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While the core molecular mechanisms driving Env-receptor interactions and subsequent membrane fusion are well characterized, the precise nanoscale spatial reorganization of these co-receptors at the viral binding site remains poorly defined. In this study, we employed single-particle tracking total internal reflection fluorescence (SPT-TIRF) microscopy to quantitatively analyze nanoscale organizational changes of CXCR4 on the surface of human CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells following binding by X4-tropic HIV-1. Our data reveal that both recombinant X4-gp120 and virus-like particles expressing physiological levels of X4 Env proteins (gp120 and gp41) promote CXCR4 clustering, a phenomenon linked to cell infection. Furthermore, these ligands induced oligomerization of CXCR4\n                    <jats:sup>R334X<\/jats:sup>\n                    , a naturally occurring mutant associated with WHIM syndrome that supports HIV-1 infection, but fails to oligomerize in response to CXCL12. Our findings establish a link between CXCR4 clustering and HIV-1 infection, enhancing our understanding of the initial events in viral attachment and entry. These results further suggest that HIV-1 depends on a specific spatial arrangement of co-receptors, distinct from that induced by their natural chemokine ligands, highlighting the critical role of cell-surface receptor spatial organization in dictating cellular function.\n                  <\/jats:p>","DOI":"10.7554\/elife.110354","type":"journal-article","created":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T16:30:17Z","timestamp":1776184217000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.110354","source":"Crossref","is-referenced-by-count":0,"title":["HIV-1 envelope glycoprotein modulates CXCR4 clustering and dynamics on the T cell membrane"],"prefix":"10.7554","volume":"15","author":[{"given":"Adriana","family":"Quijada-Freire","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/015w4v032","id-type":"ROR","asserted-by":"publisher"}],"name":"Chemokine Signaling group, Department of Immunology and Oncology, Centro Nacional de 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