ENH tighten up the mouse discussion

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Nathan Dwarshuis 2021-08-03 13:27:50 -04:00
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@ -2544,6 +2544,45 @@ CONCLUSIONS: We developed a simplified, semi-closed system for the initial selec
revised = {2008-07-28}, revised = {2008-07-28},
} }
@Article{Lozza2008,
author = {Laura Lozza and Laura Rivino and Greta Guarda and David Jarrossay and Andrea Rinaldi and Francesco Bertoni and Federica Sallusto and Antonio Lanzavecchia and Jens Geginat},
journal = {European Journal of Immunology},
title = {The strength of T cell stimulation determines {IL}-7 responsiveness, secondary expansion, and lineage commitment of primed human {CD}4+{IL}-7Rhi T cells},
year = {2008},
month = {jan},
number = {1},
pages = {30--39},
volume = {38},
doi = {10.1002/eji.200737852},
publisher = {Wiley},
}
@Article{Lanzavecchia2005,
author = {Antonio Lanzavecchia and Federica Sallusto},
journal = {Current Opinion in Immunology},
title = {Understanding the generation and function of memory T cell subsets},
year = {2005},
month = {jun},
number = {3},
pages = {326--332},
volume = {17},
doi = {10.1016/j.coi.2005.04.010},
publisher = {Elsevier {BV}},
}
@Article{Corse2011,
author = {Emily Corse and Rachel A. Gottschalk and James P. Allison},
journal = {The Journal of Immunology},
title = {Strength of {TCR}{\textendash}Peptide/{MHC} Interactions and In Vivo T Cell Responses},
year = {2011},
month = {apr},
number = {9},
pages = {5039--5045},
volume = {186},
doi = {10.4049/jimmunol.1003650},
publisher = {The American Association of Immunologists},
}
@Comment{jabref-meta: databaseType:bibtex;} @Comment{jabref-meta: databaseType:bibtex;}
@Comment{jabref-meta: grouping: @Comment{jabref-meta: grouping:

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@ -2465,6 +2465,9 @@ however, to our knowledge this is the first system that specifically drives
naïve/memory and CD4+ T cell formation in a scalable, potentially naïve/memory and CD4+ T cell formation in a scalable, potentially
bioreactor-compatible manufacturing process. bioreactor-compatible manufacturing process.
% DISCUSSION assuage krish by showing that in the isotype control fig that IL2
% doesn't activation T cells: https://www.jimmunol.org/content/jimmunol/191/12/5822.full.pdf
Memory and naïve T cells have been shown to be important clinically. Compared to Memory and naïve T cells have been shown to be important clinically. Compared to
effectors, they have a higher proliferative capacity and are able to engraft for effectors, they have a higher proliferative capacity and are able to engraft for
months; thus they are able to provide long-term immunity with smaller months; thus they are able to provide long-term immunity with smaller
@ -4148,8 +4151,6 @@ other groups in regard to the final tumor burden.
\section{discussion} \section{discussion}
% FIGURE add CD45RA to this to rule out one of the alternative possibilities
% explaining this data
\begin{figure*}[ht!] \begin{figure*}[ht!]
\begingroup \begingroup
@ -4163,7 +4164,8 @@ other groups in regard to the final tumor burden.
\subcap{fig:mouse_summary_1}{the first mouse experiment} and \subcap{fig:mouse_summary_1}{the first mouse experiment} and
\subcap{fig:mouse_summary_2}{the second mouse experiment}. The y axis \subcap{fig:mouse_summary_2}{the second mouse experiment}. The y axis
maximum is set to the maximum number of cells injected between both maximum is set to the maximum number of cells injected between both
experiments (\num{1.25e6}). experiments (\num{1.25e6}). Note that the \gls{car} was quantified using a
separate panel than the rest of the markers.
} }
\label{fig:mouse_summary} \label{fig:mouse_summary}
\end{figure*} \end{figure*}
@ -4180,32 +4182,32 @@ tumor burden was higher than DMS groups across all the total T cell doses tested
here. More interestingly, when only CAR-expressing T cell doses between bead and here. More interestingly, when only CAR-expressing T cell doses between bead and
DMS groups were compared, DMS group had significantly higher survival effects DMS groups were compared, DMS group had significantly higher survival effects
over similar or higher CAR expression T cell doses from bead group. All these over similar or higher CAR expression T cell doses from bead group. All these
results suggest that the higher proportion of memory T cells in DMS groups results suggest that the T cells in DMS groups (compared to bead group) resulted
(compared to bead group) resulted in highly effective CAR-T cells that can in highly effective CAR-T cells that can efficiently kill tumor cells.
efficiently kill tumor cells as recently reported in
literature\cite{Fraietta2018, Sommermeyer2015}.
