ADD methods for aim 3

tex/thesis.tex

@@ -84,6 +84,7 @@
 \newacronym{a2b1}{A2B1}{integrin $\upalpha$1$\upbeta$1}
 \newacronym{a2b2}{A2B2}{integrin $\upalpha$1$\upbeta$2}
 \newacronym{til}{TIL}{tumor infiltrating lymphocytes}
+\newacronym{nsg}{NSG}{NOD scid gamma}
 
 %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
 % SI units for uber nerds
@@ -1555,6 +1556,41 @@ provide these benefits.
 
 \section{introduction}
 \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{discussion}