MLAB 1235 - Immunology/Serology

After studying all materials and resources presented in the course, the student will be able to:

Syllabus Objectives for Immunology (MLAB1235.401):

Chapter One Objectives:

1. Discuss how immunology as a science began with the study of immunity.
2. Describe what is meant by an attenuated vaccine.
3. Explain how the controversy over humoral versus cellular immunity contributed to expanding knowledge in the field of immunology.
4. Contrast innate and adaptive immunity.
5. Describe the types of white blood cells (WBCs) capable of phagocytosis.
6. Discuss the roles of macrophages, mast cells, and dendritic cells in the immune system.
7. Identify the two primary lymphoid organs and discuss their overall importance to immunity.
8. List four secondary lymphoid organs and discuss their overall importance to immunity.
9. Describe the function and architecture of a lymph node.
10. Compare a primary and a secondary follicle.
11. Define “cluster of differentiation” (CD).
12. Differentiate the roles of T cells and B cells in the immune response.
13. Discuss how natural killer (NK) cells differ from T lymphocytes.

Chapter Two Objectives:

1. Differentiate between the external and internal defense systems.
2. Give examples of several external defense mechanisms.
3. Describe how normal flora act as a defense against pathogens.
4. Define “pathogen-associated molecular pattern” (PAMP) and provide some examples.
5. Discuss the role of pattern recognition receptors (PRRs) in both the innate and adaptive immune responses.
6. Describe the function of Toll-like receptors (TLRs) in the immune system.
7. Discuss the role of acute-phase reactants in the innate immune system.
8. Explain how each of the following acute-phase reactants contributes to innate immunity: C-reactive protein (CRP), serum amyloid A (SAA), complement, alpha₁- antitrypsin (AAT), haptoglobin, fibrinogen, and ceruloplasmin.
9. Determine the significance of abnormal levels of acute-phase reactants.
10. Describe the process of inflammation.
11. List the steps in the process of phagocytosis.
12. Discuss the intracellular mechanism for destruction of foreign particles during the process of phagocytosis.
13. Explain the importance of phagocytosis in both innate and adaptive immunity.
14. Explain how natural killer (NK) cells recognize target cells.
15. Describe two methods that NK cells use to kill target cells.
16. Discuss the defining characteristics and functions on innate lymphoid cells (ILCs).

Chapter Three Objectives:

1. Define and characterize the nature of immunoglobulins.
2. Differentiate an immunogen from an antigen.
3. Discuss several biological properties of individuals that influence the nature of the immune response.
4. Descript four important traits of immunogens that affect their ability to stimulate a host response.
5. Define haptens and describe some of their characteristics.
6. Describe the relationship between the epitope and an antigen.
7. Discuss the functions of adjuvants.
8. Differentiate heterophile antigens from alloantigens and autoantigens.
9. Explain what a haplotype is in regard to inheritance of major histocompatibility complex (MHC) antigens.
10. Describe the differences in the structure of class I and class II MHC molecules.
11. Contrast the transport of antigen to cell surfaces by class I and class II MHC molecules.
12. Describe the role of transporters associated with antigen processing (TAPs) in selecting peptides for binding to class I MHC molecules.
13. Discuss the differences in the sources and types of antigens processed by class I and class II MHC molecules.
14. Explain the clinical significance of the class I and class II MHC molecules.

Chapter Four Objectives:

1. Compare and contrast adaptive immunity and innate immunity.
2. Discuss the role of the thymus in T-cell maturation.
3. Describe the T-cell receptor (TCR) for antigen.
4. Explain how positive and negative selection contribute to T-cell development.
5. List and describe five different subsets of CD4 T helper (Th) cells.
6. Describe the maturation of a B cell from the pro–B-cell stage to the plasma cell stage.
7. Contrast the antigen-independent and antigen-dependent phases of B-cell development.
8. Explain how cytotoxic T cells recognize and kill target cells.
9. Discuss the role of class I major histocompatibility complex (MHC) and class II MHC molecules in the presentation of antigens to T cells.
10. Compare and contrast the immune response to T-dependent antigens with the response to T-independent antigens.
11. Discuss how Th cells influence the B-cell antibody response.
12. Explain the importance of T cells and B memory cells to the adaptive immune response.
13. Indicate surface markers characteristic of T and B lymphocytes in various stages of development.

