The quest for an HIV cure heavily depends on our ability to accurately map how the virus establishes reservoirs in the body. Today, we know that a major part of the HIV reservoir is located in gut, however, most research is still performed on blood. In addition, only a limited number of studies examined tissue-resident memory T cells (Trms), the predominant lymphocyte subset in tissues, as a cellular reservoir for HIV. Therefore, we aim to characterize CD4 Trms in gut as an HIV reservoir, study how this reservoir is formed, and explore opportunities to eradicate it. To this end, we will map interactions between HIV-infected cells and immune cells to better profile how pro-inflammatory environments are maintained in people living with HIV (PLWH). In our highly innovative approach, we will use Visium spatial transcriptomics combined with HIV RNA/DNAscope, which is the only contemporary technique that allows to collect structural and transcriptomic information at the single-cell level from an individual tissue slide. We will perform these analyses on cohorts of acutely and chronically infected PLWH, from which gut tissue is obtained before and after therapy initiation, controlled by a group of HIV-negative individuals. To dive deep into these unique atlases and answer mechanistical questions, we will make use of explant models that will be HIV infected. These insights will contribute to development of HIV cures and interventions to relieve patients from inflammatory disease.