Effects of Cytokines on the Binding of Leukocytes to Cultured Rat Hepatocytes and on the Expression of ICAM-1 by Hepatocytes

Adhesions of leukocytes to hepatocytes andsinusoidal endothelial cells mediates the induction andprogression of hepatic injury. However, in contrast toendothelial cells, information regarding the regulation of interactions between leukocytes andhepatocytes is limited. In the present study, weinvestigated the effect of inflammatory mediatorsincluding lipopolysaccharide (LPS), staphylococcalenterotoxin B (SEB), interferon- (IFN-), tumornecrosis factor- (TNF-), andinterleukin-1 (IL-1) on the adhesion ofpolymorphonuclear leukocytes or lymphocytes to primarycultured rat hepatocytes, and on the expression of intercellular adhesionmolecule-1 (ICAM-1) gene in hepatocytes. Bothpolymorphonuclear leukocyte and lymphocyte adhesion tohepatocytes were enhanced after exposure of hepatocytes to IFN- and TNF-, but not afterexposure to LPS, SEB or IL-1. The adhesion inducedby either IFN- or TNF- was inhibited bymonoclonal antibodies against ICAM-1 or lymphocytefunction-associated antigen-1 (LFA-1). Nonstimulated hepatocytesexpressed faintly ICAM-1 mRNA, which increased slightlyduring the culture period. ICAM-1 mRNA expression wasup-regulated to a greater extent by incubating hepatocytes with IFN- or TNF-,and peaked after 12 hr of incubation with TNF-and after 24 hr with IFN-. These results indicatethat IFN- and TNF- induce the expressionof ICAM-1 on parenchymal hepatocytes and that theLFA-1-ICAM-1 pathway plays an important role in theinteraction between hepatocytes and neutrophils orlymphocytes.     

Serum cytokines in patients with rheumatoid arthritis

Serum cytokines such as interleukin 1 (IL-1), interferon (IFN-), and tumor necrosis factor (TNF) were measured in 40 patients with rheumatoid arthritis (RA). In the 40 patients studied, serum IL-1 was detected in 5 patients, IFN- in 10 patients, and TNF in 20 patients. The IL-1-positive group showed increased values of activity indices compared to the IL-1-negative group. Values of serum IFN- correlated well with the number of peripheral blood lymphocytes and CD3⁺ cells and with the percentage of CD3⁺ CD26⁺ cells. Values of serum TNF correlated positively with the number of peripheral blood monocytes and the percentage of CD3⁺ HLA-DR⁺ and CD3⁺ CD25⁺ cells. These results indicated that serum IL-1 in RA patients reflects the activity of RA, while the serum IFN- and TNF in RA patients may be related to circulating activated lymphocytes and monocytes, respectively.     

γ/δ Receptor-expressing T-cell clones from a cutaneous T-cell lymphoma suppress hematopoiesis

Summary Recently we described a cutaneous T-cell lymphoma expressing the / T-cell receptor [5]. The patient suffering from this lymphoma showed low numbers of myeloid and T cells in peripheral blood, while B and NK cells were relatively increased. In vitro culture of the patient's bone marrow (BM) cells revealed a significant suppression of myeloid/monocyte colony formation (GM-CFU) compared with normal controls. This was not due to infiltration of the BM with lymphoma cells. We speculated that a soluble factor either secreted or induced by the lymphoma cells might be responsible for the marked suppression of hematopoiesis in this patient. From a skin biopsy with infiltrating / T-lymphoma cells we established T-cell clones bearing the / T-cell receptor and resembling the phenotype of the lymphoma cells. The supernatant (SN) of these / T-cell clones reduced the number of colonies in a CFU-GM assay (using normal control BM) in comparison to SN of / T-cell clones established from the same biopsy. This suppression was seen mainly on day 7 of culture and was not neutralized by the addition of placenta-CM. The main mediator of this suppression seems to be IFN-,since it was detectable in high amounts in the SN of these / T-cell tumor clones as well as in the serum of the patient. In addition, anti-IFN- antibodies can reverse the T-cell SN-mediated suppression of CFU-GM. We conclude that high serum levels of interferon-, which is secreted in high amounts from / T-cells grown from a biopsy of a cutaneous lymphoma, can suppress hematopoiesis.Abbreviations TCR T-cell receptor - IFN- interferon- - SN supernatant - placenta CM placenta conditioned medium - BM bone marrow - CFU-GM myeloid/monocyte colony formation - NK cells natural killer cells - Ab antibody M. Wilhelm was supported by theDeutsche Forschungsgemeinschaft (DFG Wi 728-2)     

