module 5- vaccines and translational immunology Flashcards
immunological techniques
- ELISA
- FLOW cytometry
- monoclonal antibodies
what is ELISA
enzyme-linked immunosorbent assay
what does ELISA do?
based on the principle of antigen-antibody interactions
- can be codified to detect and quantify substances such as peptides, proteins, antibodies, hormons +
what does flow cytometry do
is a method of detechting and quantifying different cell types in a mixed cell suspension
what does a monoclonal antibodies do
- the production of monoclonal antibodies is a technique that was developed by george kohler and cesar milsteins in 1975
- are antibodies that are produced by a single clone of a B-cell that are specific for a single epitope
epitope
the portion of the antigen that is recognized and bound by an antibody
what is ELISA highly specific
- based on the principle of antigen-antibody interactions
- antigen binding site is very specific to one antigen
how does ELISA work? step 1
- the bottom of the wells are coated with an antigen that is specifically recognized by the antibody you wish to measure (refereed to as the primary antibody in this procedure )
how does ELISA work? step wash (1)
the wells are washed to remove any excess antigen not attached to the bottom of the well
how does ELISA work? step 2
the sample containing the antibody to be measured (serum) is added to the well. the primary antibodies if present will bind to the antigen attached to the bottom of the well
how does ELISA work? step wash (2)
the wells are washed again to remove excess primary antibody not attached to the bound antigen as well as any other sample components that might interfere with subsequent steps
how does ELISA work? step 3
an enzyme-conjugated secondary antibody is added to the well
- this secondary antibody will bind to the FC portion of the primary antibodies already present in the well
- secondary antibody used specifically recognizes antibodies from a particular animalr
how does ELISA work? step wash (3)
the wells are washed to remove any excess secondary antibody not attached to the primary antibody
how does the ELISA work? step 4
- substrate of the enzyme attached to the secondary antibody is added to the well
- the reaction substrate (a chromogen) and the enzyme produces a coloured product which can be measured by absorbance
chromogen
a substance that can be readily converted into a dye or other coloured compound
what is flow cytometry?
a technique designed to detect and quantigy different immune cells in a mixed cell suspension
what does the flow cytometry measure
- physical properties of a cell
- detect specific antigens on or inside a cell
- the total number of cells in the suspension, the suspension and the overall composition of the suspension can be readily determined
- used to determine complete blood counts (CBC)
how does flow cytometry work?
- narrow stream of cells in single file is passed through a laser light source
- the way the laser light is scattered is unique to each cell type; this can be detected and analyzed
- measuring FCS allows for the discrimination of cells by size
- FCS intensity is proportional to the diameter of the cell
- SSC provides information about the internal complexity of cell
what else can flow cytometry be used to determine?
the proportion of cells expressing a particular antigen (cells labelled with specific antibody)
- antibody is coupled wit ha flurorescent marker (can be excited by a light of a specific wavelength
- fluorescent marker emits a light with a characteristic different wavelength (only cells expressing the antibody in question will emit light of this specific wavelength
what is the production of monoclonal antibodies used for?
- research
- diagnosis
- therapeutic purposes
what does monoclonal antibodies measure
- not a specific measurement it is a tool
- there are clinical application of monoclonal antibodes
what are the clinical applications of monoclonal antibodies?
immunotoxins and radiolabelled antibodies
immunotoxins
- consist of a tumor-specific monoclonal antibody attached to a deadly toxin
- technique still under investigation but a long-term objective is to use immunotoxins to target and eliminate tumor cells and treat cancer
radiolabeled antibodies
monoclonal antibodies tagged with a radioactive isotope can be used to diagnose tumors earlier than other methods
- can bind to antigens on a tumor thereby allowing the precise location of a tumor to be visualized
how does monoclonal antibodies work?
