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V designed and performed the experiments, analyzed the data and wrote the manuscript. B and J. L performed microscopy experiments. W provided the Thy1. I and N. Related to Figure 1: A Scheme of immunization with heat killed Klebsiella pneumoniae serotype K2 on day 0 and 7.

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V designed and performed the experiments, analyzed the data and wrote the manuscript. B and J. L performed microscopy experiments. W provided the Thy1. I and N. Related to Figure 1: A Scheme of immunization with heat killed Klebsiella pneumoniae serotype K2 on day 0 and 7. One out of 2 independent experiments is shown. Both groups of mice were treated with tamoxifen. The bar graphs on the right shows the frequency of exTH17 cells in the lung of non-immunized and immunized mice Day Each dot represents one mouse.

Representative dot plots are shown n: 3 mice per group. One out of 3 experiments is shown. Data is cumulative of 4 experiments for C, and 2 experiments in D. Dot plots showing blood full chimerism 3 weeks after surgery. A, B Dot plots show the percentage and B absolute numbers of Th17 and exTh17 after infection at day 7 and day C Histograms show the percentage of skin exTh17 cells that express CD left graph, and CD69 right graph at day 7 blue histogram and day 30 red histogram after infection.

Grey histograms represents negative control. Representative dot plots are shown n:3 mice per group. The graph shows the absolute number of spleen left and dLN right exTH17 cells in the isotype ctrl treated group black circles and in the anti-IL-7 treated group red circles.

Two combined experiments are shown. Mann-Whitney U Test ns non significant. B The Box plots show the specific genes for each cluster, with cluster labels on the x-axis; normalized expression values on the y-axis. The same color codes as in Figure 4F were used. Three markers are shown for each cluster. Showing one out of 2 representative experiments. Mann-Whitney U test, ns non-significant.

Each dot plot represents one mouse. Mann- Whitney U test. Mann- Whitney U test, ns non-significant. However, the cellular origin of CD4 TRM cells, and their contribution to host defense remains elusive. Conversely, using parabiosis and depletion studies, we demonstrated that exTH17 TRM cells play an important role in bacterial clearance. Thus, we delineate the origin and function of airway CD4 TRM cells during bacterial infection, offering novel strategies for targeted vaccine design.

Introduction Adaptive immunity provides life-long protection from infection by establishing an enduring reservoir of memory cells.

Upon infection CD4 and CD8 T cells undergo clonal expansion followed by a subsequent contraction, and ultimately memory formation Kaech, Wherry et al.

While central memory T TCM cells have long been known to mainly reside and circulate through secondary lymphoid tissues and effector memory TEM cells to circulate through non lymphoid tissues, the identification of TRM cells within peripheral organs has opened new concepts about tissue-specific protective immunity Schenkel and Masopust To address these questions, we devised an experimental plan combining the use of two complementary fate reporter mouse models, next generation T cell receptor TCR and single-cell sequencing approaches.

We consider that understanding CD4 TRM cell development and function is imperative, particularly as they may represent the key target cells to design specific and efficacious vaccination against the growing number of antibiotic resistant bacterial strains emerging in the clinic.

This might also help to reduce antibiotic use, avoiding the evolution of antibiotic resistant-bacteria For instance, there is currently no targeted treatment strategy for carbapenem resistant Enterobacteriaceae like Klebsiella pneumoniae Kp. This is a leading cause of nosocomial and community-acquired gram-negative bacterial pneumonia, which results in a severe pyrogenic infection with high mortality rates Falagas, Tansarli et al.

Considering all this, we hypothesized that CD4 TRM cells derive from the first wave of effector cells generated during the first encounter with a pathogen. Furthermore, since CD4 TRM cells localize at the site of immunization, we also hypothesized that some of them acquire a poised while others a more plastic status Lee, Turner et al.

Here, by using an immunization-infection model with different serotypes of Kp, we show that a significant fraction of the lung long-lived CD4 TRM cells exTH17cells derive from specific-effector TH17 cells.

Results TRM cells can derive from effector cells and protect against carbapenem resistant Klebsiella pneumoniae infection We started by characterizing the kinetics of the development of lung- TRM cells. To this end, wild type wt mice were immunized twice with heat killed serotype 2 Klebsiella pneumoniae Figure S1A and the presence of CD4 TRM cells was evaluated at multiple time points.

An in vivo antibody Ab labeling technique was used to differentiate between circulatory and lung infiltrating CD4 T cells Anderson, Mayer-Barber et al. Lung infiltrating CD4 T cells began accumulating as early as day 5 post-immunization and persisted through day Figure S1B.

Similar to classical CD8 and CD4 memory formation, lung infiltrating CD4 cells underwent robust expansion upon immunization, followed by an acute contraction phase. We next aimed to further characterize the origin of TRM CD4 cells, a point that has still remained elusive. TH17 cells have previously been shown to provide protection against Kp infection Ye, Rodriguez et al.

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Summary Steroid hormones are essential for the survival of all mammals. The testes revealed seminiferous tubules, stroma, rete testis with interstitial fibrosis and reduced number of germ cells. The mitochondria were not close to the mitochondria-associated ER membrane MAM , and cells were filled with the microfilaments. Our result revealed that absence of pregnenolone is associated with organelle stress, leading to altered protein organization that likely created steric hindrance in testicular cells. Learning points: Testes revealed seminiferous tubules, stroma, rete testis with interstitial fibrosis and reduced number of germ cells; Testicular mitochondrial size was small with little SCC expression within the mitochondria; Absence of pregnenolone is associated with organelle stress. Background Prenatal stress affects many aspects of reproduction, including sexual differentiation of the brain, development of the reproductive organs, puberty onset, reproductive behavior, gonad function and concentrations of reproductive hormones 1. In addition, the physiological consequences of stressors in a pregnant female may be transmitted to her developing fetuses, including alterations in the opioid system, as well as causing epigenetic changes to placental gene expression and alterations in the maternal hypothalamic—pituitary—adrenal HPA axis.

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