Cytokine Production by Intestinal Intraepithelial Lymphocyte Subsets in Celiac Disease

One of the earliest signs of mucosal immune activation in celiac disease (CD) is an increase in the intraepithelial lymphocyte (IEL) count in the small intestinal epithelium. Though most of those IELs express T cell receptor (TcR)- chains, CD is characterized by an increase in TcR- ⁺ IELs and by the loss of CD3^– IELs. There is currently little evidence that these changes in IEL subset distribution are of relevance in the pathogenesis of CD. We aimed to determine the pattern of cytokine production by IEL subsets isolated from duodenal biopsy specimens from control subjects and CD patients at different stages of the disease. We quantified the capacity of IEL subsets to produce IFN-, TNF-, IL-2, IL-4, and IL-10 by intracellular staining by flow cytometry. All IEL subsets studied displayed a type I cytokine profile in both CD and control subjects, with TcR-⁺ IELs being the main IFN- producers. Untreated CD exhibited a trend toward a superior accumulation of IFN- per cell but a reduced proportion of INF-⁺ cells in vitro in association with a significantly increased apoptotic rate of IELs. IL-4 was almost undetectable in all cases and IL-10 showed a tendency to increase in treated and silent celiac patients. IEL subsets have a similar Th1 profile in controls and CD patients, and the superior in vitro apoptosis of IELs from CD patients may reflect their superior in vivo activation. The induction of IL-10-dependent regulatory Tr1 responses may be of potential clinical significance in this disease and merits further investigation.     

Coexpression of matrilysin and laminin-5 gamma2 chain may contribute to tumor cell migration in colorectal carcinomas

We attempted to examine the correlation between matrilysin and laminin-5 2 chain expression with reference to the number of dedifferentiation units along the entire invasive front (tumor budding). Immunostaining for hMMP-7 and laminin-5 2 chain was performed in 50 T1 colorectal carcinomas, and immunoreactivity was evaluated at the invasive front of the tumor. On hematoxylin–eosin sections, the number of tumor budding was counted. The localization of matrilysin tended to be widespread compared with that of laminin-5 2 chain. Matrilysin and laminin-5 2 chain expression were positive in 28 (56%) and 15 (30%) tumors respectively. There was a significant correlation between matrilysin and laminin-5 2 chain expression (P = 0.02). Matrilysin(+)/laminin-5 2 chain(+) tumors had a significantly greater amount of tumor budding than matrilysin(–)/laminin-5 2 chain(–) tumors (P = 0.003) or matrilysin(+)/laminin-5 2 chain(–) tumors (P = 0.03). In conclusions, coexpression of matrilysin and laminin-5 2 chain may contribute to tumor cell migration in colorectal carcinomas.     

Majority of Gliadin-Specific T-Cell Clones from Celiac Small Intestinal Mucosa Produce Interferon-γ and Interleukin-4

An abnormal mucosal cell-mediated immune response plays a fundamental role in the pathogenesis of celiac disease. To characterize locally infiltrating T cells, gliadin-specific T-cell clones were isolated from two treated celiac patients. Mucosal biopsies were cultured in vitro for 24 hr with a peptic-tryptic digest (PT) of gliadin. T-cell clones (TCC) were then isolated by limiting dilution. The production of interferon- (IFN-) and interleukin-4 (IL-4) was evaluated by ELISA in culture supernatants obtained after a short incubation with anti-CD3 and PMA, or with antigen. Twenty-two TCC were specific for gliadin and/or PT. All were CD3⁺, CD4⁺, CD8⁺, TCR ⁺. In one such clone the PT-specific response was inhibited by an anti-DQ, but not by an anti-DR antibody. Of the five gliadin-specific TCC examined, four produced IL-4 and high levels of IFN-; the remaining one initially produced only IL-4, but subsequently also IFN-. All clones obtained from the celiac mucosa, including the gliadin-specific ones, produced high levels of IFN-, in most cases with IL-4. This cytokine profile could explain most of the immunological features of the celiac mucosa.     

