MICR 270 Module 5: Vaccines and Translational Immunology PDF

MICR 270 INFECTION, IMMUNITY, AND INFLAMMATION MODULE 05 VACCINES AND TRANSLATIONAL IMMUNOLOGY Please note: This course was designed to be interacted and engaged with using the online modules. This Module Companion Guide is a resource...

MICR 270 INFECTION, IMMUNITY, AND INFLAMMATION MODULE 05 VACCINES AND TRANSLATIONAL IMMUNOLOGY Please note: This course was designed to be interacted and engaged with using the online modules. This Module Companion Guide is a resource created to complement the online slides. If there is a discrepancy between this guide and the online module, please refer to the module. How can you help protect the integrity and quality of your Queen’s University course? Do not distribute this Module Companion Guide to any students who are not enrolled in MICR 270 as it is a direct violation of the Academic Integrity Policy of Queen’s University. Students found in violation can face sanctions. For more information, please visit https://www.queensu.ca/academic- calendar/health-sciences/bhsc/. MODULE 05 COMPANION GUIDE MICR 270 TABLE OF CONTENTS INTRODUCTION..................................................................................................................................................... 4 Introduction....................................................................................................................................................... 4 Course Toolbox.................................................................................................................................................. 4 Learning Outcomes........................................................................................................................................... 5 Module Assignments........................................................................................................................................ 5 Journal Article Assignment........................................................................................................................... 5 Scientific Poster Assignment........................................................................................................................ 5 Module Outline.................................................................................................................................................. 6 SECTION 01: Immunological Techniques and Technologies............................................................................ 7 Introduction to Immunological Techniques................................................................................................... 7 Enzyme-Linked Immunosorbent Assay (ELISA)............................................................................................. 7 Question: Enzyme-Linked Immunosorbent Assay (ELISA)....................................................................... 8 Enzyme-Linked Immunosorbent Assay (ELISA) – How Does it Work?..................................................... 8 Enzyme-Linked Immunosorbent Assay (ELISA) – What Does it Measure?............................................. 8 Video: Enzyme-Linked Immunosorbent Assay (ELISA)............................................................................. 9 Flow Cytometry.................................................................................................................................................. 9 Question: Flow Cytometry..........................................................................................................................10 Flow Cytometry – How Does it Work?.......................................................................................................10 Flow Cytometry – What Does it Measure?................................................................................................11 Clinical Application - Flow Cytometry and Disease Diagnosis...............................................................11 Video: Flow Cytometry................................................................................................................................11 Monoclonal Antibodies...................................................................................................................................12 Question: Monoclonal Antibodies.............................................................................................................12 Monoclonal Antibodies – How Does it Work?..........................................................................................12 Monoclonal Antibodies – What Does it Measure?...................................................................................13 Clinical Application – Monoclonal Antibodies and Cancer.....................................................................13 Video: Monoclonal Antibodies...................................................................................................................14 SECTION 02: Vaccines.........................................................................................................................................15 Question: Types of Immunity.........................................................................................................................15 Question: Vaccines..........................................................................................................................................15 Types of Vaccines............................................................................................................................................16 Live-Attenuated Vaccine.............................................................................................................................16 INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 1 MODULE 05 COMPANION GUIDE MICR 270 Killed-Inactivated Vaccine..........................................................................................................................17 Toxoid Vaccine.............................................................................................................................................17 Subunit Vaccine...........................................................................................................................................18 mRNA Vaccines................................................................................................................................................18 mRNA Vaccine Mechanism.............................................................................................................................19 Antivirals Medications Against COVID-19.....................................................................................................19 Evolution of