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PubMed1989

High interleukin-6 serum levels and increased production by leucocytes in alcoholic liver cirrhosis. Correlation with IgA serum levels and lymphokines production.

Hypergammaglobulinaemia and enhanced serum IgA levels are common in alcoholic liver cirrhosis. Interleukin-6 (IL-6), which is identical to B cell differentiation factor BSF2 and is implicated in various autoimmune diseases, has been studied in patients with alcoholic liver cirrhosis. Increased serum levels and spontaneous or induced production of IL-6 by peripheral blood monoclonal cells have been found. IL-6 production correlates closely with IgA serum levels and negatively with impaired interleukin-2 and interferon gamma production. This abnormality could be related to overproduction of immunoglobulins and immune disturbances observed in this disease.

Female↗
PubMed1989

Regulation of murine lymphokine production in vivo. Ultraviolet radiation exposure depresses IL-2 and enhances IL-4 production by T cells through an IL-1-dependent mechanism.

The exposure of experimental animals to the inflammatory effects of ultraviolet radiation (UVR) is known to cause depressions in their ability to initiate and effectuate various types of cellular immune responses. Contact-type and delayed-type hypersensitivity, plus the ability to generate protective forms of anti-viral and anti-tumor immunity, are all affected by the prior exposure of normal animals to the effects of this physical agent. Presently, the cellular and molecular mechanism(s) responsible for mediating the changes in immune function observed in UVR-exposed animals is not fully understood. Herein we report that one reproducible consequence of exposing normal mice to low doses of UVR is a dramatic change in the pattern of lymphokines secreted by their activated T cells. Lymphocytes isolated from UVR-exposed donors produce/secrete greatly reduced levels of the T cell lymphokines IL-2 and IFN-gamma activation in vitro with protein Ag of the polyclonal T cell stimulant anti-CD3. The secretion of IL-4 by these lymphocyte cultures, however, is consistently elevated in comparison to normal controls. Further studies determined that a similar change in lymphokine production was induced when mice were treated with either bacterial LPS or rIL-1 beta, a cytokine known to be elevated in vivo after UVR or LPS exposure. The ability of IL-1 to facilitate a change in the capacity of T lymphocytes to produce/secrete lymphokines after in vitro activation does not appear to represent a direct effect of this cytokine on lymphocyte or accessory cell targets because addition of IL-1 beta to cultures of Ag-primed lymphocytes obtained from normal donors was incapable of altering the pattern of lymphokine production. Collectively, our present results add further support to the hypothesis that UVR-induced elevations in endogenous IL-1 are, in part, responsible for the immunomodulatory effects of UVR. These findings provide compelling evidence that UVR, plus other agents capable of endogenously stimulating the production of IL-1, may function to alter the expression of different effector mechanisms in vivo. This could be facilitated through selective reductions in lymphokines produced by Th-1-type cells (IL-2 and IFN-gamma) and a simultaneous augmentation in a lymphokine produced by Th-2-type cells (IL-4).

Animals↗
PubMed1989

Cis-urocanic acid stereospecifically modulates human monocyte IL-1 production and surface HLA-DR antigen expression, T-cell IL-2 production and CD4/CD8 ratio.

UV irradiation is known to photoisomerize epidermal trans-urocanic acid (trans-UCA) to cis-urocanic acid (cis-UCA), which has been postulated to be involved in local and systemic downregulation of immune responses. We have earlier shown that cis-UCA suppresses interleukin 1 (IL-1) production in human epidermal cells. To study the possible effects of UCA isomers on human peripheral blood lymphoid cells, these cells were cultured in the presence of either UCA stereoisomer, and a number of immunological parameters were assayed. Cis-UCA (100 micrograms/ml) caused a significant downregulation of monocyte IL-1 production, and diminished monocyte HLA-DR expression. Cis-UCA also caused a significant reduction in the CD4/CD8 ratio. Furthermore, T-cells preincubated with cis-UCA caused a significant downregulation of purified protein derivative-induced interleukin 2 production by autologous T-cells. The trans isomer had no effect in any of these in vitro tests. The reported stereospecific effects of cis-UCA are compatible with the postulated function of this chemical as an UV-induced, low-molecular-weight immunomediator substance.

Cells, Cultured↗
PubMed1989

Regulation of isotype production by IL-4 and IL-5. Effects of lymphokines on Ig production depend on the state of activation of the responding B cells.