Establishment of murine macrophage hybridoma clones capable of acquiring tumoricidal activity upon activation with recombinant interferon-γ and lipopolysaccharide

Summary Murine macrophage hybridoma clones were established by fusing glycogen-elicited peritoneal exudate cells (glycogen-PEC) derived from C3H/HeN mice and the hypoxanthine-aminopterin-thymidinesensitive murine macrophage cell line, J774.3-2. The macrophage hybridomas were further screened for the capacity to acqurie tumoricidal activity upon stimulation with lipopolysaccharide (LPS) and recombinant interferon- (IFN-) using murine mammary adenocarcinoma MM48 cells as targets, and three macrophage hybridoma clones, KM-1, KM-2, and KM-3, were established. With concomitant stimulation with LPS, IFN- activated these hybridomas dose dependently to exhibit high tumoricidal activity, whereas single stimulation with either INF- or LPS, even with higher concentrations, did not activate the macrophage hybridomas. This contrasted with the activation of glycogen-PEC for eliciting tumoricidal activity with a single stimulation with LPS (>1 ng/ml) or IFN- (>10 IU/ml). Thus, the macrophage hybridoma clones established here represent inflammatory macrophages which require both IFN- and LPS for their activation.This work was supported by Grants-in-aid from the Ministry of Education, Science and Culture and the Ministry of Health and Welfare, Japan     

Antiproliferative effects of interferonγ in combination withα-difluoromethylornithine on human carcinoma cell cultures

The antiproliferative effects of human recombinant interferon (IFN) in combination with -difluoromethylornithine (DFMO) or as single agents were assessed on human cell cultures derived from carcinomas of the breast (MCF-7), the ovary (EFO-27) or the kidneys (EGI-4). Results were obtained in proliferation assays by direct cell counting. The cell lines differed considerably in their sensitivities to the antiproliferative effect of IFN as compared by the 50% inhibition doses of the growth (ID₅₀). In contrast to the findings with IFN, similar antiproliferative effects resulted from the application of comparable doses of DFMO. While IFN induced cytotoxic effects in EGI-4 cells, DFMO produced only cytostatic actions in the cell lines analyzed. Synergistic growth inhibition resulted from the combined application of IFN and DFMO in EFO-27 cell cultures. This finding was most pronounced after treatment with IFN or DFMO doses below the respective ID₅₀ values. However, antagonistic effects occurred in cells of the line EGI-4 after DFMO had been combined with IFN at concentrations below the cytotoxic dose range. Within the sensitivity of our proliferation assay, no synergistic interactions were found in MCF-7 cell cultures. In the cell lines tested, no relation between the sensitivity for the single agents and the effectivity of the drug combination was identified. Despite promising synergistic effects in the moderately IFN-sensitive ovarian carcinoma cell line EFO-27, the efficacy of the IFN/DFMO combination was restrained by possible antagonistic effects as demonstrated in the highly IFN-sensitive EGI-4 renal carcinoma cell cultures. We conclude that the differential interaction patterns in the cell cultures analyzed preclude general suggestions for clinical studies using IFN and DFMO.Abbreviations IFN interferon - DFMO difluoromethylornithine This work was part of the doctoral thesis of Mariam Klouche     

Comparison of the effects of ascorbyl γ-linolenic acid and γ-linolenic acid in the correction of neurovascular deficits in diabetic rats

Summary Essential fatty acid metabolism is impaired by diabetes mellitus and -linolenic acid rich treatments such as evening primrose oil correct deficits in nerve conduction and endoneurial blood flow in diabetic rats. Other mechanistically unrelated treatments, such as antioxidants and aldose reductase inhibitors have a similar effect and there may be positive interactions with multiple treatments. Our aim was to compare the efficacy of a novel essential fatty acid derivative, ascorbyl -linolenic acid, with that of -linolenic acid in correcting diabetic neurovascular deficits. Eight weeks of diabetes caused 20.4 and 48.2% reductions in sciatic motor conduction velocity and nutritive endoneurial blood flow, respectively. Treatment was given for the last 2 weeks with -linolenic acid (100 mg · kg^–1 · day^–1) either in pure form or as ascorbyl -linolenic acid, an equivalent dose of ascorbate (21 mg · kg^–1 · day^–1) or jointly with ascorbate and -linolenic acid. Conduction velocity was corrected by 39.8, 87.4, 13.2 and 66.8% with -linolenic acid, ascorbyl -linolenic acid, ascorbate and -linolenic acid plus ascorbate, respectively. Corresponding ameliorations of the nutritive blood flow deficit were 44.0, 87.4, 13.2 and 65.7%. For the -linolenic acid plus ascorbate combination, and especially for ascorbyl -linolenic acid, the magnitude of correction for conduction velocity and blood flow was greater than expected for simple addition of ascorbate and -linolenic acid, indicating a synergistic interaction. Thus, with an efficacy 40 times that of evening primrose oil in rats, ascorbyl -linolenic acid may be a suitable candidate for clinical trials of diabetic neuropathy.Abbreviations ARI Aldose reductase inhibitor - GLA -linolenic acid - NCV nerve conduction velocity - NO nitric oxide - ROS reactive oxygen species - PG prostaglandin     

