ADD methods for aim 3

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Nathan Dwarshuis 2021-07-25 23:09:00 -04:00
parent 537d942b5e
commit 436eccc733
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@ -84,6 +84,7 @@
\newacronym{a2b1}{A2B1}{integrin $\upalpha$1$\upbeta$1} \newacronym{a2b1}{A2B1}{integrin $\upalpha$1$\upbeta$1}
\newacronym{a2b2}{A2B2}{integrin $\upalpha$1$\upbeta$2} \newacronym{a2b2}{A2B2}{integrin $\upalpha$1$\upbeta$2}
\newacronym{til}{TIL}{tumor infiltrating lymphocytes} \newacronym{til}{TIL}{tumor infiltrating lymphocytes}
\newacronym{nsg}{NSG}{NOD scid gamma}
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% SI units for uber nerds % SI units for uber nerds
@ -1555,6 +1556,41 @@ provide these benefits.
\section{introduction} \section{introduction}
\section{methods} \section{methods}
\subsection{CD19-CAR T cell generation}
% TODO describe how T cells were grown for this aim
% TODO describe how the luciferase cells were generated (eg the kwong lab)
\subsection{\invivo{} therapeutic efficacy in NSG mice model}
% TODO use actual product numbers for mice
All mice in this study were male \gls{nsg} mice from Jackson Laboratories. At
day 0 (-7 day relative to T cell injection), 1e6 firefly luciferase-expressing
\product{Nalm-6 cells}{ATCC}{CRL-3273} suspended in ice-cold PBS were injected
via tail vein into each mouse. At day 7, saline or CAR T cells at the indicated
doses from either bead or DMS-expanded T cell cultures (for 14 days) were
injected into each mouse via tail vein. Tumor burden was quantified
longitudinally via IVIS Spectrum In Vivo Imaging System (Perkin Elmer). Briefly,
200ug/mice luciferin at 15 mg/ml in PBS was injected intraperitoneally under
isoflurane anesthesia into each mouse and waited for at least 10 minutes before
imaging. Mice were anesthetized again and imaged using the IVIS. Mice from each
treatment group/dose were anesthetized, injected, and imaged together, and
exposure time of the IVIS was limited to avoid saturation based on the signal
from the saline group. IVIS images were processed by normalizing them to common
minimum and maximum photon counts and total flux was estimated in terms of
photons/second. Endpoint for each mouse was determined by IACUC euthanasia
criteria (hunched back, paralysis, blindness, lethargy, and weight loss).
Mice were euthanized according to these endpoint criteria using carbon dioxide
asphyxiation.
\subsection{statistics}
For the \invivo{} model, the survival curves were created and statistically
analyzed using GraphPad Prism using the Mantel-Cox test to assess significance
between survival groups.
\section{results} \section{results}
\section{discussion} \section{discussion}