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10.3389/fimmu.2021.666991 http://scihub22266oqcxt.onion/10.3389/fimmu.2021.666991 34276657!8278574!34276657     free     free     free Deprecated: Implicit conversion from float 225.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 225.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Front+Immunol 2021 ; 12 (ä): 666991 Nephropedia Template TP {{tp|p=34276657|t=2021. Reliable Estimation of CD8 T Cell Inhibition of In Vitro HIV-1 Replication |pdf=|usr=}}{{34276657}} gab.com Text Reliable Estimation of CD8 T Cell Inhibition of In Vitro HIV-1 Replication JO: Front Immunol http://www.kidney.de/pget.php?&tw=1&pmid=34276657 #FOAvip The HIV-1 viral inhibition assay (VIA) measures CD8 T cell-mediated inhibition of HIV replication in CD4 T cells and is increasingly used for clinical testing of HIV vaccines and immunotherapies. The VIA has multiple sources of variability arising from in vitro HIV infection and co-culture of two T cell populations. Here, we describe multiple modifications to a 7-day VIA protocol, the most impactful being the introduction of independent replicate cultures for both HIV infected-CD4 (HIV-CD4) and HIV-CD4:CD8 T cell cultures. Virus inhibition was quantified using a ratio of weighted averages of p24+ cells in replicate cultures and the corresponding 95% confidence interval. An Excel template is provided to facilitate calculations. Virus inhibition was higher in people living with HIV suppressed on antiretroviral therapy (n=14, mean: 40.0%, median: 43.8%, range: 8.2 to 73.3%; p < 0.0001, two-tailed, exact Mann-Whitney test) compared to HIV-seronegative donors (n = 21, mean: -13.7%, median: -14.4%, range: -49.9 to 20.9%) and was stable over time (n = 6, mean %COV 9.4%, range 0.9 to 17.3%). Cross-sectional data were used to define 8% inhibition as the threshold to confidently detect specific CD8 T cell activity and determine the minimum number of culture replicates and p24+ cells needed to have 90% statistical power to detect this threshold. Last, we note that, in HIV seronegative donors, the addition of CD8 T cells to HIV infected CD4 T cells consistently increased HIV replication, though the level of increase varied markedly between donors. This co-culture effect may contribute to the weak correlations observed between CD8 T cell VIA and other measures of HIV-specific CD8 T cell function. Twit Text FOAVip Reliable Estimation of CD8 T Cell Inhibition of In Vitro HIV-1 Replication ????Front Immunol http://www.kidney.de/pget.php?&tw=1&pmid=34276657 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=34276657 #FOAvip English Wikipedia <ref>{{Cite pmid|34276657}}</ref> Reliable Estimation of CD8 T Cell Inhibition of In Vitro HIV-1 Replication #MMPMID34276657 Xu Y; Weideman AM; Abad-Fernandez M; Mollan KR; Kallon S; Samir S; Warren JA; Clutton G; Roan NR; Adimora AA; Archin N; Kuruc J; Gay C; Hudgens MG; Goonetilleke N Front Immunol 2021[]; 12 (ä): 666991 PMID34276657show ga The HIV-1 viral inhibition assay (VIA) measures CD8 T cell-mediated inhibition of HIV replication in CD4 T cells and is increasingly used for clinical testing of HIV vaccines and immunotherapies. The VIA has multiple sources of variability arising from in vitro HIV infection and co-culture of two T cell populations. Here, we describe multiple modifications to a 7-day VIA protocol, the most impactful being the introduction of independent replicate cultures for both HIV infected-CD4 (HIV-CD4) and HIV-CD4:CD8 T cell cultures. Virus inhibition was quantified using a ratio of weighted averages of p24+ cells in replicate cultures and the corresponding 95% confidence interval. An Excel template is provided to facilitate calculations. Virus inhibition was higher in people living with HIV suppressed on antiretroviral therapy (n=14, mean: 40.0%, median: 43.8%, range: 8.2 to 73.3%; p < 0.0001, two-tailed, exact Mann-Whitney test) compared to HIV-seronegative donors (n = 21, mean: -13.7%, median: -14.4%, range: -49.9 to 20.9%) and was stable over time (n = 6, mean %COV 9.4%, range 0.9 to 17.3%). Cross-sectional data were used to define 8% inhibition as the threshold to confidently detect specific CD8 T cell activity and determine the minimum number of culture replicates and p24+ cells needed to have 90% statistical power to detect this threshold. Last, we note that, in HIV seronegative donors, the addition of CD8 T cells to HIV infected CD4 T cells consistently increased HIV replication, though the level of increase varied markedly between donors. This co-culture effect may contribute to the weak correlations observed between CD8 T cell VIA and other measures of HIV-specific CD8 T cell function. |Antiviral Agents/therapeutic use[MESH] |CD4-Positive T-Lymphocytes/immunology[MESH] |CD8-Positive T-Lymphocytes/*immunology[MESH] |Case-Control Studies[MESH] |Cells, Cultured[MESH] |Coculture Techniques[MESH] |Cross-Sectional Studies[MESH] |HIV Core Protein p24/immunology[MESH] |HIV Seropositivity/blood/drug therapy/*immunology/virology[MESH] |HIV-1/*immunology[MESH] |Host Microbial Interactions/*immunology[MESH] |Humans[MESH] |Treatment Outcome[MESH]Reliable Estimation of CD8 T Cell Inhibition of In Vitro HIV-1 Replica(epmc).pdf DeepDyve Pubget Overpricing