The effects of IL-4 and IL-5 on the production of Ig of different isotypes was investigated. We compared B cells from spleen and from Peyer's patches either stimulated with LPS or without added polyclonal stimulation. We also compared high density (small) and low density (large) B cells. The effect of lymphokines depended on the size and source of the B cells as well as on whether LPS was added. As expected, small B cells from either lymphoid compartment responded to LPS alone and IL-4 suppressed IgM and IgG3 production and enhanced IgG1. In contrast, when large B cells were examined, the suppressive effects of IL-4 were much less apparent but the enhancement of IgG1 was still marked. IL-5 alone had only minimal effects in LPS-stimulated cultures but the combination of IL-4 plus IL-5 appeared to overcome much of the IL-4-mediated suppression of IgM, and IgA production was enhanced. In the absence of LPS, a quite different profile is seen. First, small B cells make little if any response. Second, there is dramatic synergy between IL-4 and IL-5 in the response of large B cells, which is independent of isotype. Third, IL-4 does not suppress any isotype in the absence of LPS. Fourth, IL-4 plus IL-5 stimulate large Peyer's patch B cells to produce 10 times more IgA but three times less IgM than large spleen B cells. Fifth, Th2 cells directly stimulate both large and small B cells.

Adjuvants, Immunologic↗
PubMed1986

Increased production of peroxidation products associated with cardiac operations. Evidence for free radical generation.

We investigated the degree and time course of neutrophil sequestration into human lungs during cardiac operations. At the same time, measurement of the concentration of peroxidation products in the plasma was used as an index of oxidant free radical activity. The study was performed in two groups of patients. Group A (n = 11) had studies extending over the entire operative period and showed a highly significant sequestration of neutrophils into the lung, together with a highly significant (p less than 0.001) rise in peroxidation products from 2.8 +/- 0.12 nmol/ml(mean +/- standard error of the mean)before bypass to a peak of 5.05 +/- 0.13 nmol/ml at the end of bypass. As these changes occurred only during the time after release of the aortic cross-clamp, we investigated this period in more detail in a second group of patients (Group B, n = 7). Results from this group showed that significant release of peroxidation products occurred at the same time as pulmonary neutrophil sequestration. This study has produced evidence of increased oxidant activity in the lung associated with cardiac operations. Nevertheless, it is not known whether the neutrophils sequestered into the lung alone induced the increased activity. Similarly, whether neutrophil-derived oxidant species are the sole cause of lung tissue injury remains unproved.

Cardiopulmonary Bypass↗
PubMed1985

Problems involved in the large scale production of biological products, such as beta-interferon, using diploid fibroblast cells as substrate.

The large-scale and continuous cycle production of Hu beta-IFN serves to indicate that with expansion of cell cultures, aspects such as the reliability of the growth media and the possibility of contamination of cultures may become critical and limiting factors for the production itself. Methods of growth media preparation are described, with particular stress on the problems related to bacterial contamination and water quality. Our production method involves the expansion of cellular cultures using roller bottles, and a final step using the microcarrier technique. The necessity of collecting the cultures in only two fermentors increases the possibilities of contamination. Methods employed in order to minimize these problems are discussed.

Cells, Cultured↗
PubMed1984

Production of arachidonic acid lipoxygenase products during platelet-neutrophil interactions.

We studied interactions of human platelets and neutrophils with particular reference to the arachidonic acid pathway. Suspensions of [3H]-arachidonate-labeled platelets and unlabeled neutrophils were stimulated with ionophore A23187. We detected several radioactive arachidonate metabolites, which are not produced by platelets alone. This included [3H]-labeled leukotriene B4 (LTB4), dihydroxyeicosatetraenoic acid (DHETE) and 5-hydroxyeicosatetraenoic acid (5-HETE). When the platelet product, [3H]12-HETE, was added to ionophore-stimulated neutrophils, DHETE was formed. In addition, when [3H]5-HETE, a neutrophil arachidonate product, was added to stimulated platelets, DHETE was the major metabolite. We, therefore, suggest that upon stimulation, the platelet derived arachidonate can serve as precursor for the neutrophil-derived eicosanoids LTB4 and 5-HETE and that the platelet-derived product 12-HETE can be metabolized to DHETE by stimulated human neutrophils. It would appear that the platelet and neutrophil lipoxygenase pathways take part in cell-cell interactions - an observation which suggests a role for the neutrophils which are present in hemostatic plugs, thrombi and inflammatory processes.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1981

Regulation of T cell growth factor production: arrest of TCGF production after 18 hours in normal lectin-stimulated mouse spleen cell cultures.

