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\begin{document}

\title{Clonality, trafficking, and molecular alterations among Hprt mutant T
lymphocytes isolated from control mice versus mice treated with
N-ethyl-N-nitrosourea}



\author[1]{Stephen Judice}%
\author[2]{Hillary Sussman}%
\author[3]{Dale Walker}%
\author[1]{J. O'Neill}%
\author[1]{Richard Albertini}%
\author[1]{Vernon Walker}%
\affil[1]{University of Vermont}%
\affil[2]{Cold Spring Harbor Laboratory Press}%
\affil[3]{The Burlington HC Research Group, Inc.}%


\vspace{-1em}



  
  \date{May 19, 2024}


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\selectlanguage{english}
\begin{abstract}
Mutations in T lymphocytes (T-cells) are informative quantitative
markers for environmental mutagen exposures, but risk extrapolations
from rodent models to humans also requires understanding how T-cell
development and proliferation kinetics impact mutagenic outcomes. Rodent
studies have shown that patterns in chemical-induced mutations in the
hypoxanthine-guanine phosphoribosyltransferase (Hprt) gene of T-cells
differ between lymphoid organs. The current work was performed to obtain
knowledge of the relationships between maturation events during T-cell
development and changes in chemical-induced mutant frequencies over time
in differing immune compartments of a mouse model. A novel RT?PCR based
method was developed to determine the specific T-cell receptor beta
(Tcrb) gene mRNA expressed in mouse T-cell isolates, enabling sequence
analysis of the PCR product that then identifies the specific
hypervariable CDR3 junctional region of the expressed Tcrb gene for
individual isolates. Characterization of spontaneous Hprt mutant
isolates from the thymus, spleen, and lymph nodes of control mice for
their Tcrb gene expression found evidence of in vivo clonal
amplifications of Hprt mutants and their trafficking between tissues in
the same animal. Concurrent analyses of Hprt mutations and Tcrb gene
rearrangements in different lymphoid tissues of control versus
N-ethyl-N-nitrosourea-exposed mice permitted elucidation of the
localization and timing of mutational events in T-cells, establishing
that mutagenesis occurs primarily in the pre-rearrangement replicative
period in pre-thymic/thymic populations. These findings demonstrate that
chemical-induced mutagenic burden is determined by the combination of
mutagenesis and T-cell clonal expansion, processes with roles in immune
function and the pathogenesis of autoimmune disease and cancer.%
\end{abstract}%



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\textbf{Hosted file}

\verb`Mouse TCR and ENU pprs cmb_final s tables and figures.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

\textbf{Hosted file}

\verb`Mouse TCR and ENU pprs cmb_tables.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

\textbf{Hosted file}

\verb`Fig 2.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

\textbf{Hosted file}

\verb`Fig 3.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

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\verb`Fig 4.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

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\verb`Fig 5.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

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\verb`Fig 6.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

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\verb`Fig 7.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

\textbf{Hosted file}

\verb`Fig 1.doc`  available at \url{https://authorea.com/users/660427/articles/663973-clonality-trafficking-and-molecular-alterations-among-hprt-mutant-t-lymphocytes-isolated-from-control-mice-versus-mice-treated-with-n-ethyl-n-nitrosourea}

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