Cytokine production by the human bladder carcinoma cell line T24 in the presence of bacillus Calmette-Guerin (BCG)

Summary The study was initiated as an in vitro approach to the situation existing during intravesical bacillus Calmette-Guerin (BCG) instillation in patients with superficial bladder cancer. Cytokine secretion of a human bladder carcinoma cell line T24 treated with BCG was investigated. A 24-h treatment of T24 cells with BCG resulted in a tenfold higher secretion of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF) when compared with T24 cells treated with Escherichia coli, Streptococcus faecalis or a cell wall preparation of Nocardia rubra (N-CWS). No secretion of IL-1 and IL-2 was detected. Pre-exposing T24 cells to BCG for various periods of time indicated that a minimum exposure time of 0.5–1 h was required to upregulate IL-6 and TNF production. Extending the BCG pre-exposure time to 2 and 3 h further increased the rate of cytokine production. No significant difference was found, however, between the rate of secretion initiated after a 2-h or 3-h pre-exposure period. The amounts of these cytokines secreted in the presence of BCG-conditioned medium did not differ significantly from the constitutively secreted amounts, excluding an effect of products possibly secreted by BCG on the upregulation of IL-6 and TNF. In addition, upregulation of cytokine production appeared to be dependent on the concentration of BCG. The results suggest that cytokines may be produced by urothelial tumor cells after intravesical instillation in patients with superficial bladder cancer, which may play a role in the mode of action of BCG.     

人内皮细胞和4种细胞因子对正常人和再生障碍性贫血患者粒系造血的影响

通过人脐静脉内皮细胞为铺层的双层细胞培养,观察内皮细胞和几种细胞因子对正常人和再障患者粒系造血的影响。方法以人内皮细胞作铺层,作体外骨髓CFU-GM半固体双层培养,并在培养体系中加入细胞因子GM-CSF、IL-2、IL-6和LAK细胞上清,观察内皮细胞和细胞因子对正常人和再障患者粒系造血的影响。结果在有内皮细胞的培养体系中CFU-GM的产率均显著高于无内皮细胞的相应培养体系。在无内皮细胞时,IL-6和LAK上清显示出与GM-CSF相似的粒系造血刺激作用。对15例初诊慢性再障患者骨髓粒系祖细胞培养结果表明,无内皮细胞存在时,无论向培养体系中加入何种细胞因子,均无集落生成,而在有内皮细胞的双层培养中,部分患者骨髓可见CFU-GM集落生成。结论内皮细胞对正常和再障粒系造血均有明显影响,并提示再障造血障碍有微环境因素。     

重症肝炎免疫治疗与细胞因子、T细胞亚群的调控关系

了解重症肝炎患者经Ｔα１免疫治疗前后外周血中、ｓＩＬ－２Ｒ,ＩＬ－４,ＩＬ－６内毒素及Ｔ细胞亚群的变化。方法采用ＥＬＩＳＡ方法或全成基质偶氮显色法或链菌素亲生物素过氧化酶连接法检测上述指标。结果治疗前各项指标较正常值均有显著变。,后除ｓＩＬ－２Ｒ和ＣＤ＾＋４外,其他指标基本恢复到正常水平。     

