ENH split Cd19 car fig
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@ -1937,7 +1937,6 @@ appeared that the \gls{mab} binding was quadratically related to biotin binding
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critical to controlling the amount and \glspl{mab} and thus the amount of signal
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critical to controlling the amount and \glspl{mab} and thus the amount of signal
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the T cells receive downstream.
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the T cells receive downstream.
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% FIGURE combine this DOE figure into one interaction plot
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\begin{figure*}[ht!]
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\begin{figure*}[ht!]
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\begingroup
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\begingroup
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@ -2367,7 +2366,7 @@ generate \gls{car} T cells\cite{Tumaini2013, Lamers2014}. We added a
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\SI{24}{\hour} transduction step on day 1 of the \SI{14}{\day} expansion to
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\SI{24}{\hour} transduction step on day 1 of the \SI{14}{\day} expansion to
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insert an anti-CD19 \gls{car}\cite{Milone2009} and subsequently measured the
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insert an anti-CD19 \gls{car}\cite{Milone2009} and subsequently measured the
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surface expression of the \gls{car} on day 14
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surface expression of the \gls{car} on day 14
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(\cref{fig:car_production_flow_pl,fig:car_production_endpoint_pl}). We noted
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(\cref{fig:car_cd19_flow,fig:car_cd19_endpoint}). We noted
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that there was robust \gls{car} expression in over \SI{25}{\percent} of expanded
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that there was robust \gls{car} expression in over \SI{25}{\percent} of expanded
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T cells, and there was no observable difference in \gls{car} expression between
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T cells, and there was no observable difference in \gls{car} expression between
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beads and \glspl{dms}.
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beads and \glspl{dms}.
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@ -2380,7 +2379,7 @@ appearance of CD107a on CD8+ T cells. CD107a is found on the inner-surface of
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cytotoxic granules and will emerge on the surface after cytotoxic T cells are
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cytotoxic granules and will emerge on the surface after cytotoxic T cells are
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activated and degranulate. Indeed, we observed degranulation in T cells expanded
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activated and degranulate. Indeed, we observed degranulation in T cells expanded
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with both beads and \glspl{dms}, although not to an observably different degree
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with both beads and \glspl{dms}, although not to an observably different degree
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(\cref{fig:car_production_flow_degran,fig:car_production_endpoint_degran}).
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(\cref{fig:car_degran_flow,fig:car_degran_endpoint}).
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Taken together, these results indicated that the \glspl{dms} provide similar
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Taken together, these results indicated that the \glspl{dms} provide similar
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transduction efficiency compared to beads.
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transduction efficiency compared to beads.
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@ -2389,7 +2388,7 @@ for CCL21; since \glspl{dms} produced a larger percentage of naïve and memory T
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cells (which have CCR7, the receptor for CCL21) we would expect higher migration
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cells (which have CCR7, the receptor for CCL21) we would expect higher migration
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in \gls{dms}-expanded cells vs.\ their bead counterparts. Indeed, we noted a
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in \gls{dms}-expanded cells vs.\ their bead counterparts. Indeed, we noted a
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significantly higher migration percentage for T cells grown using \glspl{dms}
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significantly higher migration percentage for T cells grown using \glspl{dms}
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versus beads (\cref{fig:car_production_migration}). Interestingly, there also
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versus beads (\cref{fig:car_degran_migration}). Interestingly, there also
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appeared to be a decrease in CCL21 migration between transduced and untransduced
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appeared to be a decrease in CCL21 migration between transduced and untransduced
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T cells expanded using beads, but this interaction effect was only weakly
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T cells expanded using beads, but this interaction effect was only weakly
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significant (p = 0.068). No such effect was seen for \gls{dms}-expanded T cells,
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significant (p = 0.068). No such effect was seen for \gls{dms}-expanded T cells,
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@ -2399,25 +2398,38 @@ showing that migration was likely independent of \gls{car} transduction.
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\begin{figure*}[ht!]
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\begin{figure*}[ht!]
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\begingroup
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\begingroup
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\includegraphics{../figures/car_production.png}
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\includegraphics{../figures/car_cd19.png}
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\phantomsubcaption\label{fig:car_production_flow_pl}
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\phantomsubcaption\label{fig:car_cd19_flow}
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\phantomsubcaption\label{fig:car_production_endpoint_pl}
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\phantomsubcaption\label{fig:car_cd19_endpoint}
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\phantomsubcaption\label{fig:car_production_flow_degran}
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\phantomsubcaption\label{fig:car_production_endpoint_degran}
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\endgroup
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\phantomsubcaption\label{fig:car_production_migration}
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\caption[\glspl{dms} lead to efficient CD19 transduction]
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{\glspl{dms} lead to efficient CD19 transduction.
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\subcap{fig:car_cd19_flow}{Representative flow cytometry plot for
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transduced or untransduced T cells stained with \gls{ptnl}.}
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\subcap{fig:car_cd19_endpoint}{Endpoint plots with \gls{anova} for
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transduced or untransduced T cells stained with \gls{ptnl}.}
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All data is from T cells expanded for \SI{14}{\day}.
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}
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\label{fig:car_production}
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\end{figure*}
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\begin{figure*}[ht!]
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\begingroup
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\includegraphics{../figures/car_degranulation.png}
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\phantomsubcaption\label{fig:car_degran_flow}
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\phantomsubcaption\label{fig:car_degran_endpoint}
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\phantomsubcaption\label{fig:car_degran_migration}
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\endgroup
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\endgroup
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\caption[\glspl{dms} produce functional \gls{car} T cells]
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\caption[\glspl{dms} produce functional \gls{car} T cells]
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{\glspl{dms} produce functional \gls{car} T cells.
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{\glspl{dms} produce functional \gls{car} T cells.
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\subcap{fig:car_production_flow_pl}{Representative flow cytometry plot for
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\subcap{fig:car_degran_flow}{Representative flow plot for
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transduced or untransduced T cells stained with \gls{ptnl}.}
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\subcap{fig:car_production_endpoint_pl}{Endpoint plots with \gls{anova} for
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transduced or untransduced T cells stained with \gls{ptnl}.}
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\subcap{fig:car_production_flow_degran}{Representative flow plot for
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degenerating T cells.}
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degenerating T cells.}
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\subcap{fig:car_production_endpoint_degran}{Endpoint plots for transduced or
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\subcap{fig:car_degran_endpoint}{Endpoint plots for transduced or
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untransduced T cells stained with \cd{107a} for the degranulation assay.}
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untransduced T cells stained with \cd{107a} for the degranulation assay.}
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\subcap{fig:car_production_migration}{Endpoint plot for transmigration assay
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\subcap{fig:car_degran_migration}{Endpoint plot for transmigration assay
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with \gls{anova}.} All data is from T cells expanded for \SI{14}{\day}.
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with \gls{anova}.} All data is from T cells expanded for \SI{14}{\day}.
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}
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}
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\label{fig:car_production}
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\label{fig:car_production}
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