Vaccines......................................................................................................................................20 A Modern-day Vaccine Success Story: HPV VLP Vaccines...........................................................................20 A Modern-day Vaccine Success Story: Ebola Vaccines................................................................................21 A Modern-day Vaccine Failure Story: Genital Herpes Vaccine...................................................................22 Phases of Vaccine Development...................................................................................................................23 Lab Studies...................................................................................................................................................23 Preclinical Studies.......................................................................................................................................24 Clinical Phase I.............................................................................................................................................24 Clinical Phase II............................................................................................................................................24 Clinical Phase III...........................................................................................................................................24 Health Canada Approval............................................................................................................................25 Vaccination in Canada....................................................................................................................................25 Vaccine Trends in Canada..............................................................................................................................25 Question: Development of Vaccines.............................................................................................................26 Question: Moving Forward with Vaccine Development.............................................................................27 Challenges with Vaccine Development.........................................................................................................27 Vaccine Development Challenges - Influenza Virus....................................................................................28 To Vaccinate or Not to Vaccinate? Maybe It’s Not All About You...............................................................29 Vaccinating Against Measles..........................................................................................................................30 SECTION 03: Cancer And The Immune System................................................................................................31 Cancer in Canada............................................................................................................................................31 Introduction to Cancer...................................................................................................................................32 Tumours...........................................................................................................................................................33 The Resilience of Cancer Cells.......................................................................................................................34 The Special Case of HeLa cells.......................................................................................................................34 The HeLa Cell Line - Who is Henrietta Lacks?...........................................................................................35 The HeLa Cell Line - Research Breakthroughs.........................................................................................36 The Cancer-Immunity Cycle...........................................................................................................................37 INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 2 MODULE 05 COMPANION GUIDE MICR 270 Tumour Immunosurveillance vs. Immunoediting.......................................................................................38 Cancer Immunoediting...................................................................................................................................38 Evasion of the Immune Response.................................................................................................................39 Question: The Immune System and Cancer................................................................................................39 Overview of Cancer Immunotherapy............................................................................................................40 Question: Cancer Immunotherapy...............................................................................................................40 Introduction to Cancer Immunotherapy......................................................................................................40 How would using immunotherapy as a cancer treatment affect the cancer-immunity cycle?.............41 Tumour Infiltrating Lymphocytes..................................................................................................................41 Immunological Classification of Tumours....................................................................................................42 A New Immunotherapy Tool – The Immunoscore......................................................................................42 Looking Forward: Immunotherapy Approaches Based On Tumour Immune Microenvironment........43 CONCLUSION.......................................................................................................................................................44 Module Summary............................................................................................................................................44 Concept Map: List of Immunology Terminology..........................................................................................44 Learning Outcomes.........................................................................................................................................44 Module Assignments......................................................................................................................................45 Journal Article Assignment.........................................................................................................................45 Scientific Poster Assignment......................................................................................................................45 