Alain Crusiaux

Release of tumor necrosis factor, interleukin-2, and gamma-interferon in serum after injection of OKT3 monoclonal antibody in kidney transplant recipients

High levels of tumor necrosis factor-alpha, interleukin-2, and gamma-interferon appeared in the circulation of kidney transplant recipients after the first injection of the monoclonal antibody OKT3. This initial injection was systematically followed by fever. The three cytokines were released in all patients (n = 9), with peak serum levels of tumor necrosis factor occurring 1 hr after OKT3 injection and those of interleukin-2 and gamma-interferon after 2 hr. Cytokines were not released after the second and third OKT3 injections, when CD3+ cells had disappeared from blood. These findings suggest that circulating cytokines are released by T cells after activation by OKT3. These cytokines are probably involved in the systematic reactions observed after injection of OKT3.

Tumor necrosis factor α and interleukin 6 plasma levels in infected cirrhotic patients

Abstract Background: Patients with liver cirrhosis disclose both increased production and decreased metabolism of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6). The present study analyzes the characteristic pattern of these cytokines during sepsis in cirrhotics. Methods: TNF-α and IL-6 plasma levels, measured during 15 days from the onset of cirrhotic decompensation or of the septic event, were compared between 14 infected patients with liver cirrhosis, 18 uninfected decompensated cirrhotic patients, and 35 septicemic patients devoid of liver disease. Cytokines were measured using immunoassays. Results: In infected cirrhotics, initial levels of both TNF-α and IL-6 were significantly higher than in noninfected cirrhotic patients ( P P Conclusions: Both the profoundly increased initial levels of TNF-α and IL-6 and their persistence over days after sepsis onset seem characteristic of the cirrhotic patients. The exact relationship between prolonged exposure to TNF-α and poor prognosis in these patients is unknown, but it might represent a unique opportunity for the use of anti-TNF-α antibodies during sepsis.

Interleukin-6 : an early marker of bacterial infection in decompensated cirrhosis

Fifty-seven patients with decompensated cirrhosis were studied prospectively to assess the sensitivity and specificity of early clinical or biological signs of bacterial infection. Among them, 19 had proven infection on admission (7 spontaneous bacterial peritonitis, 5 bacteraemia, 3 urinary tract infections, 2 pneumonia, 1 dental abscess and 1 cholangitis). Fever, polymorphonuclear cell count, fibrinogen and C-reactive protein levels were found to be of little or no help in diagnosing bacterial infection on admission. Interleukin-6 plasma levels were, however, significantly different between infected (median: 1386 pg/ml, range: 237-20,000) and non-infected patients (median: 34 pg/ml, range: 0-4500, p < 0.00001). Levels above 200 pg/ml were always found in infected patients, giving a sensitivity of 100% and a specificity of 74%. C-reactive protein correlated weakly with interleukin-6 levels, indicating a defective acute-phase response in cirrhosis. Tumor necrosis factor alpha plasma levels were less sensitive (95%) and specific (68%) for the diagnosis of bacterial infection at a threshold of 50 pg/ml, but were more closely related to a poor patient outcome. In decompensated cirrhosis, interleukin-6 plasma levels on admission provided the most sensitive and specific tool for the diagnosis of bacterial infection.

Anaphylactic shock caused by immunoglobulin E sensitization after retreatment with the chimeric anti-interleukin-2 receptor monoclonal antibody basiliximab.

Background. Repeated administration of chimeric or humanized monoclonal antibodies is generally well tolerated. Anti-idiotypic sensitization is rare and is considered to be of no clinical significance. We observed a child who experienced anaphylactic shock when he received a second course of basiliximab at the time of his second renal transplantation. We therefore searched for the presence of anti-basiliximab immunoglobulin (Ig) E in this patient. Methods. Serum levels of anti-basiliximab IgE, assay of the anti-murine reactivity of circulating anti-basiliximab IgE, and assays for serum anti-mouse antibodies and global anti-basiliximab anti-idiotypic antibodies were carried out by enzyme-linked immunosorbent assay. Anti-basiliximab IgE antibodies on circulating basophils were evaluated by the ability of the patient’s blood to produce leukotrienes in vitro after exposure to basiliximab. Results. Sequential assays of serum samples by enzyme-linked immunosorbent assay indicated that anti-basiliximab IgE antibodies appeared after the second basiliximab course. There was no IgE reactivity toward a control murine IgG2a monoclonal antibody (mAb), indicating that the IgE response was directed exclusively against basiliximab idiotypes. There was no IgE reactivity against the humanized anti–interleukin-2 receptor mAb daclizumab, which was derived from a distinct parental murine mAb. Patient basophils harvested months after the anaphylactic shock produced leukotrienes in vitro on exposure to basiliximab. Conclusions. Patients exposed to chimeric antibodies may develop an anti-idiotypic IgE response that can trigger anaphylactic shock on further exposure. Specific anti-idiotypic IgE may be bound to basophils even in the absence of circulating IgE.

beta IFN- Interferes with the Differentiation of Dendritic Cells from Peripheral Blood Mononuclear Cells: Selective Inhibition of CD40-Dependent Interleukin-12 Secretion

We studied the effects of interferon-beta (IFN-beta) on the differentiation of dendritic cells (DC) obtained by culturing plastic-adherent peripheral blood mononuclear cells (PBMC) from a total of 30 healthy volunteers in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). First, we found that the addition of IFN-beta at the initiation of the culture did not modify DC morphology but caused a reproducible and statistically significant upregulation of HLA-DR, CD86, and CD80 surface expression. CD1a expression was significantly reduced, and CD40 expression was unchanged. We then determined the influence of IFN-beta on the production of cytokines by DC. DC differentiated in the presence of IFN-beta secreted significantly less IL-12 (p40 and p70) both spontaneously and on activation by fibroblasts transfected with the CD40L gene. This effect of IFN-beta was dose dependent and selective, as it was not observed for IL-6, IL-8, and tumor necrosis factor-alpha (TNF-alpha). As a consequence, DC differentiated in the presence of IFN-beta induced significantly less IFN-gamma secretion by alloreactive T cells, whereas they were more efficient than control DC in eliciting IL-5 secretion. We conclude that the direct action of IFN-beta on DC causes inhibition of their ability to secrete IL-12 in response to CD40 ligation and to elicit Th1 type responses.

