AbstractAlthough effector CD4⁺ T cells readily respond to antigen outside the vasculature, how they respond to intravascular antigens is unknown. Here we show the process of intravascular antigen recognition using intravital multiphoton microscopy of glomeruli. CD4⁺ T cells undergo intravascular migration within uninflamed glomeruli. Similarly, while MHCII is not expressed by intrinsic glomerular cells, intravascular MHCII-expressing immune cells patrol glomerular capillaries, interacting with CD4⁺ T cells. Following intravascular deposition of antigen in glomeruli, effector CD4⁺ T-cell responses, including NFAT1 nuclear translocation and decreased migration, are consistent with antigen recognition. Of the MHCII⁺ immune cells adherent in glomerular capillaries, only monocytes are retained for prolonged durations. These cells can also induce T-cell proliferation in vitro. Moreover, monocyte depletion reduces CD4⁺ T-cell-dependent glomerular inflammation. These findings indicate that MHCII⁺ monocytes patrolling the glomerular microvasculature can present intravascular antigen to CD4⁺ T cells within glomerular capillaries, leading to antigen-dependent inflammation.