Chapter Five Objectives:

1. Diagram the general structure of an immunoglobulin and recognize its major components.
2. Identify the electrophoretic fraction of serum that contains the majority of immunoglobulins.
3. Differentiate between isotypes, allotypes, and idiotypes.
4. Differentiate between the light chains and heavy chains of immunoglobulins and indicate the Greek letters that denote each type.
5. Discuss the effects of treating an immunoglobulin molecule with papain, pepsin, or mercaptoethanol.
6. Describe the major characteristics of the five immunoglobulin classes found in humans.
7. Relate the differences in the structures of the five immunoglobulin classes to their functions.
8. Discuss how the immunoglobulin G (IgG) subclasses differ in functional capability.
9. Describe the functions of the J chain and the secretory component (SC) and indicate in which immunoglobulin classes they are found.
10. Discuss how immunoglobulin D (IgD) differs from other immunoglobulin types.
11. Identify the types of cells that immunoglobulin E (IgE) binds to in allergic reactions.
12. Compare and contrast the primary and secondary antibody responses to antigen.
13. Describe the genes that code for the immunoglobulin proteins, and explain how they combine to code for a unique antibody molecule.
14. Explain how immunoglobulin class switching occurs on a genetic level.
15. Explain how the clonal selection hypothesis contributes to antibody specificity.
16. Outline the traditional process of mouse monoclonal antibody production.
17. Discuss some clinical and research applications of monoclonal antibodies.

Chapter Six Objectives:

1. Define cytokine.
2. Define and describe the term cytokine storm and relate its medical importance.
3. Distinguish between autocrine, paracrine, and endocrine effects of cytokines.
4. Define pleiotropy as it relates to cytokine activities.
5. Explain the functions of interleukin-1 (IL-1) in mediating the immune response.
6. Explain the effects of tumor necrosis factors (TNFs).
7. Discuss how interleukin-6 (IL-6) affects inflammation and other activities of the immune system.
8. Determine the role of chemokines in the chemotaxis of white blood cells (WBCs).
9. Compare the functions of type 1 and type 2 interferons (IFNs).
10. Describe the actions of interleukin-2 (IL-2) on its target cells.
11. Discuss the biological roles of the hematopoietic growth factors.
12. Discuss cytokines involved in differentiation of T helper (Th) cell subpopulations: Th1, Th2, Th17, and T regulatory (Treg).
13. Explain the biological role of colony-stimulating factors (CSFs).
14. Describe current types of anti-cytokine therapies.
15. Describe clinical assays for cytokines.

Chapter Seven Objectives:

1. Describe the roles of the complement system.
2. Differentiate between the classical, alternative, and lectin pathways and indicate the proteins and activators involved in each.
3. Discuss the formation of the three principal units of the classical pathway: recognition, activation, and membrane attack units.
4. Explain how C3 plays a key role in all three pathways of complement activation.
5. Discuss regulators of the complement system and their roles in inhibiting the complement pathways.
6. Describe deficiencies of complement components and the diseases they cause.
7. Describe potential disease manifestations of improper complement regulation.
8. Differentiate tests for functional activity of complement from measurement of individual complement components.
9. Analyze laboratory findings to indicate disease implications in relation to complement abnormalities.
10. Discuss clinical applications of complement therapeutics and their effects on laboratory tests for complement.

Chapter 8 Objectives: We are skipping this chapter!

