Transfer of allergic encephalomyelitis with spleen cells from donors sensitized with myelin basic protein in incomplete Freund's adjuvant

Myelin basic protein (BP) emulsified in incomplete Freund's adjuvant (BP/IFA) is relatively nonencephalitogenic in Lewis rats. Furthermore, repeated injections of BP/IFA prevent subsequent induction of experimental allergic encephalomyelitis (EAE) by BP emulsified in complete Freund's adjuvant (BP/CFA). In spite of this, spleen cells from rats injected repeatedly with BP/IFA transfer EAE after they are cultured with BP almost as effectively as BP/CFA spleen cells. However, unlike the latter, BP/IFA spleen cells do not proliferate in response to BP in culture. Furthermore, BP/IFA spleen cells are unable to transfer EAE after culture with concanavalin A (Con A), in contrast to BP/CFA spleen cells. Both populations of spleen cells undergo a strong proliferative response to Con A in culture. For BP/IFA cells, at least, a proliferative response to BP in vitro is not a prerequisite for enhanced transfer of EAE in Lewis rats.     

Limited Digestion of Guinea Pig Myelin Basic Protein and Its Carboxy-Terminal Fragment (Residues 89–169) with Staphylococcus aureus V8 Protease

Staphylococcus aureus V8 protease has been reported to have a strict specificity for cleavage of the Glu-X bond in ammonium bicarbonate (pH 7.9). With myelin basic protein and one of its major peptic fragments (residues 89-169) as substrates, selective cleavage of Asp(32)-Thr(33), Asp(37)-Ser(38), and Glu(118-Gly(119) bonds was observed, as well as the unusual cleavage of the Gly(127)-Gly(128) bond. The Asp-Glu and Glu-Asn bonds in the sequence of Gln-Asp-Glu-Asn-Pro(81-84) were resistant to V8 protease attack. The following peptides were identified as products of limited cleavage of basic protein by V8 protease: (1-32), (1-37), (33-169), (38-169), (33-118), (38-118), (33-127), (38-127), (119-169), and (128-169). Cleavage of the peptic peptide (89-169) yielded fragments (89-118), (89-127), (119-169), and (128-169). All peptides were identified by amino acid analysis, as well as NH2- and COOH-terminal analyses. Time course studies with basic protein showed that V8 protease initially attacked the bonds between Asp(32) and Thr(33) and Asp(37) and Ser(38). With peptide (89-169) the initial cleavage was between Glu(118) and Gly(119). Peptides (89-118) and (89-127) were encephalitogenic in the Lewis rat. The activity of these peptides in the rat confirms the presence of a minor encephalitogenic site in guinea pig basic protein. Peptide (89-127) was encephalitogenic in the guinea pig, as expected, because it contains the intact encephalitogenic site. V8 protease digestion of basic protein yields some interesting new fragments, not previously available for biologic studies.     

Enhanced transfer of experimental allergic encephalomyelitis with strain 13 guinea pig lymph node cells: requirement for culture with specific antigen and allogeneic peritoneal exudate cells

Previously, we reported that transfer of experimental allergic encephalomyelitis (EAE) with sensitized peritoneal exudate cells (PEC) in strain 13 guinea pigs is markedly enhanced if the cells are first cultured with specific antigen, myelin basic protein (BP). These cells also undergo considerable antigen-specific proliferation. In contrast, the data reported here show that lymph node cells (LNC) from sensitized animals display neither enhanced transfer nor antigen-specific proliferation after culture with BP. Enhanced transfer is obtained, however, if a second nonspecific signal is available. This second signal is provided by the presence of normal allogeneic strain 2 PEC in culture. After culture with BP and strain 2 PEC, 2.5 to 5 x 10(7) strain 13 LNC transfer disease reproducibly, in contrast with approximately 1 x 10(9) previously required for successful transfer. Addition of allogeneic or syngeneic PEC without antigen does not lead to enhanced transfer by LNC. Culture with normal syngeneic PEC plus BP oly infrequently enhances transfer by LNC. The intense mixed lymphocyte reaction (MLR) induced by addition of strain 2 PEC to strain 13 LNC precludes the use of 3H-TdR incorporation for detection of proliferation by EAE effector cells. However, inhibition of transfer with low doses of mitomycin C (2 to 5 micrograms/ml) pluse the fact that EAE effector cells are found almost exclusively in the light fraction of BSA gradients after (but not before) culture suggests that the latter are induced to proliferate in culture.     

Elektronenmikroskopische Untersuchungen anPaulinella chromatophora Lauterborn,einer Thekamöbe mit blau-grünen Endosymbionten (Cyanellen)

Zusammenfassung Die Feinstruktur der Cyanellen vonPaulinella chromatophora sowie die Bildung und Struktur der Kieselschuppen, die das Gehäuse dieser Thekamöbe aufbauen, wurden untersucht.Die beiden wurstförmigen Cyanellen besitzen eine 6–13 nm dicke Wandschicht. Sie liegen eingeschlossen in Vesikeln im Cytoplasma des Wirtes. Das Chromatoplasma der Cyanelle enthält 15–20 konzentrisch angeordnete Thylakoide, Plastoglobuli und Phycobilisomen. Das Centroplasma enthält polyedrische Körper.Das Gehäuse der Thekamöbe besteht aus verkieselten rechteckigen Schuppen, die sehr regelmäßig zum Gehäuse zusammengefügt sind. Die Schuppen haben eine komplizierte Feinstruktur. Sie entstehen, vielleicht unter Mitwirkung von Mikrotubuli, vor der Zellteilung in Vesikeln, die wahrscheinlich aus Zisternen des einzigen Dictyosomes der Thekamöbe hervorgehen. Dieses Dictyosom liegt dem Zellkern am aboralen Pol derPaulinella an. Hexagonale Körper (Virionen?) werden aus dem Zellkern und dem Cytoplasma des Wirtes beschrieben.

