Star Research Presentations: Basic Science of Sepsis
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DIFFERENTIAL GENE EXPRESSION IN ADULT
VERSUS PEDIATRIC SEPTIC SHOCK
Michael Bonk1, Nuala Meyer1, Jason Christie2, John Reilly1
Learning Objectives: Sepsis, the dysregulated host response to
infection, is a major cause of ICU mortality. Peripheral blood
gene expression (GEx) may offer novel insights into host
response. Previous studies have combined pediatric and adult
sepsis populations to maximize power, however the degree of
transcriptional regulatory overlap between septic adults and
children is unknown. We hypothesized that there would be both
shared and unique transcriptional regulation between pediatric
and adult septic shock populations.
Methods: We utilized microarray data from two prospective cohort
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studies stored on NCBI’s Gene Expression Omnibus. GSE26440
included 82 whole blood samples from pediatric patients within
24 hours of septic shock onset, and 21 samples from healthy con-
trols. GSE57065 included 28 samples from adult ICU patients 24
hours after developing septic shock and 25 healthy controls. All
samples were analyzed with Affymetrix Human Genome U133
Plus 2.0 Array. Data was processed with R Bioconductor packages
oligo, limma, and biomaRt. Genes were considered differentially
expressed with a fold change > 1.5 and false discovery rate < 0.05.
The differential GEx of the adult and pediatric subjects were com-
pared and examined for enriched biological pathways utilizing the
Database for Annotation, Visualization and Integrated Discovery
(DAVID).
Results: We observed 3882 differentially expressed genes in pe-
diatric patients with septic shock vs controls, and 4103 genes in
adult patients vs controls. 2770 genes were commonly differen-
tially expressed (p < 0.05). Gene ontology analysis of the shared
differentially regulated transcripts identified pathways related
to innate and adaptive immune response. The unique pediatric
differentially regulated transcripts identified pathways related to
inflammation, apoptosis, and B cell signaling. The unique adult
transcripts identified pathways related to cell division and repair.
Conclusions: We identified a distinct set of differentially expressed
genes common to adult and pediatric septic shock, as well as a
smaller set of genes that are uniquely differentially expressed in
each patient population. Our work validates the emphasis on both
innate and adaptive immune response during sepsis and high-
lights potential age-specific mechanisms during sepsis that war-
rant further investigation.
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University of Pennsylvania Perelman School of Medicine, Philadelphia, PA,
1 1
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University of Pennsylvania School of Medicine, Philadelphia, PA
2
www.ccmjournal.org
Copyright © 2018 by the Society of Critical Care Medicine and Wolters Kluwer Health, Inc. All Rights Reserved.
CD4 T CELL MEMORY IMPROVES INNATE
IMMUNITY IN THE CECAL LIGATION AND
PUNCTURE MODEL OF SEPSIS
Matthew Taylor1, Mabel Abraham2, Clifford Deutschman3
Learning Objectives: Relative to laboratory mice kept in specific-
pathogen free facilities (naïve mice), there are significantly more
memory CD4 and CD8 T cells and fewer naïve T cells in mice
exposed to frequent antigenic stimulation (immune educated
mice). This difference should profoundly alter responses to
inflammatory insults as the activation and effector potential of
memory T cells differs from naïve T cells. We hypothesize that
the presence of a robust population of educated T cells will alter
innate immune function following CLP, a well-accepted murine
model of sepsis.
Methods: Administration of a CD3 activating antibody induced
T cell memory (immune education) in C57Bl/6, CD8 knockout
(KO) or CD4 KO mice. This approach increased the ratio of CD4
and CD8 memory to naïve T cells without changing total T cell
number. 35 days after treatment, mice underwent CLP. Animals
were euthanized 18 hrs post-CLP. Bacterial cultures were obtained
from abdominal washings. Cells harvested from liver and spleen
cells were characterized using flow cytometry. Neutrophils and
monocytes from peritoneum, liver and spleen were analyzed ex
vivo or stimulated with LPS and elicited cytokine production was
measured. Cytokine levels in serum were determined using ELISA.
Significance (Student’s t-test, P < 0.05) was determined.
Results: Relative to naïve controls, CLP in Immune educated
mice increased 1) hepatic neutrophil accumulation, 2) stimulated
production of IL1β and TNFα in monocytes and neutrophils, 3)
numbers of IFNγ- and TNFα-producing splenic CD4 T cells and
4) serum IFNγ and IL12. Peritoneal bacterial counts were lower in
educated mice. The same findings were observed following CLP
in educated CD8 KO mice, which have only CD4 T cell memory.
However, these changes were not present in CD4 KOs.
Conclusions: Immune education alters the innate immune re-
sponse to CLP. These changes are specific to CD4, but not CD8,
memory T cells. Decreased peritoneal bacterial counts suggest that
immune education improved immune function. Thus, immune
education may profoundly alter findings in animal models of in-
flammatory disorders. In addition, memory T cells may represent
a novel therapeutic target in sepsis.
Cohen Children’s Medical Center, New Hyde Park, NY, 2Feinstein Institute for
Medical Research, Northwell Health, Manhasset, NY, 3N/A, New Hyde Park, NY
Critical Care Medicine • Volume 47 • Number 1 (Supplement)