Complete..........................................................................................................................................................46 INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 3 MODULE 05 COMPANION GUIDE MICR 270 INTRODUCTION Please see the online learning module for the full experience of interactions within this document. INTRODUCTION This content was retrieved from Introduction, Slide 1 of 5 of the online learning module. Many practical applications and useful technologies have directly resulted from fundamental studies in immunology. In Module 5, you will learn about two widely used laboratory techniques for measuring immune responses and about monoclonal antibody production, one example of a valuable technology arising from immunological research. Vaccines are probably the best-known practical applications of immunology. Common types of vaccines are discussed along with the process of vaccine development. The module will end by overviewing cancer and cancer immunology as well as how immunotherapy can be used to treat cancer. The concepts explored in Module 5 will be assessed through the Module 5 Quiz and will help you understand and present your journal article, if you are assigned to choose an article for this module. For this module, continue working on your Scientific Poster assignment. COURSE TOOLBOX This content was retrieved from Introduction, Slide 2 of 5 of the online learning module. During the course, you will encounter some features that will help you enhance your understanding of course concepts. You will find the following icons throughout the modules: Orange Bolded Words Throughout the modules you will find hover definitions*. Every time you see this feature, make sure you hover over the text to see a definition or additional information. Review more information. When you click on this icon, it will reveal more details about a word or a concept covered in the course. Reveal part of an analogy created to help you understand the complexity of the immune system. Prision Break: The immune system response to an infection can be compared to someone trying to break out of prison. Each time you see this icon, click to learn more about what these components or processes would be in the analogy of a prison breakout. Definition*: Hover definition: I’m some additional information! INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 4 MODULE 05 COMPANION GUIDE MICR 270 LEARNING OUTCOMES This content was retrieved from Introduction, Slide 3 of 5 of the online learning module. At the end of Module 5, you will be able to: Describe the basic principles and applications of two techniques for measuring an immune response. Describe the technology behind monoclonal antibody production and the various uses of monoclonal antibodies. Explain the basis of four types of vaccines and discuss their utility in protecting us against various classes of infectious agents. Describe the stages of vaccine development and discuss some of the challenges faced during vaccine development. Describe the role of the immune response in cancer and discuss how immunotherapy has been established as a major pillar of cancer management. MODULE ASSIGNMENTS This content was retrieved from Introduction, Slide 4 of 5 of the online learning module. These assignments are associated to Module 5. JOURNAL ARTICLE ASSIGNMENT - Refer to page 5 SCIENTIFIC POSTER ASSIGNMENT - Refer to pages 5-6 MODULE 5 QUIZ Visit your course page to view the details of these assignments. Note on Activities Throughout the Module: Note that text responses and interactions will not be graded unless otherwise notified. However, they are recorded in the module and viewable by your instructors. JOURNAL ARTICLE ASSIGNMENT Subpage of Introduction, Slide 4 of 5 – Journal Article Assignment 1/1 In your group from the previous Journal Article Assignment, the seminar Leader for this module will choose one of the provided articles to write a summary about and create a narrated PowerPoint presentation summarizing the most important aspects of the article as it relates to module content. The seminar Leader will share this summary and presentation with their group. The seminar Peer Reviewers will be responsible for reading the article summary, watching the narrated presentation, and providing constructive feedback using a provided rubric to grade the presentation. The seminar Leader will be given the opportunity to provide commentary on the feedback received. Visit your course page for more details. SCIENTIFIC POSTER ASSIGNMENT Subpage of Introduction, Slide 4 of 5 – Scientific Poster Assignment 1/1 INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 5 MODULE 05 COMPANION GUIDE MICR 270 The purpose of this assignment is to work in pairs to evaluate scientific research that is specific to an infectious disease and relate it your current understanding of the host immune mechanisms used to control the pathogen. Each group must create a scientific poster about their assigned disease with the goal of effectively teaching a peer about the immunological underpinnings of this disease. Once your poster is compiled, create and record 5 to 10-minute oral summary of your poster that guides the viewer through each section. This should be similar in style to if you were presenting at a poster fair and geared towards informing a peer in this course. MODULE OUTLINE This content was retrieved from Introduction, Slide 5 of 5 of the online learning module. MODULE OUTLINE Section 01: Immunological Techniques and Technologies Section 02: Vaccines Section 03: Cancer and the Immune System INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 6 MODULE 05 COMPANION GUIDE MICR 270 SECTION 01: IMMUNOLOGICAL TECHNIQUES AND TECHNOLOGIES INTRODUCTION TO IMMUNOLOGICAL TECHNIQUES This content was retrieved from Section 01, Slide 1 of 4 of the online learning module. In the field of immunology, multiple techniques and tools have been developed in order to understand, identify, and help fight infections and diseases. These techniques have been essential to our understanding of the local and systemic effects of disease in the body. Review a short introduction to the three techniques that will be covered in more detail throughout this section. ELISA Enzyme-linked immunosorbent assay, or ELISA, is a fundamental tool of clinical immunology based on the principle of antigen-antibody interaction. Flow Cytometry Flow cytometry is a method of detecting and quantifying different cell types in a mixed cell suspension. A flow cytometer analyzes cell suspensions and translates this information into datasets. Monoclonal Antibodies The production of monoclonal antibodies is a technique that was developed by Georges Kohler and César Milstein in 