- produced in lab by hybridomas, immortal cells that produce unlimited quantities of one identical antibody
- hybridomas are the result of fusion between a plasma cell and canerous (myeloma) cell (share properties of both)
hybridoma cell
a perpetual source of antibodies against one antigen
types of vaccines
- live attenuated vaccine
- killed -inactivated vaccine
- toxoid vaccine
- subunit vaccine
what is a vaccine
- is a type of biological preparation which provides active artificial immunity to a particular disease-causing agent
- dependent on the nature of the disease and how the immune system recognizes and response to the infection
characteristics of live-attenuated vaccine
- contains a modified strain of the disease-causing agent which has lost its pathogenic ability
- retains it capacity to replicate within the host
pathogenic
ability of an organism to harm the host by causing a disease
advantages of live-attenuated vaccine
- provides a prolonged exposure to the disease-causing agent
- suitable to generate cell-mediated immunity
disadvantages of live-attenuated vaccine
- potential to revert to a virulent form
- requires specific storage and transport conditions
examples of live-attenuated vaccine
- small pox vaccine
- oral poliovirus vaccine (sabin)
- measles vaccine
characteristics of killed-inactivated vaccine
- contains a strain of the disease-causing agent that has been inactivated by heat, chemcials or radiation
- has the ability to generate an immune response, but unable to replicate
advantages of killed-inactivated vaccine
- safer option as it cannot mutate back to a virulent form
- easy to store and transport
disadvantages of killed-inactivated vaccine
- generally requires multiple booster doses to maintain immunity
- generally must be administered by injection
examples of killed-inactivated vaccine
- rabies vaccine
- flu vaccine
characteristic’s of toxoid vaccine
- contains an inactivated toxin which is a product from the pathogen that is causing the disease
advantages of toxoid vaccine
- safe as it is not living organism that can divide, spread and/or revert
- stable as they are less susceptible to changes in temp, humidity and light
disadvantages of toxoid vaccine
- may requie several doses and usually need an adjuvant
adjuvant
a substance that enhances the bodys immune response to an antigen
examples of toxoid vaccine
- tetanus vaccine
- diphtheria vaccine
characteristics of subunit vaccine
contains only a small part of fragment of the disease-causing agent
advantages of subunit vaccine
- safest type of vaccine, can be used on everyone, including immunocompromised, pregnant, and elderly populations
disadvantages of subunit vaccine
- rarely successful at inducing long-lasting immunity, which means it will require multiple booster doses to maintain immunity and might need to be conjugated to a carrier
carrier
- a stronger antigen than the desired target antigen
- by covalently attaching a strong antigen to a poor antigen, the overall immunological response is strengthened and hopefully, response to the poor antigen is also improved
example of subunit vaccine
hepatitis B vaccine
about mRNA vaccines
- most recent vaccine type that have changes the fueld of vaccinoogy
- known for their use against SARS-CoV-2 (covid)
- used in several formulations to fight the virus (boosters, vaccines)
the princple of the assay (mRNA)
relies on the use of mRNA to produce viral proteins and recruit immune cells to respond to the antigenic target
- proteins then displayed on surface of an APC to induce B-cells and T-cell immunity
RNA and DNA based vaccines
involve making genetic material only
- they do not require the use of the whole virus
- vaccine production of mRNA vaccine
- mRNA is made in lab from a DNA template of the virus
- it encodes an antigen of the virus
(for Covid, the mRNA encodes the spike protein on the surface of the virus) - the mRNA is incorporated into a formulation that can be administered as a vaccine
mRNA vaccine mechanisms
- vaccine production
- host cell
- APC
- immune response
- host cell
once inside the body, mRNA enters the host cell and uses host cell machinery to produce the spike protein
- APC
- newly formed spike exits the cell and is recognized by an APC
- APC internalized the spike protein and processes it into a peptide (antigen)
- APC then displays the antigen on the surface of the cell via MHC
- immune response
- the antigen is recognized by a T-cell helper, which initiates an immune response
- b cells produce antibodies that stop the virus from infecting cells
- t-cell destroys cells infected with the virus
antivirals medications against COVID-19
(only treated it once you already tested positive)
- does not prevent covid
what are the antiviral medications for covid?
polyermase inhibitor
protease inhibitor
polyermase inhibitor
- the first antiviral meducation
- is a enzyme that plays a crucial role in viral replication and transcription
molnupiravir- is a polymerase inhibitor used to treat covid. it increases the frequency of viral RNA mutations and impairs replication’s of the virus
protease inhibitor
protease cuts proteins into smaller, more workable pieces
- often adminsitered in combination
- nirmatrelvir stops protease from cutting viral proteins into functional peices
- ritonvair protects nirmatrelivir from destrcution by the body and allows it to keep working
- stops it from replicating and infecting other cells
evolution of vaccines
Dr. Edward Jenners use of extracts prepared from cowpox lesions as a means of protecting his patients against smallpox infection
- vaccination derived from the latin word for cow and was first coined by louis pasteur in 1881 to honor jenners accomplishment
- jenns was 100 years before discovery of virus by Dmitry Ivanovsky 1892
vaccine success story: HPV VLP vaccines
Cervical cancer and genital warts are caused by infection with certain types of HPV.
