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F E Maly

Publications and source records attributed to F E Maly.

At least 37 records · Page 2Linked to original sources
PubMed1998

Analysis of specific immunoglobulin G subclass antibodies for serological diagnosis of Echinococcosis by a standard enzyme-linked immunosorbent assay.

The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3.

Animals↗
PubMed1998

Strongly enhanced serum levels of vascular endothelial growth factor (VEGF) after polytrauma and burn.

Angiogenesis is a key component of the repair mechanisms triggered by tissue injury. Vascular endothelial growth factor (VEGF) is an important mediator of angiogenesis, as it acts directly and specifically on endothelial cells. VEGF produced locally in regenerating tissue may spill over into the systemic circulation, and measuring levels of circulating VEGF may allow monitoring of angiogenesis. To determine whether circulating VEGF is increased after severe injury, we measured concentrations of VEGF in serial serum samples of 23 mechanical burn patients, 55 patients with multiple trauma and 56 healthy normal controls, using a newly established ELISA assay. In burn patients, serum VEGF was increased on day 1 (369.4 +/- 88.0 pg/ml) and on day 3 (452.0 +/- 65.3 pg/ml), reached highest levels on day 14 (1809.5 +/- 239.7 pg/ml) and was still elevated on day 21 post-burn (1339.8 +/- 208.7 pg/ml) (mean +/- SEM, p < 0.01), when compared with healthy controls (82.2 +/- 10.8 pg/ml (mean +/- SEM)). Likewise, in trauma patients, serum VEGF showed a trend towards elevated values on the day of admission (186.9 +/- 43.9 pg/ml) and on day 3 after injury (193.2 +/- 62.1 pg/ml). Thereafter, serum VEGF increased further (day 7,507.0 +/- 114.7 pg/ml), peaked on day 14 (742.4 +/- 151.8 pg/ml) and was still elevated on day 21 after injury (693.1 +/- 218.6 pg/ml (mean +/- SEM, p < 0.01)). No significant correlation was observed between peak serum VEGF and initial severity of mechanical (Injury Severity Score) or burn injury (percentage of body surface burned). However, in both burn and trauma patients, the subgroup of patients with uncomplicated healing showed significantly higher increases of serum VEGF than the subgroup who developed severe complications during the post-traumatic course, such as sepsis, adult respiratory distress syndrome or multiple organ failure (p < 0.05). Thus, markedly enhanced levels of serum VEGF are present one to three weeks after trauma or burn injury. Further, occurrence of severe complications during the post-traumatic period is associated with lesser increases of serum VEGF.

Adolescent↗
PubMed1997

[Cellular antigen stimulation test (CAST)--applicability in the diagnosis of insect toxin allergies].

In the CAST, blood leukocytes are prestimulated with the cytokine IL-3 and exposed to allergen(s). Mainly basophiles react by synthesizing sulfidoleukotrienes (sLT), namely LTC4 and its metabolites LTD4 and LTE4. They are detected by ELISA technique. 66 patients with suspected allergy to hymenoptera venoms were evaluated. Allergic reactions comprised all severity grades (I-IV ref. H. L. Mueller). The workup included skin tests (up to 1 microgram/ml, Pharmacia), measurement of specific serum IgE with RAST-CAP (Pharmacia), and CAST with three concentrations of bee (Apis mellifera) and wasp (Vespula spec.) venom (Aquagen ALK). A control group of 13 nonallergic, RAST-CAP negative individuals was established. CAST demonstrated pronounced variations of individual s-LT synthesis in patients and controls. The use of elevated cut-offs improved the results, while the use of three allergen concentrations did not increase reliability. Diagnostic accuracy of CAST was evaluated by comparison with skin tests as the "gold standard". With bee venom and a cut-off at 3 standard deviations, a sensitivity and specificity of 73%/71% was obtained. Wasp venom showed better results at 2 standard deviations, with a sensitivity and specificity of 68%/100%. CAST results were not influenced by the severity of the allergic reaction nor by the time lapse since the sting had occurred. In conclusion, CAST is a valuable test in insect sting allergy. However, established methods such as skin tests and RAST-CAP achieve better results. CAST should therefore be considered a supplementary method in cases where established methods fail. It remains to be shown whether CAST can be used as a parameter for monitoring patients who have undergone immunotherapy.

