Antibody-mediated protection of plants against infection from potyviruses

Transgenic plants expressing single-chain antibodies have been produced to investigate the feasibility of antibody-mediated broad-spectrum protection against plant virus infections. This study indicates that protection against a wide range of plant viruses can be achieved in transgenic plants expressing a single antibody construct.

Use of ultraviolet C (UVC) radiation to inactivate infectious hematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV) in fish processing plant effluent

We determined the stability of infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) suspended in either fish processing plant effluent blood water (EBW) or culture media and examined the effectiveness of UVC radiation to inactivate IHNV and VHSV suspended in both solutions. Without exposure to UVC, IHNV and VHSV were maintained in 4°C blood water for up to 48 hours without significant reduction in virus titer. However when exposed to UVC radiation using a low pressure mercury vapour lamp collimated beam, IHNV and VHSV were inactivated, and the efficacy of UVC radiation was dependent upon the solution and virus type being treated. A 3-log reduction for VHSV and IHNV in culture media was achieved at 3.28 and 3.84 mJ cm-2, respectively. The UV dose needed for a 3-log reduction of VHSV in EBW was 3.82 mJ cm-2. However, exposure of IHNV in EBW to the maximum UVC dose tested (4.0 mJ cm-2) only led to a 2.26-log-reduction. Factors such as particle size, and possible association of viruses with suspended EBW particulate, were not investigated in this study, but may have contributed to the difference in UVC effectiveness. Future work should emphasize improved filtration methods prior to UV treatment of processing plant EBW at an industrial scale.

The raft-promoting property of virion-associated cholesterol, but not the presence of virion-associated brij 98 rafts, is a determinant of human immunodeficiency virus type 1 infectivity

Lipid rafts are enriched in cholesterol and sphingomyelin and are isolated on the basis of insolubility in detergents, such as Brij 98 and Triton X-100. Recent work by Holm et al. has shown that rafts insoluble in Brig 98 can be found in human immunodeficiency virus type 1 (HIV-1) virus-like particles, although it is not known whether raft-like structures are present in authentic HIV-1 and it is unclear whether a virion-associated raft-like structure is required for HIV replication. Independently, it was previously reported that virion-associated cholesterol is critical for HIV-1 infectivity, although the specific requirement of virion cholesterol in HIV-1 was not examined. In the present study, we have demonstrated that infectious wild-type HIV-1 contains Brij 98 rafts but only minimal amounts of Triton X-100 rafts. To directly assess the functional requirement of virion-associated rafts and various features of cholesterol on HIV-1 replication, we replaced virion cholesterol with exogenous cholesterol analogues that have demonstrated either raft-promoting or -inhibiting capacity in model membranes. We observed that variable concentrations of exogenous analogues are required to replace a defined amount of virion-associated cholesterol, showing that structurally diverse cholesterol analogues have various affinities toward HIV-1. We found that replacement of 50% of virion cholesterol with these exogenous cholesterol analogues did not eliminate the presence of Brij 98 rafts in HIV-1. However, the infectivity levels of the lipid-modified HIV-1s directly correlate with the raft-promoting capacities of these cholesterol analogues. Our data provide the first direct assessment of virion-associated Brij 98 rafts in retroviral replication and illustrate the importance of the raft-promoting property of virion-associated cholesterol in HIV-1 replication.

Use of the wound-inducible NtQPT2 promoter from Nicotiana tabacum for production of a plant-made vaccine

The wound-inducible quinolinate phosphoribosyl transferase promoter from Nicotiana tabacum (NtQPT2) was assessed for its capacity to produce B-subunit of the heat-labile toxin (LTB) from enterotoxigenic Escherichia coli in transgenic plant tissues. Comparisons were made with the widely used and constitutive Cauliflower Mosaic Virus 35S (CaMV35S) promoter. The NtQPT2 promoter produced somewhat lower average concentrations of LTB protein per unit weight of hairy root tissue but allowed better growth thereby producing similar or higher overall average yields of LTB per culture batch. Transgenic tobacco plants containing the NtQPT2-LTB construct contained LTB protein in roots but not leaves. Moreover, wounding NtQPT2-LTB transgenic plants, by removal of apices, resulted in an approximate 500% increase in LTB levels in roots when analysed several days later. CaMV35S-LTB transgenic plants contained LTB protein in leaves and roots but wounding made no difference to their LTB content.