The detailed kinetics of TCGF accumulation in Con A-stimulated spleen cell cultures shows a maximum at 24 hr, with a subsequent decrease in activity. This decrease is not due to the appearance of inhibitory substances "masking" TCGF activity. Pulse experiments show that the rate of TCGF production falls sharply after 18 hr and is completely arrested after 24 hr of Con A stimulation. The arrest in TCGF production is the result neither of culture depletion in medium components nor of limiting accessory cell function or inactivation of the lectin, and it thus seem to be the result of inactivation of TCGF-producing T cells. This regulation is not the result of a TCGF-mediated feedback mechanism but rather of lectin-induced suppressive cells that appear in culture after 24 hr and turn off de novo production of TCGF in fresh cultures.

Animals↗
PubMedDate not supplied

[Mass spectrometric data on some drugs, their biotransformation products and intermediate products of illegal synthesized narcotics].

Additional mass spectral data of some rather unusual drugs and their biotransformation-products are presented updating the three large and authoritative mass spectra data collections for forensic/clinical toxicological purposes published by Finkle, Foltz and Saferstein et al. Furthermore mass-spectra data of chemicals, intermediates, by-products and final products of illegal drug syntheses are summarized in one chapter.

Biotransformation↗
PubMedDate not supplied

Lipases production by Bacillus circulans under mesophilic and osmophilic conditions. Factors affecting lipases production.

A facultative osmophilic bacterium able to produce lipases (isolated from bee honey) was identified as Bacillus circulans. Factors affecting lipases production by such isolate were studied. Dox's liquid medium supplied with 2 and/or 30% sucrose was used as a basal medium, under static culture condition, for such a purpose. Results have showed that the optimal sucrose concentration at which the isolate produced its maximal yield of lipases was 30% after an incubation period of 24 hours at 40 degrees C, and this was corresponding to an incubation period of 48 hours at 40 degrees C in presence of 2% sucrose. The optimal pH value was 7 in the first case while, 6.5 in the second. Furthermore production of extracellular lipases by such isolate was achieved in presence and absence of tributyrin as a carbon source instead of sucrose when supplied to Dox's liquid medium. Also the production of extracellular lipases by such isolate was not enhanced in presence of tributyrin as a supplement to Dox's liquid medium when supplied with 2 and/or 30% sucrose as carbon sources. This might suggest that the extracellular lipases of such isolate is constitutive in nature.

Bacillus↗
PubMedDate not supplied

Endogenous nitric oxide production by human monocytic cells regulates LPS-induced TNF production.

The ability to produce nitric oxide (NO) of human monocytes macrophages is object of debate. While studying the regulation of tumor necrosis factor (TNF) synthesis induced by endotoxin (LPS) in a human cell line of monocyte origin (THP-1) and in human peripheral blood mononuclear cells (PBMC) we found an indirect evidence of such production. We showed that L-N-monomethyl-arginine (L-NMMA), an inhibitor of NO synthase, and hemoglobin, a chelator of NO, are able to significantly reduce TNF synthesis, indicating that NO production is induced by LPS and contributes to the induction of TNF. Since NO is a known cytostatic agent, we also studied the cytostatic effect of LPS, and demonstrated that it is reverted by L-NMMA. Although we were unable to show any nitrites/nitrates accumulation in the culture media, taken together our data give an indirect evidence of a physiologically relevant LPS-induced NO production in human monocytes-macrophages.

Arginine↗
PubMed1994

Rehabilitation technology product taxonomy: a conceptual tool for analysing products and extracting demand determinants.

This paper proposes and describes a conceptual tool, namely the rehabilitation (assistive) technology (RT) product taxonomy, as a framework for analysing rehabilitation technology products and extracting demand determinants. The ultimate objective of this work is to provide demand and supply-related actors with a meaningful tool for identifying and focusing on specific aspects, which may be directly or indirectly related to existing or new products, so that demand and consumption allocation decisions can be targeted, evaluated and/or predicted.

Equipment and Supplies↗
PubMed1993

Cytokine production by T-cell lines derived from tumor-infiltrating lymphocytes from patients with ovarian carcinoma: tumor-specific immune responses and inhibition of antigen-independent cytokine production by ovarian tumor cells.

