Rainova Iskra Georgieva, Harizanov Rumen Nenkov, Todorova Yana Dimitrova, Vanyova Videnova Mihaela, Kaneva Eleonora Marinova, Enikova Raina Borisova, Tsvetkova Nina Dimitrova
Department of Parasitology and Tropical Medicine, National Centre of Infectious and Parasitic Diseases (NCIPD), Sofia, Bulgaria.
Department of Immunology, National Centre of Infectious and Parasitic Diseases, Sofia, Bulgaria.
Interdiscip Perspect Infect Dis. 2025 Jul 17;2025:9514227. doi: 10.1155/ipid/9514227. eCollection 2025.
The intracellular parasite stimulates the human immune system, resulting in the activation of both cellular and humoral immune responses. In HIV-infected individuals, latent 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 -specific IFN-γ producing memory T cells in patients with both toxoplasmosis and HIV. ELISA was used to test 104 blood samples from HIV + individuals for 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 were selected to stimulate CD4+ and CD8+ T cells when performing the ELISPOT. 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. 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.
这种细胞内寄生虫会刺激人体免疫系统，导致细胞免疫和体液免疫反应的激活。在感染HIV的个体中，潜伏感染可能会重新激活，从而导致弓形虫性脑炎（TE）。检测此类患者体内的特异性记忆T细胞将预防弓形虫病相关并发症的风险。ELISPOT可评估CD4+和CD8+ T细胞对抗原的反应，并有助于识别同时患有弓形虫病和HIV的患者中产生特异性干扰素-γ的记忆T细胞。采用酶联免疫吸附测定法（ELISA）检测了104份来自HIV阳性个体的血液样本中的抗体。从弓形虫病阳性的HIV感染患者的血液样本中分离出外周血单核细胞，用于分析T细胞免疫反应。在进行ELISPOT检测时，选择来自该病原体的肽段来刺激CD4+和CD8+ T细胞。在全部患者中检测到29例（27.6%）有弓形虫病的血清学数据。在有和没有弓形虫病的HIV阳性患者之间，观察到CD4+ T细胞计数存在显著差异。7例感染HIV且患有弓形虫病的患者CD4+/CD8+ T细胞比值较低。在对这7例患者进行16 - 20小时的肽段刺激ELISPOT检测以研究特异性产生干扰素-γ的细胞的存在后，未检测到分泌干扰素-γ的细胞。随后，采用了一种改良方法，即对免疫细胞进行5天的刺激。在这种刺激结束时，所有来自感染HIV且患有弓形虫病患者的样本ELISPOT检测均为阳性，平均每孔分别有32个和45个斑点。监测感染HIV、患有弓形虫病和免疫缺陷的患者很重要。这有助于预防诸如TE等并发症。可能需要一种改良的ELISPOT方案来确定免疫受损患者的特异性细胞介导反应。