In 2002, a clinical trial showed that a Virus-Like Particle (VLP) vaccine could protect against the HPV type most often linked to cervical cancer.
The vaccine showed 100% effectiveness in this trial, and later larger trials continued to show similar high efficacy rates, almost 100%.
VLPs
VLPs (Virus-Like Particles) are made of the structural proteins of HPV. These proteins self-assemble into virus-like structures that look and act like the real virus but don’t contain viral DNA, so they can’t cause infection.
what are the three HPV vaccines currently authorized
- cervrix
- gardasil
- gardasil 9 (9 means can protect against 9 different types of HPV)
success story: ebola vaccines
- ebola virus (EBOV) and marburg virus (MARV) are part of the filoviridae family of viruses that cause hemorrhagic fever with high mortality rate in humans and nonhuman primates
- these virus produce transmembrane glycoproteins thought to play a role in the virulence of these viruses
2000 ebola
Winnipeg inadvertently discovered that mice infected with EBOV and MARV glycoproteins became protected against infection from live virus
- development of petenially viable vaccine using glycoproteins from EBOV and putting them into a live attenuated recombination vesicualr stomatitis virus (VSV) that expresses the transmembrane glycoproteins of EBOV and MARV
2005 ebola vaccines
canadia research team tested this vaccine the 2000 vaccine on macaques (old world monkeys) and found it to be 100% effective against EBOV infection
failure story: genital herpes vaccine
a subunit vaccine was developed which was composed of a prominent structural protein of HSV-2 (the type most commonly associated with genital infections)
- the vaccine showed promise in early trials with 70% efficacies in women who were seronegative for both HSV-1 and HSV-2 at the beginning of the trial
- larger trial was 20% efficacy and no protection from HSV-2
what did the genital herpes vaccine cause
researchers in the field to consider developing live attenuated vaccines for genital herpes and has spurred basic research efforts aimed at better understanding and augmenting immune responses in the genital tract
what is genital herpes
- caused by infection with herperes simplex virus (HSV) types 1 or 2
- are permanent
- require lifelong mamgement
- serious health threat to new borns through vertical transfer
- emotional distress in infected individuals
- increase susceptibility to and spread of HIV infection
vertical transfer
is the passage of a disease-causing agent from mother to baby during the period immediately before and after birth
seronegative
giving a negative result for a test of blood serum for the presence of a host antibody response against a foreign particle/pathogen, such as HSV-1 and HSV-2 virus
- informs you of 1. IgG vs IgM levels, indicating when the host was exposed to the pathogen 2. if the host has ever been exposed to a pathogen 3. if host has ever been vaccinated
phases of vaccine development
- lab studies
- preclinical
- clinical phase I
- clinical phase II
- clinical phase III
- health Canada approval
- lab studies
- identify the infectious agent causing the disease and select a strain (or subtype)
- that is relevant to the target population
- dependent on research carried out in the laboratory utilizing assays, which may involve exhaustive screening to identify a suitable antigen
- creation of vaccine concept
what else does lab studies involve
- developing and testing the manufacturing process of the vaccine according to GOOD MANUFACTURING PRACTICE standards
- preclinical
- involves research carried out in animal models to evaluate the pharmacological aspects of the product
clinical phase I
- due to rigorus testing requirements, only a very small % of vaccines progress to licensing, making the costs of vaccine research and development extremely high
- invloves 3 phases spanning over servela years
phase I clinical trails for vaccines
involve small scale trials in humans to asses vaccine safety by evaluating local and systemic reactions after administration
- provides preliminary data on the immunogenicity of the vaccine and the immune response it evokes
clinical phase II
- occur at bigger scale (50-500) to collect data on the safety, side-effects, and efficacy of the vaccine
- evaluates the dosage requirements of the vaccine
- mointor the effects of increasing vaccine dosage and conduct challenge tests to define the optimal dose and vaccination schedule
clinical phase III
- involve multiple geographic sites with many hundreds of subjects (300-30000) to evaluate efficacy under natural disease condition
- researchers are required to demonstrate the efficacy in target populations and complete a safety assessment for the vaccine
- if successful manufactur will then apply to the regulatory authorities for a license to market
health canada approval
- is the regulatory authority in Canada responsible for ensuring the quality, safety, and efficacy of all biologic drugs, including vaccines for human use
- regulation is necessary
- vaccine candidates must be submitted to health Canada to be considered for approval with sufficient scientific and clinical evidence to show that it is safe
vaccination in Canada
- routine
- responsible for decline in vaccine preventable disease
- free of charge through provincal immunization
what is the canadian immunization guide
a comprehensive resource on immunization