Chapter 9 Objectives:

1. Describe how whole blood is processed to obtain serum for serological testing.
2. Explain the difference between a volumetric pipette and a graduated pipette.
3. Define the following: serial dilution, solute, diluent, and compound dilution.
4. Describe how accurate measurements are made using volumetric pipettes and graduated pipettes.
5. Given the volumes of diluent and solute, calculate the dilution obtained.
6. Calculate the amount of diluent or solute needed to prepare a specific dilution of a serum sample.
7. Explain how an antibody titer is determined.
8. Calculate the dilution factor required to change a given dilution to a specified new dilution.
9. Given a starting dilution and a dilution factor, determine the new dilution obtained.
10. Calculate the final dilution of a sample, given the initial dilution and all subsequent dilutions.
11. Determine how to make a specific percentage solution from a concentrate.
12. Calculate the diagnostic sensitivity and specificity of a serological test, and discuss the significance of these parameters.
13. Discuss the clinical significance of positive and negative predictive values.

Chapter 10 Objectives:

1. Discuss affinity and avidity and their influence on antigen–antibody reactions.
2. Describe how the law of mass action relates to antigen–antibody binding.
3. Define precipitation and agglutination and differentiate between the two types of reactions.
4. Describe the relative concentrations of antigen and antibody in the three zones of the precipitin curve and discuss why optimal precipitation occurs in the zone of equivalence.
5. Differentiate between immunoturbidimetry and nephelometry and discuss the role of each in the measurement of precipitation reactions.
6. Compare single diffusion to double diffusion.
7. Summarize the principle of the endpoint method of radial immunodiffusion (RID).
8. Determine the relationship between two antigens by looking at the pattern of precipitation resulting from Ouchterlony immunodiffusion.
9. Describe the principle of immunofixation electrophoresis (IFE).
10. Recognize how immunoglobulin M (IgM) and immunoglobulin G (IgG) differ in their ability to participate in agglutination reactions.
11. Describe physiological conditions that can be altered to enhance agglutination.
12. Define and give an example of each of the following: (a) direct agglutination, (b) passive agglutination, (c) reverse passive agglutination, (d) agglutination inhibition, and (e) hemagglutination inhibition.
13. Describe the principle of measurement used in particle-enhanced turbidimetric inhibition immunoassay (PETINIA).
14. Identify conditions that must be met for optimal results in agglutination testing.

Chapter 11 Objectives:

1. Describe the difference between competitive and noncompetitive immunoassays.
2. Distinguish between heterogeneous and homogeneous immunoassays.
3. Describe major characteristics of colorimetric, chemiluminescent, radioactive, and fluorescent labels.
4. Explain the principle of competitive binding in immunoassay design.
5. Recognize common enzymes used in enzyme immunoassays (EIAs).
6. Explain the principle of biotin-streptavidin labeling and its applications in immunoassays.
7. Outline the steps of a noncompetitive indirect enzyme-linked immunosorbent assay (ELISA) to detect an antibody in a patient sample.
8. Outline the steps of a sandwich, capture, or immunometric assay to detect an antigen in a patient sample.
9. Discuss clinical applications for homogeneous EIAs.
10. Describe basic principles and clinical uses for rapid immunoassays.
11. Compare and contrast the analytical sensitivity of immunoassays using different labels.
12. Describe the difference between direct and indirect immunofluorescence techniques.
13. State the principles of the enzyme-multiplied immunoassay technique (EMIT), cloned enzyme donor immunoassay (CEDIA), multiplex immunoassay (MIA), and fluorescence polarization immunoassay (FPIA).
14. Review advantages and disadvantages of immunoassays, based on each method design.
15. Choose an appropriate immunoassay for a particular analyte.
16. Discuss interfering factors and other technical concerns that may contribute to false-positive or false-negative immunoassay results.

Chapter 12 Objectives

1. Know the characteristics and differences of DNA and RNA.
2. Define a mutation.
3. Explain the basics of electrophoresis.
4. Know uses of electrophoresis.
5. Explain the basics of PCR.

Chapter 13 Objectives: We are skipping this chapter!