Severe malaria in African adults living in a seasonal endemic area

Objective This study investigates severe malaria in african adults living in a seasonal endemic area.Design: A prospective study of all adults admitted with severe malaria over 2 consecutive seasons: October 1990 till January 1991 and October 1991 till January 1992.Setting ICU (15 beds) of Hôpital Principal, Dakar, Sénégal.Patients 23 patients: 14 men and 9 women with a mean age of 30±3 years were included in the study; all fulfilled the 1990 WHO criteria for severe malaria.Results At admission, 12 patients were comatose (Glasgow Coma Scale<10), 7 had generalized convulsions. Parasitaemia was 135±52×10⁹/l. Biological indications of severity were as follows: hypophosphataemia <0.8 mmol/l in 14 cases, serum creatine phosphokinase >500 IU/l in 15 cases; and PaO₂<70 mmHg in 5 cases. Serum TNF levels, measured in 16 cases, were increased at 298.4±63.5 pg/ml, serum levels of IL-6 and IL-2SR were also elevated: 609.5±304.2 pg/ml and 297.6±35.6 pg/ml respectively. Circulating IgM and IgG antibodies were found in 14 out of 16 patients. Serum levels of TNF, IL-6 and IL-2SR correlated positively with each other. TNF and IL-2SR were also positively correlated to parasitaemia. Intravenous therapy with quinine at loading dose was favorable in 19 patients. Four patients died during the study, 3 from multiple organ failure.Conclusions: This work demonstrated that severe malaria in a seasonal endemic area displays original clinical features with a high rate of either cerebral malaria or multiple organ failure.     

Microheterogeneity of acute phase proteins in patients with clinically active and clinically nonactive osteoarthritis

Summary Microheterogeneity of two acute phase glycoproteins, -1-acid glycoprotein (AGP) and -1-antichymotryspin (ACT), concentrations of AGP, ACT, and C-reactive protein (CRP), and levels of three cytokines: interleukin 1 (IL-1-), interleukin 6 (IL-6), and tumor necrosis factor (TNF-) were determined in 61 serum samples and 7 synovial fluids (SFs) obtained from patients (n=61) with osteoarthritis. Using affinity immunoelectrophoresis with concanavalin A (conA), a significant decrease in the reactivity of AGP and ACT with this lectin was found in patients with clinically active osteoarthritis when compared to those with clinically nonactive disease (p<0.001 and p<0.05, respectively). There was no increase in the concentration of AGP, ACT, and C-reactive protein (CRP) in the sera examined. In particular, no increase in the serum level of these proteins was found in the patients with clinically active disease. Low concentrations of IL-6 and TNF- were found in most sera and SFs examined. In 6 out of 7 SFs available, IL-6 concentrations were higher than in the respective serum samples but for TNF- the same could be shown in one case only. Low concentrations of IL-1- were found in 4 serum samples obtained from patients with clinically active osteoarthritis and in no SF specimen studied. In the entire group, serum level of TNF- correlated weakly with the AGP and ACT reactivity coefficients with conA (r=0.3634, p<0.005 and r=0.3324, p<0.02, respectively).Our findings suggest that there are changes in the microheterogeneity of acute phase glycoproteins in some patients with osteoarthritis similar to those observed in rheumatoid arthritis and other chronic inflammations. Possible mechanisms of the involvement of cytokines in the regulation of glycosylation of acute phase glycoproteins in osteoarthritis are discussed.     

T-lymphocyte immunointerferon receptors in patients with multiple sclerosis

Multiple sclerosis (MS) is a T-cell-mediated autoimmune demyelinating disease of the central nervous system (CNS), associated with an altered immunoregulation. Interferon (IFN)-, also known as immune IFN, is a cytokine with several effects on the immune system. Specific IFN- receptors have been found on human lymphocytes, as well as on other cell types (e.g. gliocytes), even in the CNS. The aim of the present study was to evaluate IFN- binding on peripheral blood T-lymphocytes from MS patients, compared with those from healthy subjects. Thirty-two patients were selected according to the classical criteria for definite MS; as controls, 21 healthy subjects were studied. We have found that T-lymphocytes from MS patients bear a significantly smaller amount of IFN- receptors than those from controls [B _max: 568, 18 vs 708, 14 (mean, SE) receptors/cell]. Such IFN- binding sites are of the same type in patients and healthy subjects [K _d: 1.0, 0.05 vs 0.9, 0.02 (mean, SE) nM]. These findings are discussed in terms of immunopathogenesis of MS, since it has been reported that activated T-lymphocytes have decreased amounts of IFN- receptors.     