Effects of Cytokines, Without and with Helicobacter pylori Components, on Mucus Secretion by Cultured Gastric Epithelial Cells

Cytokines are suspected to play a crucial rolein the pathogenesis of Helicobacter pylori associatedgastric diseases. Hence, considerable attention has beenpaid to the actions of cytokines on gastric cells. We examined the effects of cytokines onmucus secretion by gastric epithelial cells, without orwith H. pylori components. Mucus secretion by culturedgastric epithelial cells was assessed as secretion of [³H]glucosamine-prelabeledhigh-molecularweight glycoproteins. Interleukin(IL)-1 and IL-6 significantly stimulated mucussecretion, but other cytokines such as IL-7, IL-8,IL-10, interferon (IFN)- and tumor necrosis factor (TNF)- had noeffect. H. pylori lysate caused a decrease in both basaland stimulated secretion of mucus. In addition,IFN- significantly potentiated the lysate-induced reduction of basal and stimulated secretion.Cell viability was not affected by any of treatments.These results indicate that IL-1 and IL-6stimulate mucus secretion, while IFN- potentiatesH. pylori-decreased secretion by gastric epithelialcells.     

Compound heterozygosity for a β∘-thalassemia (frameshift codons 38/39; -C) and a nondeletional swiss type of HPFH (A→C at NT -110, Gγ) in a Czechoslovakian family

Summary We have analyzed the levels and composition of the fetal hemoglobin (Hb F) in several members of a Czechoslovakian family with a heterozygosity for a newly discovered -thalassemia (codons 38/39; -C), or for a newly detected nondeletional hereditary persistence of fetal hemoglobin (a form of Swiss-HPFH with an AC mutation at nucleotide –100 5 to the Cap site of G), or with a compound heterozygosity for these two conditions. The Hb F level in the -thalassemia heterozygotes averaged 0.3% with low G values ( 28%) and relatively high AT values ( 50%), that in the two Swiss-HPFH heterozygotes averaged 0.8% with 95% G, while that of the compound heterozygote was 3.1% with 95% G. The low Hb F levels were determined with a recently published cation exchange high-performance liquid chromatography (HPLC) procedure that is accurate at the 0.1%–0.2% Hb F level [3]. This method, together with a reversed-phase HPLC procedure, made it possible to detect this unusual type of nondeletional G-HPFH and provided the data indicating that the increased Hb F in the compound heterozygote was derived mainly from the chromosome with the HPFH determinant.This study was supported in part by USPHS Research Grant HLB-41544     

Serum concentrations of resistin-like molecules β and γ are elevated in high-fat-fed and obese db/db mice, with increased production in the intestinal tract and bone marrow

Aims/hypothesis Resistin and the resistin-like molecules (RELMs) comprise a novel class of cysteine-rich proteins. Among the RELMs, RELM and RELM are produced in non-adipocyte tissues, but the regulation of their expression and their physiological roles are largely unknown. We investigated in mice the tissue distribution and dimer formation of RELM and RELM and then examined whether their serum concentrations and tissue expression levels are related to insulin resistance.Methods Specific antibodies against RELM and RELM were generated. Dimer formation was examined using COS cells and the colon. RELM and RELM tissue localisation and expression levels were analysed by an RNase protection assay, immunoblotting and immunohistochemical study. Serum concentrations in high-fat-fed and db/db mice were also measured using the specific antibodies.Results The intestinal tract produces RELM and RELM, and colonic epithelial cells in particular express both RELM and RELM. In addition, RELM and RELM were shown to form a homodimer and a heterodimer with each other, in an overexpression system using cultured cells, and in mouse colon and serum. Serum RELM and RELM levels in high-fat-fed mice were markedly higher than those in mice fed normal chow. Serum RELM and RELM concentrations were also clearly higher in db/db mice than in lean littermates. Tissue expression levels revealed that elevated serum concentrations of RELM and RELM are attributable to increased production in the colon and bone marrow.Conclusions/interpretation RELM and RELM form homo/heterodimers, which are secreted into the circulation. Serum concentrations of RELM and RELM may be a novel intestinal-tract-mediating regulator of insulin sensitivity, possibly involved in insulin resistance induced by obesity and a high-fat diet.     

