Inactivation of respiratory syncytial virus by zinc finger reactive compounds

<p>Abstract</p> <p>Background</p> <p>Infectivity of retroviruses such as HIV-1 and MuLV can be abrogated by compounds targeting zinc finger motif in viral nucleocapsid protein (NC), involved in controlling the processivity of reverse transcription and virus infectivity. Although a member of a different viral family (<it>Pneumoviridae</it>), respiratory syncytial virus (RSV) contains a zinc finger protein M2-1 also involved in control of viral polymerase processivity. Given the functional similarity between the two proteins, it was possible that zinc finger-reactive compounds inactivating retroviruses would have a similar effect against RSV by targeting RSV M2-1 protein. Moreover, inactivation of RSV through modification of an internal protein could yield a safer whole virus vaccine than that produced by RSV inactivation with formalin which modifies surface proteins.</p> <p>Results</p> <p>Three compounds were evaluated for their ability to reduce RSV infectivity: 2,2'-dithiodipyridine (AT-2), tetraethylthiuram disulfide and tetramethylthiuram disulfide. All three were capable of inactivating RSV, with AT-2 being the most potent. The mechanism of action of AT-2 was analyzed and it was found that AT-2 treatment indeed results in the modification of RSV M2-1. Altered intramolecular disulfide bond formation in M2-1 protein of AT-2-treated RSV virions might have been responsible for abrogation of RSV infectivity. AT-2-inactivated RSV was found to be moderately immunogenic in the cotton rats <it>S.hispidus </it>and did not cause a vaccine-enhancement seen in animals vaccinated with formalin-inactivated RSV. Increasing immunogenicity of AT-2-inactivated RSV by adjuvant (Ribi), however, led to vaccine-enhanced disease.</p> <p>Conclusions</p> <p>This work presents evidence that compounds that inactivate retroviruses by targeting the zinc finger motif in their nucleocapsid proteins are also effective against RSV. AT-2-inactivated RSV vaccine is not strongly immunogenic in the absence of adjuvants. In the adjuvanted form, however, vaccine induces immunopathologic response. The mere preservation of surface antigens of RSV, therefore may not be sufficient to produce a highly-efficacious inactivated virus vaccine that does not lead to an atypical disease.</p

Histology can be predictive of the clinical course of a primary aneurysmal bone cyst.

INTRODUCTION: Aneurysmal bone cyst is a benign lesion occurring in young patients which frequently recurs after treatment. Biopsy is mandatory for the diagnosis of a putative aneurysmal bone cyst as this lesion can be secondary to another underlying process including a malignant bone tumour. The histopathological features of aneurysmal bone cysts have been examined with the goal of finding relevant criteria for predicting favourable evolution or recurrence of the disease. PATIENTS AND METHODS: Twenty-one biopsies of surgically treated aneurysmal bone cysts, from 21 patients, were analysed. Histomorphometry by two different methods (3,000- and 200-point-counting) and by two observers was performed to quantify the percentage of each tissue type in the cyst (cellular, fibrillar, osteoid). A healing index was developed by calculating a ratio of osteoid and fibrillar material divided by cellular tissue. Biopsies were also examined using two immunostains, cluster of differentiation 68 (CD68) and proliferating cell nuclear antigen (PCNA). RESULTS: The final outcome was healing for 16 aneurysmal bone cysts (healing group) and recurrence for the five others (recurrence group), after a mean follow-up of 4.43 years. The two groups differed significantly in the proportion of their cellular content and their healing index. The ratio of CD68 negative to CD68 positive cells was also significantly different between the two groups. CONCLUSION: Biopsy should be considered as a helpful prognostic factor for aneurysmal bone cyst

Anti-human immunodeficiency virus-gag CD8(+) memory T cells generated in vitro from Listeria-immunized mice

The goal of vaccination is the generation of immune memory, an immune state that permits rapid and intense recall responses to a pathogen. Considerable effort is being made to understand the nature of memory T cells. We report here that by extending the length of in vitro culture following a single restimulation with specific peptide, preparations of highly enriched, highly active antigen-specific CD8(+) memory T cells could be obtained. These cultures were begun with splenocytes from mice primed by infection either with an attenuated strain of Listeria monocytogenes or vaccinia virus, both expressing the human immunodeficiency virus-1-gag gene. In the cultures, antigen-specific cytotoxic T lymphocyte (CTL) activity reached a maximum at about 9 days and thereafter fell to negligible values. Concomitant with the fall of CTL activity, however, we observed enrichment for a subset of CD11a(high) antigen-specific gag-tetramer(pos) CD8(+) T cells. The cells showed little or no 4-hr CTL activity, but had high delayed (18-hr) CTL activity, and very high cytolytic activity after restimulation. They rapidly expressed interferon-γ production. Their growth and survival after sorting was completely dependent on interleukin-2 or -15. As few as 5000 of the fluorescence-activated cell sorting-purified cells protected recipients against challenge 3 months after transfer. In response to the challenge, the cells repopulated lymphoid and non-lymphoid organs and showed a sizeable increase in number. The cells therefore demonstrate high protective activity for long periods of time. These cultured cells are thus a potential source of enriched natural memory T cells for reperfusion studies and in which the mechanisms that underlie the generation, differentiation and persistence of memory can be examined