IL‐5 in post‐traumatic eosinophilic pleural effusion

Thoracic trauma or pneumothorax can result in pleural fluid eosinophilia. In this study we investigated the role of the eosinophilopoietic cytokine IL‐5 in three cases of post‐traumatic eosinophilic pleural effusions (EPE). Using a specific immunoenzymatic assay, significant levels of IL‐5 were found in EPE (range 100–3000 pg/ml), while IL‐5 was undetectable (<25 pg/ml) in corresponding serum samples and in non‐eosinophilic pleural fluids. IL‐5 present in pleural fluids was found bioactive in a proiiferative assay using a mouse CTLL‐2 cell line transfected with the cDNA corresponding to the a chain of the human IL‐5 receptor. Using a reverse polymerase chain reaction (PCR) method, we found IL‐5 mRNA expression within pleural mononuclear cells from patients with EPE, but not in corresponding peripheral blood mononuclear cells (PBMC), confirming that IL‐5 is synthesized locally in the pleural cavity. In the two cases in which pleural CD4+ cells were purified, these cells were identified as the major source of IL‐5. Taken together, these data indicate that the development of post‐traumatic EPE is related to a local secretion of IL‐5 by CD4+ cells present in the pleural cavity.

Interleukin-5 production by T lymphocytes in atheroembolic disease with hypereosinophilia

1. Bennett JE. Chemotherapy of systemic mycoses (first of two parts). N Engl J Med 1974;290:30-2. 2. Murray HW. Allergic reactions to amphotericin B [Letter]. N Engl J Med 1974;290:693. 3. Lorber B, Cutler C, Barry WE. Allergic rash due to amphotericin B [Letter]. Ann Intern Med 1976;84:54. 4. Clements JS, Peacock JE. Amphotericin B revisited: reassessment of toxicity. Am J Med 1990;88:22N-27N. 5. Gigliotti F, Shenep JL, Lott L, Thornton D. Induction of prostaglandin synthesis as the mechanism responsible for the chills and fever produced by infusing amphotericin B. J Infect Dis 1987;156:784-9.

IL-6 and TNFα in ascitic fluid during spontaneous bacterial peritonitis

To the Editor: Spontaneous bacterial peritonitis (SBP) is one of the major complications of portal hypertension (1). Symptomatology is characterized by fever, abdominal pain, or simply unexplained clinical deterioration. In order to be effective, antibiotic therapy must be initiated prior to the bacterial diagnosis. The latter is frequently hampered by the low number of bacteria in ascitic fluid (2). Laboratory methods enabling early diagnosis are thus mandatory. Leukocyte and granulocyte counts (1, 3) or arterial ascitic fluid pH gradient measurement (1, 4) are currently among the most widely used. In the absence of infection, macrophages represent the predominant cell type in the ascitic fluid. These cells are an important source of proinflammatory mediators such as tumor necrosis factor e~ (TNFc0 or interleukin 6 (IL-6). These cytokines may be implicated in a variety of systemic manifestations (5-7). We have studied TNFo~ and IL-6 levels in the plasma and ascitic fluid of six patients with alcoholic liver cirrhosis (ALC) and ascites, considered as controls, and in four patients with ALC complicated by SBP caused by either E. coli (three cases) or Streptococousfaecalis (1 case). In each case, SBP was successfully treated with ampicillin and gentamycin intravenously, started immediately after the first sampling. For patients with SBP, ascitic fluid samples were taken daily during five days, conserved on ice, and immediately centrifuged. TNF~ in plasma and ascitic fluid were measured by an immunoradiometric assay (Ire-Medgenix, Fleurus, Belgium) and IL-6 by a bioassay using the IL-6-dependent 7TD1 cell line (8). In control ALC subjects, monokine concentrations were low but slightly elevated in ascitic fluid compared to plasma (P < 0.05 for IL-6, NS for TNFa). In contrast, a tremendous increase of IL-6 and TNFa was initially present in ascitic fluid of patients with SBP (Figure 1). In plasma, IL-6 and TNFoL levels were only slightly and inconsistently elevated among these patients. Following successful treatment, the monokine levels returned to normal values within five days while the patients clinically recovered. As rapid new tests such as ELISA will soon be available for their measurement, IL-6 and TNFa might become useful markers both for the diagnosis of SBP and for monitoring treatment. On the other hand, these inflammatory mediators have been implicated in the pathogenesis of septic shock and in the cytopathic effects associated with infections (5-7). Their marked overproduction in ascitic fluid might reflect an important pathogenic mechanism involved in the severity of such infection and the subsequent poor prognosis associated with the decompensation of the underlying liver disease (9, 10). J. DEVIERE, MD J. CONTENT, MD A. CRUSlAUX, MS E. DUPONT, MD Departments of Gastroenterology and Immunology Universitd Libre de Bruxelles HOpital Erasme Institut Pasteur du Brabant Brussels, Belgium