Tobacco mosaic virus in the lungs of mice following intra-tracheal inoculation.

Plant viruses are generally considered incapable of infecting vertebrates. Accordingly, they are not considered harmful for humans. However, a few studies questioned the certainty of this paradigm. Tobacco mosaic virus (TMV) RNA has been detected in human samples and TMV RNA translation has been described in animal cells. We sought to determine if TMV is detectable, persists, and remains viable in the lung tissues of mice following intratracheal inoculation, and we attempted to inoculate mouse macrophages with TMV. In the animal model, mice were intratracheally inoculated with 10(11) viral particles and were sacrificed at different time points. The virus was detected in the mouse lungs using immunohistochemistry, electron microscopy, real-time RT-PCR and sequencing, and its viability was studied with an infectivity assay on plants. In the cellular model, the culture medium of murine bone marrow derived macrophages (BMDM) was inoculated with different concentrations of TMV, and the virus was detected with real-time RT-PCR and immunofluorescence. In addition, anti-TMV antibodies were detected in mouse sera with ELISA. We showed that infectious TMV could enter and persist in mouse lungs via the intratracheal route. Over 14 days, the TMV RNA level decreased by 5 log(10) copies/ml in the mouse lungs and by 3.5 log(10) in macrophages recovered from bronchoalveolar lavage. TMV was localized to lung tissue, and its infectivity was observed on plants until 3 days after inoculation. In addition, anti-TMV antibody seroconversions were observed in the sera from mice 7 days after inoculation. In the cellular model, we observed that TMV persisted over 15 days after inoculation and it was visualized in the cytoplasm of the BMDM. This work shows that a plant virus, Tobacco mosaic virus, could persist and enter in cells in mammals, which raises questions about the potential interactions between TMV and human hosts.

Tobacco mosaic virus in cigarettes and saliva of smokers.

BACKGROUND: Tobacco mosaic virus (TMV) has been described as viable in cigarettes or cigar tobacco. It has been cultured about 50 years ago from sputa and thoracentesis fluids of cigarette smokers with a history of pulmonary disease and from lung cancerous matter. In addition, TMV RNA has been recovered recently from human stools while tobacco DNA was recovered from smokers' bronchoalveolar lavages. OBJECTIVES: We assessed the prevalence, titers, and infectivity of TMV in tobacco cigarettes and in the saliva of smokers and non smokers. STUDY DESIGN: Tobacco cigarettes from 10 packs of different brands were purchased. Saliva was collected from 12 smokers and 15 non-smokers. Cigarettes and saliva samples were tested for the presence of TMV RNA using a home-made quantitative real-time RT-PCR assay. TMV RNA quantification was enabled by using dilutions of purified TMV. TMV viability was tested by inoculating leaves of Nicotiana tabacum Xanthi (NtX). RESULTS: All 47 smoking cigarettes of six brands were TMV RNA-positive (mean titer, 9.5 log(10)copies/cigarette); TMV was found viable in 53% of them. In addition, 20/44 (45%) saliva from 12 smokers compared to 0/16 saliva from 15 non-smokers tested positive for TMV RNA (mean titer, 3.8 log(10)copies/ml) (p=0.001). CONCLUSIONS: Our results indicate that the TMV genome may get access to the human body by direct exposure through smoking. Although plant viruses are considered not pathogenic for animals, these data prompt to study if TMV RNA is present and induces a modification of the transcriptional program in lung cells of cigarette smokers.