Adolescent↗
PubMed1996

Hypoxic induction of gene expression in chronic granulomatous disease-derived B-cell lines: oxygen sensing is independent of the cytochrome b558-containing nicotinamide adenine dinucleotide phosphate oxidase.

Reduced oxygenation of a variety of cells results in transcriptional upregulation of several genes, including the hematopoietic hormone erythropoietin, the angiogenic vascular endothelial growth factor (VEGF), and glycolytic enzymes such as aldolase. Recently, the heme protein cytochrome b558 of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex has been proposed as a key component of the oxygen-sensing mechanism. Cytochrome b558 consists of the p22phox and gp91phox subunits and is essential for superoxide generation in phagocytes and B lymphocytes. Mutations in these subunits result in cytochrome b558-negative chronic granulomatous disease (cytb- CGD), an inherited disorder in humans characterized by reduced microbicidal activity due to deficient superoxide generation. To test whether NADPH oxidase is involved in oxygen sensing, we exposed wild-type B-cell lines as well as cytb- CGD-derived B cell lines, deficient in either p22phox or gp91phox, to hypoxia (1% oxygen) or CoCl2 (100 mumol/L) and compared the mRNA levels of VEGF and aldolase with the untreated controls. Northern blot analysis revealed unimpaired basal and inducible expression of VEGF and aldolase mRNA in all four cytb- CGD-derived B-cell lines compared with wild-type cells. Furthermore, reconstitution of cytochrome b558 expression in cytb- CGD-derived B cells by transfection with p22phox or gp91phox expression vectors did not modify VEGF and aldolase mRNA expression. Thus, cytochrome b558 of the NADPH oxidase complex appears not to be essential for hypoxia-activated gene expression and can be excluded as a candidate for the putative universal oxygen sensor.

B-Lymphocytes↗
PubMed1994

Activated or dominant inhibitory mutants of Rap1A decrease the oxidative burst of Epstein-Barr virus-transformed human B lymphocytes.

Rap1A is a GTP-binding protein of the Ras superfamily that is highly abundant in phagocyte membranes. Although Rap1A copurifies with cytochrome b558, a component of the superoxide-generating NADPH oxidase complex of human phagocytes and B lymphocytes, the involvement of Rap1A in the regulation of the oxidative burst in these cells has not been clearly established. Therefore, we have stably transfected human Epstein-Barr virus-transformed B lymphocytes that possess an activable NADPH oxidase complex with cDNAs for mutants of Rap1A "locked" in a GTP-bound (63E) and GDP-bound (17N) state. Both the 17N and 63E mutants of Rap1A inhibited phorbol ester-stimulated O2-. production by 50 and 80%, respectively, while transfection with cDNA for wild-type Rap1A had no effect on the respiratory burst. No effects of the Rap1A mutants on cell viability, proliferation, expression of cell-surface markers, or phorbol 12-myristate 13-acetate-stimulated interleukin-8 generation were detected. These data demonstrate that Rap1A is a regulator of O2-. formation in intact cells. Furthermore, the inhibitory effect of both GTP- as well as GDP-bound mutants indicates that Rap1A functions in a dynamic cycle as opposed to a unidirectional pathway, as is the case for the other NADPH oxidase regulatory GTP-binding protein, Rac.

Antigens, Surface↗
PubMed1994

Colonic epithelial cell lines as a source of interleukin-8: stimulation by inflammatory cytokines and bacterial lipopolysaccharide.

