From model cell line to in vivo gene expression: disease-related intestinal gene expression in IBD.

Crohn's disease (CD) and ulcerative colitis (UC) are subforms of inflammatory bowel diseases (IBD). Genetic and environmental factors influencing the onset and course of the diseases have been recently identified. This study uses a two-step approach to detect genes involved in the pathogenesis of IBD by microarray analysis and real-time PCR (RT-PCR). In a first step, microarray expression screening was used to obtain tumour necrosis factor-alpha (TNF-alpha) induction profiles of two human cell lines to represent the tissue cell types involved in IBD. In a second step, a subset of differentially expressed genes was examined by real-time PCR in intestinal biopsy samples of normal controls (NC) compared with UC and CD patients, as well as to a cohort of patients suffering from intestinal diseases other than IBD. Data were obtained from 88 CD, 88 UC, 53 non-IBD patients (inflammatory control), DC and 45 NC individuals. The experimental design enabled the identification of disease-specific expressed genes. DnaJ (Hsp40) homologue, subfamily B, member 5 (DNAJB5) was downregulated in intestinal biopsy samples of the UC cohort compared with NC. A difference in JUNB expression levels was observed by comparing biopsy samples from inflamed and non-inflamed areas of UC patients. Transcript expression differences between IBD and control cohorts were found by examining histamine N-methyltransferase (HNMT), interleukin-1A (IL-1A) and proplatelet basic protein (PPBP) expression. The experimental procedure represents an approach to identify disease-relevant genes, which is applicable to any disease where appropriate model systems are available.

Effect of surface modifications of intraocular lenses on the adherence of Staphylococcus epidermidis.

The development of polymers with different surface properties and surface modifications of intraocular lenses (IOL) should reduce foreign body reactions after implantation by reducing the surface hydrophobicity of the lenses. It was examined how far such surface variations influenced the adhesiveness of bacteria. The most common organism isolated from cases of postoperative endophthalmitis is Staphylococcus epidermidis. For this reason, three strains of this species, the type strain ATCC 14990 and two clinical isolates (8687, 6579 I), with different hydrophobic surfaces, were studied. IOL made of PMMA, silicone, and a copolymer as well as PMMA lenses with modified surfaces (unpolished, polished, silanized, and heparinized) were used. Bacteria were radiolabelled with 3H-thymidine and the adherent bacteria were calculated per mm2 of lens surface. The three strains adhered better to the unpolished surface of silicone than to PMMA. Treatment of PMMA surface by polishing diminished the differences between the strains. An influence of hydrophobic interactions on the adherence of S. epidermidis ATCC 14990 was demonstrated. The adherence of this hydrophobic type strain was clearly reduced by heparinization of the PMMA surface. In contrast, the hydrophilic catheter isolate 6579 I adhered better to modified surfaces. This strain differed clearly in its PFGE pattern from both hydrophobic strains. Hydrophobic interactions play a role in the bacterial adherence to intraocular lenses in vitro and in vivo. Modifications of polymer surfaces, however, can result in rather different effects depending on the bacterial surface composition and properties.

[Adherence of staphylococci of different hydrophobicity. Study of various intraocular lenses]. / Adhärenz von Staphylokokken unterschiedlicher Hydrophobizität. Untersuchung an verschiedenen Intraokularlinsen.

BACKGROUND: A major goal in research on intraocular lenses (IOL) is the development of new polymers and modifications to reduce foreign-body reactions after implantation. This effect may be achieved by a reduction in the surface hydrophobicity of the polymers. To illustrate the influence of surface modifications on bacterial adhesiveness, the most often isolated organism in "low-grade" postoperative endophthalmitis, Staphylococcus epidermidis, was used. MATERIALS AND METHODS: For this reason three strains of this species, the type strain ATCC 14990 and two clinical isolates (8687, 6579 I) with different hydrophobic surface properties were studied. IOL, used in the experiments were either made of PMMA or silicone with modified surfaces (unpolished, polished, heparinized). The adhesiveness of H3-thymidin-labeled bacteria was calculated/mm2 of lens surface. Each experiment was performed in triplicate and repeated three times. RESULTS: The hydrophobic-type strain showed stronger adherence to unpolished PMMA surface (8000 bacteria per mm2) compared to the polished (5200 bacteria/mm2). In contrast, the hydrophilic strain adhered with 2000 bacteria/mm2 to the unpolished and with 4200 bacteria/mm2 to the polished surface. Polishing PMMA lenses diminished the differences between the three strains. However, surface passivation of silicone lenses increased the adhesion rate of the hydrophilic strain up to 9600 bacteria/mm2. Treatment of PMMA lenses with heparin increased the adhesiveness of the hydrophilic strain and reduced the adhesion rate of the hydrophobic type strain to 250 bacteria/mm2. CONCLUSIONS: It was demonstrated that bacterial adherence to IOL also involves hydrophobic interactions. Obviously, however, that adherence reflects a complex of interactions between the two surfaces.