The ability of T-cell lines derived from tumor-infiltrating lymphocytes (TIL) from patients with ovarian carcinoma to produce interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and IL-6 in response to autologous or allogeneic ovarian carcinoma tumor cells was determined. These T-cell lines were derived in low concentrations of recombinant IL-2 (rIL-2), 200 IU/ml. Certain of these T-cell lines (3 of 7) exhibited antigen-independent (spontaneous) production of moderate to low concentrations of IFN-gamma (75 to 112 pg/ml), TNF-alpha (62-88 pg/ml), and IL-6 (38 to 690 pg/ml), in culture medium alone in the absence of rIL-2, tumor cells or polyclonal activators. Addition of 20 IU/ml of rIL-2 to the cultures resulted in a significant increase of the antigen-independent production of IFN-gamma (range 75-6480 pg/ml) by all seven T-cell lines and TNF-alpha (44-490 pg/ml) by 5 of 7 T-cell lines. Certain T-cell lines (4 of 7) exhibited antigen-independent production of IL-6 (38-690 pg/ml), which was not affected by the addition of 20 IU/ml of rIL-2. Certain TIL-derived T-cell lines (2 of 7) produced IFN-gamma, TNF-alpha, and IL-6 in response to stimulation with autologous ovarian tumor cells, in the presence of 20 IU/ml of rIL-2. Irradiated autologous ovarian tumor cells alone or in the presence of rIL-2 did not produce any detectable levels of IFN-gamma, TNF-alpha, or IL-6 during the 6 day culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, CD↗
PubMed1996

Expression of human Pit-1 product in the human pituitary and pituitary adenomas. Immunohistochemical studies using an antibody against synthetic human Pit-1 product.

OBJECTIVE: Pit-1, a member of a family of the POU-domain DNA binding factors, has been known as a pituitary-specific transcriptional factor that regulates functional differentiation toward somatotrophs, lactotrophs, and thyrotrophs in the rodent pituitary gland. The aim of this study is to elucidate the role of human Pit-1 (hPit-1) protein in the differentiation of human pituitary adenomas, using immunohistochemistry. DESIGN: Anti-human Pit-1 polyclonal antibody against synthetic peptide was applied to perform the avidin-biotinperoxidase complex method on paraffin sections of 75 surgically obtained pituitary adenomas and 12 nontumorous human pituitaries obtained at autopsy. RESULTS: In normal human pituitary glands, many cells were positive for hPit-1 product in the nuclei of somatotrophs, lactotrophs, and somatomammotrophs. Among 75 pituitary adenomas, hPit-1 product was expressed in 55 adenomas (73.3%). All (100%) of the growth hormone-positive and thyrotroph cell adenomas were positive for hPit-1 product. Unexpectedly, expression of hPit-1 was found in a limited number of adrenocorticotropic hormone-producing adenomas and clinically nonfunctioning adenomas, including gonadotroph cell adenomas, although localization was found only in occasional cells. CONCLUSIONS: The invariable expression of hPit-1 protein in growth hormone-positive adenomas and thyroid-stimulating hormone-positive adenomas may suggest the role of Pit-1 protein in specific differentiation of the adenoma cells. The expression of hPit-1 in various other types of adenomas may indicate the involvement of other unidentified transcription factors or specific mediators that have roles in these differentiations. Our observation may provide some insight into the origin of cell types of some clinically nonfunctional adenomas.

Adenoma↗
PubMed1996

Uncoupling IL-2 production from apoptosis and TNF production by changing the signal through the TCR.

T cells may discriminate between stimuli in a variety of ways, including the presence of cytokines or other costimulatory signals, the type of Ag (peptide, superantigen, or allorecognition), or the magnitude of the signal through the TCR. We have used anti-CD3 stimulation of T hybridomas to examine signals generated through the TCR in the absence of exogenous APCs. Soluble whole anti-CD3, but not F(ab')2 anti-CD3, was able to stimulate the T hybridomas to produce IL-2. Plastic-bound anti-CD3, in contrast, stimulated TNF production, G1 arrest, and apoptosis by the T hybridoma. Engagement of the CD4 coreceptor on these cells had no effect on the overall pattern of signaling observed. Although TNF production was correlated with apoptosis, anti-TNF treatment did not prevent cell death or G1 arrest. The response of the T hybridoma to both forms of anti-CD3 included significant IL-2 production even at the lowest dose tested. However, soluble anti-CD3 at the highest dose tested elicited only minor apoptosis, while plastic-bound anti-CD3 elicited significant apoptosis even at the lowest dose. The difference in response was not evident at the level of phosphotyrosine proteins two min after cross-linking of the TCR.

