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Our story

Riley Ennis is a sophomore at Dartmouth College and founder of Immudicon. The company began from an idea Riley had to model the horseshoe crab immune system in order to teach the body to recognize and destroy the cancer.

Immudicon In the News

WE ARE TIRED OF THE TREATMENT KILLING THE PATIENT

Treat

DIAGNOSE
Pipeline overview

Treat (MacroVax)

Macrovax
Mannose Binding Lectin

CpG and GM-CSF Coated Antigens

PLGA

OR

Mannose Binding Lectin Fusion Moiety

Targets Tumor Microenvironment

Depresses IL10 Associated with Immune Inhibition

Key Advantages

Activates Both Th1 and Th2 Immunity via Compliment Pathways

Mannose Binding Lectin Serves as the Basis of the Human Immune System

Current Research:
Research has been conducted for three years at Georgetown (Dr. Anita Sidhu and Dr. Chris Albanese), Childrens National Medical Center (Dr. Anthony Sandler and Dr. Larry Mahan), and the Dartmouth Hitchcock Medical Center (Dr. Barjor Gimi). The next couple of slides outline some of the data collected.

MacroVax is Phagocytosed by Both Mouse and Human APCs


Macrophages Dendritic Cells

Epifluorescent image of MacroVax- treated dendritic cells and macrophages. The vaccine complex is shown here to bind to and be phagocytosed by these antigen presenting cells. Actin (Red), Nuclei (Blue), Vaccine Complex (Green). 1200X magnification.

Confocal Image of Human Dendritic Cell Proves MacroVax Enters APCs Directly

MERGE: Confocal image of MacroVax-treated immature human dendritic cells. Actin (Red), Nuclei (Blue), Conjugated Nanoparticles (Green).

Mannose Binding Lectin Blocking Antibody Decreased Phagocytosis of MacroVax Indicated by Lower Levels of Intracellular GFP
10 9 8 7 6 5 4 3 2 1 0
0 10 20 30 40 50 Concentration of Blocking Antibody (ug) 60 70

GFP Concentration (ng/ul)

Mouse macrophages were treated with a Mannose Binding Lectin Antibody (Genetex) and then treated with GFP-tagged MacroVax to study dose-response of MacroVax with respect to phagocytosis. Using a GFP Quantification Assay (BioVision) intracellular GFP

MacroVax Activates Human Dendritic Cells


Untreated Treated

Epifluorescent image of vaccine treated dendritic cells. The MacroVax complex is shown here to bind to and be phagocytosed by these antigen presenting cells. Actin (Red), Nuclei (Blue), Vaccine Complex (Green). 1200X magnification, scale bar = 50 um.

MacroVax Treated APCs Express Activation Biomarker CD86

Vehicle (PLGA Only)

Vaccine-Treated

Flow cytometry was performed on macrophages treated with either PLGA (vehicle) particles or the vaccine complex for three hours. The two markers of interest used were CD11b (X axis: macrophage marker) and CD86 (Y axis: activation marker).

MacroVax Upregulates Expression of Activation Cytokines and Downregulates Expression of Deactivation Cytokines in Human Dendritic Cells

In human dendritic cells the expression or transcription levels of certain chemical signals called cytokines was studied. Dendritic cells were either untreated or vaccine-treated for three hours and then subject to mRNA extraction and RT-PCR to study DNA activities posttreatment. *=P <0.001, Students T-test ** = P <0.01, Students T-test

MacroVax Induces Secretion of Activation Cytokine TNF- and Depresses Secretion of Deactivation Cytokine IL-10
IL-10 Levels in Vaccine Treated Macrophages
2000 1800 450 400

TNF- Levels in Vaccine Treated Macrophages

1600
1400 1200 1000 800 600 400 200 0 1 2 3 4

350
300

Pg/ml

Pg/ml

250 200 150 100 50 0 1 2 3 4

Legend: 1- PLGA + Lysate 2- PLGA + Immunostimulatory Agents + Lysate 3- Microbial PLGA + CpG + GM-CSF + Lysate 4- MacroVax

