Further development of a recombinant feline herpesvirus type 1 expressing the Gag protein of feline immunodeficiency virus

Summary.  We have previously reported the construction of a recombinant feline herpesvirus type 1 (FHV-1), designated C7301ddlTK-gag, expressing the Gag precursor protein of feline immunodeficiency virus (FIV). In this study, we report the construction of a further recombinant FHV-1 (ddlTK(gBp)-gag) which carries an FHV-1 gB promoter sequence upstream of the FIV gag gene of C7301ddlTK-gag. Strong expression of the FIV Gag protein by ddlTK(gBp)-gag was confirmed by immunoblot analyses and enzyme-linked immunosorbent assays. Although C7301ddlTK-gag and ddlTK(gBp)-gag failed to induce anti-FIV Gag antibodies in cats, we confirmed the infectivity and stability of these recombinants in cats. Received January 14, 2000 Accepted August 4, 2000     

Immunoglobulin superfamily molecules in the nervous system.

Among the various types of membrane molecules involved in cell-cell interactions in the nervous system, we have focused in this review upon membrane proteins belonging to the immunoglobulin superfamily (IgSF). IgSF molecules are distinctive in that: (1) a large percentage of known neural adhesion molecules belongs to the IgSF; (2) they are homologous in structure (Ig domain), yet exhibit large variation of function in cell-cell interactions. The structure of IgSF molecules is briefly summarized in Section II, and each member of the IgSF which has been found in the nervous system is reviewed in Section III. In Section IV, we have discussed possible properties of yet-unknown nervous system IgSF molecules, on the assumption that nervous system IgSF molecules thus far discovered comprise only a small portion of those existing. Discussion is based upon an analogy with the immune system and upon knowledge of cell-cell interactions in the development of the nervous system. Our principal aims in this review are to summarize knowledge of neural IgSF molecules and to discuss the possibility that some IgSF molecules may encode in their structures instructions for recognizing, or for being recognized by, target neural cells. Further growth of knowledge of IgSF molecules may yield insights into the patterns of cell-cell interactions underlying the formation of neuronal circuits during development.     

Preparation of monoclonal antibody to hepatocellular membranes and its application to induction of liver cell membrane damage

Monoclonal antibody (MoAb) to rat liver plasma membranes was prepared by hybridization of mouse immune lymphocytes with mouse myeloma cells, and was identified by the immunodiffusion method in a fraction of IgM secreted from the hybridoma thus obtained. In indirect immunofluorescence tests, specific fluorescence was detected only on the surface of rat hepatocytes, but neither on the cells from other organs of the rat nor on the hepatocytes of other species of animals, suggesting that the antibody may be organ- and species-specific. When the primary culture rat hepatocytes, labelled with isotopic chromium (51Cr), were treated with the MoAb together with complement, a specific release of 51Cr from the cells was found shortly after treatment, accompanied with bubbling of the cell membranes, and a significant release of 51Cr was observed at an MoAb concentration of 15 micrograms/ml or more. Without complement, or with inactivated complement, these reactions were not observed. These facts suggest strongly that the cell surface of the hepatocytes was damaged by the MoAb in the presence of complement.     

Topographical gradient in expression of R2D5 antigen in superior olivary nuclei and hippocampal dentate gyrus of the cat.

An immunohistochemical analysis of the cat central nervous system revealed that a monoclonal antibody which recognizes a soluble cytosolic protein, R2D5, bound two regions in a prominent spatial gradient. In the medial and lateral superior olivary nuclei of the brainstem, R2D5 immunoreactivity appeared as a gradient across a population of topographically ordered principal neurons. The spatial gradient corresponded to the tonotopic organization in the superior olivary nuclei: i.e., R2D5 immunoreactivity tended to occur more frequently and intensely in low-frequency neurons than in high-frequency neurons. Granule cells in the hippocampal dentate gyrus also had a pronounced spatial gradient in R2D5 immunoreactivity expression, and this gradient corresponded to the septotemporal axis of the hippocampus. Granule cells of the temporal (ventral) portions of the hippocampus were labeled intensely with R2D5 antibody, while those located in progressively more septal (dorsal) portions had gradually less immunoreactivity. These results suggest that in both the superior olivary nuclei and the hippocampal dentate gyrus, neurons differ in intrinsic properties by their position along specific axes. They suggest also that the hippocampus has an intrinsic functional organization related to the spatial gradient along its septotemporal axis.     

Histologic transformation in extranodal non-Hodgkin's lymphoma

Histologic transformation in the course of non-Hodgkin's lymphoma (NHL) has been reported to occur in 18 to 30% of the cases. Less favorable prognosis in cases with initial low grade malignancy followed by emergence of high grade malignancy has been previously described. In the previous literature, the histologic transformation has been examined mostly in nodal NHL. In the present study, histologic transformation in the course was investigated on 20 cases with early extranodal NHL. All these 20 cases were diffuse lymphomas, and were composed of 7 cases with low grade malignancy and 13 cases with high grade malignancy. Histologic transformation was not observed in any of these cases. These findings indicate that the frequency of histologic transformation is much lower in extranodal NHL than in nodal NHL. The prognostic significance of these findings is also discussed.     

Continuous measurements of Na, Li, and Cl in the perfused salivary gland by use of NMR

The present study was undertaken to measure tissue contents of Na, Li, and Cl non-invasively in the isolated perfused organ by nuclear magnetic resonance (NMR) using a broadband tunable probe. The NMR signals of 23Na, 7Li, and 35Cl from the isolated perfused rat mandibular gland were collected continuously and each spectrum was obtained for every 15 or 20 s. The Na concentration in the perfusate was varied by replacement with Li, and the resulting changes were monitored by measuring the signal intensities of the electrolytes. The time constant for Na exchange was slower following complete removal of extracellular Na than following its half replacement, suggesting that the Na extrusion by Na+/K+ ATPase was reduced by lowering the extracellular Na level. The time constant for Li exchange was slower than that for Na exchange. The level of Cl was nearly constant during experiment, except for a very slow increase in Cl, possibly resulting from increasing edema and/or intracellular Li storage.     

Identification of a novel allele HLA-B*7805 in a Japanese female

A new HLA-B*78 allele, B*7805, was identified in a healthy Japanese female. The results of her serological HLA class I typing showed an unusual Bw4/Bw6 pattern with strongly positive reactivity to anti-Bw6, i.e. A24, -, B52, -, Bw4, Bw6. In DNA typing, she was typed as A*24, -, B*52, B*78-like, Cw1202, -, (Bw4, Bw6). Cloning and sequencing of exon 2 and exon 3 of her B locus genes revealed a new allele B*7805. The cloned B*7805 differed from B*78021 by three nucleotide substitutions in exon 2 at position 259 (A to G), 261 (C to G) and 272 (A to C), and contained sequences defining Bw6 motif in the region of codon 77 to 83.     

Voltammetry in unanesthetized rat: Increases of striatal dopamine turnover after unilateral haloperidol injection into the substantia nigra

In freely moving rats, effects of unilateral haloperidol injection into the substantia nigra were monitored with in vivo voltammetry in the bilateral striata. The electrochemical responses at 120 mV versus Ag-AgCl, reflecting mainly a level of 3,4-dihydroxyphenylacetic acid (DOPAC), increased both in the striata within 1.5 h after 5 μg of haloperidol treatment. In the experiments of high-performance liquid chromatography with electrochemical detection, the ratio of DOPAC to dopamine in the striata significantly increased at 2.75 h after drug treatment. These data support the idea that unilateral injection of haloperidol into the substantia nigra in freely moving rats increases dopamine turnover in the bilateral striata.