Cytokines produced by intestinal epithelial cells may function as signals to neighbouring immune and inflammatory cells. We investigated production of the neutrophil and T-lymphocyte chemotactic cytokine interleukin-8 (IL-8) by intestinal epithelial cells using four colonic adenocarcinoma cell lines, T84, CaCo-2, HT29 and SW620, as a model system. These cell lines secreted substantial amounts of IL-8 if stimulated with IL-1 beta, tumour necrosis factor-alpha (TNF-alpha) or interferon-gamma (IFN-gamma), except CaCo-2 cells, which responded only to IL-1 beta. Bacterial lipopolysaccharide (LPS) was also an efficient stimulus of IL-8 release in SW620 and HT29 cells, whereas T84 and CaCo-2 cells were completely unresponsive to LPS, IL-8 secretion was greater at 4 hr after stimulation and was accompanied by induction of IL-8 messenger RNA. In T84 cells IFN-gamma and epidermal growth factor (EGF) stimulated IL-8 secretion synergistically with TNF-alpha, whereas in SW620 cells this synergism occurred only between IFN-gamma and TNF-alpha. IL-4, IL-10 and transforming growth factor-beta (TGF-beta), which can down-regulate IL-8 production in macrophages, had no effect on IL-8 generation by our cell lines. Adenocarcinoma cell culture supernatants also induced rapid transients of intracellular calcium in neutrophils. Depending on cell line and stimulus, supernatant bioactivity was completely or partially abrogated by neutralizing antibodies to IL-8, indicating that the cell lines investigated also generate other neutrophil-activating factors. IL-8 and possibly other chemokines generated by colonic adenocarcinomas may help to attract tumour-infiltrating leucocytes. Possibly, normal intestinal epithelial cells also have the potential to secrete this potent chemoattractant and thus might contribute to inflammatory responses of the intestinal mucosa, for example in inflammatory bowel disease.

Colon↗
PubMed1993

Restitution of superoxide generation in autosomal cytochrome-negative chronic granulomatous disease (A22(0) CGD)-derived B lymphocyte cell lines by transfection with p22phax cDNA.

The respiratory burst oxidase of phagocytes and B lymphocytes is a multicomponent enzyme that catalyzes the one-electron reduction of oxygen by NADPH. It is responsible for the O2-production that occurs when these cells are exposed to phorbol 12-myristate 13-acetate or physiologic stimuli, such as phagocytosis in phagocytes or cross-linking of surface immunoglobulin in B lymphocytes. The activity of this enzyme is greatly diminished or absent in patients with chronic granulomatous disease (CGD), an inherited disorder characterized by a severe defect in host defense against bacteria and fungi. In every CGD patient studied so far, an abnormality has been found in a gene encoding one of the four components of the respiratory burst oxidase: the membrane proteins p22phox or gp91phox which together form the cytochrome b558 protein, or the cytosolic proteins p47phox or p67phox. Autosomal recessive cytochrome-negative CGD (A22(0) CGD) is associated with mutations in the gene coding for p22phox. We report here that the capacity for O2- production and cytochrome b558 protein expression were restored to Epstein-Barr virus-transformed B lymphocytes from two A22(0) CGD patients by transfection with an expression plasmid containing a p22phox cDNA. No detectable O2- was generated by untransfected p22phox-deficient lymphocytes. The genetic reconstitution of the respiratory burst in A22(0) CGD B lymphocytes by transfer of the wild-type p22phox cDNA represents a further step towards somatic gene therapy for this subgroup of A22(0) CGD. This system will also be useful for expression of genetically engineered mutant p22phox proteins in intact cells, facilitating the structure-function analysis of cytochrome b558.

B-Lymphocytes↗
PubMed1993

The effect of endotoxin and tumour necrosis factor on erythrocyte and leucocyte deformability in vitro.

Microcirculatory disorders are a common finding in sepsis. We have analysed the influence of two factors released in sepsis, endotoxin and tumour necrosis factor (TNF), on rheological properties of blood cells. The deformability of mixed cell suspensions, isolated erythrocytes, mononuclear cells, or polymorphonuclear leucocytes exposed to endotoxin and TNF in vitro was assessed by filtration through pores of different sizes. Mixed blood cell suspensions showed an increase in cell rigidity when incubated with 100 ng/ml endotoxin. The filtration resistance of isolated erythrocytes, mononuclear or polymorphonuclear leucocytes was not affected by endotoxin. Incubation with TNF in physiological concentrations increased the rigidity of mixed blood cells and of isolated polymorphonuclear leucocytes in a dose- and time-dependent manner, while erythrocytes and mononuclear leucocytes remained unaffected. Polymorphonuclear cells showed decreased deformability associated with shape changes (polarized and non-polar cells with surface protrusions and a shift of F-actin into protrusions). The decrease in deformability was reversed by cytochalasin B or xanthin derivatives such as pentoxifylline. We conclude that TNF decreases the passive deformability of polymorphonuclear leucocytes, which may affect the microcirculation in sepsis. The reversibility with xanthin derivatives may represent a new therapeutic approach for the high morbidity and mortality in sepsis.

