Antiretroviral treatment can reduce the death rate of human immunodeficiency virus (HIV) infection, and its effectiveness is maximized at the early stage of HIV infection. The present protocol demonstrates an early stage high-content HIV diagnosis based on multicolor concurrent monitoring of CD4, CD8, and CD3 coreceptors and F-actin cytoskeleton using quantum dot (Qdot)-antibody conjugates at the single cell level. Artificial HIV infection of peripheral blood mononuclear cells (PBMCs) can be achieved by treating PBMCs with gp120. Using the present methodology, we can determine the CD4-CD8 ratios of normal PBMCs and artificial HIV-infected PBMCs. In addition, this protocol enables monitoring of structural changes of actin filament alignments in PBMCs bound to gp120 proteins using the multicolor single cell imaging system. Overall, this approach presents a new model for accurate early stage HIV diagnosis. Simultaneously the approach provides information on actin cytoskeleton and subtypes of PBMCs as well as their CD4-CD8 ratios.
Keywords: HIV diagnosis; High-content imaging; PBMCs and actin filament; Quantum dots.