Chapter 14 Objectives:

1. Explain the concept of hypersensitivity.
2. Differentiate between the four types of hypersensitivity reactions in terms of antibody involvement, complement involvement, antigen triggers, and timing of the response.
3. Associate specific examples of clinical manifestations with each type of hypersensitivity.
4. Discuss the immunologic mechanisms involved in each of the four types of hypersensitivity reactions.
5. Provide examples of preformed and newly synthesized mediators released from immunoglobulin E (IgE)-sensitized mast cells and basophils and discuss their effects.
6. Discuss the influence of genetic and environmental factors on susceptibility to type I hypersensitivity responses.
7. Discuss the types of reactions that can result from latex sensitivity and their clinical manifestations.
8. Explain the underlying mechanisms of pharmacological therapy, monoclonal anti-IgE therapy, and allergy immunotherapy (AIT) in the treatment of allergies.
9. Discuss the procedure, clinical applications, advantages, and limitations of skin testing for type I hypersensitivity.
10. Discuss the principles and clinical applications of allergen-specific and total-IgE testing.
11. Explain how hemolytic disease of the fetus and newborn (HDFN) arises.
12. Explain the principles and clinical uses of the direct antiglobulin test (DAT) and the indirect antiglobulin test (IAT).
13. Discuss the principle of cold agglutinins testing and associate the presence of a positive result with specific disorders.
14. Discuss how skin testing for delayed hypersensitivity is performed, its clinical applications, and how to interpret the results.
15. Explain the principle of interferon gamma release assays (IGRA) and as well as their clinical utility, advantages, and limitations.

Chapter 15 Objectives:

1. Explain the mechanisms of central and peripheral tolerance that are essential in preventing the development of autoimmunity.
2. Discuss genetic and environmental factors that are thought to contribute to the development of autoimmunity.
3. Explain the relationship between microbial infections and the development of autoimmune disease.
4. Distinguish between organ-specific and systemic autoimmune diseases, providing examples of each and their associated target tissues.
5. Discuss the immunopathology and clinical manifestations of each of the following diseases: systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), granulomatosis with polyangiitis (Wegener’s granulomatosis), Graves’ disease, Hashimoto’s thyroiditis, type 1 diabetes mellitus, celiac disease, autoimmune hepatitis (AIH), primary biliary cirrhosis, multiple sclerosis (MS), myasthenia gravis (MG), and anti-glomerular basement membrane disease.
6. Associate each of the diseases listed in Learning Outcome 5 with its corresponding autoantibodies and laboratory findings.
7. Identify autoantibodies associated with Sjogren’s syndrome, systemic sclerosis (SSc), mixed connective tissue disease (MCTD), and the inflammatory myopathies.
8. Explain the principles of laboratory methods used to screen for and confirm the presence of anti-nuclear antibodies (ANAs).
9. Describe common immunofluorescence patterns seen in the indirect immunofluorescence (IIF) test for ANAs and their clinical significance.
10. Describe the c-ANCA and p-ANCA patterns seen in the IIF test for ANCAs and their clinical significance.
11. Discuss the composition and clinical significance of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP).

Chapter 16 Objectives:

1. List the histocompatibility systems relevant to clinical transplantation.
2. Compare the mechanisms of direct and indirect alloantigen recognition.
3. Distinguish between allograft, autograft, xenograft, and syngeneic graft (isograft).
4. Compare the immunologic mechanisms involved in hyperacute, acute, and chronic graft rejection.
5. Identify risk factors for graft-versus-host disease (GVHD) and the types of grafts in which this mechanism of rejection could occur.
6. List the major classes of immunosuppressive agents and their effects on the immune system.
7. Explain the principles of laboratory methods for human leukocyte antigen (HLA) typing.
8. Describe laboratory methods for detecting and identifying HLA antibodies (i.e., antibody screening, identification, and crossmatching).
9. Identify common tests used to monitor patients post-transplant.
10. Deduce the suitability of a possible donor for a transplant recipient, based on results of HLA typing and antibody identification.
11. Describe the nomenclature used for HLA antigens and alleles.