1975. Monoclonal antibodies are antibodies that are produced by a single clone of a B-cell that are specific for a single epitope*. Definition*: Epitope: The portion of the antigen that is recognized and bound by the antibody ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) This content was retrieved from Section 01, Slide 2 of 4 of the online learning module. An ELISA is a broadly-applicable technique which can be modified to detect and quantify substances such as peptides, proteins, antibodies, hormones, and other molecules. Learn more about ELISAs. Reflection Question - Refer to page 8 How does it work? - Refer to pages 8 What does it measure? - Refer to pages 8-9 Video of the Technique - Refer to page 9 INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 7 MODULE 05 COMPANION GUIDE MICR 270 QUESTION: ENZYME -LINKED IMMUNOSORBENT ASSAY (ELISA) Subpage of Section 01, Slide 2 of 4 – Reflection Question 1/1 Various types of ELISAs exist, however they are all very specific to the substance that needs to be detected or quantified. Using what you have learned in this course so far, why do you think an ELISA test is highly specific? Feedback: As mentioned on the introduction slide of this section, an ELISA is based on the principle of antigen-antibody interaction. Recall that the antibody binding site is very specific to one antigen (or epitope). ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) – HOW DOES IT WORK? Subpage of Section 01, Slide 2 of 4 – How does it work? 1/1 Learn the steps of an indirect ELISA. 1. The bottom of the wells are coated with an antigen that is specifically recognized by the antibody you wish to measure (referred to as the primary antibody in this procedure). The wells are washed to remove any excess antigen not attached to the bottom of the well. 2. The sample containing the antibody to be measured (for example, a patient's serum) is added to the well. The primary antibodies, if present, will bind to the antigens attached to the bottom of the well 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. 3. An enzyme-conjugated secondary antibody* is added to the well. This secondary antibody will bind to the F c portion of the primary antibodies already present in the well. The secondary antibody used specifically recognizes antibodies from a particular animal (e.g. anti-human, anti-rat, anti-rabbit, etc). The wells are washed to remove any excess secondary antibody not attached to the primary antibody. 4. The substrate of the enzyme attached to the secondary antibody is added to the well. The reaction of the substrate (a chromogen*) and the enzyme produces a coloured product which can be measured by absorbance. Definitions*: Enzyme-conjugated secondary antibody: A secondary antibody specifically binds to the primary antibody. In this example, the secondary antibody used has an enzyme attached to it. Chromogen: A substance that can be readily converted into a dye or other coloured compound. ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) – WHAT DOES IT MEASURE? INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 8 MODULE 05 COMPANION GUIDE MICR 270 Subpage of Section 01, Slide 2 of 4 – What does it measure? 1/1 An ELISA will measure a coloured reaction product by absorbance with the help of a machine called a spectrophotometric plate reader. The data measured correlates with the presence of an antibody or an antigen. This information can be used, for example, to detect the presence of a viral disease. Within the numerous variants of ELISA, we will focus on one type called the indirect ELISA. INDIRECT ELISA Detects or quantifies antibody. For example, to determine the presence of serum antibodies against HI V. Another type of ELISA. To compare and contrast with the indirect ELISA, there is another type called the sandwich ELISA which detects or quantifies an antigen. Note that this type of ELISA is explained in more detail in BCHM 270 - Biochemical Basis of Health and Disease. When conducting immunological research, most often one is interested in the level of antibodies present against a certain antigen, which is why the indirect ELISA is emphasized in this course. In contrast, in the field of biochemistry, the sandwich ELISA test is typically used to identify and measure molecules, which in this case are antigens. Detection vs. Quantification. Detection only confirms the presence or the absence of a substance, while quantification evaluates the concentration. To quantify a substance, one needs to develop a specific tool for each set of experiments called a standard curve, which represents the measurement of absorbance of known concentrations of the substance. The sample concentration can be determined by comparing the measured absorbance of the sample against the absorbance of the standard curve. VIDEO: ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) Subpage of Section 01, Slide 2 of 4 – Video of the Technique 1/1 Enzyme-Linked Immunosorbent Assay (ELISA) Watch this video to visualize the steps of the indirect ELISA. Keep your focus on the components used in the test and each of their roles. Page Link: https://www.youtube.com/embed/RRbuz3VQ100 FLOW CYTOMETRY This content was retrieved from Section 01, Slide 3 of 4 of the online learning module. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 9 MODULE 05 COMPANION GUIDE MICR 270 Flow cytometry is a technique designed to detect and quantify different immune cells in a mixed cell suspension. Learn more about flow cytometry. Reflection Question - Refer to page 10 How does it work? - Refer to pages 10-11 What does it measure? - Refer to page 11 Clinical Application - Refer to page 11 Video of the Technique - Refer to page 12 QUESTION: FLOW CYTOMETRY Subpage of Section 01, Slide 3 of 4 – Reflection Question 1/1 Flow cytometry is now an indispensable tool in immunology and cell biology. Using what you have learned so far in the course, how do you think flow cytometry can be used in the diagnosis of HIV/AIDS? Feedback: Recall that the diagnosis of AIDS is made based upon the number of CD4+ T-cells found a patient’s serum (below 200 cells/m m3). Flow cytometry can be used to identify and count these specific T-cells in a blood sample. FLOW CYTOMETRY – HOW DOES IT WORK? Subpage of Section 01, Slide 3 of 4 – How does it work? 1/1 In flow cytometry, a narrow stream of cells in single file is passed through a laser light source. The difference in wavelength of the light and the size of the cell scatters the light which can be detected and analyzed. The way 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. FSC intensity is proportional to the diameter of the cell. SSC provides information about the internal complexity (i.e. granularity) of a cell. Flow cytometry can also be used to determine the proportion of cells