T helper 17 (Th17) cells develop in response to T cell receptor signals (TCR) in the presence of specific environments, and produce the inflammatory cytokine IL17A. These cells have been implicated in a number of inflammatory diseases and represent a potential target for ameliorating such diseases. The kinase ITK, a critical regulator of TCR signals, has been shown to be required for the development of Th17 cells. However, we show here that lung inflammation induced by Saccharopolyspora rectivirgula (SR) induced Hypersensitivity pneumonitis (SR-HP) results in a neutrophil independent, and ITK independent Th17 responses, although ITK signals are required for γδ T cell production of IL17A. Transcriptomic analysis of resultant ITK independent Th17 cells suggest that the SR-HP-induced extrinsic inflammatory signals may override intrinsic T cell signals downstream of ITK to rescue Th17 responses in the absence of ITK. These findings suggest that the ability to pharmaceutically target ITK to suppress Th17 responses may be dependent on the type of inflammation.

Endogenous proteases are associated with the ovulation process and inflammation, among other human physiological processes. Still their role in the transition from normal tissue to cancer development remains uncertain. For example, the Protease activated receptor 2 (PAR‐2) is a G Protein Couple receptor expressed in mostly in epithelial cells, including the female reproductive system. Previous work from our lab showed that the PAR‐2 antagonist‐FSLLRY‐NH2 inhibited the cell‐mediated inflammation in the airways. The aim of this study is to evaluate the effect of the PAR‐2 antagonist when a normal cell progresses to a cancerous one, using trypsin as a gold standard enzyme for Par‐2 activation. In this case, the transition from normal endometrium, to endometriosis, ovarian. We hypothesized that PAR‐2 antagonist will decrease the activation of its receptor reducing the transition to endometriosis and ovarian cancer. To test this, we performed western blot to determine PAR‐2 expression in endometriosis (12Z) cells, human endometrial stromal cells (HESC) cells, ovarian cancer (A2780) cells, and breast cancer (MCF‐7) cells. In addition, we quantified the activation of PAR‐2 receptor using a Fluorogenic intracellular calcium assay in human normal endometrial (uterine) epithelial cells (HEUEC), ovarian cancer (Caov‐3), and MCF‐7 cells as internal positive control. We also performed the BrdU proliferation assay on these cells. Our results showed a higher expression (p<0.05) of PAR‐2 in A2780 cells (ovarian aggressive cancer) when compared with the other cells lines. The intracellular calcium assay resulted in a significant reduction (p<0.05) once exposed to the PAR‐2 antagonist in the HEUEC cells and Caov‐3 when compared to cells treated with trypsin (p<0.05). In contrast, there was not significant difference in the MCF‐7 cell line upon exposure to treatments. We observed a significant decrease in MCF‐7 proliferation when treated with the PAR‐2 antagonist in comparison to trypsin treatment. Our results suggested that PAR‐2 antagonist can be used as treatment or adjunct therapy for specific female reproductive cancers. Next steps of this study will include the active full‐length PAR‐2 recombinant and test other concentrations of the antagonist to determine the LD50 in the selected cells. Proliferating markers will be measured for all the previous cell lines as well for endometrial cancer.

Hypersensitivity pneumonitis (HP) is an extrinsic allergic alveolitis disease that is constantly affecting the farmers. It is triggered by the continuous exposure of environmental antigens, specifically antigens that grow in hot humid conditions in hay like Saccharopolyspora rectivirgula (SR). While a role of T cells has been established in the development of this diseases, the role of the innate immune system is still largely unknown in the development of lung inflammation leading to HP. To study this disease, we used a mouse model of HP, induced by SR, along with Rag−/− mice which only have an innate immune system and lack an adaptive immune system (and thus have no T cells). We hypothesized that the development of Hypersensitivity pneumonitis requires the presence of T (and B) cells, at the same time, there will be a lower percentage and number of inflammatory cells in the lungs of Rag−/− mice upon exposure to SR. WT and Rag−/− mice were exposed to SR. Lungs, spleen and lymph nodes where collected to analyze their response. We examined the percentage and number of myeloid and Innate Lymphoid Cells (ILC), as well as their ability to produce cytokines. We found that there was no significant difference in the number of Innate Lymphoid cells and myeloid cells between SR‐exposed WT and Rag−/− mice. These results may be due to the low concentration of bacteria used for these experiments. In future experiments, we propose to use a higher concentration of SR to induce inflammation. Support or Funding Information This work was supported in part by grants from the NIH‐AI38570, NIH‐AI20701, and NIH‐AI129422. NIH‐NIGMS #2R25GM096955 (RISE). VIP‐Veterinary Investigation Program.