Molecular weight analysis of Fcγ-binding proteins of lymphoid leukemia,myeloid leukemia,and hairy-cell leukemia

Summary The molecular weights of EDTA-mercaptoethanol-soluble Fc-affined proteins isolated from chronic lymphocytic leukemia of the B type, prolymphocytic leukemia of the B type, chronic myeloid leukemia and hairy-cell leukemia were compared. SDS polyacrylamide gel electrophoresis of the Fc-binding material obtained from all six cases of B type leukemia revealed a single peak with an apparent molecular weight of 28,000. The Fc-affined material isolated from the cells of two cases of chronic myeloid leukemia showed two peaks, one with an apparent molecular weight of 42,600 and one with an apparent molecular weight of 18,800. The Fc-affined material isolated from the cells of two cases of hairy-cell leukemia electrophoresed in the form of a closely spaced double peak. One component of the double peak had an apparent molecular weight of 28,000 and thus corresponds to the Fc-binding material of leukemic B cells. The second component had a slightly lower molecular weight. The latter component is not present on either leukemic B cells or myeloid cells. The results indicate that the EDTA-mercaptoethanol-soluble Fc-affined proteins of different types of cells differ in molecular weight, and thus in molecular structure.Supported by the Deutsche Forschungsgemeinschaft, SFB 111, project nos. D8 and D7     

Laminin-5 gamma2 chain expression as a possible determinant of tumor aggressiveness in T1 colorectal carcinomas

This study was undertaken to clarify the associations between laminin-5 2 chain expression, and tumor budding and lymph node metastasis or local recurrence (locoregional failure) of 50 T1 colorectal carcinomas immunohistochemically. Fifteen (30%) of 50 lesions were positive for laminin-5 2 chain expression. By univariate analysis, less histological differentiation (P = 0.02), nonpolypoid growth pattern (P = 0.03) and tumor budding (P < 0.001) were associated with laminin-5 2 chain positivity. By multivariate analysis, tumor budding alone was significantly associated with laminin-5 2 chain positivity (P = 0.006), and correlation between nonpolypoid growth pattern and laminin-5 2 chain positivity lost its significance (P = 0.09). Tumor budding (P = 0.004) and laminin-5 2 chain expression (P = 0.001) were significantly associated with locoregional failure. Laminin-5 2 chain expression may contribute to the formation of budding tumor cells at the invasive front, and immunostaining of this adhesion molecule may be useful in identifying high-risk patients for locoregional failure in T1 colorectal carcinomas.     

Regulation of the release of tumour necrosis factor (TNF)α and soluble TNF Receptor by γ irradiation and interferon γ in Ewing's sarcoma/peripheral primitive neuroectodermal tumour cells

This study analyses the production of tumour necrosis factor (TNF) and soluble TNF receptor (sTNF-R) before and after exposure to irradiation and interferon (IFN) in 12 cell lines derived from Ewing's sarcoma (ES)/peripheral primitive neuroectodermal tumours (pPNET). Supernatants from ES/pPNET cell cultures were tested in a TNF-specific amplified enzyme-linked immunosorbent assay (ELISA), a bioassay, and sTNF-R_p55 and sTNF-R_p75 ELISA. The tumour cell lines released minimal amounts of TNF, prominent amounts of sTNF-R_p55 (7/12 cell lines) and no sTNF-R_p75. Exposure to irradiation (5 Gy) either induced (3/12) cell lines) or up-regulated (3/12 cell lines) TNF release without changing sTNF-R_p55 and sTNF-R_p75 levels. Priming of cultures with recombinant human IFN (rhIFN) markedly enhanced TNF secretion in the radiation-responsive cell lines and had no influence on sTNF-R_p55 and sTNF-R_p75 levels. rhIFN affected the magnitude rather than the sensitivity of the radiation response. The TNF secreted was bioactive, as shown by its cytotoxic effect of WEHI-164 cells, and neutralization of its activity by anti-TNF monoclonal antibody. Herbimycin A (a tyrosine-specific protein kinase inhibitor) but not calphostin C (a protein kinase C inhibitor), H89 (a protein kinase A inhibitor), AACOCF3 (a specific inhibitor of phospholipase A₂) and MK-886 (a specific inhibitor of 5-lipoxygenase) abrogated -irradiation-stimulated TNF release. The antioxidantsN-acetylcysteine, nordihydroguaiaretic acid and mepacrine dose-dependently inhibited -irradiation-mediated TNF production. Collectively our findings indicate that IFN priming potentiates the secretion of bioactive TNF by ES/pPNET cells in response to irradiation without affecting sTNF-R release. The data suggest a requirement for protein tyrosine kinase activity and a role for reactive oxygen species in the -irradiation-mediated intracellular signalling pathway leading to TNF production.     

