This study examined how glioblastoma tumors influence microglia cells' expression of IL-1B and IL-1RA genes through secreted factors. Results showed that glioma cell conditioned media induced both IL-1B and IL-1RA expression in microglia. Inhibiting CSF-1R or CCR1 signaling in microglia enhanced IL-1B expression while repressing IL-1RA, suggesting glioma signals cause an anti-inflammatory microglia phenotype. Future work will assess how these changes in IL-1 family expression impact microglia polarization.
Proteomics Exploration of Chronic Lymphocytic Leukemia_Crimson PublishersCrimsonpublishersCancer
Chronic Lymphocytic Leukemia (CLL) is an adult heme malignancy characterized by the presence of mature-appearing CD5+ B cells in the blood, bone marrow, and secondary lymphoid organs [1]. In the United States, there will be an estimate of 20,720 new cases and 3,930 deaths according to the American Cancer Society statistics. Symptoms include swollen lymph nodes, frequent infections, and fatigue which negatively impacts the quality of life of people affected [1]. CLL is heterogeneous in its progression and clinical outcomes. Factors that contribute to the heterogeneity include the immunoglobulin heavy chain (IGHV) status and chromosomal aberrations [2,3]. There are two subtypes of CLL: Unmutated(U-CLL) and Mutated CLL(M-CLL). 40% and 60% of patients are diagnosed with unmutated and mutated CLL. U-CLL is characterized by the presence of CLL cells that have less than two percent of their IGHV mutated, whereas M-CLL cells have more than two percent mutated [4]. U-CLL is the more aggressive phenotype [2]. These cells have increased responsiveness to antigens that bind the B cell receptor (BCR) versus M-CLL cells [5]. M-CLL is the more indolent phenotype. Increased BCR signaling results in increased cell survival and proliferation [5].
Dr William Barnes - The I Factor - Inflammation, Immunity, IllnessDr William Barnes
Immunity and Inflammation
Inflammation and Macrophages
Macrophage Pathology
Foam Cell, Viruses, TAMs
The Brune Theory & Nitric Oxide
Macrophages in Disease States
Treatment Strategies
Colds and Influenza
Treatment Strategies
Endothelial Cell Mediated Delay of Blood Brain Barrier Recovery Following Tra...Arthur Stem
TBI is the leading cause of death among young adults and children in the developed world, accounting for over 50,000 deaths per year. [12] TBI results in a sleuth of poor health outcomes, including hemorrhaging, seizures, neural edema, neural inflammation, and cognitive and emotional disabilities. All of these outcomes are a direct result of fundamental degradation of the BBB over a time course post TBI. [1] [12] The BBB is an integral structure that forms around the microvascular of the cerebral cavity. Endothelial cells form the basal membrane through which strictly controlled movement of molecules is observed between the extravascular and intravascular space across this basal membrane. This basal membrane is maintained by endothelial cells, having tight junctions between them to make up the pores through which transport of molecules can occur between the brain and microvasculature. These tight junctions are maintained through cross-talk between the endothelial cells and supporting neurons such as astrocytes and pericytes. [2] A multitude of proteins make up the tight junctions between the endothelial cells, including six main scaffolding structures Claudins 1, 3, and 5, ZO-1, Occludins, and Cadherins. [3] VEGF release following trauma induces endothelial cells to release matrix metalloproteinases (MMPs), in particular MMP9, which can catalyze the N-terminal amino acids that compose the tight junction protein ZO-1. [10] [11] MMP9 when in circulation is also known to activate tumor necrosis factor alpha (TNFɑ) which in turn upregulates transcription of MMP9, creating a positive feedback loop. [11] The management of MMP production is three fold, transcription, proenzyme activation, and substrate inhibition. [11] In our study, it is proenzyme activation via TP that is the focus and how that affects the overall transcription levels of the tight junction proteins within the endothelial cells and astrocytes.
Recent publication showing an interesting approach to identify potential cancer cells and related cell signalling inhibitors in development of anticancer drugs.
1. IL-1B and IL-1RA Expression in Glioma Stimulated Microglia Cells
Sara Maass, Thuy-Dung Ngo, and Victor Suarez
Advisor: Dr. Salvatore Coniglio
New Jersey Center for Science, Technology, & Mathematics
Kean University
Introduction
Glioblastoma is an extremely deadly brain cancer.
Glioblastoma tumors recruit macrophages which in turn
promote cell invasion and immune escape. In the brain
macrophages are called microglia. Macrophages play an
important role in the bodies immune system. There are
two major types of macrophage states: M1 and M2. The
M1 response induces inflammation while M2 reduces
inflammation and promotes tumor spread. Interleukins
also play a role in the immune response. The IL-1 family
induces fever and inflammation. IL-1B tends to be pro-
inflammatory while IL-1RA acts as an antagonist of IL-1B
function and therefore down regulates inflammation. In
this study, we examined the effect of glioma cell
conditioned media on IL-1B family member genes in
microglia using Quantitative Real Time PCR(QRT-PCR)
and immunofluorescence. The role of CSF-1R and CCR1
signaling in this process was also assessed using small
pharmacological inhibitors of these receptors. The
expression of CCR1 on glioma cells and
microglia/macrophages was determined using
immunofluorescence.
Methods
Results
Results cont.
Conclusion
Using Quantitative Real Time PCR (QRT-PCR)
and immunofluorescence we have observed that
conditioned media from the murine glioma cell
line GL261 is able to induce both IL-1B and IL-
1RA expression in microglia. CSF-1R and CCR1
inhibition enhances IL-1B while repressing IL-1RA
expression in glioma stimulated microglia. This
follows the idea that the CSF-1 pathway causes
microglia to become immuno suppressive.
We would like to thank Dr. Salvatore Coniglio
and the fellow research students in our lab for all
of their help and support. Thank you Dr. James
Merritt for providing the CCR1 antagonists. Also,
thank you to New Jersey Center for Science,
Technology, & Mathematics and Kean University
for giving us this research opportunity.
Figure 1: Expression of CCR1 receptor in GL261
glioblastoma cells using immunofluorescence staining.
4 ug/ml aCCR12ary Alone
Figure 2: Expression of CCR1 Receptor in THP1
Macrophages differentiated with PMA express using
immunofluorescence staining.
OVERLAY
Figure 3: Expression of CCR1 in MG Microglia
using immunofluorescence staining.
Phalloidin (Actin) CCR1
Future Works
Acknowledgements
We would like to assess the consequences of the
shift in IL-1 family expression on tumor associated
macrophages/microglia polarity by measuring
changes in M1 vs M2 markers.
QRT-PCR
2ary Alone 4 ug/ml aCCR1
Immunofluorescence
Figure 4: QRT-PCR analysis of JnJ CSF-1R inhibitor in untreated
and glioma conditioned cells. A) IL-1B expression increased
with JnJ. B) IL-1RA expression reduced with JnJ. C) IL-18
expression slightly increased.
0123456
IL-1B
JnJ: - - +
Glioma Cond. Media
A IL-1RA
JnJ: - - +
Glioma Cond. Media
B
0123456
Untx Untx
00.20.40.60.811.2
IL-18
JnJ: - - +
Untx Glioma Cond. Media
C
IL-1B IL-1RA
+ GL261 Cond. Media + GL261 Cond. Media
DMSO DMSO CCR1 inhibitors DMSO DMSO CCR1 inhibitors
Figure 5: QRT-PCR analysis of CCR1 inhibition. CCR1
inhibition enhances IL-1B and represses IL-1RA gene
expression in glioma stimulated microglia.
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