The ELISPOT assay is increasingly used for assessing cellular immune responses in clinical trials of HIV-1 or cancer vaccines. However, to date, data from clinical trials do not consistently show that immune responses...The ELISPOT assay is increasingly used for assessing cellular immune responses in clinical trials of HIV-1 or cancer vaccines. However, to date, data from clinical trials do not consistently show that immune responses are correlated with clinical endpoints. This is due in part to the lack of assay standardization and validation across laboratories and therefore, a quality control panel is required to establish competency and comparability amongst different laboratories. In this study peripheral blood mononuclear cells (PBMCs) from healthy individuals were screened and frozen in liquid nitrogen. The recovery and viability of the PBMCs and the frequencies of interferon (IFN)-γ-secreting cells after CEF peptide pool stimulation were detected after various intervals in seven different laboratories. The recovery and viability did not differ significantly after different intervals. Although the frequencies of IFN (interferon)-γ-secreting cells among thawed PBMCs (peripheral blood mononuclear cells) fluctuated after CEF peptide pool stimulation at different intervals, they were not significantly decreased compared with those among fresh PBMCs. However, the viabilities, recoveries and frequencies of IFN-γ-secreting cells differed significantly among the seven laboratories. Our results indicate that cryopreserved PBMCs could be used as a quality control panel for ELISPOT. However, the procedures for ELISPOT need to be standardized amongst different laboratories.展开更多
Breast cancer is the most common cancer among women worldwide[1].In 2018,2.09 million new breast cancer cases were identified globally,accounting for 11.6% of all cancers in that year[2].The breast cancer incidence in...Breast cancer is the most common cancer among women worldwide[1].In 2018,2.09 million new breast cancer cases were identified globally,accounting for 11.6% of all cancers in that year[2].The breast cancer incidence in Asian women has also increased rapidly over the past 30 years.展开更多
Background The immunological differences between children and adults with AIDS in China are not well documented. Th1/Th2 cytokines and chemokines are two types of immune factors intimately involved in disease progress...Background The immunological differences between children and adults with AIDS in China are not well documented. Th1/Th2 cytokines and chemokines are two types of immune factors intimately involved in disease progression of HIV-1 infection. This study aimed to identify changes in plasma levels of Th1/Th2 cytokines inerleukin （IL）-18, IL-16, IL-10 and chemokines regulated on activation, normal T cell expressed and secreted （RANTES）, stromal cell-derived factor-1 （SDF-1） and monocyte chemoattractant protein-1 （MCP-1） in HIV-l-infected children and adults in China. Methods Seventy-five children with AIDS and 35 adult AIDS patients were recruited and clinical data were collected. CD4^＋ T lymphocyte counts were measured by flow cytometery and plasma HIV RNA levels were measured by quantitative RT-PCR. Plasma levels of IL-18, IL-10, IL-16, RANTES, MCP-1, SDF-1a and SDF-1β were quantified by enzyme-linked immunosorbent assay. The levels of β2-microglobulin （β2-MG） and soluble Fas （sFas） were measured to validate the level of humoral and cellular immune activation. Results The mean levels of all cytokines in pediatric and adult AIDS patients were significantly higher than in their healthy controls （P 〈0.01）. The mean levels of these cytokines were higher in pediatric patients than in adult patients （P 〈0.05, except for SDF-la and β2-MG）. Some of the cytokine levels in patients younger than 6 years old was higher than in older children and adults with AIDS （IL-10, IL-18, SDF-la, MCP, RANTES and sFas, P 〈0.05）. Levels of IL-18, IL-10, RANTES and β2-MG of pediatric patients increased as the levels of viral load increased （P 〈0.05）. Conclusions Abnormal immune activation can be measured in Chinese pediatric and adult patients with AIDS, and is higher in children than in adult patients. The cytokines levels coincide with disease progression of AIDS, but have no direct relationship with total CD4^＋ T cell count.展开更多
文摘The ELISPOT assay is increasingly used for assessing cellular immune responses in clinical trials of HIV-1 or cancer vaccines. However, to date, data from clinical trials do not consistently show that immune responses are correlated with clinical endpoints. This is due in part to the lack of assay standardization and validation across laboratories and therefore, a quality control panel is required to establish competency and comparability amongst different laboratories. In this study peripheral blood mononuclear cells (PBMCs) from healthy individuals were screened and frozen in liquid nitrogen. The recovery and viability of the PBMCs and the frequencies of interferon (IFN)-γ-secreting cells after CEF peptide pool stimulation were detected after various intervals in seven different laboratories. The recovery and viability did not differ significantly after different intervals. Although the frequencies of IFN (interferon)-γ-secreting cells among thawed PBMCs (peripheral blood mononuclear cells) fluctuated after CEF peptide pool stimulation at different intervals, they were not significantly decreased compared with those among fresh PBMCs. However, the viabilities, recoveries and frequencies of IFN-γ-secreting cells differed significantly among the seven laboratories. Our results indicate that cryopreserved PBMCs could be used as a quality control panel for ELISPOT. However, the procedures for ELISPOT need to be standardized amongst different laboratories.
基金supported by Beijing Center for Diseases Prevention and Control&Center of Preventive Medical Research[Grant#2019-BJYJ-05]。
文摘Breast cancer is the most common cancer among women worldwide[1].In 2018,2.09 million new breast cancer cases were identified globally,accounting for 11.6% of all cancers in that year[2].The breast cancer incidence in Asian women has also increased rapidly over the past 30 years.
文摘Background The immunological differences between children and adults with AIDS in China are not well documented. Th1/Th2 cytokines and chemokines are two types of immune factors intimately involved in disease progression of HIV-1 infection. This study aimed to identify changes in plasma levels of Th1/Th2 cytokines inerleukin （IL）-18, IL-16, IL-10 and chemokines regulated on activation, normal T cell expressed and secreted （RANTES）, stromal cell-derived factor-1 （SDF-1） and monocyte chemoattractant protein-1 （MCP-1） in HIV-l-infected children and adults in China. Methods Seventy-five children with AIDS and 35 adult AIDS patients were recruited and clinical data were collected. CD4^＋ T lymphocyte counts were measured by flow cytometery and plasma HIV RNA levels were measured by quantitative RT-PCR. Plasma levels of IL-18, IL-10, IL-16, RANTES, MCP-1, SDF-1a and SDF-1β were quantified by enzyme-linked immunosorbent assay. The levels of β2-microglobulin （β2-MG） and soluble Fas （sFas） were measured to validate the level of humoral and cellular immune activation. Results The mean levels of all cytokines in pediatric and adult AIDS patients were significantly higher than in their healthy controls （P 〈0.01）. The mean levels of these cytokines were higher in pediatric patients than in adult patients （P 〈0.05, except for SDF-la and β2-MG）. Some of the cytokine levels in patients younger than 6 years old was higher than in older children and adults with AIDS （IL-10, IL-18, SDF-la, MCP, RANTES and sFas, P 〈0.05）. Levels of IL-18, IL-10, RANTES and β2-MG of pediatric patients increased as the levels of viral load increased （P 〈0.05）. Conclusions Abnormal immune activation can be measured in Chinese pediatric and adult patients with AIDS, and is higher in children than in adult patients. The cytokines levels coincide with disease progression of AIDS, but have no direct relationship with total CD4^＋ T cell count.