- based on recommedendations by the national advisory committee on immunization
- critical role with herd immunity
vaccine trends
- measles vaccine
- rubella vaccine
- diptheria vaccine
- tetanus toxoid
- varicella vaccine (chicken pox)
measles vaccine, when was it introduced
1960-1968
rubella vaccine, when was it introduced
1958-1968
diphtheria vaccine, when was it introduced
1926
routine with it in 1930
tetanus vaccine, when was it introduced
1940
varicella vaccine, when was it introduced
1998
challenges with vaccine development
cost
cold chain
continuous monitoring
the gold standard
cost challenge
- development cost a lot
- leads to premature abandonment of clinical research
(especially is disease threat is restricted to developing countries)
what would reducing the cost of a vaccine mean
ensuring that a larger pop can afford the vaccine and can be used in developing countries
the cold chain
- live attenuated vaccines, the most effective as producing long term memory
- needs to be stroes and transported in a specific condition
- if broken, vaccine risks losing potency or unusebale
continuous monitoring
- efficacy of a vaccine is directly related to specific antigen against which the immune response has developed
- if disease causing agents evlove by changing or losing their antigenic determines could lose its efficacy
antigenic determinants
antigen molecule which can be recognized and bound by an antibody
if a vaccine is based on variable antigen
the genetic variation of the pathogen must be continually monitored
the gold standard
if vaccine is approved, that doesnt mean work is done
- new strategies are developed to create better vaccine by using recombinant vector or adjuvant
recombinant vector
vaccine using an attenuated virus or bacteria to introduce microbial or viral DNA into the body
adjuvant
substance added to a vaccine to enhance bodys immune response
what is the gold standard conundrum
when a new version of vaccine is developed, must be as good or better then the old vaccine
neuraminidase antigen
is a surface protein that removes sialic acid from cell surfaces, and enables new viral copies to infect and spread to other cells
how many NA have researhcers found?
11, each with sequence variability’s in their receptor binding sites
hemagglutinin antigen
is a surface protein that recognizes and binds sialic acid on cell surface glycoproteins
what do HA surface interactions lead to
lead to endocytosis of the virus and HAs are activated to fuse the endomsome and viral membrane
herd immunity
is a form of indirect protection from an infectious disease that occurs when a large proportion of the population is immune to an infectious agent
what does herd immunity do
provides protein for people who are and who aren’t immunized as it slows or completely stops the spread of the disease
who is herd immunity rly important for
immunocompromised people and people who cannot be vaccinated
how is herd immunity measured
calculating what is called the basic reproduction number Ro
what is Ro
is the average arising from a typical case in a totally susceptible population
- determines the herd immunity threshold and therefore the immunization coverage to eliminate the disease
as Ro increases
higher immunization coverage is required to achieve herd immunity
cancer in canada
- high prevalence making it important social and health issue
what does the Canadian cancer society estimate
one in 2 Canadian will develop cancer in their lifetime
cancer in 2017
reported estimated that 206,200 Canadians were diagnosed with cancer and 80,800 died
how is cancer characterized
by an error in either one of these processes
- cancer cell VS normal cell
- tumour
- cancer and immune system
- cancer immunotherapy
cancer cell vs normal cell
- cancer cells: do not need specific growth factors to divide. they do not response to the signals that cause normal cells to stop diving
- cancer cells viewed as self cells
- results in alteration in DNA, which induce cell transformation. often due to exposure of carinogenics chemicals or radiation
tumour
cancer calls will continue to divide and grow, ultimately forming a tumour
- abnormal mass of tissue
cancer and the immune system
- cancer immunity cycle
- cancer immunoediting
- immune escape mechanisms
- tumour immune microenvironment
cancer immmunotherapy
- eithers based on secreted or cellular components of the immune system are (immunotherapies)
- aimed at enhancing host anti-tumour immune responses and have joined the pillars of cancer treatment
types of tumours
benign
malignant
benign tumour
is not cancerous
- unable to grow indefinitely or invade surrounding tissues
malignant tumour
is cancerous
- abilitiy to metatasize
- grow and become progressively invasive
metastasis
- is the colonization by tumor cells of sites different from their primary site of origin
- small cluster of cancerous cells will break off from the tumor and invade the surrounding blood or lymphatic vessels, traveling to different areas, where they
resilience of cancer cells
- is a cells ability to cope with environmental disturbances, and possibly adapt for future exposure
- making it harder to find treatment
resilience and stress
stress is an umbrella term for the harsh environmental exposures on tumor cells
HeLa cells
biomedical research originated from this
- the resilience allowed for immortality, thus making them a important tool
who is henrietta?