Endotoxins↗
PubMed1992

O2- production by B lymphocytes lacking the respiratory burst oxidase subunit p47phox after transfection with an expression vector containing a p47phox cDNA.

The respiratory burst oxidase of phagocytes and B lymphocytes is a complicated enzyme that catalyzes the one-electron reduction of oxygen by NADPH. It is responsible for the O2- production that occurs when these cells are exposed to phorbol 12-myristate 13-acetate or other appropriate stimuli. The activity of this enzyme is greatly decreased or absent in patients with chronic granulomatous disease, an inherited disorder characterized by a severe defect in host defense against bacteria and fungi. In every chronic granulomatous disease patient studied to date, an abnormality has been found in a gene encoding one of four components of the respiratory burst oxidase: the membrane protein p22phox or gp91phox, or the cytosolic protein p47phox or p67phox. We report here that O2- production was partly restored to phorbol 12-myristate 13-acetate-stimulated Epstein-Barr virus-transformed B lymphocytes from a patient with p47phox-deficient chronic granulomatous disease by transfection with an expression plasmid containing a p47phox cDNA inserted in the sense direction. No detectable O2- was produced by untransfected p47phox-deficient lymphocytes or by p47phox-deficient lymphocytes transfected with an antisense plasmid. The finding that O2- can be produced by p47phox-deficient B lymphocytes after the transfer of a p47phox cDNA into the deficient cells suggests that this system could be useful for studying the function of mutant p47phox proteins in whole cells.

Amino Acid Sequence↗
PubMed1992

B lymphoblasts show oxidase activity in response to cross-linking of surface IgM and HLA-DR.

Human B lymphocytes express components of the superoxide generating system of phagocytes, NADPH oxidase. We studied regulation of this 'B-cell oxidase' during in vitro blast transformation, using Lucigenin-amplified chemiluminescence (CL) to detect superoxide release. While freshly isolated tonsil B lymphocytes showed no CL responses, culture with phorbol myristate acetate (PMA) and ionomycin induced susceptibility to CL triggering by anti-IgM and anti-HLA-DR. Maximal effects were observed after 3 days of culture with 0.4 ng/ml PMA + 1 microgram/ml ionomycin. Cells from such B lymphoblast cultures showed no CL responses to opsonized zymosan. In contrast, peripheral blood mononuclear cells, where monocytes are the predominant oxidant source, showed CL responses to opsonized zymosan but not to anti-IgM and anti-HLA-DR, either before or after culture with PMA + ionomycin. Culture of B cells with the surface immunoglobulin cross-linking agent staphylococcus aureus Cowan I also led to emergence of a CL, response to anti-IgM, which was enhanced by interferon-gamma. Interestingly, markedly fewer B blasts than freshly isolated B lymphocytes expressed cytochrome b-558 surface antigen. Thus, the B-cell oxidase is up-regulated during blast transformation and can be triggered via surface IgM and HLA-DR; however, this appears to be restricted to a subset of B lymphoblasts.

B-Lymphocytes↗
PubMed1991

Evidence that exposure to fibrinogen or to antibodies directed against Mac-1 (CD11b/CD18; CR3) modulates human monocyte effector functions.