expressing a particular antigen. In this case, cells are labelled with a specific antibody. The antibody is coupled with a fluorescent marker (or ‘stained’). This fluorescent marker can be excited by a light of a specific wavelength. In turn, the fluorescent marker emits a light with a characteristic different wavelength. Only cells expressing the antibody in question will emit light of this specific wavelength. The proportion of cells with a particular antigen can also be measured using a flow cytometer. The cell suspension is treated with an antibody that recognizes the antigen and is coupled to a fluorescent molecule (also termed as ‘stained’). During flow cytometry, a laser light source is used that specifically INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 10 MODULE 05 COMPANION GUIDE MICR 270 excites the fluorescent molecule, causing light of a specific colour to be emitted only by those cells that have bound the antibody. FLOW CYTOMETRY – WHAT DOES IT MEASURE? Subpage of Section 01, Slide 3 of 4 – What does it measure? 1/1 Flow cytometry is used to measure physical properties of a cell. It can also be used to detect specific antigens on or inside a cell. As each cell in a mixed suspension is assessed as an individual detection event, the total number of cells in the suspension, the number of cells of a particular type in the suspension, and the overall composition of the suspension can be readily determined. Flow cytometers are used to determine complete blood counts (CBC), which you learned about in Module 4. By measuring size, shape and granularity, distinct cell types can be identified in a mixed cell suspension. CLINICAL APPLICATION - FLOW CYTOMETRY AND DISEASE DIAGNOSIS Subpage of Section 01, Slide 3 of 4 – Clinical Application 1/1 Flow cytometry can be used to diagnose cancer in a variety of ways. The main diagnostic tests focus on detecting DNA aneuploidy*, analyzing cell cycles and the immunophenotypical characterization. Dr. Graeme Quest is a Hematopathologist, Director of Transfusion Medicine & Immunology and an assistant professor at Queen’s University. He is also a Deputy Director of Flow Cytometry and works extensively with the technology to study diseases and disorders of the blood. Watch Dr. Quest explain how he uses flow cytometry in his clinical work. Definition*: Aneuploidy: The presence of an abnormal number of chromosomes. Page Links: https://player.vimeo.com/video/287704947 Reference: Orfao, A., Ciudad, J., Gonzalez, M., Lopez, A., del mar Abad, M., Paz Bouza, J. I.,... San Miguel, J. F. (1995). Flow cytometry in the diagnosis of cancer. Scandinavian Journal of Clinical & Laboratory Investigation, 55(S221), 145-152. doi:10.3109/00365519509090577 VIDEO: FLOW CYTOMETRY Subpage of Section 01, Slide 3 of 4 – Video of the Technique 1/1 INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 11 MODULE 05 COMPANION GUIDE MICR 270 To further your understanding of flow cytometry, watch a visualization of the theory and processes involved Keep your focus on the components of the technique and each of their roles and functions. Page Link: https://www.youtube.com/embed/EQXPJ7eeesQ MONOCLONAL ANTIBODIES This content was retrieved from Section 01, Slide 4 of 4 of the online learning module. The production of monoclonal antibodies is a technique designed to be used in research, diagnosis, and for therapeutic purposes. Learn more about monoclonal antibodies. Reflection Question - Refer to page 12 How does it work? - Refer to pages 12-13 What does it measure? - Refer to page 13 Clinical Application - Refer to pages 13-14 Video of the Technique - Refer to page 14 QUESTION: MONOCLONAL ANTIBODIES Subpage of Section 01, Slide 4 of 4 – Reflection Question 1/1 Monoclonal antibodies serve a variety of purposes in immunology and in many other scientific fields. Using what you have learned throughout the course so far, what do you think is the advantage of using monoclonal antibodies in contrast to polyclonal antibodies (which are derived from numerous different B-cells)? Feedback: One advantage of monoclonal antibodies over polyclonal antibodies is that they are a homogeneous population of antibodies with exquisite specificity for a single antigen whereas polyclonals could recognize multiple antigens. This property reduces, for example, the non-specific background staining that is often seen with polyclonal antibodies MONOCLONAL ANTIBODIES – HOW DOES IT WORK? Subpage of Section 01, Slide 4 of 4 – How does it work? 1/1 The development of a method to produce monoclonal antibodies proved to be an extremely useful and versatile technology. George Kohler and César Milan shared the 1984 Nobel Prize in Physiology or Medicine for their development of this technology alongside theoretical immunologist Niels Jerne. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 12 MODULE 05 COMPANION GUIDE MICR 270 Monoclonal antibodies are produced in the laboratory by hybridomas, immortal cells that produce unlimited quantities of one identical antibody. Hybridomas are the result of fusion between a plasma cell and a cancerous (or myeloma) cell. Hybridomas share properties of both plasma cells and myeloma cells. Learn about the properties of each cell. Myeloma cell Immortal growth; that is, divides indefinitely. Plasma Cell Produce specific antibodies against one antigen. Hybridoma cell A perpetual source of antibodies against one antigen. MONOCLONAL ANTIBODIES – WHAT DOES IT MEASURE? Subpage of Section 01, Slide 4 of 4 – What does it measure? 1/1 A monoclonal antibody is not a specific technique of measurement, rather it is a tool useful in many applications. Clinical applications of monoclonal antibodies include immunotoxins and radiolabelled antibodies. Learn more about each clinical application. Immunotoxins Immunotoxins consist of a tumour-specific monoclonal antibody attached to a deadly toxin. This technique is still under investigation, but a long-term objective is to use immunotoxins to target and eliminate tumour cells and treat cancer. Radiolabelled antibodies Monoclonal antibodies tagged with a radioactive isotope (radiolabelled) can be used to diagnose tumours earlier than other methods. Radiolabelled antibodies can bind to antigens on a tumour thereby allowing the precise location of a tumour within the body to be visualized. 3D representation of a radiolabelled antibody showing the four strands of polypeptide chains. CLINICAL APPLICATION – MONOCLONAL ANTIBODIES AND CANCER Subpage of Section 01, Slide 4 of 4 – Clinical Application 1/1 Since the development of monoclonal antibodies, the number of potential applications for this discovery keeps growing. Monoclonal antibodies can be produced for defence against specific diseases and even cancer. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 13 MODULE 05 COMPANION GUIDE MICR 270 Watch an explanation of how monoclonal antibodies can be used to enhance the activity of cytotoxic T- lymphocytes (CT L S) in targeting cancer. Page Link: https://www.youtube.com/embed/5AXApBbj1ps VIDEO: MONOCLONAL ANTIBODIES Subpage of Section 01, Slide 4 of 4 – Video of the Technique 1/1 Monoclonal Antibodies To further your understanding of monoclonal antibodies, watch the video to visualize how they are produced. Keep your focus on what makes a hybridoma and how monoclonal antibodies are produced. Page Link: https://www.youtube.com/embed/uuT08OT3wTc INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 14 MODULE 05 COMPANION GUIDE MICR 270 SECTION 02: VACCINES QUESTION: TYPES OF IMMUNITY This content was retrieved from Section 02, Slide 1 of 19 of the online learning module. Immunity is the ability of an organism to resist a particular infection through the action of specific antibodies or sensitized white blood cells. Immunity can also be described as the state of being insusceptible to a particular disease. Recall that you learned about the four major types of immunity in Module 3 in the context of their ability to generate immunological memory. Associate the type of immunity to its correct description by using the dropdown menu. Options: Acquired by infection by a pathogen, possibly leading to symptoms/a disease state, Acquired by injection of serum containing antibodies, Acquired by vaccination, Acquired from placental transfer of antibodies during pregnancy or breastfeeding Natural Passive Immunity Artificial Passive Immunity Natural Active Immunity Artificial Active Immunity Feedback: Natural Passive Immunity Acquired from placental transfer of antibodies during pregnancy or breastfeeding Artificial Passive Immunity Acquired by injection of serum containing antibodies Natural Active Immunity Acquired by infection by a pathogen, possibly leading to symptoms/a disease state Artificial Active Immunity Acquired by vaccination QUESTION: VACCINES This content was retrieved from Section 02, Slide 2 of 19 of the online learning module. Answer the following question about vaccination. Explain why vaccines are preventative in protecting against infection? Feedback: Vaccines are made up of fragments of the pathogen or the entire pathogen that is modified (killed or attenuated), allowing the immune response to develop without contracting the disease. This results in the formation of an immune response against the pathogen without exposure to the pathogen, so that when the vaccinated person encounters the natural pathogen for the first time, their immune system is already ‘trained’ to act against that pathogen. Vaccination allows the immune system the opportunity for ‘target practice’. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 15 MODULE 05 COMPANION GUIDE MICR 270 TYPES OF VACCINES This content was retrieved from Section 02, Slide 3 of 19 of the online learning module. A vaccine is a type of biological preparation which provides active artificial immunity to a particular disease-causing agent. The type of vaccine is largely dependent on the nature of the disease-causing agent and how the immune system recognizes and responds to the infection. We will focus on four types of vaccines that currently exist. Learn more about these types of vaccine. Live-attenuated vaccine - Refer to pages 16-17 Killed-inactivated vaccine - Refer to page 17 Toxoid vaccine - Refer to pages 17-18 Subunit vaccine - Refer to page 18 Unfortunately there is no magic formula for making an effective vaccine. Different types of vaccines have been developed to counter the varied characteristics of pathogens and the ways the immune system recognizes and responds to subsequent infection. The objective with a vaccine is to develop an effective immune response and produce immunological memory so the body will be able to counter the fully active and virulent (infectious) form of the disease-causing agent. LIVE-ATTENUATED VACCINE Subpage of Section 02, Slide 3 of 19 – Live-attenuated vaccine 1/1 Characteristics Contains a modified strain of the disease-causing agent which has lost its pathogenic* ability, but retains its capacity to replicate within the host. Examples Smallpox vaccine Oral poliovirus vaccine (Sabin) Measles vaccine Advantages and Disadvantages ADVANTAGES Provides a prolonged exposure to the disease-causing agent, and is suitable to generate cell- mediated immunity. DISADVANTAGES INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 16 MODULE 05 COMPANION GUIDE MICR 270 Potential to revert to a virulent form. Requires specific storage and transport conditions (e.g. refrigeration). Definition*: Pathogenic: Ability of an organism to harm the host by causing a disease. KILLED-INACTIVATED VACCINE Subpage of Section 02, Slide 3 of 19 – Killed-inactivated vaccine 1/1 Characteristics Contains a strain of the disease-causing agent that has been inactivated by heat, chemicals, or radiation. It has the ability to generate an immune response, but it is unable to replicate. Examples Rabies vaccine Flu (influenza) vaccine Advantages and Disadvantages ADVANTAGES Safer option as it cannot mutate back to a virulent form. Easy to store and transport. DISADVANTAGES Generally requires multiple booster doses* to maintain immunity. Generally must be administered by injection Definition*: Booster doses: Extra administration of a vaccine after a primary dose has been given. TOXOID VACCINE Subpage of Section 02, Slide 3 of 19 – Toxoid vaccine 1/1 The tetanus vaccine is an example of a toxoid vaccine. Learn about this type of vaccine. Characteristics Contains an inactivated toxin which is a product from the pathogen that is causing the disease. Examples Tetanus vaccine INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 17 MODULE 05 COMPANION GUIDE MICR 270 Diphtheria vaccine Advantages and Disadvantages ADVANTAGES Safe as it is not a living organism that can divide, spread and/or revert. Stable as they are less susceptible to changes in temperature, humidity, and light. DISADVANTAGE May require several doses and usually need an adjuvant*. Definition*: Adjuvant: A substance that enhances the body’s immune response to an antigen. SUBUNIT VACCINE Subpage of Section 02, Slide 3 of 19 – Subunit vaccine 1/1 The hepatitis B vaccine is an example of a subunit vaccine. Learn about this type of vaccine. Characteristics Contains only a small part or fragment of the disease-causing agent. Examples Hepatitis B vaccine Advantages and Disadvantages ADVANTAGES The safest type of vaccine - can be used on everyone, including immunocompromised, pregnant, and elderly populations. DISADVANTAGE Rarely successful at inducing long-lasting immunity, which means it will require multiple booster doses to maintain immunity and might even