- African American women from virgina, mother of 5
- unwitting donor of cell known as HeLa cell line
- had carcinoma
- samples of her ovarian were taken without the consent and knowledge
what as the finding of henrietta?
these cells were grown in a lab becoming the first line of human cell culture to ever survive outside the human body
HeLa cell line- researhc break throughs
- recombinant protein production
- HPV vaccine
- Understanding virology
- toxicity testing
- monoclonal antibody production
- polio vaccine
- genome sequencing
- telomerase activity
recombinant protein production
- bacteria cells can be used for protein production they lack mechanics to produce more complex proteins
- the HeLA line allowed researchers to overcome this
HPV vaccine
- found that HeLa cells were infected with HPV-18 virus, and decades of characterizing HPV-18 led to the development of HPV vaccines
understanding virology
- experimental viral infection on HeLa cells allowed researchers to characterize how specific virus can evade the immune system, such as CD4-Tcell receptor utilization by HIV
toxicity testing
hepatocytes were used for testing, but were too unstable
- the reilient HeLa cells are now used, important in drug development and discovery
monoclonal antibody production
- can be produced using hybridoma crosses of HeLa and other animal cells
- have many application such as medical diagnoses and cancer therapy
polio vaccine
developed in 1950s but HeLa cells were the only human cells that could be used to test the vaccine
genome sequencing
HeLa cells fused with mouse cells became the first hybrid cells, fusion of 2 cells types
- aided in the emergence of the human genome project
telomerase activity
1996, Gregg morin isolated telomerase from HeLa cells, which were previously only found in animal embryos
- supported that both embryo and cancer cells utilize telomerase to rapidly divide
the cancer immunity cycle
describes how the immune system balances the recognition of, and defense against, non-self molecules and the prevention of autoimmunity
step 1: of cancer immunity cycle
release of cancer cell antigens (caner cell death)
release of cancer cell antigens (caner cell death)
- antigens are released by mutated cancer cells, indicating that they are not healthy cells
- immune system can recognize these
step 2: of cancer immunity cycle
: cancer antigen presentation (dendritic cell/APCS)
cancer antigen presentation (dendritic cell/APCS)
the cells of the immune system capture the released antigens and travel to the lymph nodes where they find T-cells
step 3: of cancer immunity cycle
priming and activation (APC and T-cell)
priming and activation (APC and T-cell)
T-cells are activated by the antigen and the immune response against the cancer cells is initiated
step 4: of the cancer immunity cycel
trafficking of t-cells to tumors (CTLs)
trafficking of t-cells to tumors (CTLs)
- the activated T-cells move through the blood vessels to the site of the tumour
step 5: of the cancer immunity cycle
infiltration of T-cells into tumors
(CTLs, endothelial cells)
infiltration of T-cells into tumors
(CTLs, endothelial cells)
once the t-cells reach the cancerous cells, they invade the tumor to attack it
step 6: of the cancer immunity cycle
recognition of cancer cells by T-cells (CTLs, cancer cells)
recognition of cancer cells by T-cells (CTLs, cancer cells)
t-cells recognize cancer cells because of the antigens they had previously released
step 7: of cancer immunity cycle
killing of cancer cells (immune and cancer cells)
killing of cancer cells (immune and cancer cells)
t-cells initiate a pathway that results in cancer cell death
what is tumor immunosurveillance
states that tumor cells are identified and kept under control by the immune system of healthy individuals
- however, cancer cells can evade recognition or the magnitude of the anti-tumour immune response is not good to kill of the cancer cells
immunoediting
is a dynamic process which describes the connection between the tumor cells and the immune system in the context of immunosurveillance and tumor progression
what is cancer immunoediting
- cancer evolution there is communication between them and immune cells
- interaction between tumor cells and immune cells, led to tumor progession
what are the 3 processes of immunoediting
- elimination
- equilibrium
- escape
what are the steps of cancer immunoediting
phase 1: elimination
phase 2: equilibrium
phase 3: escape
