First-line dolutegravir/lamivudine penetrates lymph nodes and reduces HIV reservoirs comparably to triple therapy.

Dual antiretroviral therapy with dolutegravir and lamivudine (DTG/3TC) is a recommended first-line regimen for people with HIV, but whether reduced-drug therapy maintains adequate antiviral pressure within lymph node (LN) reservoirs remains uncertain. In the DUALITY clinical trial, peripheral HIV-1 reservoir markers declined similarly in participants initiating DTG/3TC or dolutegravir-based triple therapy. Here we investigated viral persistence and antiretroviral drug distribution in LN using multimodal imaging. In 39 participants undergoing inguinal LN excision at baseline or during the first year of treatment, HIV-1 DNA and RNA were detected by DNAscope/RNAscope and antiretroviral spatial distribution was mapped by mass spectrometry imaging. HIV-1-infected cells declined rapidly after treatment initiation, with comparable reductions between treatment groups. Antiretrovirals were widely detected within LN tissue regions susceptible to containing HIV-positive cells. These findings provide spatial evidence that DTG/3TC achieves effective LN penetration and supports effective antiviral exposure within a key anatomical HIV reservoir.