Laboratory investigations on "postvaccinall rabies" caused by live sheep-brain vaccine.

The viral ethiology of postvaccinal complications among 30 dogs vaccinated by live antirabies vaccine (Umeno-Doi type, sheep brain vaccine) was fully confirmed. Three lots of virulent vaccine were inoculated subcutaneously into groups of "Wistar" rats according to the different schemes. Between the 1st and 12th day after the end of the vaccination there were no isolations of fixed virus in direct and blind i.c. passages of suspensions made from the thalamus area on succkling mice and rats. Also the viral antigen in the CNS of vaccinated but apparently healthy rats was undetectable. The "postvaccinal rabies" with the next isolation of fixed r.v. in the CNS was developed experimentally in rats only following the subcutaneous injection of the crude sheep-brain's and spinal cord's suspensions--composing the materials to production of antirabies vaccine.     

Genetic live vaccines mimic the antigenicity but not pathogenicity of live viruses.

The development of an effective HIV vaccine is both a pressing and a formidable problem. The most encouraging results to date have been achieved using live-attenuated immunodeficiency viruses. However, the frequency of pathogenic breakthroughs has been a deterrent to their development. We suggest that expression libraries generated from viral DNA can produce the immunologic advantages of live vaccines without risk of reversion to pathogenic viruses. The plasmid libraries could be deconvoluted into useful components or administered as complex mixtures. To explore this approach, we designed and tested several of these genetic live vaccines (GLVs) for HIV. We constructed libraries by cloning overlapping fragments of the proviral genome into mammalian expression plasmids, then used them to immunize mice. We found that inserting library fragments into a vector downstream of a secretory gene sequence led to augmented antibody responses, and insertion downstream of a ubiquitin sequence enhanced cytotoxic lymphocyte responses. Also, fragmentation of gag into subgenes broadened T-cell epitope recognition. We have fragmented the genome by sequence-directed and random methods to create libraries with different features. We propose that the characteristics of GLVs support their further investigation as an approach to protection against HIV and other viral pathogens.     

Long live the Queen's subjects.

In 1952, the Queen congratulated 255 people on their hundredth birthdays and 1135 couples on their sixtieth wedding anniversaries. By 1996, these numbers had risen to 5218 and 11,688, respectively. Semilogarithmic plots, normalized to constant numbers of births and marriages, show steady exponential rises in the number of centenarians with a doubling time of 11 years, and of diamond weddings with a doubling time of 19 years. An alternative plot of the numbers of those reaching a hundred between 1910 and 1990, based on registers of births and deaths and normalized to constant births, shows an annual rise of only 1% from 1910 to 1946, followed by a steady exponential rise with a doubling time of 12 years, closely matching that of 11 years derived from the Queen''s figures. The exponential rise in the number of those born from 1846 onwards living to a hundred precedes by many years the general rise in the expectation of life at birth and the general drop in mortality from infectious diseases, but it coincides with the beginning of a steady rise in real wages. Another important factor may be improved medical treatment at old age from 1946 onwards.     

Dynamics of the cytoskeleton in live cells.

Actin filaments, microtubules, and intermediate filaments, have all been found to be dynamic structures in living cells. Recent studies have shed important light on the assembly, disassembly, and mobility of these structures. In addition, a growing emphasis has been placed on the regulation of cytoskeletal activities by various signal transduction pathways.     

A radio-controlled live trap.

A live trap, designed to drop a net by radio control, was constructed to capture small wild mammals and birds. Advantages of the radio-controlled trap included capabilities to select and trap only the species desired, collect animals immediately after capture, eliminate injuries during capture and operate from a protected area a distance from the trap.     

Gene expression and the development of live enteric vaccines.

Rationally attenuated live Salmonella vaccines provide good protection against homologous challenge and can act as carriers of heterologous antigens. However, the optimum expression system for each heterologous antigen will need to be established individually. This will ensure that the antigen in question is produced at appropriate levels, in the correctly folded conformation, within or at the surface of the carrier cell, and can therefore elicit the optimal immune response.     

Structure and dynamics of the fusion pore in live cells.

Atomic force microscopy reveal pit-like structures typically containing three or four, approximately 150 nm in diameter depressions at the apical plasma membrane in live pancreatic acinar cells. Stimulation of secretion causes these depressions to dilate and return to their resting size following completion of the process. Exposure of acinar cells to cytochalasin B results in decreased depression size and a loss in stimulable secretion. It is hypothesized that depressions are the fusion pores, where membrane-bound secretory vesicles dock and fuse to release vesicular contents. Zymogen granules, the membrane-bound secretory vesicles in exocrine pancreas, contain the starch digesting enzyme, amylase. Using amylase-specific immunogold labeling, localization of amylase at depressions following stimulation of secretion is demonstrated. This study confirms depressions to be the fusion pores in pancreatic acinar cells. High-resolution images of the fusion pore in live pancreatic acinar cells reveal the structure in much greater detail than has previously been observed.     

Development of a live attenuated varicella vaccine.

The Oka strain of varicella virus, isolated in our labolatory, was serially cultivated in guinea-pig embryo cultures (GPEC), and a considerable amount of cell-free virus was obtained from infected cell. GPEC passage virus at the 6th passage level was used in a small scale field trial. Susceptible children of 1 to 10 years old were injected subcutaneously with 100 to 1,000 PFU of virus. No clinical reactions due to the vaccination were observed in any children, and a high rate of antibody response was obtained with viral doses of more than 200 PFU. Attenuated virus obtained by passage in GPEC was propagated in human diploid (WI-38) cells, and it was also effective in inducing an immune response without clinical reactions. The results show that the Oka strain of varicella virus passaged in GPEC and human diploid (WI-38) cells may be used safely and effectively as a live attenuated vaccine.     

Encephalitis after administration of live measles vaccine.

In a previously well child with no evidence of pre-existing immunologic defect a fatal encephalitis developed 10 days after administration of measles vaccine. There was pathologic evidence of an early viral encephalitis characterized by perivascular mononulcear infiltrates. Although the virus was not recovered, the diagnosis of a measles virus infection and encephalitis is supported by the postmortem findings of Warthin-Finkeldey cells in lymphoid tissues, an intranuclear inclusion in the brain and histologic changes of encephalitis.