Prognostic significance of laminin-5 gamma2 chain expression in colorectal carcinoma: immunohistochemical analysis of 103 cases

PURPOSE: The laminin-5 2 chain plays an important role in cell migration during tumor invasion and tissue remodeling. Although this chain has been reported to be expressed in tumor-stroma interface of colorectal carcinoma, prognostic significance of its expression has not been elucidated in these tumors, so we investigated the clinicopathologic significance of Laminin-5 2 chain expression in colorectal carcinoma. METHODS: Laminin-5 2 chain expression was investigated immunohistochemically in 103 colorectal carcinoma patients with Stage II, III, and IV disease. The patients were categorized into three groups according to the number of immunopositive tumor cells in the sections containing the maximum diameter of the tumor as follows: +, less than 20 tumor cells were positive; ++, 20 to 500 tumor cells were positive; +++, more than 500 tumor cells were positive. RESULTS: Laminin-5 2 chain expression was observed in cytoplasm of tumor cells, especially those in the invasive front of the tumor penetration. Eighteen (17 percent) of tumors showed +, 60 (58 percent) showed ++, and 25 (24 percent) showed +++. The increased number of immunopositive tumor cells was significantly associated with synchronous liver metastasis (P = 0.048). The univariate (P = 0.036) and multivariate (P = 0.001) analysis of the patients survival revealed that the prognosis became significantly poorer in patients with the increased number of immunopositive tumor cells. CONCLUSIONS: Increased laminin-5 2 chain immunoreactivity, suggesting a high invasive potential of tumor cells, was a significant poor prognostic indicator for the patients with colorectal carcinoma.     

Lymphocyte apoptosis and macrophage function: correlation with disease activity in systemic lupus erythematosus

Increased lymphocyte apoptosis and defects in macrophage removal of apoptotic cells have been suggested to contribute to the development of systemic lupus erythematosus (SLE). The aim of this study was to investigate the relationship between peripheral lymphocyte apoptosis, macrophage function as determined by the serum levels of neopterin and interferon- (IFN-), and SLE disease activity. Peripheral apoptotic lymphocytes (AL) were detected by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry. Serum levels of neopterin and IFN- were measured by enzyme-linked immunosorbent assay (ELISA). SLE disease activity was determined using the systemic lupus activity measure (SLAM) and the serum titer of anti-dsDNA antibodies. The percentage of AL in the peripheral blood of active SLE patients was significantly higher (13.07±7.39%, n=30) than that of the inactive SLE patients (4.08±3.55%, n=8, p<0.01) and normal controls (5.13±3.37%, n=11, p<0.01). Serum levels of neopterin in active SLE patients were significantly higher (1.39±1.10 g/dl, n=22) than in controls (0.26±0.19 g/dl, n=20, p<0.01). Serum levels of IFN- in active SLE patients were elevated (58.97±34.52 ng/l, n=15) when compared with controls (28.06±2.35 ng/l, n=16, p<0.05). The percentage of AL correlated significantly with serum levels of neopterin (r=0.446, p<0.05, n=22) and SLAM score (r=0.533, p<0.001, n=38), but not with the serum levels of IFN-. The SLAM score also correlated with the serum levels of neopterin (r=0.485, p<0.05, n=22), but not with those of IFN-. Our study supported the hypothesis that increased lymphocyte apoptosis has a pathogenic role in SLE. The increased levels of serum neopterin may suggest an attempt of the patients macrophage system to remove the apoptotic cell excess. Since serum levels of neopterin correlated with the overall lupus disease activity, they may be regarded as an index of SLE disease activity.     

