CD4+ T lymphocytes were infected for three days with an HIV virus at an MOI of 0.01 using ViroMag Magnetofection reagent.
This paper shows the efficiency of ViroMag from OZ Biosciences to enhance infection at low MOI in primary CD4+ T lymphocytes.article reference: AIDS. 2014 Apr 21. [Epub ahead of print]
Natural killer cell and T-cell subset distributions and activation influence susceptibility to perinatal HIV-1 infection.
Gasper MA, Kunwar P, Itaya G, Lejarcegui N, Bosire R, Maleche-Obimbo E, Wamalwa D, Slyker J, Overbaugh J, Horton H, Sodora DL, John-Stewart G, Lohman-Payne B.
Abstract
OBJECTIVE: To determine neonatal immunologic factors that correlate with mother-to-child-transmission of HIV-1.
DESIGN: This
case-control study compared cord blood natural killer (NK) and T-cell
populations of HIV-1 exposed infants who subsequently acquired infection
by 1 month (cases) to those who remained uninfected by 1 year of life
(controls). Control specimens were selected by proportional match on
maternal viral load.
METHODS: Cryopreserved cord blood mononuclear cells (CBMCs) were thawed and stained for multiparameter flow cytometry to detect NK and T-cell subsets and activation status. CBMCs were also used in a viral suppression assay to evaluate NK cell inhibition of HIV-1 replication in autologous CD4 T cells.
RESULTS: Cord blood from cases contained a skewed NK cell repertoire characterized by an increased proportion of CD16CD56 NK cells. In addition, cases displayed less-activated CD16CD56 NK cells and CD8 T cells, based on HLA-DRCD38 costaining. NK cell suppression of HIV-1 replication ex vivo correlated with the proportion of acutely activated CD68CD16CD56 NK cells. Finally, we detected a higher proportion of CD27CD45RA effector memory CD4 and CD8 T cells in cord blood from cases compared with controls.
CONCLUSION: When controlled for maternal viral load, cord blood from infants who acquired HIV-1 had a higher proportion of CD16CD56 NK cells, lower NK cell activation and higher levels of mature T cells (potential HIV-1 targets) than control infants who remained uninfected. Our data provide evidence that infant HIV-1 acquisition may be influenced by both innate and adaptive immune cell phenotypes and activation status.
METHODS: Cryopreserved cord blood mononuclear cells (CBMCs) were thawed and stained for multiparameter flow cytometry to detect NK and T-cell subsets and activation status. CBMCs were also used in a viral suppression assay to evaluate NK cell inhibition of HIV-1 replication in autologous CD4 T cells.
RESULTS: Cord blood from cases contained a skewed NK cell repertoire characterized by an increased proportion of CD16CD56 NK cells. In addition, cases displayed less-activated CD16CD56 NK cells and CD8 T cells, based on HLA-DRCD38 costaining. NK cell suppression of HIV-1 replication ex vivo correlated with the proportion of acutely activated CD68CD16CD56 NK cells. Finally, we detected a higher proportion of CD27CD45RA effector memory CD4 and CD8 T cells in cord blood from cases compared with controls.
CONCLUSION: When controlled for maternal viral load, cord blood from infants who acquired HIV-1 had a higher proportion of CD16CD56 NK cells, lower NK cell activation and higher levels of mature T cells (potential HIV-1 targets) than control infants who remained uninfected. Our data provide evidence that infant HIV-1 acquisition may be influenced by both innate and adaptive immune cell phenotypes and activation status.
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