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Updated: Jun 15, 2026

Single-Cell Characterization of Calcium Influx and HIV-1 Infection using a Multiparameter Optofluidic Platform
Published on: May 18, 2021
Blood cell membrane fluidity and intracellular Ca2+ changes in antiretroviral-naïve and -treated HIV-1-infected
Nuno C Santos1, J Martins e Silva, Teresa Freitas
1Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Lisboa, Portugal. nsantos@fm.ul.pt
Insights
Highly active antiretroviral therapy (HAART) partially reverses HIV-1-induced changes in blood cell properties. While lymphocyte alterations persist, erythrocyte membrane fluidity improves, but intracellular calcium levels remain affected in treated patients.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Human Immunodeficiency Virus type 1 (HIV-1) infection significantly alters lymphocyte and erythrocyte intracellular calcium concentration ([Ca(2+)](int)) and membrane fluidity.
- These cellular changes were previously observed in patients prior to antiretroviral therapy.
Purpose of the Study:
- To evaluate the impact of highly active antiretroviral therapy (HAART) on lymphocyte and erythrocyte [Ca(2+)](int) and membrane fluidity in HIV-1-infected patients.
- To compare cellular parameters between untreated patients, HAART-treated patients, and healthy controls.
Main Methods:
- Blood samples were collected from HIV-1-infected patients (pre- and post-HAART) and control subjects.
- Intracellular calcium ([Ca(2+)](int)) was measured using fura-2.
- Membrane fluidity was assessed using fluorescence spectroscopy with TMA-DPH and DPH probes.
Main Results:
- Both untreated and HAART-treated patients showed increased lymphocyte [Ca(2+)](int) and decreased membrane fluidity compared to controls, with no significant difference between patient groups.
- HAART reversed the altered membrane fluidity in erythrocytes.
- Decreased erythrocyte [Ca(2+)](int) in untreated patients was not restored by HAART.
Conclusions:
- HAART partially reverses HIV-1-induced changes in lymphocyte and erythrocyte properties.
- Persistent alterations in lymphocytes and erythrocytes may facilitate HIV-1 propagation and viral reservoir maintenance.
- Observed changes might be linked to HIV Nef protein activity and endocytosis-mediated viral entry.
Abstract:
We previously showed that lymphocytes and erythrocytes of HIV-1-infected patients, prior to antiretroviral therapy, presented significant changes in intracellular calcium concentration ([Ca(2+)](int)) and membrane fluidity. The present study evaluates the same parameters after response to highly active antiretroviral therapy (HAART). Blood samples were collected from patients prior to and after antiretroviral therapy, and from control subjects. Membrane fluidity and [Ca(2+)](int) were assessed by fluorescence spectroscopy measurements, using three different probes: TMA-DPH and DPH for membrane fluidity, and fura-2 for Ca(2+). When compared with the control group, both untreated and treated patients presented increased lymphocyte [Ca(2+)](int) and decreased lymphocyte membrane fluidity, without significant differences between the two groups of patients. On the contrary, the therapy reversed the membrane fluidity variations observed in erythrocytes. The decreased erythrocyte [Ca(2+)](int) of untreated patients was not reversed by HAART. AIDS patients present changes in lymphocyte (mostly noninfected) and erythrocyte properties, partially reversed by HAART, consistent with a process of facilitated propagation of the infection to new cells, stimulation of virion production, and maintenance of a reservoir of erythrocyte-bound infectious virus. These observations can be related with the action of the HIV Nef protein in the cell's proteins and lipid composition, as well as with the recently observed cell infection by HIV-1 via endocytosis.
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