need to be conjugated to a carrier*. Definition*: Carrier: A carrier is 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, the immunological response to the poor antigen is also improved. mRNA VACCINES INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 18 MODULE 05 COMPANION GUIDE MICR 270 This content was retrieved from Section 02, Slide 4 of 19 of the online learning module. mRNA vaccines are the most recent vaccine type that have changed the field of Vaccinology. They are known for their use against SARS-CoV-2, the virus causing COVID-19, and are used in several formulations to fight the virus (e.g., bivalent vaccine, boosters). Formulations of mRNA vaccines are also being investigated for other infectious diseases (e.g., HIV, influenza) and non-infectious clinical conditions (e.g., pancreatic cancer, heart tissue regeneration). The principle of the assay relies on the use of mRNA to produce viral proteins and recruit immune cells to respond to the antigenic target. The proteins are then displayed on the surface of an antigen presenting cells to induce B-cells (i.e., antibodies) and T-cell immunity. You will learn about the mechanism of the mRNA vaccine on the next slide. mRNA VACCINE MECHANISM This content was retrieved from Section 02, Slide 5 of 19 of the online learning module. RNA -and DNA -based vaccines involve making genetic material only; they do not require the use of the whole virus. The mRNA vaccine for COVID-19 uses lab-made SARS- CoV-2 messenger RNA (mRNA) to trigger an immune response. Learn about the mechanism behind the mRNA vaccine for COVID-19. 1. Vaccine Production mRNA is made in the lab from a DNA template of the virus. The mRNA encodes an antigen of the virus. For the COVID-19 vaccine, 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. 2. Host Cell Once inside the body, the mRNA enters the host cell and uses host cell machinery to produce the spike protein. 3. APC The newly formed spike protein exits the cell and is recognized by an antigen presenting cell (APC). The APC internalizes the spike protein and processes it into a peptide (or antigen). The APC then displays the antigen on the surface of the cell via the major histocompatibility complex (MHC). 4. Immune Response The antigen is recognized by a T-helper cell, which initiates an immune response. B cells produce antibodies that stop the virus from infecting cells. T cells destroys cells infected with the virus. ANTIVIRALS MEDICATIONS AGAINST COVID -19 This content was retrieved from Section 02, Slide 6 of 19 of the online learning module. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 19 MODULE 05 COMPANION GUIDE MICR 270 Once you’ve already been infected with the SARS-CoV-2 virus, it can be treated with antiviral medication. This medication is taken orally and does not prevent COVID-19 (like the vaccine does), it only treats it once you are already positive. There are two main antiviral medications used to treat COVID-19. Learn about these two antiviral medications. Polymerase Inhibitor The first antiviral medication is polymerase inhibitors. Polymerase is an enzyme that plays a central role in viral replication and transcription. Molnupiravir is a polymerase inhibitor used to treat COVID-19. It increases the frequency of viral RNA mutations and impairs replication of the virus. Protease Inhibitor The second antiviral medications involves a protease inhibitor. Proteases cut proteins into smaller, more workable pieces. Protease inhibitors are often administered in combination. For example, nirmatrelvir stops protease from cutting viral proteins into functional pieces. Ritonavir protects nirmatrelvir from destruction by the body and allows it to keep working. Since these drugs disrupt the assembly of the virus, they can’t replicate and infect other cells. EVOLUTION OF VACCINES This content was retrieved from Section 02, Slide 7 of 19 of the online learning module. Vaccines, recognized by the British Medical Journal as one of the greatest medical advances in history, have saved hundreds of millions of lives since their introduction. One of the greatest vaccine success stories, which you learned about in Module 1, was Dr. Edward Jenner’s use of extracts prepared from cowpox lesions as a means of protecting his patients against smallpox infection. The term vaccination is, in fact, derived from the Latin word for cow (vacca) and was first coined by Louis Pasteur in 1881 to honour Jenner’s accomplishment. Remarkably, Jenner’s discovery of using vaccination for protection against a viral disease came nearly 100 years prior to the discovery of viruses by Dmitry Ivanovsky in 1892. Watch a short video about the evolution of vaccine research. Page Link: https://www.youtube.com/embed/exxmQO5z8U8 A MODERN-DAY VACCINE SUCCESS STORY: HPV VLP VACCINES This content was retrieved from Section 02, Slide 8 of 19 of the online learning module. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 20 MODULE 05 COMPANION GUIDE MICR 270 Cervical cancer and genital warts are caused by prior infection with certain types of human papillomavirus (HPV). In 2002, a landmark clinical trial demonstrated that a virus like particle (VLP) vaccine could protect against infection by a type of HPV often associated with cervical cancer. The efficacy* reported in this clinical trial was 100% and subsequent larger scale clinical trials continued to report efficacies approaching 100%. VLPs are composed of the structural proteins of HPV, which can self assemble into particles that resemble the natural virus both structurally and immunologically. As they do not contain viral DNA , VLP s are not infectious. There are currently three HPV vaccines authorized for use in Canada: Cervarix, Gardasil, and Gardasil 9 (9 refers to the fact that this vaccine protects against 9 different types of HPV - some of which are the causative agents of other cancers [penile, neck, etc.]). How safe are HPV VLP vaccines? The Centers for Disease Control (CDC) in the U.S. continually monitors vaccine safety. Their monitoring of HPV VLP vaccines indicates that they are very safe. Consider this early post-licensure safety data gathered by the CDC for Gardasil and reported by the World Health Organization: of the approximately 40 million doses of vaccine distributed in the U.S. there were 20,096 adverse event reports, 92% of which were classified as non-serious. Review the information sheet about papilloma virus vaccine. If you want to know more For more information about HPV vaccines view information provided by the Public Health Agency of Canada. Definition*: Efficacy: With respect to vaccines, efficacy is a measure of the ability of a vaccine to reach a defined endpoint, generally the prevention of disease development. An efficacy of 100% means that all subjects enrolled in a clinical trial were protected from disease development. Page Links: https://www.who.int/vaccine_safety/initiative/tools/HPV_vaccine_rates_information_sheet_1217.pdf https://www.canada.ca/en/public-health/services/infectious-diseases/sexual-health-sexually- transmitted-infections/hpv-prevention-vaccines-questions-answers.html A MODERN-DAY VACCINE SUCCESS STORY: EBOLA VACCINES This content was retrieved from Section 02, Slide 9 of 19 of the online learning module. Ebola virus (EBOV) and Marburg virus (MARV) are part of the filoviridae family of viruses that cause hemorrhagic fever with high mortality rates in humans and nonhuman primates. These virus produces transmembrane glycoproteins (GPs) thought to play a role in the virulence of these viruses. