Regulation of the CD2 alternate pathway of T cell activation by CD3. Evidence for heterologous desensitization

Normal resting T cells were stimulated through the alternate CD2 pathway. A CD3 mAb VIT3 completely blocked their proliferative response. The time interval for 50% inhibition lasted for 24 h after the onset of CD2 stimulation. Mitogen-activated cloned long term cultured T cells could also be stimulated via CD2. This proliferative response was again inhibitable by VIT3, indicating that CD3 regulates the CD2 pathway not only in resting cells, but also in lymphocytes actively involved in an Ir. T cells were further loaded with Quin2 and their free cytoplasmic Ca2+ levels were monitored in response to CD3 and CD2 stimulation. Antibodies directed against both surface R triggered a rapid elevation of Ca2+ levels. Both responses were abrogated when the cells had been treated overnight with VIT3. The free cytoplasmic Ca2+ levels of VIT3-pretreated cells, however, were not higher than those of control cells. These results point to a functional interaction between CD3 and CD2 possibly at the level of signal transducing proteins. Finally, cholera toxin was found to inhibit the Ca2+ response in Jurkat T cells. Both the CD3 and CD2 stimulation were sensitive to cholera toxin, indicating that a GTP-binding protein may be involved in signal transduction for both surface structures.     

Amianthoid (asbestoid) transformation: electron microscopical studies on aging human costal cartilage

The present study reports on the fine structure of human costal cartilage at different ages in order to obtain information on the morphogenesis of amianthoid fibers. Our results reveal an overall increase of collagen fibril diameter with increasing age, even in areas with no signs of amianthoid transformation. Ultrastructural evidence is presented that this increase in diameter is due to a gathering of the preexisting collagen fibrils. The age-related change in collagen fibril diameter is paralleled by changes in the composition and ultrastructural appearance of cartilage proteoglycans (as revealed by acridine orange staining). Acridine-orange-positive filaments indicative for proteoglycans are markedly reduced in size with advancing age in centrally located regions of costal cartilage. Treatment with testicular hyaluronidase previous to acridine-orange staining leaves these small proteoglycan filaments unaffected. By contrast, the filaments visible after acridine-orange staining in the extracellular matrix near to the perichondrium are susceptible to hyaluronidase treatment. Infrequently, a sharp increase in collagen fibril diameter can be observed in territorial matrix areas of degenerating chondrocytes. This observation is conspicuous at ages of 10 and 20 years. Amianthoid transformation is characterized by the appearance of collagen fibrils strictly arranged in parallel. These amianthoid fibers are embedded in a matrix rich in small acridine-orange-positive filaments similar to the proteoglycan filaments observed in centrally located matrix regions. It can be concluded that extensive remodelling not only of the collagen fibrils but also of the cartilage proteoglycans is involved in the development of amianthoid transformation.     

Kinetics of activation antigen expression by in vitro-stimulated human T lymphocytes

In this study a panel of monoclonal antibodies was used to investigate the kinetics of the appearance of activation-linked surface determinants as well as cytoplasmic and nuclear determinants in human T cells following lectin stimulation. Well known activation markers, such as Ia/DR, transferrin receptor, IL-2 receptor, T10, and gp24, were compared and investigated together with the T13 structure, recently found in this laboratory. T13, not demonstrable on resting T cells, could be seen within 24 hr after lectin stimulation. Kinetics of the appearance were similar to IL-2 receptor and transferrin receptor expression. Ia/DR synthesis was investigated separately for each polypeptide and the cytoplasmic invariant gamma-chain expression could be demonstrated for the first time with a gamma-chain-specific monoclonal antibody VIC-Y1. Moreover, gamma-chain synthesis seems to precede alpha- and beta-chain occurrence in human T cells. In addition, data from quantitative studies on antigenic densities are presented.     