In vitro effects of solubilized HLA-DR--role in immunoregulation?

To test the hypothesis that soluble HLA-DR antigens (sHLA-DR), binding to the T-cell receptor (TCR) and/or CD4 structures, compete with and abrogate functions of their cell-bound counterparts, we studied effects of detergent-solubilized, affinity-purified HLA-DR molecules on the DNA synthesis, IL-2, and IL-1 secretion by human peripheral blood mononuclear cells (PBMC). While resting T cells did not show any response, there was a dose-dependent suppression of T-cell responses induced by mitogen (phytohemagglutinin, PHA), recall antigen (purified protein derivative of tuberculin, PPD), or HLA class II alloantigens (Daudi cells). In the PHA system, sHLA-DR affected DR-identical and DR-disparate PBMC with equal efficiency, suggesting a nonspecific interference with accessory functions of cell-bound HLA class II molecules. A competitive ligation of lymphocyte (rather than of monocyte) CD4 is suggested based on the failure of sHLA-DR (i) to potentiate inhibitory effects of anti-CD4 mAb, (ii) to suppress mitogen responses after depletion of CD4+ lymphocytes and also following separate monocyte (vs lymphocyte) pretreatment, and (iii) to induce a reproducible IL-1 secretion inhibition. In the PPD system, suppressive sHLA-DR effects on autologous or DR-identical PBMC significantly exceeded that on DR-disparate PBMC, and in the MLR, third party allogeneic sHLA-DR was exceeded in its suppressive potency both by sHLA-DA pertinent to responder cells and by sHLA-DR pertinent to stimulator cells. These additional specific effects may result from competition (with cell-bound class II restriction and recognition determinants) at the TCR level rather than from peptide competition at the antigen-presenting cell level. Interference by sHLA-DR with the primary and/or accessory signaling may offer new therapeutic strategies in allotransplantation and autoimmunity.

Detection of mannose-binding proteins on mouse lymphocytes.

D-Mannose influences both the allogeneic stimulation of mouse spleen cells and the induction of cytotoxic T-lymphocytes. At low concentrations (10(-3) -4 x 10(-3) M), D-mannose increases [3H]thymidine incorporation and induction of cytotoxic cells. Higher concentrations lead to an inhibition of the allogeneic response. To test the involvement of mannose receptors, unstimulated and allogeneically stimulated spleen cells were lysed and the proteins electrophoretically separated. After blotting, the D-mannose-binding structures were identified by means of a biotinylated mannose-neoglycoprotein and avidin peroxidase. Two mannose-binding proteins of unstimulated mouse spleen lymphocytes with a molecular mass of 29 and 31 kDa were detected. In vitro allogeneically stimulated spleen lymphocytes show three other mannose-binding proteins with a molecular mass of 53, 65, and 76 kDa.

[Interleukin 1 (IL 1) in gingival fluid from patients with plaque induced gingivitis]. / Der Interluekin-1 (IL 1)-Gehalt in der Sulkusflüssigkeit von Patienten mit plaqueassoziierter Gingivitis.

The level of interleukin 1 in gingival fluid is determined in 17 volunteers with healthy gingiva and in 58 patients with gingivitis of different severity. Continously increased levels of local IL 1 are found in connection with increased plaque accumulation in the dentogingival sites and with exacerbating gingival inflammation. The relevance of IL 1 in gingival fluid as a sensitive indicator of plaque-associated gingivitis is discussed.

Are soluble monocyte-derived HLA class II molecules candidates for immunosuppressive activity?

Supernatants of human blood monocyte cultures suppressed PHA responses (IL-2 synthesis, IL-2R expression, DNA synthesis) of autologous and allogeneic lymphocytes. The main suppressive activity was found in the 65-kDa (and 23-kDa) range. It could be incompletely neutralized by mAb specific for a non-polymorphic HLA DR determinant and could also be adsorbed to and eluted from an anti-DR immunoabsorbent column. On blots of monocyte lysates and monocyte culture supernatants, the mAb RoDR recognized antigens of nearly the same Mr. The hypothesis that soluble HLA DR alpha beta heterodimers or beta chains are likely candidates for the suppressor factor was confirmed by analogous effects of purified HLA DR molecules. We favor a model in which soluble MHC class II molecules (in contrast to surface-bound ones) may interfere with the association and cross-linking processes necessary for T cell activation by competing for CD4 binding sites.

[Studies on the local defense reactions of the nasal mucosa in employees in Rostock harbor exposed to large amounts of dust]. / Untersuchungen zur lokalen Abwehrleistung der Nasenschleimhäute bei extrem staubexponierten Werktätigen im Seehafen Rostock (SHR).