Chapter 17 Objectives:

1. Describe the characteristics that differentiate cancer cells from normal cells and the process by which malignant cells are thought to develop.
2. Differentiate between various types of tumor antigens and recognize examples of each.
3. Summarize the uses of tumor markers in screening for cancer, diagnosing malignancy, detecting prognosis, and monitoring patient responses to treatment.
4. Identify the characteristics that should be possessed by an ideal tumor marker and explain how nonideal features can affect the clinical utility of a marker.
5. Explain the principles of immunohistochemistry as they apply to tumor marker detection.
6. Distinguish the clinical applications of each of the following tumor markers: alpha-fetoprotein (AFP), cancer antigen 125 (CA125), carcinoembryonic antigen (CEA), human chorionic gonadotropin (hCG), and prostate-specific antigen (PSA).
7. Contrast the advantages and limitation of immunoassays for tumor markers.
8. Describe the innate and adaptive immune responses that play a role in defense against tumors and how they contribute to immunosurveillance.
9. Discuss the process of immunoediting and how this relates to mechanisms of tumor escape from the immune system.
10. Cite the overall goal of immunotherapy and provide the specific examples of active, passive, and adoptive immunotherapy for cancer.
11. Discuss principles and applications of molecular tests for cancer diagnosis.

Chapter 18 Objectives:

1. Differentiate leukemias, lymphomas, and plasma-cell dyscrasias, and provide examples of each type of malignancy.
2. Describe some of the cellular properties and genetic changes that occur during malignant transformation of hematologic cells.
3. Differentiate between Hodgkin lymphomas and various types of non-Hodgkin lymphomas in terms of their cellular origin and characteristic surface markers.
4. Associate specific CD markers with selected hematologic malignancies.
5. Correlate clinical manifestations and laboratory results with multiple myeloma.
6. Specify the ways in which laboratory tests can be used to diagnose and follow the progression of immunoproliferative disorders.

Chapter 19 Objectives:

1. Differentiate between primary immunodeficiency diseases and secondary immunodeficiency diseases
2. Indicate the general immunologic defects associated with each of the nine categories of primary immunodeficiency diseases.
3. Associate examples of specific immunodeficiencies with each category.
4. Describe the types of infections typically associated with defects in the B-cell, T-cell, myeloid, or complement systems.
5. Recognize the association between immunodeficiency states and the risk of developing malignancy.
6. Explain the immunologic defects and clinical manifestations associated with selected primary immunodeficiency diseases.
7. Select appropriate laboratory tests to screen for and confirm the presence of specific congenital immunodeficiencies.
8. Correlate laboratory results with the presence of different types of primary immunodeficiency diseases.

Chapter 20 Objectives:

1. Differentiate between commensalistic, mutualistic, and parasitic relationships.
2. Differentiate between pathogenicity, virulence, and infectivity.
3. Cite structural features of bacteria that contribute to increased virulence.
4. Describe host defenses against bacteria and the means by which bacteria can evade the immune system.
5. Compare and contrast endotoxins and exotoxins with respect to biological activity, immunogenicity, and the genetic encoding for the production of the two toxin categories.
6. Cite the five general laboratory means of detecting the causative agent of a bacterial infection.
7. Explain the principle of lateral flow immunochromatographic assays (LFA).
8. List the exotoxins produced by Group A streptococci and the roles they play in contributing to the virulence of Streptococcus pyogenes.
9. Describe the symptoms and pathogenesis of acute rheumatic fever and poststreptococcal glomerulonephritis.
10. Explain the principle, interpretation, and clinical significance of the anti-streptolysin O (ASO), anti-deoxyribonuclease B (anti-DNase B), and streptozyme tests.
11. Recognize the role Helicobacter pylori plays in gastrointestinal ulcers and the virulence factors that contribute to infection by this organism.
12. Discuss the various types of tests that may be performed to detect H. pylori infection.
13. Describe the respiratory and dermatological manifestations of Mycoplasma pneumoniae infections.
14. Discuss the use of serology for the diagnosis of M. pneumoniae infections, including the clinical value of detecting cold agglutinins.
15. Describe the epidemiology of Rocky Mountain spotted fever (RMSF) with respect to etiologic agent, transmission, and pathogenesis.
16. Select the appropriate serological and molecular techniques to diagnose Rocky Mountain spotted fever.

