Pathogen Cell-to-Cell

Variability Drives Heterogeneity

in Host Immune Responses

Avraham, R., Haseley, N., Brown, D., Penaranda, C., Jijon, Humberto B.,
Trombetta, John J., Satija, R., Shalek, Alex K., Xavier, Ramnik J., Regev, A., &
Hung, Deborah T. (2015). Pathogen Cell-to-Cell Variability Drives
Heterogeneity in Host Immune Responses. Cell, 162(6), 1309–1321.
https://doi.org/10.1016/j.cell.2015.08.027 [1]
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Context and framework
Is the variability in pathogen cell to cell leads to different heterogeneity
in responses of host cell?

• Previous research done on the field

• Interaction between host and Field of research: microbiology, immunology,
pathogen leads to different chemistry and biology interference
outcome of the pathogen’s
infection [2]
• S. Typhimurium is the most
studied pathogen [3]
• Different response to PAMPs
shows difference response to
infefction [4] Iwasaki, Medzhitov (6)
• Live bacteria provides a broader
range of interactions [5]

BIG QUESTION

How and what interactions

that get changed affect the
out comes of an infection?
METHODS AND TECHNOLOGY
GFP
Gene set rich analysis (GSEA)
• Two type of dye, GFP and pHrodo
• GFP is a type of dye that binds to protein • This technology helps testing the
• pHrodo is a red dye that binds to the enrichment of gene variable, this
bacterial cell wall that increases technology helps identifying the pathway
fluorescence activity while in the that takes parts with infected macrophages
macrophage lysosome. that varies after host cell’s exposure to
• If bacteria is death, it only shows red pathogens
colour
• If bacteria is alive, it has a mixture of red
and green.
• This method is used to determine
whether the host cell dies or not after
the experiment

https://www.gsea msigdb.org/gsea/index.jsp [7]

 Hypothesis 1
LPS modification by PhoPQ determines the
induction of type I IFN responses

Fig 5A, a heat map shows Fig 5B, a Z score

the responses of type I IFN
responses of LPS from
PhoPc and LPS from WT
strain.
tests illustrates how
Fig 5C +D, graphic illustration of
type I IFN responses
how cell responds to LPS modified
toward modified LPS
from PhoPQ
by PhoPQ strain

Data shows that when the LPS is undergone modification by LPS, the cell have a higher level of
expression.
Hypothesis 2
Component of TLr4/LPS signaling participates
in different activation of gene cluster III

Fig S3A, heat map

shows how cluster I, II
and III behave after 4
hours of exposing to S.
Typhimurium

In conclusion, gene cluster I and II

is activated by Myd88 and Trif but
for cluster III, it is regulated only
Fig S3C, a by Trif *Myd88 and Trif are
illustration the adaptor proteins [8]
gene expression
of each cluster
 EXPERIMENTAL DESIGN
Cons
• Did not indicate any types of
replicates when doing the
experiment
• No real “methods” section,
have a experiment
procedure but still vague
• Discussion barely refers to
the table and figures
Pros
• Have a proper index at the
end that explaining terms
and how they come get
the resources for their
work
• Have proper Z score test to
back up the alternate
hypothesis
• Use Heat map and density
plot so it is easier to
visualize the data

• Summary
• The experiment aims to study how the host cell-pathogen encounters can
have heterogerneity.
• The data from the experiment shows that the LPS modified by PhoPQ strain
leads to the heterogenous type I IFN responses. And this modification only
occurs in a set sub of bacteria that infects during infection
Future work, importance and model
• Importance: The study has showcased the relationship between pathogen
infection and host cell. It uses specifically S. Typhimurium and study how it
interacts with macrophage derived from bone marrow of mouse. It also highlight
the ability of immune cell responds to difference pathogen
• Next step : understanding bacterial heterogeneity so it can be used to drive to
different responses in host cell and used to determine the outcome of an
infection
• Experiment 1: Experiment 2
• Model: compare the effect of LPS modified by LpxC. (9) • Model : study the effects of how
Negative control: LPS are not modified by LpxC modified LPS on lymphocyte
Positive control: LPS are modified by LpxC
responses (10)
• representativecontrol: responses of
Experiment
lymphocyte
2 test group, one has LPS modified by LpxC and one does
not. Biological replicate. Measure the expression level of • Experiment : let the PhoPQ
cell after the exposure to S. Typhimurium modifies the LPS.
• Measure how the
• results of the modification affect
the response of lymphocytes in
host cell