Defense mechanisms of the nasal mucosa are characterised by resistance and immunity processes. Secretory IgA seems to be very important for immune response. The aim of our work was to clarify whether the concentration of S-IgA in nasal secretion was influenced by extreme exposure of dock workers to dust. We investigated the nasal fluid of both the septum and the turbinate mucosa of 75 dock workers compared with 12 controls. A decrease of S-IgA concentration in the nasal fluid was seen after the work. This decrease differed for the various types kinds of dust and was significant (p less than 0.05) for apatite, urea and potash. A reduction of S-IgA was also seen after exposure for a longer time (several years). Dockers with repeated infections do not have decreased S-IgA values. These investigations make a contribution to the immune response in the nasal mucosa.

Relations between the classification of endogenous psychoses by Leonhard, the doctrine of pathoclisis by Vogt and the neurological conception of system diseases.

On the basis of 30 years of cowork with Leonhard it is attempted to enlighten the theoretical background of his classification of endogenous psychoses. Inspired by the conceptions of Wernicke and Kleist he explained the differentiation of symptomatology by the model of system diseases applying this in completing and improving the psychopathological classification. His ideas about the origin of a differentiated vulnerability of brain structures are very similar to the doctrine of pathoclisis of Vogt, although Leonhard did not use this term.

[Nosologic classification of Fazio-Londe disease]. / Uber die nosologische Einordnung der Fazio-Londe-Krankheit.

The observation of a progressive bulbar paralysis with lethal exit in a 20 years old patient, whose mother had died in the age of 29 years after a similar course of disease, is coordinated as Fazio: Londe-disease. But peculiarities are mentioned, that refer to traits of the special form of progressive bulbar paralysis described by Kennedy and of Kugelberg-Welander-disease. A genetic basis of variability is supposed.

Effect of recombinant human tumor necrosis factor on tumor and spleen in mice.

The ability of recombinant human tumor necrosis factor (rH-TNF-alpha) to induce regression of sarcoma 180 in vivo was evaluated. The tumor was cured by TNF in the course of 4 weeks. TNF inhibited proliferation of sarcoma 180 cells in vitro, which suggests a direct effect of TNF on tumor cells in vivo. In parallel to the TNF effect on tumor growth, some cell parameters in spleen were investigated. Activation of splenic macrophages was enhanced in vitro. This result suggests that macrophages may participate in the host defense against the tumor. In the first phase of therapy, TNF did not affect the proliferation of splenocytes but increased the transition of G0 into G1 cells. Furthermore, TNF normalized the tumor-induced increase of null cells in tumor-bearing mice. All parameters investigated in spleen reached normal values at the time of tumor regression. Our results suggest that various mechanisms may be involved in TNF-induced regression of sarcoma 180.

[Functional capacity of T cell subpopulations in mice. II. Effect of allogeneically stimulated T-cell subpopulations on the primary MLR]. / Untersuchungen zur funktionellen Kapazität von T-Zell-Subpopulationen der Maus. II. Einfluss von allogen stimulierten T-Zell-Subpopulationen auf die primäre MLR.

The effect of allogeneically stimulated lymphocytes or their subpopulations respectively on a primary allogeneic MLC was studied. It was shown that this effect depends either on the amount of the transferred cells or on the composition of the culture medium. In contradiction to the common view that allogeneically stimulated lymphocytes act as suppressor cells, a significant stimulating effect, mediated by these cells, could be demonstrated under respective conditions. Also in the experiments with the subpopulations of allogeneically stimulated lymphocytes the effects of these cells are rather influenced by the conditions then determined by a restricted functional repertoire of the cells.

[Psychiatry and psychotherapy in the many-sided contributions of Alexander Mette]. / Psychiatrie und Psychotherapie im vielseitigen Schaffen von Alexander Mette.

Appreciating the personality and work of Alexander Mette, who founded together with Karl Bonhoeffer this journal, especially his psychiatric and psychotherapeutic inclinations as well as his influences on the development of our discipline are taken into consideration in the frame of his many-sided activities as a man of letters and cultural policy.

[Contribution of E. W. Schmidt to the development of Soviet neurology and its international relations]. / Der Beitrag von E. W. Schmidt zur Entwicklung der sowjetischen Neurologie und ihrer internationalen Verbindungen.

A short retrospect, occasioned by the death on July 13, 1985, of E. W. Schmidt, of his career as director of the Neurological Research Institute at the Academy of Medical Sciences of the USSR, and his influence as scientist, medical man, and humanist on the development of neurology in the Soviet Union and its representation on the international scene.

Suppression of GvHR by in vitro allogeneically stimulated lymphocytes: the role of lymphoblasts in the regulation of a primary T-cell response.

Graft versus host reaction (GvHR) induced in 10-day-old F1 mice by in vitro allogeneically or mitogenically stimulated spleen cells is lower than that induced by unstimulated fresh spleen cells. In vitro stimulated lymphoblasts are unable or only slightly able to induce a GvHR. An active suppression by the blasts is not involved. Since lymphoblasts after in vivo stimulation show an increased ability to elicit a GvHR it is concluded that in vitro and in vivo stimulated lymphoblasts have different properties. A different homing cannot be excluded after transfer to the mouse.