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The gut as a site of HIV immunopathogenesis: study of the microbiome, mucosal immunology and epithelial barrier structure in antiretroviral naïve and treated individuals
Italy (SISM) - University of Milan, Milan
Laboratory of Infectious Diseases, Department of Health Sciences, San Paolo Hospital, Via di Rudinì 8, 20142, University of Milan, Italy
Prof. Antonella d’Arminio Monforte and Prof. Giulia Marchetti
Camilla Tincati, M.D., Ph.D; Elvira Stefania Cannizzo, Ph.D; Esther Merlini, Ph.D.; Giuseppe Ancona, M.D.; Federico Cazzaniga BSc. E-mail: firstname.lastname@example.org; email@example.com; firstname.lastname@example.org; Phone: +39 028184 3061
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
The gastrointestinal tract is a major site of HIV immunopathogenesis. HIV, in fact, infects CD4+ T-cells which reside in the gut, causing not only T-cell apoptosis but also profound changes in the structure of the epithelial barrier, composition of the microbiome and function of mucosal immunity. Taken together, these alterations lead to the translocation of bacterial bioproducts from the gut lumen to the systemic circulation, causing a state of “T cell hyperactivation”, known to impact on further CD4+ T-cell loss and clinical outcome. Antiretroviral therapy suppresses viral replication in the peripheral blood, yet has a partial effect in restoring the composition of the microbiota, structure of the gut barrier and function of the local immune system. In particular, whether timing of antiretroviral therapy may impact on the above-mentioned parameters in currently unknown.
What is the aim of the project?
Aim of our research project is to better understand whether early treatment of HIV disease may restore the gut microbiome, structure of the epithelial barrier and function of the mucosal immune system.
What techniques and methods are used?
Immunohistochemistry on tissue biopsies; flow cytometry on tissue biopsies and peripheral blood mononuclear cells (PBMCs); ELISAs on plasma
What is the role of the student?
- The student will mainly observe
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Students will master the principal method for PBMCs separation from whole-blood (Ficoll-Histopaque technique) and gut biopsies. Further, they will gain insights into the basic principles of flow cytometry: cell cultures; fluorochrome-antibody staining and acquired events; compensation; plot interpretation. Students will also learn how to master the ELISA technique (standard-curve interpretation). Finally, students will perform preliminary statistical analyses of their results and learn how to make figures of their work, which will be presented in the course of lab meetings.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Preliminary readings on the principal laboratory techniques will be provided What is the role of the student and what is expected from him/her during the research exchange? (minimum 3 lines) Students will separate peripheral blood mononuclear cells (PBMCs) from fresh, whole-blood and gut tissue biopsies of HIV-infected subjects. Cells will then be used for flow cytometric experiments. Students will also separate plasma from peripheral blood which will be used for cytokine quantification and microbial translocation measurements by ELISA. Immunohistochemistry on tissue biopsies will be performed in collaboration with the pathology lab to study the gut junctional complex.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare an abstract - The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system - Students in biomedical fields
- Tincati C et al. AIDS Res Ther 2016; Gut barrier structure; mucosal immunity and intestinal microbiota in the pathogenesis and treatment of HIV infection. doi: 10.1186/s12981-016-0103-1
- Tincati C et al. AIDS 2016; mpaired gut junctional complexes feature latetreated individuals with suboptimal CD4+ T-cell recovery upon virologically suppressive combination antiretroviral therapy. doi: 10.1097/QAD.0000000000001015
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