Functional analyses of natural killer cells in macaques infected with neurovirulent simian immunodeficiency virus.

Clearance of HIV and SIV from the peripheral blood by the cellular immune system lessens the viral burden in infected individuals and may have an impact on virus infection of the CNS and the development of CNS lesions. However, the role of immune responses in preventing or limiting CNS infection has not been clearly defined. We investigated the role of natural killer cells in the outcome of SIV infection of macaques as a model for humans with AIDS and HIV encephalitis. In our study, six pig-tailed macaques were infected with the neurovirulent virus, SIV/17E-Fr, and the immunosuppressive virus, SIV/ DeltaB670, in a model system that causes rapid progression to AIDS and a high frequency of CNS lesions. NK lytic activity in each macaque was monitored longitudinally. In addition, we enumerated NK cells and tested macaque PBMC for the ability to lyse SIV-infected target cells. We found that there was a significant inverse correlation (P=0.02) between the robustness of NK response and the development of CNS lesions. Animals lacking strong NK cell responses developed more severe CNS lesions than those with robust NK responses did. Furthermore, pre-infection levels of NK activity were predictive of CNS lesion severity. The macaque with the most robust pre-infection NK activity developed no CNS lesions. In these infected macaques, NK activity was shown to be directed against SIV-infected cells. We extended these in vivo findings to delineate precisely which cell type was mediating this SIV-directed lysis. We used both macaque and human cells to demonstrate that the population that mediated anti-SIV and anti-HIV cytolytic effects was NK cells. Furthermore, we showed that this anti-SIV and anti-HIV cytolytic effect was directed at the envelope protein and not gag proteins. Thus, NK cells have the capacity to recognize and lyse cells expressing SIV and HIV antigens. These data support a role for NK cells in the modulation of CNS disease.

CD56 identifies monocytes and not natural killer cells in rhesus macaques.

BACKGROUND: CD56 is a lineage-specific marker of human natural killer (NK) cells. There are conflicts in the literature regarding the role of CD56 as a marker of NK cells in non-human primates. In the present study, we examined the role of CD56 in identifying rhesus NK cells. METHODS: The immunophenotype of normal macaque and human NK cells was analyzed by two- and three-color flow cytometry. Flow cytometric cell sorting was subsequently used to deplete or purify NK cells; the resulting cell populations were then used in standard chromium release assays of NK lytic function. RESULTS: In peripheral blood mononuclear cells of the rhesus macaque, CD56 was expressed primarily on cells with the light scatter and immunophenotypic profile of monocytes. Flow cytometric depletion of rhesus CD56(+) monocytic cells did not diminish functional activity against K562 cells, whereas depletion of CD8(+) or CD16(+) lymphocytes completely abrogated functional activity. Three-color flow cytometric analysis of CD8(+), CD16(+) lymphocytes showed that they expressed other markers (CD2, CD7, TIA-1) associated with NK cells, but notably, not CD56. CONCLUSIONS: These studies demonstrate that CD56 is not suitable as a marker of NK cells in the rhesus macaque.

Time dependence of 2.5% nitric acid solution as an etchant on human dentin and enamel.

Although nitric acid is a component in some new bonding systems, the action of nitric acid as an etchant for the improvement of adhesion of bonding systems for resin composites to dentin and enamel has not been reported. A determination of the extent of etching on both dentin and enamel using 2.5% HNO3 solution at various application time periods was the purpose of this study. Extracted human molars were cleaned and sectioned so that flat samples of dentin and enamel would be produced. Surfaces were abraded with 320-grit aluminum-oxide paper, washed with distilled water for 10 s, and blown with air for 10 s. Duplicate samples of dentin and enamel were treated with a drop of 2.5% HNO3. Application periods varied by 10-second intervals, from 10 s up to 60 s. After being rinsed with distilled water and dried, the sections were routinely processed for observation by SEM. The micrographs of the treated surfaces showed various degrees of etching and erosion proportional to the length of application time. The 30-second application revealed a well-etched surface with minimal erosion.

Pulpal and micro-organism responses to two experimental dental bonding systems.

Several new bonding systems have been reported that promote strong adhesion. This in vivo study involves treatment with two experimental bonding systems of Class V cavity preparations in the teeth of three Macaca fascicularis primates and reports the pulpal responses and degree of micro-organism invasion associated with each treatment. In each monkey, the teeth in the upper left quadrant were treated with the experimental solution, containing 2.5% aluminum nitrate + 1.5% oxalic acid + 4.9% NPG, followed by application of PMDM and Silux XL composite. The lower right quadrant was treated with the experimental solution, containing 5.7% NPG + 2.4% nitric acid, followed by PMDM and Silux XL composite. The upper right and lower left quadrants were treated with clinical materials to establish positive and negative controls. After four, 25, and 59 days, the teeth were removed and underwent routine histological and bacteriological evaluation. Slight pathological conditions were noted for superficial and deep responses, but all values approached 0.0 by the 59th day. Micro-organisms were seen under only 12% of the restorations. Both experimental systems appear to be safe for human clinical trials.

Dentine and enamel bonding agents.

Previous studies have shown that sequential use of aqueous FO (ferric oxalate containing a small concentration of HNO3), acetone solutions of NPG (N-phenylglycine), and PMDM (the reaction product of pyromellitic dianhydride and 2-hydroxyethylmethacrylate) yields strong adhesive bonding of composite resins to both dentine and enamel. The purpose of this study was to determine if aluminum ions could be substituted for ferric ions and if the procedure could be simplified. Aqueous solutions containing aluminum oxalate and aluminum nitrate, followed in sequence by acetone solutions of NPG and PMDM, gave strong tensile adhesive bond strengths between a composite and extracted human teeth. Comparable values have been obtained with FO, NPG and PMDM. Aluminum oxalate solutions containing no nitrate gave lower bond strengths, as was the case with FO. Aqueous solutions of acidified aluminum oxalate can dissolve NPG, thereby allowing a simplification of the procedure. Tested for comparison, commercially available dentine bonding agents gave lower average bond strengths on dentine than did some of the experimental materials.

Bond strength of a luting composite to dentin with different bonding systems.

This study evaluated the tensile bond strength of Conclude luting composite to dentin under different environmental conditions with Scotchbond dental adhesive, the FNP system, and unfilled BIS-GMA resin used as bonding agents. Although the bonding systems reportedly yield high tensile bond values to enamel, large differences were observed in this study for bond values to dentin. Only the FNP bonding system produced consistently high tensile load values to dentin under the three environmental conditions tested.