\documentclass[10pt]{article}

\usepackage{fullpage}
\usepackage{setspace}
\usepackage{parskip}
\usepackage{titlesec}
\usepackage[section]{placeins}
\usepackage{xcolor}
\usepackage{breakcites}
\usepackage{lineno}
\usepackage{hyphenat}





\PassOptionsToPackage{hyphens}{url}
\usepackage[colorlinks = true,
            linkcolor = blue,
            urlcolor  = blue,
            citecolor = blue,
            anchorcolor = blue]{hyperref}
\usepackage{etoolbox}
\makeatletter
\patchcmd\@combinedblfloats{\box\@outputbox}{\unvbox\@outputbox}{}{%
  \errmessage{\noexpand\@combinedblfloats could not be patched}%
}%
\makeatother


\usepackage{natbib}




\renewenvironment{abstract}
  {{\bfseries\noindent{\abstractname}\par\nobreak}\footnotesize}
  {\bigskip}

\titlespacing{\section}{0pt}{*3}{*1}
\titlespacing{\subsection}{0pt}{*2}{*0.5}
\titlespacing{\subsubsection}{0pt}{*1.5}{0pt}


\usepackage{authblk}


\usepackage{graphicx}
\usepackage[space]{grffile}
\usepackage{latexsym}
\usepackage{textcomp}
\usepackage{longtable}
\usepackage{tabulary}
\usepackage{booktabs,array,multirow}
\usepackage{amsfonts,amsmath,amssymb}
\providecommand\citet{\cite}
\providecommand\citep{\cite}
\providecommand\citealt{\cite}
% You can conditionalize code for latexml or normal latex using this.
\newif\iflatexml\latexmlfalse
\providecommand{\tightlist}{\setlength{\itemsep}{0pt}\setlength{\parskip}{0pt}}%

\AtBeginDocument{\DeclareGraphicsExtensions{.pdf,.PDF,.eps,.EPS,.png,.PNG,.tif,.TIF,.jpg,.JPG,.jpeg,.JPEG}}

\usepackage[utf8]{inputenc}
\usepackage[ngerman,greek,english]{babel}



\usepackage{float}








\begin{document}

\title{Components from spider venom activate macrophages against glioblastoma
cells: new potential adjuvants for anticancer immunotherapy}



\author[1]{Catarina Raposo}%
\author[1]{Jaqueline Munhoz}%
\author[1]{Gabriela Peron}%
\author[1]{Amanda Bonfanti}%
\author[1]{Janine  Oliveira}%
\author[2]{Thomaz A.A. da Rocha‐e‐Silva Rocha‐e‐Silva}%
\author[3]{Rafael Sutti}%
\author[1]{André  Bombeiro}%
\author[1]{Natália Barreto}%
\author[4]{Ghanbar Chalbatani}%
\author[5]{Elahe  Gharagouzloo}%
\author[3]{João  Vitorino-Araujo}%
\author[1]{Liana Verinaud}%
\affil[1]{UNICAMP}%
\affil[2]{Faculdade Israelita de Ciências da Saúde Albert Einstein}%
\affil[3]{Faculdade de Ciências Médicas da Santa Casa de São Paulo}%
\affil[4]{Tehran University of Medical Sciences School of Medicine}%
\affil[5]{Tehran University of Medical Sciences}%


\vspace{-1em}



  
  \date{May 05, 2024}


\begingroup
\let\center\flushleft
\let\endcenter\endflushleft
\maketitle
\endgroup





\selectlanguage{english}
\begin{abstract}
Immunomodulation has been considered an important approach in the
treatment of various types of malignant tumors. However, adaptive immune
cells have received more attention, while the modulation of innate
immune cells is still an underexplored tool. A recent study by our group
demonstrated that the Phoneutria nigriventer spider venom (PnV)
administration increased the circulating NK cells and monocytes and the
infiltration of macrophage in glioblastoma, in addition to decreasing
the tumor size in a preclinical model. The hypothesis that PnV would be
modulating the innate immune system led us to the main objective of the
present study: to elucidate the effects of PnV and its purified
fractions on cultured macrophages. After differentiation from bone
marrow precursors, cells were pre-activated with IFN-\selectlanguage{greek}γ \selectlanguage{english}and treated as
follow: Control (untreated); LPS (1 \selectlanguage{greek}μ\selectlanguage{english}g/mL); PnV (14 \selectlanguage{greek}μ\selectlanguage{english}g/mL);
PnV-fractions F1, F2 and F3 (1 \selectlanguage{greek}μ\selectlanguage{english}g/mL) or PnV-subfractions (1 \selectlanguage{greek}μ\selectlanguage{english}g/mL).
Results showed that PnV and the three fractions activated macrophages.
Further purification generated twenty-three subfractions named Low
Weight (LW-1 to LW-12) and High Weight (HW-1 to HW-11). LW-9 presented
the best immunomodulatory effect. Treated cells were more phagocytic,
migrated more, showed an activated morphological profile and induced an
increased cytotoxic effect of macrophages on tumor cells. However, while
M1-controls (LPS) increased IL-10, TNF-alpha and IL-6 release, PnV,
fractions and subfractions did not alter any cytokine, with the
exception of LW-9 that stimulated IL-10 production. These findings
suggest that molecules present in LW-9 have potential to be used as
immunoadjuvants in the treatment of cancer.%
\end{abstract}%



\sloppy


\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-1/Figure-1}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-2/Figure-2}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-3/Figure-3}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-4/Figure-4}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-5/Figure-5}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-6/Figure-6}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-7/Figure-7}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-8/Figure-8}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure-9/Figure-9}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Graphic-abstract/Graphic-abstract}

\end{center}
\end{figure}

\selectlanguage{english}
\FloatBarrier
\end{document}