We have recently shown that the treatment of fibrinogen-coated monocytes (MO) with anti-fibrinogen as well as the exposure of MO to surface-bound fibrinogen (Fg) or to albumin haptenized with the Fg C-gamma-terminal pentadecapeptide, induces on oxidative burst. Using chemiluminescence (CL) for indicating an oxidative burst, and the ingestion of IgG-coated erythrocytes as a test of phagocytosis, we have now studied the impact of Fg on MO effector functions. MO that had been either pretreated with Fg and washed, or that were exposed to surface-adsorbed Fg, exhibited impaired Fc receptor-mediated phagocytosis. A similar impairment was observed when MO were pretreated with activating agents such as phorbol myristate acetate, n-formyl-methionyl-leucyl-phenylalanine (fMLP) or the calcium ionophore A23187. Moreover, following exposure to Fg or IgG, MO exhibited a reduced oxidative burst upon stimulation with a variety of agents. Similarly, MO pretreated or coincubated with anti-Mac-1 exhibited a reduced oxidative burst upon stimulation. Our results raise the possibility that inflammatory mononuclear phagocytes experience a functional modulation upon encountering fibrin by interacting with specific receptors for fibrin(ogen). This type of modulation is analogous to effects induced by the triggering of Fc gamma receptors. MO showed a decreased oxidative burst when either pretreated or coincubated with anti-Mac-1 antibodies, whereas antibodies directed against other MO surface constituents had no, or a weak effect only. This is compatible with the suggestion that Mac-1 acts as a fibrin (ogen) receptor.

Antibodies, Monoclonal↗
PubMed1990

Dual antibody stimulation: role of monocyte products and activation requirements of T cell subsets.

Using the model of dual antibody stimulation (cross-linking anti-CD3 BMA030-F(ab')2 antibodies with subset specific anti-T cell antibodies, i.e., anti-CD4 or anti-CD8 antibodies) we investigated the role of monocyte products in eliciting IL2 production and asked whether T cell subsets differ in their activation requirements. We found that activation mechanisms of dual antibody stimulation were equal for CD4 and CD8 as well as CD4 CDw29 and CD4 CD45R T cell subsets: dual antibody stimulation was necessary to induce responsiveness to soluble monocyte products which trigger IL2 synthesis. IL2 production induced by soluble monocyte products was measured in CD4 and CD8 T cells as specific IL2 mRNA expression and with a biological assay. Combined actions of IL1 beta and TNF alpha were identified as active monocyte products as (a) treatment of LPS-stimulated monocyte supernatants with anti-IL1 beta and anti-TNF alpha antibodies abrogated the proliferation inducing effect of monocyte supernatants and (b) addition of rIL1 beta and/or rTNF alpha enhanced proliferation of T cells stimulated by dual antibody cross-linking. If both recombinant monokines were added simultaneously a potentiated proliferative effect was observed. Whereas rIL1 beta plus/or rTNF alpha sustained proliferation, only rIL1 beta but not rTNF alpha induced IL2 synthesis in T cell subsets stimulated by dual antibody cross-linking.

Antigens, CD↗
PubMed1990

Enhancement of mononuclear cell oxidative burst by early post-transfer sera from human patients after unsuccessful embryo transfer.

Early pregnancy sera were earlier shown to modulate T lymphocyte rosetting with sheep erythrocytes. We set out to investigate whether early pregnancy sera also modulate the state of activation of another immunologically relevant cell type, the monocyte. As an index of monocyte activation, we measured phorbol ester-triggered oxidative burst activity by a highly sensitive chemiluminescence method. Contrary to our expectations, incubation of mononuclear cells with sera taken early after embryo transfer from patients with a successfully developing pregnancy had no effect. Unexpectedly, such sera from patients from a control group of patients in whom no pregnancy developed after embryo transfer caused enhancement of mononuclear cell chemiluminescence. Stimulatory activity of these sera appeared between days 4 and 6 and was maximal between days 7 and 9 post embryo transfer. Whether this phenomenon is causally related to, or represents a consequence of the failure of embryo transfer, can currently not be decided.

Chaperonin 10↗
PubMed1990

Oxidative burst in human Epstein-Barr virus-transformed B-cell lines triggered by immobilized specific antigen.

Normal human B lymphocytes and Epstein-Barr virus-transformed B-cell lines can produce reactive oxygen species such as superoxide if treated with phorbol myristate acetate (PMA) or with the surface immunoglobulin cross-linking agents protein A and anti-immunoglobulin. Here, we investigated under which conditions specific antigen, the natural ligand of surface immunoglobulin, can stimulate an oxidative burst in monoclonal Epstein-Barr virus-transformed B-cell lines producing antibodies of known specificities. After a short lag time of 1-2 min, exposure to the specific antigen stimulated a prolonged oxidative burst (tmax 30-90 min), as measured by Lucigenin-enhanced, superoxide dismutase-inhibitable chemiluminescence, in the corresponding line only. The effect was induced in each line if the specific antigen was immobilized to a solid support. Except in one line in which antigen also stimulated an oxidative burst if presented at relatively high density on a soluble carrier, soluble antigen did not induce B-cell oxidase activation. This suggests that normal, non-transformed B lymphocytes also require interaction with relatively dense deposits of specific antigen for activation of their oxidase.

