Cell surface CD4 molecules are known to be important in several physiological responses of T lymphocytes. The use of human immunodeficiency virus (HIV) particles or purified gp120 molecules as CD4 cross-linking agents has been shown to result in a cascade of intracellular biochemical events. In addition, we and others have provided evidence suggesting that virus-mediated CD4 multimerization can lead to modulation of HIV-1 long terminal repeat-dependent activity and virus production. We were thus interested in measuring the effect of HIV-1 particles on intracellular tyrosine-phosphorylation levels, mostly of CD4-associated proteins. Using the T cell line CEM-T4, we observed that HIV-1 induces an increase in tyrosine phosphorylation of four major proteins physically complexed to the CD4 molecule. Immunoblot analysis permitted the identification of two of these proteins, p56^lckand phosphatidylinositol 3-kinase (PI 3-kinase) p85α. No concomitant variation in the level of these two CD4-associated proteins was observed after HIV-1-induced CD4 cross-linking. To our knowledge, this is the first report linking HIV-1-mediated CD4 multimerization to an increase in tyrosine phosphorylation of the PI 3-kinase complex. The four CD4-associated molecules described in this report are most likely implicated in virus-induced CD4-linked signaling events.