{"title":"利用ELISPOT研究HIV和弓形虫病患者t细胞免疫应答。","authors":"Iskra Georgieva Rainova, Rumen Nenkov Harizanov, Yana Dimitrova Todorova, Mihaela Vanyova Videnova, Eleonora Marinova Kaneva, Raina Borisova Enikova, Nina Dimitrova Tsvetkova","doi":"10.1155/ipid/9514227","DOIUrl":null,"url":null,"abstract":"<p><p><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.</p>","PeriodicalId":39128,"journal":{"name":"Interdisciplinary Perspectives on Infectious Diseases","volume":"2025 ","pages":"9514227"},"PeriodicalIF":0.0000,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12289364/pdf/","citationCount":"0","resultStr":"{\"title\":\"Study on the T-Cell Immune Response in Individuals With HIV and Toxoplasmosis Using ELISPOT.\",\"authors\":\"Iskra Georgieva Rainova, Rumen Nenkov Harizanov, Yana Dimitrova Todorova, Mihaela Vanyova Videnova, Eleonora Marinova Kaneva, Raina Borisova Enikova, Nina Dimitrova Tsvetkova\",\"doi\":\"10.1155/ipid/9514227\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><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.</p>\",\"PeriodicalId\":39128,\"journal\":{\"name\":\"Interdisciplinary Perspectives on Infectious Diseases\",\"volume\":\"2025 \",\"pages\":\"9514227\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2025-07-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12289364/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Interdisciplinary Perspectives on Infectious Diseases\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1155/ipid/9514227\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q3\",\"JCRName\":\"Immunology and Microbiology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Interdisciplinary Perspectives on Infectious Diseases","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1155/ipid/9514227","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/1 0:00:00","PubModel":"eCollection","JCR":"Q3","JCRName":"Immunology and Microbiology","Score":null,"Total":0}
Study on the T-Cell Immune Response in Individuals With HIV and Toxoplasmosis Using ELISPOT.
Introduction: The intracellular parasite Toxoplasma gondii stimulates the human immune system, resulting in the activation of both cellular and humoral immune responses. In HIV-infected individuals, latent Toxoplasma 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 T. gondii-specific IFN-γ producing memory T cells in patients with both toxoplasmosis and HIV. Patients and Methods: ELISA was used to test 104 blood samples from HIV + individuals for Toxoplasma 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 T. gondii were selected to stimulate CD4+ and CD8+ T cells when performing the ELISPOT. Results: 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. Conclusion: 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.