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ENH proofread conclusions

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Nathan Dwarshuis 2021-08-04 20:04:49 -04:00
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@ -4522,7 +4522,7 @@ porcine-derived collagen, which itself is not \gls{gmp}-compliant due to its
non-human animal origins. However, using any other source of collagen should non-human animal origins. However, using any other source of collagen should
work so long as the structure of the microcarriers remains relatively similar work so long as the structure of the microcarriers remains relatively similar
and it has lysine groups that can react with the \gls{snb} to attach \gls{stp} and it has lysine groups that can react with the \gls{snb} to attach \gls{stp}
and \glspl{mab}. Obviously these would need to be tested and verified, but they and \glspl{mab}. Obviously these would need to be tested and verified, but these
should not be insurmountable. Furthermore, the \gls{mab} binding step requires should not be insurmountable. Furthermore, the \gls{mab} binding step requires
\gls{bsa} to prevent adsorption to the non-polar polymer walls of the reaction \gls{bsa} to prevent adsorption to the non-polar polymer walls of the reaction
tubes. A human carrier protein such as \gls{hsa} could be used in its place to tubes. A human carrier protein such as \gls{hsa} could be used in its place to
@ -4531,7 +4531,7 @@ expensive. Alternatively, the use of protein could be replaced altogether by a
non-ionic detergent such as Tween-20 or Tween-80, which are already used for non-ionic detergent such as Tween-20 or Tween-80, which are already used for
commercial \gls{mab} formulations for precisely this purpose\cite{Kerwin2008}. commercial \gls{mab} formulations for precisely this purpose\cite{Kerwin2008}.
Validating the process with Tween would be the best next step to eliminate Validating the process with Tween would be the best next step to eliminate
\gls{bsa} from the process. The \gls{stp} and \glspl{mab} in this process were \gls{bsa} from the process. The \gls{stp} and \glspl{mab} in this work were
not \gls{gmp}-grade; however, they are commonly used in clinical technology such not \gls{gmp}-grade; however, they are commonly used in clinical technology such
as dynabeads and thus the research-grade proteins used here could be easily as dynabeads and thus the research-grade proteins used here could be easily
replaced. The \gls{snb} is a synthetic small molecule and thus does not have any replaced. The \gls{snb} is a synthetic small molecule and thus does not have any
@ -4545,7 +4545,12 @@ as well as it does. Several broad areas remain to be investigated, including the
role of the increased cytokine output (including \il{15} which was explored to role of the increased cytokine output (including \il{15} which was explored to
some extent in this work), the role of cells on the interior of the \gls{dms} some extent in this work), the role of cells on the interior of the \gls{dms}
relative to those outside the \gls{dms}, and the role of the physical surface relative to those outside the \gls{dms}, and the role of the physical surface
properties of the \gls{dms} (including the morphology and the stiffness). properties of the \gls{dms} (including the morphology and the stiffness). One
plausible hypothesis to be tested is that the bumpy microcarrier surface is more
like that of an \gls{apc}, which enhances immunological synapse formation and
thus activation. Another related hypothesis is that the signal strength is
lower than the beads, which leads to increased proliferation, less exhaustion,
and by extension more memory.
\subsection{additional ligands and signals on the DMSs} \subsection{additional ligands and signals on the DMSs}
@ -4553,22 +4558,23 @@ In this work we only explored the use of \acd{3} and \acd{28} \glspl{mab} coated
on the surface of the \gls{dms}. The chemistry used for the \gls{dms} is very on the surface of the \gls{dms}. The chemistry used for the \gls{dms} is very
general, and any molecule or protein that could be engineered with a biotin general, and any molecule or protein that could be engineered with a biotin
ligand could be attached without any further modification. There are many other ligand could be attached without any further modification. There are many other
ligands that could have profound effects on the expansion and quality of T cells ligands (in addition to integrin-binding domains and \il{15} complexes as
which may be utilized. The simplest next step is to simply vary the ratio of described at the end of \cref{aim2b}) that could have profound effects on the
\acd{3} and \acd{28} signal. Another obvious example is to attach expansion and quality of T cells which may be utilized. The simplest next step
\il{15}/\il{15R$\upalpha$} complexes to the surface to mimic \textit{trans} is to simply vary the ratio of \acd{3} and \acd{28} signal. Another obvious
presentation from other cell types\cite{Stonier2010}. Other adhesion ligands or example is to attach \il{15}/\il{15R$\upalpha$} complexes to the surface to
peptides such as GFOGER could be used to stimulate T cells and provide more mimic \textit{trans} presentation from other cell types\cite{Stonier2010}. Other
motility on the \glspl{dms}\cite{Stephan2014}. Finally, viral delivery systems adhesion ligands or peptides such as GFOGER could be used to stimulate T cells
could theoretically be attached to the \gls{dms}, greatly simplifying the and provide more motility on the \glspl{dms}\cite{Stephan2014}. Finally, viral
transduction step. delivery systems could theoretically be attached to the \gls{dms}, greatly
simplifying the transduction step.
\subsection{assessing performance using unhealthy donors} \subsection{assessing performance using unhealthy donors}
All the work presented in this dissertation was performed using healthy donors. All the work presented in this dissertation was performed using healthy donors.
This was mostly due to the fact that it was much easier to obtain healthy donor This was mostly due to the fact that it was much easier to obtain healthy donor
cells and was much easier to control. However, it is indisputable that the most cells and was much easier to control. However, it is indisputable that the most
relevant test cases of the \gls{dms} will be for unhealthy patient T cells, at relevant test cases of the \glspl{dms} will be for unhealthy patient T cells, at
least in the case of autologous therapies. In particular, it will be interesting least in the case of autologous therapies. In particular, it will be interesting
to see how the \gls{dms} performs when assessed head-to-head with bead-based to see how the \gls{dms} performs when assessed head-to-head with bead-based
expansion technology given that even in healthy donors, we observed the expansion technology given that even in healthy donors, we observed the