Einfluß des operationstraumas auf die NK-zell-aktivität beim ösophaguskarizinom nach transmediastinaler dissektion vs. transthorakaler en-bloc-resektion

Zusammenfassung Um den Einfluß des chirurgischen Traumas beim Ösophaguskarzinom auf das zelluläre Immunsystem zu erfassen, wurden perioperativ in einer prospektiven Studie die Aktivität der natürlichen Killerzellen sowie die Serumkonzentrationen von Interleukin-2, Interleukin-6 un TNF- bei transmediastinaler Dissektion (n=12) vs. transthorakaler En-bloc-Resektion (n=10) der Speiserörhre im Vergleich zu einer Kontrollgruppe mit thorakoabdominalen chirurgischen Eingriffen bei nicht maligner Grunderkrankung erfaßt. Die Bestimmung der NK-Zell-Aktivität erfolgte präoperativ sowie am 4. und 10. Tag postoperativ durch einen standardisierten Europiumchlorid-release-Assay unter Verwendung von K-562-Targetzellen, die Lymphokine Interleukin-2, Interleukin-6 und TNF- wurden zusätzlich am 1. und 7. Tag postoperativ mit stadardisierten ELISA-Assays bestimmt.In unserem Patientengut sank die NK-Zellaktivität am 4. postoperrativen Tag sowohl in der Kontrollgrupe al auch bei beiden Operations verfahren zur Speiseröhrenresektion signifikant (p<0,05) zum Ausgangswert: in der Kontrollgruppe durchschnittlich um 45%, nach transmediastinaler Speriseröhrendissektion (1-Höhlen-Eingriff) durchschnittlich um 34%, nauch transthorakaler En-bloc-Resektion (2-Höhlen-Eingriff) im Mittel um 63% zum präoperativen Wert. Die transthorakale En-bloc-Resektion der Speiseröhre führte durch das größere chirurgische Trauma zu einer stäkeren Abnahme der zytotoxischen Aktivität der natürlichen Killerzellen. Eine Suppression der immunologischen Tumorabwehr insbesondere in der vulnerablen perioperativen Phase kann damit indirekt das Risiko der Manifestation von hämatogene Metastasen auf dem Boden einer intraoperativen Tumorzelldissemination u. a. bedingt durch Tumormanipulation begünstigen und damit prognostisch relevant werden.

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Influence of surgical trauma on natural killer cell activity in esophageal carcinoma following transmediastinal dissection compareed with transthoracic en bloc resection

In order to assess the impact of surgical trauma involved in the therapy of esophageal carcinoma on the cellular immune system, a perspective study was performed involving perioperative hematological parameters. The activity of natural killer cells and the serum concentrations of interleuin-2, interleukin-6 and TNF- were measured in 12 cases of transmediastinal dissection and 10 cases of transthioracic en bloc esophageal resection and compared to values of a control group of thoracic and abdominal surgical patients with non-malignant maladies. Natural killer cells assume a central role in the non-specific immunological response in tumor patients. Their main function is the destruction of tumor cells via cytotoxic activities amplified by the release of interleukin-2 and TNF-. Natural killer cell activity was measured prior to surgery and on postoperative days 4 and 10 using a standardized europium chloride release assay, utilizing K 562 target cells. Lymphokines interleukin-2, interleukin-6, and THF- were also measured on postoperative days 1 and 7 using standardized ELISA assays. The activity of natural killer cells in our patient group sank significantly (P<0.05) on postoperative day 4 and likewise in the control group and both study groups, activity sank to the original values. In the control groups, natural killer cell activity averaged 45% of preoperative values, in comparison with an average of 63% following transmediastinal esophageal carcinoma resection (one cavity procedure), and transthoracic en bloc resection patients only reached 61% of preoperative values, transmediastinal dissection patients assumed 75%, and 77% was achieved by control group members. Transthoracic en bloc resection of the esophagus led to a more extreme reduction in cytotoxic cellular activity owing to the greater surgical trauma. Suppression of the immunological tumor resistance, especially in the vulnerable perisurgical pahse, can have an indirect negative effect on the manifestation risk of hematogenic metastases owing to intraoperative tumor cell dissemination resulting from tumor manipulation and may thus be prognostically relevant.