Decreased levels of interleukin-12p40 in the serum of patients with Whipple’s disease

Background An impaired production of interleukin (IL)-12 and T cell interferon- (IFN-) of in vitro stimulated monocytes has been discussed as a pathogenic factor in Whipples disease (WD). It is unclear whether this defect of cellular immunity is translated to the humoral immune system and to serum correlates.Methods We analyzed the serum of 40 patients with Whipples disease in various degrees of disease activity by sandwich enzyme-linked immunosorbent assay for differences in cytokine and cell adhesion molecule concentrations compared with age- and sex-matched controls.Results We observed a highly significant reduction of IL-12p40 levels (patients, 0.18±0.05 ng/ml (mean±SEM); controls, 3.19±0.39 ng/ml; p<0.01) in all stages of disease activity, whereas the concentration of IL-12p70 was comparable with controls. Furthermore, we observed a slight decrease in tumour necrosis factor (TNF-) concentrations in the serum of patients (patients, 6.36±0.90 pg/ml; controls, 10.5±1.23 pg/ml; p<0,05). The levels of other cytokines such as IFN-, IL-2, IL-13 and transforming growth factor , as well as soluble cell adhesion molecules lymphocyte function-associated antigen 3 and intercellular adhesion molecule 1, were not significantly different compared with controls. Levels of immunoglobulin G2 (IgG2) measured in the serum of WD patients were below normal in 24 of 29 patients and were even below the 95% confidence interval in 10 patients.Conclusion Our data demonstrate a persistent defect of the cellular immune response with decreased serum concentrations of IL-12p40 and TNF- and decreased IgG2 levels in a large group of WD patients. These data support as in vivo finding the results obtained in previous investigations with stimulated monocytes/lymphocytes. The isolated decrease in IL-12p40 may hint at possible defects in the IL-12/IFN- promoter system.     

Rheumatoid arthritis associated with transient gamma 2-heavy chain disease

Summary This paper describes the clinical history of a patient (F.-O.) with longstanding rheumatoid arthritis (RA), who subsequently developed a transient gamma 2-heavy chain disease (₂-HCD). Immunochemical studies comprised serial determinations of serum levels of intact IgG, the -HCD protein, IgA, and IgM. New applications of the rocket immunoselection and the radial immunodiffusion were used for the quantitation of the -HCD protein and intact IgG, respectively, in the presence of one another. Immunofluorescent microscopy on bone marrow cells showed cells containing -heavy chains but devoid of light chains. Protein studies of the isolated -HCD protein revealed a molecular weight of 72 000 in the dimeric form, a carbohydrate content of 9.7%, and a PCA-Val-Gln NH₂-terminal amino acid sequence. The literature on the rare coexistence of RA and -HCD in a single patient is reviewed.     

Local and Systemic Liberation of Proinflammatory Cytokines in Ulcerative Colitis

Determination of plasma and tissue cytokinelevels in inflammatory bowel disease have frequentlyresulted in conflicting data. In the present study wedetermined in patients with ulcerative colitis (UC), the levels of the proinflammatory cytokinesinterleukin (IL)-1, IL-6, interferon(IFN)-, and tumor-necrosis factor (TNF)-liberated by peripheral blood mononuclear cells (PBMC)and lamina propria mononuclear cells (LPMC) after 48-hrculture with pokeweed mitogen (PWM). IL-1, IL-6,IFN- and TNF- in the supernatant weredetected by ELISA. Results show low basal levels ofIL-1 secretion by PBMC and LPMC, and a considerableincrease after mitogen stimulation. Basal IL-6production by PBMC was higher in UC patients than incontrols [2029 pg/ml, CI₉ (–165 to4223) vs 572 pg/ml (–383 to 1527) respectively, P = 0.05] and also afterPWM activation [14,995 pg/ml (7759 -22230) vs 6598 pg/ml(3240-9956), respectively, P = 0.05]. In LPMC, nodifferences in IL-6 secretion were observed. TNF- in activated PBMC of patients with UC was notsignificantly increased in relation to control (P =0.09). No constitutive secretion of IFN- wasobserved in mononuclear cells. IFN- levelssecreted by activated LPMC were lower in patients withUC than in controls [1571 pg/ml (–108 to 3251) vs7953 pg/ml (3851-12,055), respectively, P = 0.03]. Theseresults suggest that IL-6, IL-1, and TNF- participate as mediators in the inflammatoryphenomena observed in UC. Further studies are necessaryto evaluate the role of IFN- in thiscondition.