Study on the T‐Cell Immune Response in Individuals With HIV and Toxoplasmosis Using ELISPOT

<b>Introduction:</b> The intracellular parasite <i>Toxoplasma gondii</i> stimulates the human immune system, resulting in the activation of both cellular and humoral immune responses. In HIV-infected individuals, latent <i>Toxoplasma</i> infection can reactivate, resulting in toxoplasmosis encephalitis (TE). Detection of specific memory T cells in such patients will prevent the risk of toxoplasmosis-related complications. ELISPOT assesses CD4+ and CD8+ T cell responses to antigens, and facilitates the identification of <i>T. gondii</i>-specific IFN-γ producing memory T cells in patients with both toxoplasmosis and HIV. <b>Patients and Methods:</b> ELISA was used to test 104 blood samples from HIV + individuals for <i>Toxoplasma</i> antibodies. Peripheral blood mononuclear cells were isolated from the blood samples of the toxoplasmosis-positive HIV-infected patients and used to analyze the T-cell immune response. Peptides from the <i>T. gondii</i> were selected to stimulate CD4+ and CD8+ T cells when performing the ELISPOT. <b>Results:</b> Serological data for toxoplasmosis was identified in 29 (27.6%) of the total number of patients. A significant difference was observed in the CD4+ T cell count between HIV-positive patients with and without toxoplasmosis. Seven of the HIV-infected patients with toxoplasmosis had a low CD4+/CD8+ T cell ratio. After performing a 16-20 h ELISPOT with peptide stimulation to investigate the presence of specific IFN-γ-producing cells in these seven patients, no IFN-γ-secreting cells were detected. Subsequently, a modified method was used, in which the immune cells were stimulated for a period of 5 days. At the end of this stimulation, all samples from HIV-infected patients with toxoplasmosis were ELISPOT positive, with a mean of 32 and 45 spots per well, respectively. <b>Conclusion:</b> It is important to monitor patients with HIV, toxoplasmosis, and immunodeficiency. This can help prevent complications such as TE. A modified ELISPOT protocol may be required to determine the specific cell-mediated response in immunocompromised patients.