The young investigators: A ‘day in the lab’

With Nicola Cotugno

Why are you involved in the EPIICAL project and what’s your role?

I became interested in HIV-related immunology after receiving my degree in Medicine and when, as a medical student, I began following vertically HIV infected children at the Bambino Gesù Children’s Hospital in Rome. During my residency in Pediatrics, back in 2013, I observed the first baby steps made by EPIICAL, stemming from the ideas of Paolo Palma, Paolo Rossi and Carlo Giaquinto. I immediately sensed that EPIICAL was a great opportunity to pursue my passion for Immunology Research in order to contribute to future possible strategies aimed at improving the quality of life of HIV infected children. Since then, I have been working within the Immunological Platform through the active collaboration between the Miami Center for AIDS Research (CFAR) and OPBG. Through ongoing projects and during my postdoctoral fellowship experience in Miami in 2014 and 2015 we were able build preliminary analysis to support the EPIICAL experimental plan. Today, in the 4th and last year of the project, I am now actively involved in the research and clinical activities of the Immunological Platform for the EPIICAL studies (CARMA, EARTH and HVRRICANE) through scientific discussions, SOPs validation and data dissemination.

How has your participation in EPIICAL helped prepare your career for this role in the Project?

I believe that my current research is strongly influenced by the concepts and approaches pursued by the EPIICAL consortium. Moreover, I feel that this project, more than any other in which I have participated, has given young researchers like me the opportunity to share work experiences with great scientists and top-level physicians in the field of pediatric HIV from all over the world. On the other hand, however, EPIICAL is a broad and extremely diverse consortium which aims to merge knowledge from different fields. I now feel that this complexity has greatly helped my personal growth and adaptability in future collaborative projects.

Tell us about your typical day in the lab!

There is no perfect day in the lab. But the fact that you are surrounded by young, ambitious and valuable young researchers certainly makes it better. The research group has grown incredibly since the beginning of this experience, and I am now lucky enough to follow two PhD students who are involved in HIV projects. Accordingly, the day in the lab is spent between bench work, data analysis and discussion through the weekly based data club and journal club.

The main activities of the lab work are FACS analysis, FACS sorting and multiplexed gene expression analysis. But we all have the same impression: the best is yet to come!

Which are the main results of your Work Package (Immunological Platform)?

In the last few years we have been focusing on humoral and cell immunity in early treated HIV infected children. Indeed, we recently showed how HIV serology profiles may be crucial to predict the size of viral reservoirs in vertically HIV infected children. In addition, we produced gene expression data on HIV specific B cells after in vitro stimulation in early treated patients. We believe that this data may inform future immune therapeutic strategies able to improve HIV specific responses. Indeed, sorted gp140+ B cells from these patients were analyzed for gene expression and BCR sequence. Toll Like Receptor pathways proved to be downregulated in seronegative early treated children, suggesting that targeting such molecules may improve the HIV specific immune response in these children. On the T cell side, our work showed early treated children present a functional advantage compared with late treated patients in terms of intracellular cytokine production upon in vitro stimulation (data produced by Miami CFAR). In line with this, RNA Seq (data produced at Case Western University) also pointed to specific differences between early and late treated patients. This evidence further suggests that the timing of ART initiation is a pivotal characteristic in clinical studies aiming at remission.

These differences and the selective advantage in the memory NK compartment was also recently showed by our group at the EPIICAL general assembly held in Madrid in November 2018.

What will be the next milestones of the Immunological Platform?

We are all very excited about the upcoming enrollment and experiments for the HVRRICANE project. Indeed, for the first time, this study will test the ability of a HIV DNA vaccination together with a TLR4 agonist to reduce viral reservoir in HIV infected/early treated patients. We will test how the vaccination induces HIV specific T and B cell immunity and its ability to induce neutralizing Ab as well as Ab dependent Cytotoxic activity.

The other EPIICAL projects, CARMA and EARTH, are also moving fast. In 2019 we will produce the final results for CARMA and also start receiving the EARTH samples from the African sites. Needless to say, plenty of work and excitement still in the pipeline.

What advice would you offer to current young researchers?

Follow your passion, don’t let setbacks discourage your research, be determined …and try to do something EPIICAL!!!