Human DNA sequences isolated with an immunoglobulin switch region probe: sequence,chromosomal localization,and restriction fragment length polymorphisms

Summary We have screened a human genomic DNA library with an immunoglobulin (Ig) derived switch (S) region specific probe for homologous sequences. Five Ig independent phage clones were isolated and characterized. The S sequence homologous DNA fragments are short compared to the S region sequences. Ig independent S sequences are flanked by highly repetitive DNA elements and perfect inverted repeats can be demonstrated in their close vicinity. Using subclones of S homologous sequences restriction fragment length polymorphisms were shown within DNA of different T cell leukemias. Burkitt lyphhomas, lymphoblastoid cell lines, and DNA of healthy individuals. One of the five clones isolated with the S region probe was evidently localized to chromosome 2 and/or 40 and showed a complex hybridisation pattern with several different human DNAs. S homologous sequences of another clone are most likely localized on chromosome 1. It is possible that these Ig indenpendent S sequences have arisen by amplification and transposition and that they are involved in genetic recombination.     

Eine bisher unbekannte Art der Zentriolenvermehrung

Zusammenfassung An Flimmerzellen von spätembryonalen, neugeborenen und wenige Tage alten Ratten wird die Neubildung einer großen Zahl von Zentriolen bei der Differenzierung der Zellen beschrieben. Viele Zellen zeigen im Übergangsstadium zwischen der sezernierenden Form und der Ausbildung des Flimmerapparates eine Aussat von granulären Zentriolenvorstufen; diese sind meist in Gruppen angeordnet. Von diesen granulären Vorstufen leiten größere Zwischenstufen kontinuierlich zu den ausdifferenzierten Zentriolenformen über. Da keine Beteiligung der präexistenten Zentriolen gefunden werden konnte, muß eine echte Neubildung angenommen werden.

---

Summary In the ciliated cells of embryonic (late stage of pregnancy), new-born, and a few-day-old rats, the new formation of a great number of centrioles, that takes place during the differentiation of the cells, is studied. During an intermediate stage (between the secretory form and the formation of the ciliated apparatus) a dissemination of granular precursors of centrioles takes place in many cells. Most of these pre-centrioles are arranged in groups. Various intermediate steps lead from this granular rudimentary stage to the highly differentiated centrioles. As no participation of the preexisting centrioles is observed, it is postulated that the observed phenomenon is to be regarded as a genuine new formation.

---

---

Frau Prof. C. Coronini ergebenst zum 80. Geburtstag gewidmet.

---

Assistent am Morphologischen Institut der medizinischen Fakultät der Universität Ege, Izmir-Türkei (Vorstand: Prof. Dr. I. Ulutas). Das Stipendium, das meinen Studienauf enthalt in Österreich ermöglichte, verdanke ich dem Österreichischen Bundesministerium für Unterricht.     

Die Ultrastruktur des Flimmerepithels des Nasenseptums der Ratte

Zusammenfassung Das Epithel des mittleren Abschnittes des Nasenseptums der Ratte ist gestuft hochprismatisch; es enthält 4 Zelltypen: Flimmerzellen, indifferente Zellen, Becherzellen und Ersatzzellen.Der Bau der Flimmerhaare entspricht im Prinzip dem weit verbreiteten Bauschema dieser Strukturen. Bisher wenig beachtete Details sind: eine kornartige Verdichtung an der Spitze; ein quergestreifter lateraler Sporn am Basalknötchen, der hypothetisch mit der Richtung des Flimmerschlages in Zusammenhang gebracht wird. Wurzelfäden (rootlets) im Sinne Fawcetts fehlen. Eine Präzision des Terminus rootlet im Sinne von Wurzelfäden wird vorgeschlagen.In indifferenten und Flimmerzellen wurden mitunter sehr viele Centriolen im apikalen Cytoplasma und in der oberflächlichen Grenzzone der Zellen dargestellt; ebenso Übergangsformen dieser Strukturen zu Basalknötchen inkomplett und komplett ausgebildeter Flimmerhaare.Zahlreiche Pinocytosevakuolen sprechen für eine starke Resorptionstätigkeit dieser Zellen. Auch die dünnen Cytoplasmahüllen der Flimmerhaare scheinen sich durch Ausbildung von Pinocytosevakuolen an dieser Funktion zu beteiligen. Flimmer- und indifferente Zellen weisen im übrigen ähnliche Cytoplasmastrukturen auf. An ihrer Oberfläche finden sich besonders lange Cytoplasmafortsätze für die die Bezeichnung Cytofila zur Abgrenzung gegen die viel kürzeren Mikrovilli vorgeschlagen wird.Die Strukturen der Becherzellen sind in der Regel wesentlich dichter; ihre basalen Teile sind baumwurzelartig verzweigt und in die Nachbarzellen eingesenkt; diese innige Verbindung könnte der Aufnahme resorbierter Flüssigkeit dienen. Nicht alle basalen Fortsätze erreichen die Zellbasis.Intrazelluläre Cysten verschiedener, von der Oberfläche gegen die Basis zunehmender Größe enthalten in ihrer Oberfläche Mikrovilli, Cytofila und Flimmerhaare, im Lumen Zelldetritus und undefinierbare amorphe Massen. Im Gegensatz zu den Interpretationen Miháliks wird auf Grund der eigenen Befunde am Nasenepithel der Ratte der Zusammenhang zwischen der Genese des oberflächlichen Flimmersaumes und derartigen Cysten in Frage gestellt. Möglicherweise handelt es sich dabei um pathologische Vorgänge.     