     

Neuroaxonal dystrophy in HTLV-1-associated myelopathy/tropical spastic paraparesis: neuropathologic and neuroimmunologic correlations

Summary Detailed neuropathologic and immunohistologic analysis of a case of serologically and polymerase chain reaction-confirmed human immunodeficiency virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is reported in a 73-year-old North American black woman. In addition to the usual neuropathologic features of HAM/TSP, including tractal degeneration of the spinal cord, leptomeningeal and perivascular fibrosis, perivascular demyelination and chronic inflammation, neuroaxonal spheroids were prominent in the spinal cord. Neuroaxonal dystrophy was characterized by neurofilamentous masses that were immunoreactive for phosphorylated neurofilament epitopes, but not ubiquitin. Neuroimmunologic analysis of the inflammatory reaction revealed a prevalence of CD8⁺ T cells and class I major histocompatibility molecules (MHC) (HLA-ABC and 2-microglobulin), but very few CD4⁺ T cells. Microglia were highly reactive for class II MHC (HLA-DR) and this was attributed to activation, rather than CD4 interaction, since CD4 presence was minimal. Inflammatory cytokine immunoreactivity was also detected in glia. It is concluded that the cumulative effects of cytotoxic T cell (CD8) infiltration and the possible involvement of cytokines were responsible for the unusual degree of neuroaxonal dystrophy and vascular fibrosis, as well as the observed demyelination in this case.Supported in part by grants NS 07098, NS 08952, NS 11920 and MH 47667 from the NIH; and RG 1001-G-7 from the National Multiple Sclerosis Society.     

Cytokine enhancement of monocyte/synovial cell attachment to the surface of cartilage: a possible trigger of pannus formation in arthritis

Summary When rheumatoid articular cartilage samples were incubated with normal peripheral blood monocytes and cultured synovial cells in the presence of recombinant human interleukin-1 (IL-1) in vitro, large numbers of monocytes were seen to be attached to the articular surface. Significant numbers of monocytes invaded the cartilage tissue when the rheumatoid cartilage samples were pre-incubated with 10 U/ml of IL-1. Considerable numbers of monocytes were also attached to normal cartilage when these were pre-incubated with IL-1. It is of interest that recombinant human gamma interferon (-IFN) did not enhance monocyte attachment. However, there was a significantly greater attachment of monocytes to rheumatoid than to normal cartilage. When normal cartilage was exposed to collagenase and then incubated with monocytes or synovial cells in the presence of 10 U/ml of IL-1, large numbers of cells were attached to the natural cartilage surface but not to the cut surface. These phenomena were much more intense when the rheumatoid cartilage was pre-incubated with collagenase. These results indicate that increased levels of IL-1 in the rheumatoid joint may play an important role in joint destruction by stimulation of pannus formation by inducing synovial cell attachment to the articular surface.     

中药双利肝在肝纤维化与肝硬化形成中的作用

目的探讨中药双利肝在肝纤维化与肝硬化发展中的作用。方法采用大鼠饮用TAA(0.03%)4个月复制肝硬化动物模型,从第4个月始加用中药双利肝合剂灌胃,进行内毒素水平、肿瘤坏死因子(TNF)-琢、内皮素(ET)-1及一氧化氮(NO)的检测,以及组织学观察。结果用中药双利肝治疗后,内毒素水平、TNF-琢、ET-1及NO均有明显下降;肝组织胶原蛋白含量显著减少。结论中药双利肝通过抑制细胞因子介导的细胞-细胞相互作用所引起的肝星状细胞(HSC)激活,从而间接抑制肝纤维增生。