Leu-Asn-Arg31. To examine whether this region participates in TNF receptor binding in human cell lines, four kinds of TNF-alpha mutants (Gln25 --> Glu, Gln27 --> Glu, Leu29 --> Val, and Arg31 --> Ser) were prepared using site-directed mutagenesis. One mutant, mRS31, which has a nonconserative mutation at position 31 (Arg --> Ser), showed markedly reduced binding in U-937 cells and in HL-60 cells compared with the wild-type recombinant TNF-alpha (rTNF-alpha). These two cell lines have been reported to have both type I and type II TNF receptors. mRS31 also showed reduced cytotoxicity on U-937 cells. Another mutant, mLV29, which has a conservative mutation at position 29 (Leu --> Val), showed, to a lesser extent, reduced binding in U-937 cells and HL-60 cells and reduced cytotoxic activity in U-937 cells. However, all four TNF-alpha mutants showed a similar binding in HEp-2 cells and in HeLa cells, which have been reported to have only the type I TNF receptor. These results suggest that Leu29 may be involved in direct contact with the type II receptor and that the nonconservative mutation at position 31 may induce a local conformational change in the site involved in type II TNF receptor binding.(ABSTRACT TRUNCATED AT 250 WORDS)</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 3","pages":"173-80"},"PeriodicalIF":0.0,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18692824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S Morecki, A Nagler, Y Puyesky, C Nabet, R Condiotti, M Pick, S Gan, S Slavin
{"title":"Effect of various cytokine combinations on induction of non-MHC-restricted cytotoxicity.","authors":"S Morecki, A Nagler, Y Puyesky, C Nabet, R Condiotti, M Pick, S Gan, S Slavin","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Efforts were directed to achieve an increased lymphokine-activated non-MHC-restricted killing (LAK) activity greater than that induced by rIL-2 alone. Human peripheral blood (PB) and bone marrow (BM)-derived mononuclear cells (MC) were exposed in vitro to multiple cytokine combinations, including rIL-6, rIL-7, rIFN-alpha and rIFN-gamma in the presence of either suboptimal or optimal doses of rIL-2. Our results have shown that BMMC are a potential source for induction of increased LAK activity upon exposure to multiple cytokine combinations, whereas PBMC could not be successfully stimulated under the same conditions. Fifty-five to 62% of BM-derived samples stimulated with high dose rIL-2 + rIL-7 or rIL-2 + rIL-7 + rIL-6 + rIFN-gamma exhibited a higher degree of cytotoxicity than BM samples stimulated with rIL-2 alone. Exposure of PB-derived large granular lymphocytes (LGL) to various cytokine combinations led to increased proliferation after stimulation with suboptimal dose of rIL-2 in the presence of rIL-6 and rIL-7. This increase was not observed in induction of cytotoxicity. We suggest that BMMC activated by multiple cytokine combinations could play an active role in improving antitumor response in vivo by contributing to the control of minimal residual tumor cell growth, particularly post-BM transplantation.</p>","PeriodicalId":77246,"journal":{"name":"Lymphokine and cytokine research","volume":"12 3","pages":"159-65"},"PeriodicalIF":0.0,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19333610","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}