Balance between proinflammatory IL-1, TNFα, IL-6 and regulatory TGFβ, IL-10 cytokine serum levels in chronic hemodialysis patients

Cytokines in injPammation

25 June 1997 - Poster presentations 1P.3.10.32 ) Systemic cytokine production following a minor trauma I. Grzdak. M. Zaieska. W.L. Clszewski. M. Durlik. B. Laoiewska. M. Muszvnski. W. Rowtn&i. Surg Res TRlnsplDept, kd Res & Polkad &i, Warsaw. FDland, Gen Sung Tmnsppl C/in, Med Acad, Warsaw,Poland, Dept Gen Sum, Mi/it Hosp, Wars& Poland In spite of the extensive studies of recent years and the growing understanding of different aspects of processes evoked by trauma, the exact mechanism behind the surgery-related immunodepression is not fully understood. There is a large body of evidence that many of the symptoms observed after trauma may be evoked by exaggerated cytokine production which, present in excessive amounts, may produce hemodynamic, metabolic, and immune effects. In this etudy, a systemic cytoktne response (ILla, ILIP, ILlra, 116, lL6sR, 118, ILIO, IL13, IFNy, GM-CSF, 113, ILlI) was examined in patients with cholelithiasis. undergoing elective cholecystectomy under general anesthesia, in the “silent” period dudng which no clinical signs of inflammation were present. Cytokine plasma levels were esttmated on the day preceding the operation and postoperatively, on days 1, 3, 7, 10 and 14, using an ELISA method (R&D, Genzyme, Endogen). Reeultr: No changes were found in ILlcr and ILig plasma levels during the entire observation period, whereas there was a significant increase in IL1ra values by 66.6% on day 1,95.6% on day 3,103.5% on day 7.230.4% on day 10 and 140.3% on day 14 following surgery (p < 0.05). Furthermore, a significant rise in IL6 plasma level was seen in all pattents on day 1 (by 2254.4%) on day 3 (by 694.29/o)and on day 7 (by 312.7%) after surgery (p 4 0.05). Moreover, a decrease in lL6sR level was observed on day 1 after the operation, followed by a rise, above the preoperative values, on days 10 and 14. Interestingly, an elevation in IL6 level was noted on days 1 (by 101.3%) and 3 (by 62.3%) after trauma (p < 0.05). In some patients, an increase in IL10 level was also noted on days 1 and 3 following the operation. No significant changes in mean hematopoietic cytokine plasma levels (GM-CSF, 113, ILll) were noted during the entire observation period, although in 3 patients GM-CSF level ttsed on day 3 and in 1 patient IL11 level increased on days 3 and 7 after the operation. A slight decrease in IFNy plasma levels was seen in four of eight patients on day 1 after surgery (p t 0.05). Finally, IL13 plasma levels remained undetectable during the entire observation period. Rwufts ctentonetraMd that even a minor surgical procedure as choleqstectomy with uneventful wound heating was followed by appearance in blood circulation of significant levels of cytokines between days 1-14 after surgery. Induction of the release of inflammatory cytokines to blood circulation was followed by the release of cytokines representing a down-regulatory mechanism of the cytokine circuit (IL1ra, IL10), most likely limiting the excessive inflammatory cytokine response evoked by surgery.

IP.3.10.33

~I~I~~KAs

factors in inflammatory bowel

B. Mascher ‘, A. Reil ’ , G. Jantschek’, E.F. Stange 2, M. Seyfarth ’ . ’ institute of Immunology and Transfusion Medicine, University of Ltibeck Medical School, 23538 Libeck, GermanN 2Department of l&ma/ Medicine, University of L&e& Medical School, 23538 Lobeck, Germany Introduotkm: Ettdogy and pathogenesis of inflammatory bowel disease (IBD) are still unknown. Tumor necrosis factors (TNF) are potent inflammatory mediators whose particular importance in IBD could be well undedined in an open clinical trial where 9 out of 10 patients suffering from steroid refractory Crohn’s disease (CD) could be brought into clinical and cotonoscopical remission by treatment with an anti TNF mot&human antibody (van Dullemen, Gastroenterology, 1995). Mstwialand Methods: We investigated two known di-allelic polymorphisms in the genes of TNFar (promotor. pos.306, named TNF) and TNFB (first intron, pos. 1069, called TNF B’) in 16 CD patients and 23 patients with ulcerative colitis (UC). Using PCR methods followed by Ncol-digestion the poiymotphisms could be detected via the restriction fragment length pofymotphism. As controfs (N) served 111 healthy blood donors. Moreover, we measured TNF (I levels in unstimulated sera of 10 CD and 16 UC patients using a commercial high sensitivity ELISA (R&D Systems). As controls sewed 42 of the primattly typed blood donors. Reeub Concerning the TNF B’ polymorphism we could not find any relevant differences between patients and controls. As to the TNF pc+morphism we found an elevation of the seldom TNF 2/2 phenotype in the UC group (UC 0.067 vs. N 0.016) an increase of the hetemzygous TNF l/2 in the CD group (CD 0.45 vs. N 0.24), both could not reach significance. Concerning the TNF 01levels in unstimulated sera we detected a significant increase in both of the patient groups (median level: N 0.663 p#nl vs. CD 2.547 p@ml, p < 0.001; vs. UC 1.564 pg/ml, p < 0.01). Conclueion: Since TNF (I is a strong proinflammatory mediator k can wntdbute to the chmnicity of IBD, its importance in act&e IBD is further stressed by the clinical success of a treatment wkh an antibody against this mediator. We could find elevated TNF u in unstimulated sera and varieties concerning the

413

genetical background of TNF in IBD patients. This might lead to the wndusion that elevated TNF (I in IBD patients is not only due to the chronic inflammatory process but in contrast may be primarily elevated and sustain the chronic inflammation of the bowel.