Animals↗
PubMed1995

[Study on evaluating methods for the quality control of glycoprotein products. (1). Erythropoietin products].

The ability of high performance anion exchange chromatography (HPAEC) with pulsed amperometric detection was studied for evaluation of carbohydrate moieties of erythropoietin (EPO) products. The N-linked oligosaccharides were released from EPO by the treatment with N-glycosidase F. HPAEC analysis of oligosaccharide standards revealed that elution time of sialylated oligosaccharides were dependent on the number of sialic acid, which contributed to the activity of EPO. Using HPAEC, N-linked oligosaccharides of two kinds of recombinant human EPO (rhEPO) produced in chinese hamster ovary (CHO) cells and baby hamster kidney (BHK) cells were compared. The HPAEC profiles of oligosaccharides of these EPO products indicated that there were some differences in the carbohydrate moieties between CHO rh-EPO and BHK rh-EPO. In conclusion, HPAEC method is useful to evaluate the quality of EPO products.

Animals↗
PubMed1997

Dharmendra antigen but not integral M. leprae is an efficient inducer of immunostimulant cytokine production by human monocytes, and M. leprae lipids inhibit the cytokine production.

Killed integral Mycobacterium leprae, Mitsuda antigen, and chloroform-treated M. leprae, Dharmendra antigen (Dh-Ag), have been used for the classification of leprosy patients based on cell-mediated immunity. Heat-killed M. leprae also were used as a component of the Convit vaccine. Human blood monocytes were stimulated with M. leprae or Dh-Ag and their cytokine-inducing ability was compared. Monocytes were cultured in the presence of fresh human serum because of the efficiency of cytokine induction and the phagocytosis of M. leprae have been shown to be optimal in the presence of fresh serum. M. leprae and Dh-Ag were equally phagocytosed by monocytes. Dh-Ag was more potent than M. leprae in the induction of immunostimulatory/proinflammatory cytokines, interleukin-1 (IL-1), IL-6 and tumor necrosis factor (TNF). In contrast, a comparable level of IL-1ra, an immunosuppressive cytokine, was induced by M. leprae and Dh-Ag. The lipids extracted from M. leprae induced none of these cytokines by monocytes. Nevertheless, when monocytes were pretreated with the lipids followed by stimulation with Dh-Ag, productions of IL-1, IL-6 and TNF were all inhibited in a dose-dependent manner. However, the lipids did not inhibit the cytokine production induced by other stimuli including BCG and lipopolysaccharide. Moreover the lipids did not affect the production of IL-1ra. These results suggest that the lipids from M. leprae are responsible for the poor cytokine-inducing ability of M. leprae, thus favoring their infection. These results also suggest that Dh-Ag rather than integral M. leprae may be useful as a vaccine candidate because Dh-Ag is able to induce a large amount of cytokines from monocytes.

Antigens, Bacterial↗
PubMed1997

The anti-tumor effects of IL-12 involve enhanced IFN-gamma production by anti-tumor T cells, their accumulation to tumor sites and in situ IFN-gamma production.

We investigated cellular and molecular mechanisms underlying the anti-tumor effects of IL-12. Intraperitoneal injections of rIL-12 into later stages of tumor-bearing mice induced not only a striking reversal of suppressed IFN-gamma-production by splenic T cells, but also complete regression of s.c. growing tumors. A massive infiltration of lymphoid cells was found following IL-12 treatment. Whereas fresh spleen cells obtained from IL-12-treated tumor-bearing mice failed to express IFN-gamma mRNA, significant levels of IFN-gamma mRNA were detected in the tumor mass of the same mice. The systemic administration of anti-IFN-gamma antibody (Ab) prior to IL-12 injection abrogated the anti-tumor effect of IL-12 although this Ab did neither inhibit accumulation of lymphoid cells to tumor sites nor in situ IFN-gamma expression. Importantly, while high levels of inducible nitric oxide synthase (iNOS) mRNA expression was induced in tumor masses after IL-12 treatment, its expression was completely inhibited by pretreatment with anti-IFN-gamma Ab. Thus, induction of tumor regression by IL-12 is ascribed to a series of events: a striking reversal of suppressed IFN-gamma production by anti-tumor T cells, their accumulation and IFN-gamma production at tumor sites, and manifestation of IFN-gamma activity as exemplified by iNOS expression.

Animals↗