Separating Lentiviral Vector Injection and Induction of Gene Expression in Time, Does Not Prevent an Immune Response to rtTA in Rats

BACKGROUND: Lentiviral gene transfer can provide long-term expression of therapeutic genes such as erythropoietin. Because overexpression of erythropoietin can be toxic, regulated expression is needed. Doxycycline inducible vectors can regulate expression of therapeutic transgenes efficiently. However, because they express an immunogenic transactivator (rtTA), their utility for gene therapy is limited. In addition to immunogenic proteins that are expressed from inducible vectors, injection of the vector itself is likely to elicit an immune response because viral capsid proteins will induce "danger signals" that trigger an innate response and recruit inflammatory cells. METHODOLOGY AND PRINCIPAL FINDINGS: We have developed an autoregulatory lentiviral vector in which basal expression of rtTA is very low. This enabled us to temporally separate the injection of virus and the expression of the therapeutic gene and rtTA. Wistar rats were injected with an autoregulatory rat erythropoietin expression vector. Two or six weeks after injection, erythropoietin expression was induced by doxycycline. This resulted in an increase of the hematocrit, irrespective of the timing of the induction. However, most rats only responded once to doxycycline administration. Antibodies against rtTA were detected in the early and late induction groups. CONCLUSIONS: Our results suggest that, even when viral vector capsid proteins have disappeared, expression of foreign proteins in muscle will lead to an immune respons

Mouse models of breast cancer metastasis

Metastatic spread of cancer cells is the main cause of death of breast cancer patients, and elucidation of the molecular mechanisms underlying this process is a major focus in cancer research. The identification of appropriate therapeutic targets and proof-of-concept experimentation involves an increasing number of experimental mouse models, including spontaneous and chemically induced carcinogenesis, tumor transplantation, and transgenic and/or knockout mice. Here we give a progress report on how mouse models have contributed to our understanding of the molecular processes underlying breast cancer metastasis and on how such experimentation can open new avenues to the development of innovative cancer therapy

Dispersal of conidia of Ascochyta fabae f. sp. lentis from infected lentil plants by simulated wind and rain

Splash was shown to be an effective mechanism for short-range dispersal of conidia of Ascochyta fabae f. sp. lentis, the cause of ascochyta blight of lentil. The dispersal gradients were well described by the power law model in its linear form, 1ny = 1na - b 1nx. In still air the slope of the linearized dispersal gradient, b, ranged from 2.83 to 4.07 and was steeper for 4.9 mm than for 3.9 mm incident drops. Nevertheless, for all drop sizes tested, fewer than 50% of the conidia were splashed more than 15 cm from the source. The pattern of conidium dispersal was similar for both drop sizes when horizontal windspeeds were 2.5 or 5 mis. Wind significantly decreased the value of b (range 2.35-2.43 at 25 m/s, 1.71-1.91 at 5 m/s) and increased by about 2 m the maximum distance that conidia in ballistic droplets were deposited. In addition, the experiments suggested two other potentially important mechanisms for dispersal of the pathogen over longer distances, namely conidia in small air-borne droplets and windblown leaflets.Peer reviewe