phase 4: overview
phase 1: elimination
when a tumor cells arises in a tissue, immune system quickly act to remove it
- NK cells, cytotoxic T-cells, helper- T-cells can recognize altered cell to work on eliminating
phase 2: equilibrium
- if tumor cells are not eliminated, they enter a state of equilibrium where they proliferated and is matched by cell killing by the immune system
- equilibrium phase can last for a short time or many years
phase 3: escape
- tumor cells are no longer recognized by the immune system and so avoid elimination
- are to grow uncontrolled and proliferate to form a tumor
overview of cancer immunoediting
cancer cells can overcome the immune response, proliferate, and form tumours
ways the cancer cells invade the immune system
reduced MHC expression
poor costimulatory molecules
reduced MHC expression
- tumor cells display low levels of MHC class I molecules on surface
- as cytotoxic T lymphocytes recognize antigens in the context of MHC class I molecules
poor costimulatory molecules
T-cells require both expression of MHC and costimulatory molecules to become activated
- tumor cells lack costimulatory molecules contributing to poor immunogenicity
- T-cells only partially activated
immunotherapy is able to…
attack cancerous cells throughout all organs in the body
immunotherapy allows the immune system too…
specifically target and eliminate cancer cells without damaging healthy cells, resulting in fewer side effects, than traditional cancer treatments
immunotherapy takes..
advantage of immunological memory, allowing for the possibility of long term protection
immunotherapy can..
be applied to almost all types of cancer
what are tumor infiltrating lymphocytes?
b-cells
about tumor infiltrating lymphocytes?
the type, density, and location of TILs has been suggested as a prognostic biomarker in some cancers
prognostic biomarkers
biological characteristics that are objectively measured and evaluated to predict the course of disease or a response to a therapeutic intervention among patients with same characteristics
how do tumor infiltrating lymphocytes work?
leave blood stream and migrate to infiltrate the tumor under influence of various chemotactic gradients of specific types of chemokines
what are tumor infiltrating lymphocytes a mix of ?
T and b cells
sometimes NK cells, dendritic and macrophages
TILs is ___ always reflective of their ___ and ___ significance
NOT
activity
prognostic
activation status of tumor infiltrating lymphocytes
some of the dynamic phenotypic markers expressed on these cells reflect their activation status and therefore, must accompany and interpretations on their roles as biomarkers for diagnosis
biomarkers
a measurable substance in an organism, the presence of which is indicative of some phenomenon such as disease, infection, or environmental exposure
what are solid tumors
classified as either T-cell inflamed “hot”
or T-cell non- inflamed “cold”
what are T-cell inflammed “hot” tumors
show comparatively higher immune activity compared to cold
characteristics of hot tumors
- high numbers of CD8+ TILS
- high levels of interferon (IFN) genes
- usually respond well to treatment (chemo immunotherapy)
what are T-cell non-inflamed
lower immune activity compared to hot
characteristics of cold tumors
- low numbers of CD8+ TILS
- low levels of interferon (IFN) genes
- usually inferior response to treatment (chemo and immuno)
treatment for cold and hot tumors
one could convert cold to hot by stimulating the tumor interferon activity
- this has led to exploitation of anti-tumor immunity towards development of newer treatments
a new immunotherapy tool
the immunoscore
what is the immunoscore
combining the knowledge of TILs and the immunological classification of hot and cold tumors make new and reliable prognostic biomarker for cancer diagnosis
what does immunoscore measure
- density/number of T-cells in the center (CT) and the periphery (IM) of the tumor by immunochemistry
what can immunoscore measure help with
stratify patients as having high or low risks of cancer and aid in developing treatment plans
work flow for determining an immunoscore: step 1
separating the tumor in the central and perioheral regions
work flow for determining an immunoscore: step 2
staining for T-cells and conducting digital pathology
work flow for determining an immunoscore: step 3
assigning a score to the tumor to relate it with an associated diagnosis or risk attribution