Herpes-Simplex-virus-specific, H-2Dk-restricted T lymphocytes bear receptors for H-2Dd alloantigen

Cytotoxic T lymphocytes generated in the course of an HSV-infection of CBA (H-2k) mice not only lyse syngeneic, virus-infected target cells but also cross-react with noninfected taraget cells expressing the Dd alloantigen. On the effector cell level, this alloreactivity is mediated by virus-specific CTL's that are restricted to H-2Dk determinants. On the prekiller cell level, the anti-HSV-reactive T cells exhibiting cross-reactivity for Dd alloantigen could be positively selected on H-2d spleen-cell monolayers. After differentiation into cytolytic effector cells, target cells expressing Dd alloantigens and syngeneic HSV-infected target were lysed with equal efficiency. The results imply that the phenomenon of H-2-restricted versus nonrestricted T-cell reactivity is not due to distinct T-cell subsets, but rather is dependent on the antigeneic determinants recognized.     

An evidence-based consensus for the classification of arbuscular mycorrhizal fungi (Glomeromycota)

The publication of a large number of taxon names at all levels within the arbuscular mycorrhizal fungi (Glomeromycota) has resulted in conflicting systematic schemes and generated considerable confusion among biologists working with these important plant symbionts. A group of biologists with more than a century of collective experience in the systematics of Glomeromycota examined all available molecular–phylogenetic evidence within the framework of phylogenetic hypotheses, incorporating morphological characters when they were congruent. This study is the outcome, wherein the classification of Glomeromycota is revised by rejecting some new names on the grounds that they are founded in error and by synonymizing others that, while validly published, are not evidence-based. The proposed “consensus” will provide a framework for additional original research aimed at clarifying the evolutionary history of this important group of symbiotic fungi.     

Proteomic analysis of resistance mediated by Rcm 2.0 and Rcm 5.1, two loci controlling resistance to bacterial canker of tomato

Two quantitative trait loci from Lycopersicon hirsutum, Rcm 2.0 and Rcm 5.1, control resistance to Clavibacter michiganensis subsp. michiganensis, the causal agent of bacterial canker of tomato. Lines containing Rcm 2.0 and Rcm 5.1 and a susceptible control line were compared at 72 and 144 h postinoculation, using 2-dimensional gel electrophoresis to identify proteins regulated in response to C. michiganensis subsp. michiganensis infection. A total of 47 proteins were subjected to tandem mass spectrometry. Database queries with resulting spectra identified tomato genes for 26 proteins. The remaining 21 proteins were either identified in other species or possessed no homology to known proteins. Spectra were interpreted to deduce peptide amino acid sequences that were then used to query publicly available data. This approach identified tomato genes or expressed sequence tags for 44 of the proteins analyzed. Three superoxide dismutase (SOD) enzymes were differentially regulated among genotypes, and patterns of hydrogen peroxide accumulation were genotype- and tissue-specific, indicating a role for oxidative stress in response to C. michiganensis subsp. michiganensis. Steady-state mRNA and protein levels for SOD, thioredoxin M-type, S-adenosylhomocysteine hydrolase, and pathogenesis-related proteins demonstrated similar patterns of differential regulation. Lines containing Rcm 2.0 and Rcm 5.1 accumulate different proteins and steady-state mRNAs in response to inoculation, suggesting that the two loci may confer resistance through distinct mechanisms.