T Cell Proliferation Assay and IFN-y MacroVax Induces Generation of Tumor-Specific T Lymphocytes Staining
Prime Immune System With Vaccine Treatment (Day 1) Boost Immune System With Vaccine Treatment (Day 14) Collect Mouse Spleens and Isolate Splenocytes (Day 21)

MacroVax Induces Generation of Tumor-Specific T Cell Proliferation Assay and IFN-y T Lymphocytes Staining

CD3 (Positive Control)

Survivin

Tumor Lysate (N2A)

N2A Tumor Cells

CyQuant Proliferation Assay (Invitrogen)

MacroVax Induces Generation of Tumor-Specific T Lymphocytes


Control (PLGA+Lysate) Lymphocytes Vaccine-Treated Mouse Lymphocytes

Exposed to Tumor Antigens

Exposed to Tumor Antigens

Mouse T lymphocytes collected from PLGA/lysate-treated mice (left) and vaccine-treated mice (right) spleens (3 weeks of treatment) were exposed to tumor specific antigens and analyzed for clonal expansion via a cell proliferation assay.

Untreated Lymphocytes

MacroVax Induces Generation of Tumor-Specific T Lymphocytes


Vaccinated Splenocytes

MacroVax Induces Activation of Tumor-Specific T Lymphocytes Through IFN- Production


350 300 250 pg/ml 200 150 100 50 0 Unexposed Exposed to Tumor Antigens

PBS Treated CpG Vax MacroVax (Control) Mouse Vaccine Treatments


Mouse T lymphocytes collected from PLGA/lysate-treated, CpG/PLGA/Lysate-treated, and MacroVax-treated mouse spleens (3 weeks of treatment) were exposed to tumor specific antigens and analyzed for activation via ELISA for IFN-.

DIAGNOSEME

Detecting the tumor Immune-Inhibitory cloud

Proteins indicative of immune activity are found in your sweat and we can measure these proteins with our telemedicine platform.

Preliminary Proof of concept


We built a biosensor iteration of DiagnosMe to test for inflammatory marker IL6 in sweat at the Thayer School of Engineering at Dartmouth College.

Analyzed the precision of the biosensor between measurements

Sweat Sample Concentration of IL6 (pg/ml)


14 IL6 Concentration (pg/ml) 12 10 Concentration of IL6 (pg/ml)

8
6 4 2 0

Sample 1

Sample 2

Average

Sweat Sample
1 2

Optical Density
.0975 .0801

IL6 Concentration (pg/ml)


12.88 pg/ml 10.582 pg/ml

Studied changes in IL6 levels in sick individuals

Disease Progression (Individual 1)


IL6 Concentration (pg/ml)
IL6 Concentration (pg/ml) 200 150 100 50 0 0 0.5 1 1.5 2 Optical Density 2.5 3 3.5

?
Reading 1 Reading 2
Reading 3

Disease Progression (Individual 2)


IL6 Concentration (pg/ml)

150 135 120 105 90 75 60 45 30 15 0


0 0.5

IL6 Concentration (pg/ml)

1.5 2 Optical Density

2.5

3.5

Reading 1

Reading 2

Reading 3

Studied the effects of chemical impurities such as deodorant on the biosensor readings

Effect of IL6 Concentration Due to Sample Impurities


12 IL6 Concentration (pg/ml)

10
8 6 4 2 0 Control Sunscreen Lotion Impurities Perfume Deodorant

Control

Sunscreen

Lotion

Perfume

Deodorant

Studied the effect of sweat sample volume on the accuracy of the biosensor

IL6 Concentration at Different Sample Volumes


16
IL6 Concentration (pg/ml) 14 12 10 8 6 4 2

0
300 250 200 150 Volume of Sample (l) 100 Average

300 l (Control)

250 l

200 l

150 l

1oo l