P.3.10.34

Interleukin-4 levels In peripheral blood CD8+ T cells In mild atoplc asthmatics

L. Stanciu, J. Shuts, C. Pmmwong, S. Holgate, R. Djukanovic. Univekty Medicine, Southampton, UK Introduction: We hypothesized that higher levels of in viw IL-4 in asthmatics may modulate IL-4 levels in CD6+ T cells. Materlele and Methode: The levels of IL-4 and IFN-y in freshly isoJated blood CD6+ T cell and in 20 and 46 h Con A-stimulated CD6+ T ceils in 7 mild atopic asthmatic and 7 healthy nonatopic subjects were measured. We have used the MACS system for negative selection of CD6+ T cells to obtain a purity of >90%. Using in house ELISA (sensitivity 10 pslml and respectively 5 w/ml) IL-4 and IFN-y were measured in lysates of “restina” CD6+ T cells (1 x 106 cells in 1 ml PBS/2% Tdton X-100) and in 20 and 46 h supematants of’cultured CD6+ T cells stimulated with Con A (1 x lo6 cells/ml f 10 pg Con A/ml). Results Resting CD6+ T cells in asthmatics contained -significantly more IL-4 as compared with controls (35 f 21.6 vs 16.9 f 5.5 pgI1 x l@ cells/ml). The levels of IL-4 correlated negatively with levels of IFN-y in resting CD6+ T cells in healthy controls (r = -9411, p = 0.005). In Con A stimulated CD6+ T cells the levels of IL-4 in culture supematants were only slightly increased as compared with medium, but significantly higher levels of IFN-y were found in both groups. In 46 h Con A-stimulated CD6+ T cells a strong positive correlation was found between the levels of IL-4 and IFN-y in normal controls. Conclusion: We have demonstrated that CD6+ T cells from asthmatics may be an important source of the ThP-type cytokine IL-4. This capacity appears to be acquired in vtvo and cannot be demonstrated ex viw when cells are stimulated with Con A.

P.3.10.35

Balance between prolnflammmatory IL-l, TNFa, IL-6 and reaulatorv TGFB. IL-10 cvtoklne serum levels In &ironic tiemo&ysis

paiients

M. Koziol-Montewka, A. Ksiafek. NephrOogy Department, Medical Academy in Lublin, Poland Introduction: Hemodialysis procedure and uraemia itself cause immune dysregulations, manifested as various clinical side effects and impaired efficacy of vaccination in patients wtth chronic renal failure. Increased chronic production of proinflammatory cytokjnes IL-ifi. TNFo(. and IL-6 occurs wtth some imbafante between cytokine secretion and their natural regulators releasing in these patients. The aim of our study was to evaluate TGFB and IL-10 serum levels, natural proinflammatory cytokine regulator factors, in hemodialyzed (HD), peritoneal dialyzed (CAPD) and healthy control persons. Meterlal end Methods: Thirty-five patients on chronic hemcdialysis, fifteen CAPD dialized patients and fifteen healthy controls were studied. EDTA plasma samples were obtained during stable period of treatment. IL-l@, TNFa, IL-6, TGFB and IL-10 were measured using commercial ELISA test systems, Genzyme USA, R&D Systems USA kits were used. Reeutk Monocycte preactivation markers and increased serum TNFa and IL-6 were accompanied by significantly decreased TGFB and IL-IO serum levels (70 f 9.9 pg/ml and 11.7 f 2 p@ml) in comparison to healthy controts (143.5 f 26 wrnl and 25 f 6.3 pglml) respectively. The significant negative correlation was found between TGFB and HIA-DR expression and IL-6 levels. which are regarded as the immunological activation markers. However. there was a positive correlation between IL-IO serum levels and these factors. Only the patients with good immune responsiveness to hepatitis 8, and influenza vaccination showed the same TGFB and IL-10 levels as healthy controls. Conduslon: The results of our study show the decreased levels of TGFB and IL-10 in chronic hemodialized patients thus suggesting the existence of some disturbances in the secretion of both regulatory cytokines. Dependence on immune responsiveness to vaccination points out to their participation in immune deficiency state of the patients.

P.3.10.36

Dantrolene lnhlbits 11-12, TNF-a, and nltrlc oxide production, but increases the formation of IL-10 In tindotoxemic mice

Z.H. Nemeth ’ , G. Hask6 ’ , C. Saab6 1,2,A.L. Salzman 2, ES. Vizi 1. ’ Deparbnent of Pha~acof~ Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary 2Diviskm of Critical Care, Children’s Hospital Medical Center. Cincinnati, OH, USA Introduction: Increases in intracellular calcium levels have been implicated as