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Electron microscopical investigations onPaulinella chromatophora Lauterborn, a thecamoeba containing blue-green endosymbionts (cyanelles)

Summary The ultrastructure of the sausage-shaped cyanelles and the ultrastructure and formation of the thecal scales ofPaulinella chromatophora were investigated. The cyanelles have a 6–13 nm thick wall. They are lying within vesicles in the cytoplasma of the host. The chromatoplasma has 15–20 concentrically arranged thylakoids, plastoglobuli and phycobilisomes. The centroplasma contains polyhedral bodies. The theca ofPaulinella chromatophora is composed of rectangular scales arranged in a very regular manner. These scales exhibit a very complex ultrastructure. They are produced prior to cell division in large vesicles probably derived from cisternae of the only dictyosom which is located close to the nucleus in the aboral part of the thecamoeba. Microtubules may play a role in the morphogenesis of these scales.Hexagonal particles (virions?) are described from the nucleus and the cytoplasma of some of the thecamoebae.

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FräuleinBrigitte Schendel danke ich für ihre gute technische Mitarbeit, der Deutschen Forschungsgemeinschaft danke ich für Sachbeihilfe.     

The nature of the defect in experimental allergic encephalomyelitis (EAE)-resistant Lewis (Le-R) rats

Recently, a colony of Lewis rats has been described which is resistant to experimental allergic encephalomyelitis (EAE). These rats, termed Le-R, are still histocompatible with other Lewis rats. The genetic defect which results in EAE-resistance was shown not to be linked to the RT1.B (Ir) region of the MHC. Myelin basic protein (BP)-sensitization of Le-R rats induces cells capable of mounting a proliferative response to BP in culture but incapable of transferring EAE after culture with BP. The present study demonstrates that the latter deficiency can be overcome either by incorporating lipopolysaccharide (LPS) in the BP-culture medium or by simultaneous transfer of LPS-activated antigen-nonspecific spleen cells with the BP-sensitized cells. The BP-sensitized Le-R cells fail to transfer EAE due to their inability to initiate lesions in the CNS. LPS, working through an antigen-nonspecific cell or cell products, can correct the defect in the Le-R cells such that the antigen-specific cells become capable of initiating CNS lesions which lead to development of clinical EAE.     

Adoptive transfer of experimental allergic encephalomyelitis: incubation of rat spleen cells with specific antigen.

Spleen cells from myelin basic protein (BP)-sensitized donor rats appear to be incapable of adoptively transferring experimental allergic encephalomyelitis (EAE) directly to normal recipients. It has been reported that in vitro incubation with concanavalin A (Con A) activates rat spleen cells so that they are capable of transferring EAE. We report here that incubation with specific antigen, BP, also permits transfer of disease with spleen cells. Data are presented in which activation of EAE spleen cells by Con A is compared with activation by BP. Cellular proliferation does not appear to be necessary for in vitro activation with specific antigen.     

Performance of three phase fluidized bed reactor for quinoline degradation on various supports at steady state and dynamic conditions

Quinoline degradation by Comamonas acidovorans was investigated in a three phase fluidized bed reactor at dilution rates below and above the critical value (mu(max) = 0.42 h(-1)). Quinoline was used as the sole source of carbon, nitrogen, and energy. Two attachment carriers, polyurethane foam (Bayvitec(R)) and modified cellulose (Aquacel(R)), and a gel entrapment carrier (polyvinyl alcohol) were studied and compared with regard to their effectiveness to immobilize cells. Attachment and biofilm formation was best at higher dilution rates, regardless of carrier type used. Except for the maximum biomass concentration on the carrier, Y(V) (biomass per volume of solid particles), there was no significant difference in reactor performance between the investigated carriers under stationary conditions. The highest value for Y(V) was found for the gel entrapment carrier (Y(V) = 35 g L(-1)). In a long-term run (66 days), the gel entrapment carrier established a permanent biofilm on the surface of the gel beads after 900 h of cultivation time. Complete quinoline mineralization was achieved at a dilution rate of 2.0 h(-1), which is 4.7 times higher than the critical dilution rate. Identical substrate overloads were applied to the gel entrapment and the cellulose carrier by a step increase of the quinoline feed concentration at a dilution rate of 0.8 h(-1) (D approximately 2mu(max)). The cells survived the overload, but the accumulation of quinoline and quinoline degradation products and the degradation efficiency were different for the two systems during the overload, showing the influence of the carrier type on the dynamic performance and stability of the process. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 295-303, 1997.     

Aluminum concentrations in tissues of rats: effect of soft drink packaging

Aluminum is a commonly occurring trace element for which no nutritional requirements have been set. Some non-conclusive evidence exists suggesting a need of aluminum for growth, reproduction or health of man and animals. There is concern that exposure or consumption of aluminum may be toxic to humans and animals. The objective of the current study was to compare tissue levels of aluminum of rats fed soft drinks packaged in aluminum cans, glass bottles or distilled water. Thirty male weanling rats (Sprague-Dawley) were divided into three treatment groups of 10 rats each. All rats were fed rodent chow ad libitum throughout the study. Three different fluids, i.e. distilled water, diet soft drinks from aluminum cans and diet soft drinks from glass bottles, were fed for a period of 3 weeks. Aluminum contents of tissues were measured by atomic absorption spectrophotometry. Canned soft drink fed rats had significantly higher blood, liver and bone aluminum concentration than rats that were given glass bottled soft drink. There was a 69% higher bone aluminum concentration and 16% lower femur weight in rats fed aluminum canned soft drinks when compared with rats fed with distilled water.