Acridines↗
PubMed1990

Human monocytes CD36 and CD16 are signaling molecules. Evidence from studies using antibody-induced chemiluminescence as a tool to probe signal transduction.

A panel of monoclonal antibodies (mAb) directed against human monocyte surface antigens was tested for their capacity to mediate signal transduction by measuring luminol-enhanced chemiluminescence (CL). The response patterns fell into three categories. The mAb Mo4, OKM3, OKM6 and antibodies specific for Fc receptor (FcR) type I and II did not mediate signal transduction directly or were weak triggers, but upon second-order cross-linking by goat anti-mouse immunoglobulin (Ig) F(ab')2 or rabbit anti-mouse Ig, a strong CL response was induced. The mAb recognizing different epitopes on CD11b (complement receptor type III alpha chain) were unique in their ability to induce a CL response either by direct stimulation or by second-order cross-linking. The mAb 3G8 recognizing FcR type III (FcRIII; CD16) on a monocyte subpopulation and CD36-specific monoclonals directly elicited a CL response. A response of similar magnitude was obtained with 3G8 F(ab')2 or with intact 3G8. Furthermore, elutriation-centrifugation-purified monocytes were stimulated by 3G8 to a similar extent as unseparated mononuclear cells, whereas lymphocytes did not respond. This suggests that a FcRIII-expressing monocyte subpopulation may mediate effector functions, including the generation of reactive oxygen species, via FcRIII triggering. Our finding that anti-CD36 F(ab')2 directly induces an oxidative burst is the first evidence that CD36 itself is a trigger molecule. CD36, which is thought to interact with erythrocytes infected with Plasmodium falciparum and with thrombospondin, may constitute a signal-transducing element and thus may have functions extending beyond mediation of adherence. The present CL system constitutes a simple assay for detecting mAb directed against monocyte surface signalling elements. Probing mAb for signal transduction requires suspended cells and antibodies in the fluid phase in order to avoid inadvertent FcR triggering, which may occur when cells are stimulated by surface-adherent whole antibodies.

Antibodies, Monoclonal↗
PubMed1990

Long-lasting polymorphonuclear leukocyte oxidative burst activation by products of lipopolysaccharide-treated mononuclear cells is only partially due to tumor necrosis factor.

Supernatants of human mononuclear cells cultured in the presence of LPS (LPS-MNC-SN) directly induced production of superoxide by isolated polymorphonuclear leukocytes, measured as superoxide dismutase--inhibitable cytochrome c reduction or, more sensitively, Lucigenin-enhanced chemiluminescence. Of recombinant human cytokines, only TNF and to a lesser degree also LT, but not GM-CSF, showed this activity. Neutralising antibody to TNF reduced the activity of LPS-MNC-SN to activate an oxidative burst in polymorphonuclear leukocytes by 30-50%. In contrast, killing of TNF-sensitive mouse L 929 cells by LPS-MNC-SN was completely inhibited by anti-TNF, while the L929 killing assay was at least as sensitive for TNF as the granulocyte chemiluminescence assay. Material with granulocyte chemiluminescence inducing (GCI) activity but devoid of TNF was obtained by sequential chromatography of LPS-MNC-SN on Mono Q anion exchange and phenyl sepharose hydrophobic interaction chromatography. The time course of GCI-induced superoxide generation was prolonged with a tmax of approximately 60 minutes. Subsequent separation of this GCI-active material on Superose gel filtration yielded two overlapping activity peaks with apparent molecular weights of approximately 40-50 and 200 kDa. Further, two distinct GCI activity peaks were found when Mono Q--and phenyl-sepharose--separated material was subjected to reversed phase chromatography. Thus, in addition to TNF, LPS-triggered mononuclear cells produce also other granulocyte chemiluminescence inducing mediators which appear distinct from presently known cytokines.

Antibodies↗