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 21 MODULE 05 COMPANION GUIDE MICR 270 In the early 2000s researchers in Winnipeg, Canada inadvertently discovered that mice infected with EB OV and MARV glycoproteins became protected against infection from the live virus. Researchers then developed a potentially viable vaccine using glycoproteins from EBOV and putting then into a live attenuated recombinant vesicular stomatitis virus (VSV), that expresses the transmembrane glycoproteins of EBOV and MARV. In 2005, the Canadian research team tested this vaccine on Macaques (old world monkeys) and found it to be 100% effective against EBOV infection. This vaccine was recently (May 2018) shipped to Kinshasa in the Democratic Republic of Congo to curb an emerging Ebola outbreak. Unlike the outbreak in 2016 which killed 11,000 people, the vaccine prevented the spread of EBOV in the DRC in 2018. References: http://www.sciencemag.org/sites/default/files/styles/article_main_large/public/images/si- vsvebolajpg_0.jpg?itok=3JyWF9mc&c=d5c762b61c41d41483f1c3d7e4a23004 https://globalnews.ca/news/4229827/ebola-vaccine-canada-invention/ Feldmann, H., Daddario, K. M., Geisbert, J. B., Klenk, H., Jahrling, P. B., Ströher, U.,... Grolla, A. (2005). Live attenuated recombinant vaccine protects nonhuman primates against ebola and marburg viruses. Nature Medicine, 11(7), 786-790. doi:10.1038/nm1258 https://www.cbc.ca/news/health/second-opinion-ebola-vaccine-1.4672807 A MODERN-DAY VACCINE FAILURE STORY: GENITAL HERPES VACCINE This content was retrieved from Section 02, Slide 10 of 19 of the online learning module. Genital herpes is caused by infection with herpes simplex virus (HSV) types 1 or 2. Genital herpes infections are permanent, requiring lifelong management, pose a serious health threat to newborns through vertical transfer*, cause significant emotional distress in infected individuals and increase susceptibility to, and spread of, HIV infection. An effective vaccine against genital herpes is, therefore, needed. A subunit vaccine was developed which was composed of a prominent structural protein of HSV-2 (the type most commonly associated with genital infections). This vaccine showed promise in early clinical trials, with reported efficacies of >70% in women who were seronegative* for both HSV-1 and HSV-2 at the beginning of the trial. A subsequent larger clinical trial involving 8,323 seronegative women, however, showed much lower efficacy (20%) and, curiously, no protection from infection by HSV-2. This vaccine failure has caused 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. How prevalent are HSV-2 infections? Estimates of the number of people with prevalent HSV-2 infection in 2012. These values are likely underestimates as HSV-2 infections are often asymptomatic. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 22 MODULE 05 COMPANION GUIDE MICR 270 Definitions*: Vertical Transfer: Vertical transmission 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. This informs you of three things: 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 the host has been vaccinated. Reference: Image adapted from Looker et al. (2015). https://doi.org/10.1371/journal.pone.0114989. PHASES OF VACCINE DEVELOPMENT This content was retrieved from Section 02, Slide 11 of 19 of the online learning module. The development of new vaccines, similar to the development of new drugs (covered in PHAR 100), requires adherence to a strict and rigorous quality control process before being approved for use in Canada. Learn about each stage of vaccine development. Lab Studies - Refer to pages 23-24 Preclinical - Refer to page 24 Clinical Phase I - Refer to page 24 Clinical Phase II - Refer to page 24 Clinical Phase III - Refer to pages 24-25 Health Canada Approval - Refer to page 25 LAB STUDIES Subpage of Section 02, Slide 11 of 19 – Lab Studies 1/1 The first step in the process of vaccine development is to identify the infectious agent causing the disease and select a strain (or subtype) which is relevant to the target population and will be used to produce the vaccine. This stage is largely dependent on research carried out in the laboratory utilizing assays, which may involve exhaustive screening to identify a suitable antigen and creation of a vaccine concept. This stage also involves developing and testing the manufacturing process of the vaccine according to Good Manufacturing Practice standards. For interest, read about Good Manufacturing Practices provided by the Government of Canada. INFECTION, IMMUNITY, AND INFLAMMATION | MICR 270 M05 PAGE 23 MODULE 05 COMPANION GUIDE MICR 270 Page Link: https://www.canada.ca/en/health-canada/services/drugs-health-products/compliance- enforcement/good-manufacturing-practices.html PRECLINICAL STUDIES Subpage of Section 02, Slide 11 of 19 – Preclinical 1/1 Preclinical studies involve research carried out in animal models to evaluate the pharmacological aspects of the product. This is a stage of research that occurs before clinical trials can begin. At this point, the researchers will carry out challenge studies to demonstrate the immunogenicity of the vaccine in animal models. “Immunogenicity” means that the vaccine has the ability to induce an immune response which will prevent the development of the disease in case of subsequent infection with the pathogen. Another part of this stage is to carry out safety studies to evaluate the possible toxicity of the vaccine which would prevent its use in humans. CLINICAL PHASE I Subpage of Section 02, Slide 11 of 19 – Clinical Phase I 1/1 Due to rigorous regulatory testing requirements, only a very small percentage of vaccines progress to licensing, making the costs of vaccine research and development extremely high. Clinical development involves three phases spanning over several years. You learned about the general steps of a clinical trial in PHAR 100, however this course will discuss clinical trials in the context of vaccine development. Phase I clinical trials for vaccines involve small-scale trials in humans (10 -