Ignacio Caballero

**Summary of Research Activity** I graduated in Veterinary Medicine in 2003 and joined the Animal Reproduction research group (E0A1-03) from the University of Murcia as a PhD student (2003-2007). The main goal of my research during my **PhD studies** was to improve the outcome of sperm cryopreservation and flow cytometric sex-sorting (that allows choosing the sex of the offspring) in porcine using seminal plasma and its proteins. I obtained my **first postdoctoral appointment** at Dr Piedrahita’s lab in North Carolina State University to develop transgenic pigs for animal and human medicine research. The main objective of the project was the development of “humanized” organs in the pig for “reverse” xenotransplantation. To achieve this goal the project was divided in 2 sub-objectives: 1) Development of a transgenic pig with a “humanized” immune system: I developed a number of transgenic pigs where the RAG2 (specific for B and T cells) and the NKp46 promoter (specific for NK cells) drove the expression of diphtheria toxin A in order to ablate the pig immune system. 2) Establishing a fetal injection protocol: I developed a pCAG-lacZfloxed-DTA transgenic pig. Pig foetuses were injected at 35 days of gestation with lacZ-positive bone marrow stem cells and lacZ-positive cells were detected in 2 month old pigs indicating that transgenic cells were able to engraft in the bone marrow and generate lacZ-positive hematopoietic cells. This transgenic pig lines are currently being used for engraftment of human hematopoietic stem cells.. In 2011, **I obtained a Marie Curie IIF (January 2011-January 2013) at the University of Sheffield, UK**. The project I was involved was “Regulation of innate immunity in the female reproductive tract”. Implantation of the embryo in the uterus is considered to be one of the most complex and critical steps during pregnancy. Sub-clinical infections in the female reproductive tract (FRT) contribute to implantation failure. Toll like receptors (TLRs), have been shown to have a key role in the induction of immune and inflammatory responses in mammals by activating both the nuclear factor kB transcription factor (NF-κB) and AP-1 transcription factors. TLRs have a cyclical expression during the menstrual cycle in the human endometrium. TLR expression also changes in oviductal and endometrial cell lines in response to sex hormones. In this project, we aimed to systemically investigate the effect of the embryo and sex hormones on the expression and function of TLR5 at population level and single cell level using live imaging confocal microscopy. Our results showed that: 1) the presence of the embryo is able to modulate TLR5 response by increasing NF-κB activity. Also, abnormal levels of NF-κB activity lead to implantation failure. 2) Single cell live imaging allowed us to determine for the first time dynamics of TLR5-mediated NF-κB pulsatile signalling. Moreover, we also showed that estradiol effect on NF-κB occurs at the transcription level in the nucleus rather than affect the translocation of NF-κB. In September 2013, I obtained a position as the **leader of the Porcine Mucosal Immunology team (**[**https://www6.val-de-loire.inra.fr/infectiologie-santepublique\_eng/Research/Bacteriology-teams/Porcine-mucosal-immunology**](https://www6.val-de-loire.inra.fr/infectiologie-santepublique_eng/Research/Bacteriology-teams/Porcine-mucosal-immunology)**) at INRA, France**. Our major interest lies in the use of the pig as a biomedical model in order to develop novel alternative therapies and immunostimulation strategies. Similarities between pigs and humans both anatomically and in the immune response make the pig a highly demanded animal model. In collaboration with the University of Munich we have established a transgenic pig line of CFTR knockout pigs that develop cystic fibrosis lung disease, a chronic inflammatory lung disease. We are also interested in new therapies that boost immunity through stimulation of Toll-like receptors. In summary, I published a total of **42 original articles** in international journals (and 1 submitted, h-index=18), 2 original articles in peer-reviewed national journals, 1 book and 9 book chapters. All the articles have been published in journals ranked in the highest positions of their area. I participated actively at **international and national conferences** with 52 poster presentations and oral communication. Most of them (28) have been published in international journals, such as Reproduction, fertility and development and Reproduction in domestic animals. I have been **invited speaker in 10 international conferences**.