% DISCUSSION cite a bunch of data saying memory and CD4 T cells are better in
% mice
When comparing the total number of T cells of different phenotypes, we observed When comparing the total number of T cells of different phenotypes, we observed
that when comparing low-dose \gls{dms} group to the mid- bead groups (which had that when comparing low-dose \gls{dms} group to the mid- bead groups (which had
similar numbers of \gls{car} T cells), the number of \ptmem{} T cells injected similar numbers of \gls{car} T cells), the number of \ptmem{} (both with and
was much lower in the \gls{dms} group (\cref{fig:mouse_summary_1}). This could without CD45RA) T cells injected was much lower in the \gls{dms} group
mean several things. First, the \ptmem{} phenotype may have nothing to do with (\cref{fig:mouse_summary_1}). This could mean several things. First, the
the results seen here, at least in this model. Second, the distribution of \ptmem{} phenotype may have nothing to do with the results seen here, at least
\gls{car} T cells across different subtypes of T cells was different between the in this model. While this may have been the case in our hands, this would
\gls{dms} and bead groups (with possibly higher correlation of \gls{car} contradict previous evidence suggesting that \gls{tn} and \gls{tcm} cells work
expression and the \ptmem{} phenotype). Third, the \ptmem{} phenotype may not be better in almost the same model (the only difference being Raji cells in place
precise enough, and the functional `memory' phenotype is a subset of \ptmem{} of Nalm-6 cells, both of which express CD19)\cite{Sommermeyer2015}. Second, the
which may be higher in the \gls{dms} group and explains the discrepancy between distribution of \gls{car} T cells across different subtypes of T cells was
the two methods. different between the \gls{dms} and bead groups (with possibly higher
correlation of \gls{car} expression and the \ptmem{} phenotype). It is hard to
assess this without strong assumptions as the \gls{car} was quantified using a
separate flow panel relative to the other markers.
% DISCUSSION cite why CD4 or CD8 matters in this model
We can also make a similar observation for the number of \pth{} T cells injected We can also make a similar observation for the number of \pth{} T cells injected
(\cref{fig:mouse_summary_1}). In this case, either the \pth{} phenotype doesn't (\cref{fig:mouse_summary_1}). In this case, either the \pth{} phenotype doesn't
matter in this model (or the \ptk{} population matters much more), or the matter in this model (or the \ptk{} population matters much more), or the
distribution of \gls{car} is different between CD4 and CD8 T cells in a manner distribution of \gls{car} is different between CD4 and CD8 T cells in a manner
that favors the \gls{dms} group. that favors the \gls{dms} group. While in a glioblastoma model and not a B-cell
\gls{all} model, previous groups have shown that \pthp{} T cells are important
for response\cite{Wang2018}.
When testing \gls{car} T cells at earlier timepoints relative to day 14 as used When testing \gls{car} T cells at earlier timepoints relative to day 14 as used
in the first \invivo{} experiment, we noted that none of the \gls{car} in the first \invivo{} experiment, we noted that none of the \gls{car}
@ -4221,10 +4223,11 @@ to perform better at day 6 as it held off the tumor longer, and also slowed the
tumor progression relative to the bead group at day 14 tumor progression relative to the bead group at day 14
(\cref{fig:mouse_timecourse_ivis_plots}). (\cref{fig:mouse_timecourse_ivis_plots}).
Taken together, these data suggest that on average, the \gls{dms} platform Taken together, these data suggest that the \gls{dms} platform produces T cells
produces T cells that have an advantage \invivo{} over beads. While we may not that have an advantage \invivo{} over beads. While we may not know the exact
know the exact mechanism, our data suggests that the responses are mechanism, our data suggests that the responses are unsurprisingly influenced by
unsurprisingly influenced by the \ptcarp{} of the final product. the \ptcarp{} of the final product. Followup experiments would need to be
performed to determine the precise phenotype responsible for these responses.
\chapter{conclusions and future work}\label{conclusions} \chapter{conclusions and future work}\label{conclusions}
@ -4287,13 +4290,13 @@ to control and optimize the \gls{dms} system. We determined that altering the
\gls{dms} concentration temporally has profound effects on the phenotype and \gls{dms} concentration temporally has profound effects on the phenotype and
expansion rate. This agrees with other data we obtained in \cref{aim2a} and with expansion rate. This agrees with other data we obtained in \cref{aim2a} and with
what others have generally reported about signal strength and T cell what others have generally reported about signal strength and T cell
differentiation\cite{Gattinoni2012}. We did not find any mechanistic differentiation\cite{Gattinoni2012, Lozza2008, Lanzavecchia2005, Corse2011}. We
relationship between either integrin signaling or \gls{il15} signaling. In the did not find any mechanistic relationship between either integrin signaling or
case of the former, it may be more likely that the \glspl{dms} surfaces are \gls{il15} signaling. In the case of the former, it may be more likely that the
saturated to the point of sterically hindering any integrin interactions with \glspl{dms} surfaces are saturated to the point of sterically hindering any
the collagen surface. In the case of \gls{il15} more experiments likely need to integrin interactions with the collagen surface. In the case of \gls{il15} more
be done in order to plausibly rule out this mechanism and/or determine if it is experiments likely need to be done in order to plausibly rule out this mechanism
involved at all. and/or determine if it is involved at all.
% TODO make this tighter and cite paper showing that this makes at least some % TODO make this tighter and cite paper showing that this makes at least some
% sense % sense