Immune evasion by tumors includes several different mechanisms, including the inefficiency of antigen presenting cells (APCs) to trigger anti-tumor T cell responses. B lymphocytes may display a pro-tumoral role but can also be modulated to function as antigen presenting cells to T lymphocytes, capable of triggering anti-cancer immune responses. While dendritic cells, DCs, are the best APC population to activate naive T cells, DCs or their precursors, monocytes, are frequently modulated by tumors, displaying a tolerogenic phenotype in cancer patients. In patients with cervical cancer, we observed that monocyte derived DCs are tolerogenic, inhibiting allogeneic T cell activation compared to the same population obtained from patients with precursor lesions or cervicitis. In this work, we show that B lymphocytes from cervical cancer patients respond to treatment with sCD40L and IL-4 by increasing the CD80+CD86+ population, therefore potentially increasing their ability to activate T cells. To test if B lymphocytes could actually trigger anti-tumor T cell responses, we designed an experimental model where we harvested T and B lymphocytes, or dendritic cells, from tumor bearing donors, and after APC stimulation, transplanted them, together with T cells into RAG1-/- recipients, previously injected with tumor cells. We were able to show that anti-CD40 activated B lymphocytes could trigger secondary T cell responses, dependent on MHC-II expression. Moreover, we showed that dendritic cells were resistant to the anti-CD40 treatment and unable to stimulate anti-tumor responses. In summary, our results suggest that B lymphocytes may be used as a tool for immunotherapy against cancer.

Cervical cancer continues to be a public health problem in developing countries. Previous studies have shown that cervical cancer cells display markers of aerobic glycolysis, indicating that these tumors are likely to secrete lactate. Mostly, lactate is recognized as a molecule capable of suppressing immune responses, through inhibition of T cells, Mϕs, and dendritic cells. We and others have previously shown that Mϕs are frequent cells infiltrating cervical cancers with the ability to inhibit antitumor immune responses and promote tumor growth through angiogenesis. Here, we have tested the hypothesis that lactate, secreted by cervical cancer cells, can modulate Mϕ phenotype. First, we showed higher lactate plasma concentrations in patients with increasing cervical lesion grades, with maximum concentration in the plasma of cancer patients, which supported our hypothesis. We then inhibited lactate production in tumor cell spheroids established from cervical cancer derived cell lines, using the lactate dehydrogenase inhibitor, oxamate, prior to co‐culture with monocytes. Lactate mediated part of the crosstalk between tumor cells and Mϕs, promoting secretion of IL‐1β, IL‐10, IL‐6, and up‐regulation of hypoxia induced factor‐1α expression, and down‐regulation of p65‐NFκB phosphorylation in Mϕs. We also showed that Mϕs from co‐cultures treated with oxamate were better inducers of T cell activation. Of note, experiments performed with inhibition of the monocarboxylate transporters rendered similar results. Our data confirms the hypothesis that lactate, secreted by cervical tumor cells, influences the phenotype of tumor Mϕs, promoting a suppressive phenotype.

No abstract

Rope bridges are being increasingly installed worldwide to mitigate the negative impacts of roads on arboreal animals. However, monitoring of these structures is still limited and an assessment of factors influencing the crossing behaviours is lacking. We monitored the use of a rope bridge near Busselton, Western Australia by the endangered western ringtail possums (Pseudocheirus occidentalis) in order to identify the patterns of use and factors influencing the crossings. We installed motion sensor cameras and microchip readers on the bridge to record the crossings made by individual animals, and analysed these crossing data using generalised linear models that included factors such as days since the installation of the bridge, breeding season, wind speed, minimum temperature and moonlight. Possums started investigating the bridge even before the installation was completed, and the first complete crossing was recorded only 36 days after the installation, which is remarkably sooner than arboreal species studied in other parts of Australia. The possums crossed the bridge increasingly over 270 days of monitoring at a much higher rate than we expected (8.87 ± 0.59 complete crossings per night). Possums crossed the bridge less on windy nights and warm nights probably due to the risk of being blown away and heat stress on warmer days. Crossings also decreased slightly on brighter nights probably due to the higher risk of predation. Breeding season did not influence the crossings. Pseudocheirus occidentalis habituated to the bridge very quickly, and our results demonstrate that rope bridges have a potential as an effective mitigation measure against the negative impacts of roads on this species. More studies and longer monitoring, as well as investigating whether crossing results in the restoration of gene flow are then needed in order to further assess the true conservation value of these crossing structures.

The fragmentation of habitats by roads and other artificial linear structures can have a profound effect on the movement of arboreal species due to their strong fidelity to canopies. Here, we used 12 microsatellite DNA loci to investigate the fine-scale spatial genetic structure and the effects of a major road and a narrow artificial waterway on a population of the endangered western ringtail possum (Pseudocheirus occidentalis) in Busselton, Western Australia. Using spatial autocorrelation analysis, we found positive genetic structure in continuous habitat over distances up to 600 m. These patterns are consistent with the sedentary nature of P. occidentalis and highlight their vulnerability to the effects of habitat fragmentation. Pairwise relatedness values and Bayesian cluster analysis also revealed significant genetic divergences across an artificial waterway, suggesting that it was a barrier to gene flow. By contrast, no genetic divergences were detected across the major road. While studies often focus on roads when assessing the effects of artificial linear structures on wildlife, this study provides an example of an often overlooked artificial linear structure other than a road that has a significant impact on wildlife dispersal leading to genetic subdivision.