Chapter 21 Objectives:

1. Describe identifying characteristics of Treponema pallidum, Borrelia burgdorferi, and Leptospira species.
2. Explain how syphilis, Lyme disease, and leptospirosis are transmitted.
3. Discuss the different stages of syphilis.
4. Discuss the advantages of direct fluorescent staining for T. pallidum over dark-field examination without staining.
5. Define reagin.
6. Distinguish treponemal tests from nontreponemal tests for syphilis.
7. Describe the principle of the following tests for syphilis: Venereal Disease Research Laboratory (VDRL), rapid plasma reagin (RPR), fluorescent treponemal antibody absorption (FTA-ABS), and T. pallidum particle agglutination (TP-PA).
8. Provide reasons for false-positive nontreponemal test results.
9. Compare and contrast typical results of treponemal and nontreponemal testing during the various stages of syphilis.
10. Discuss the advantages and limitations of polymerase chain reaction (PCR) and enzyme immunoassay (EIA) testing for syphilis.
11. Explain the traditional and reverse algorithms for syphilis and discuss their advantages and limitations.
12. Select appropriate laboratory methods for testing of cerebrospinal fluid (CSF) for neurosyphilis and testing for congenital syphilis.
13. Describe early and late clinical manifestations of Lyme disease.
14. Discuss the role of immunoglobulin M (IgM) and immunoglobulin G (IgG) antibody responses in the diagnosis of Lyme disease.
15. Discuss serological methods that play a key role in the laboratory evaluation of Lyme disease and describe recommended two-tier testing algorithms.
16. Discuss causes of false positives and false negatives in serological testing for Lyme disease.
17. Discuss the etiology, mode of transmission, clinical presentation, laboratory diagnosis of leptospirosis.

Chapter 22 Objectives:  We are skipping this chapter!

Chapter 23 Objectives:

1. Describe the immune defenses that are important in protecting humans from viral infections.
2. Discuss mechanisms by which viruses can escape host defenses.
3. Correlate the presence of viral immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies with their clinical significance in detecting current infections, congenital infections, or immunity to infections.
4. Discuss the role of molecular tests in diagnosing and monitoring patients with viral infections.
5. Differentiate between the different hepatitis viruses and their modes of transmission.
6. Correlate the various serological markers of hepatitis with their diagnostic significance.
7. Explain the laboratory methods that are most commonly used to screen for, confirm, or monitor hepatitis virus infections.
8. Associate the following viruses with the specific diseases they cause: Epstein-Barr virus (EBV), cytomegalovirus (CMV), varicella-zoster virus (VZV), rubella virus, rubeola virus, mumps virus, and the human T-cell lymphotropic virus type I.
9. Discuss the laboratory methods used to diagnose and monitor infections with the preceding viruses.
10. Correlate the heterophile antibody and EBV-specific antibodies with their clinical significance and describe the laboratory methods used to test for these antibodies.

Chapter 24 Objectives:

1. Describe the classification system used to identify HIV isolates.
2. Explain the conditions under which transmission of HIV can occur.
3. Describe the structure of the HIV particle, including pertinent antigens and the genes that encode them.
4. Depict the replication cycle of HIV, beginning with entry of the virus into host cells.
5. Describe the host’s immune responses to HIV and the effects of HIV on the immune system.
6. Describe the clinical manifestations of HIV infection.
7. Explain the Centers for Disease Control and Prevention (CDC) classification system for HIV infection.
8. Discuss antiretroviral treatments and the impact they have had on HIV infection.
9. Discuss the current CDC-recommended algorithm for screening for HIV infection, as well as its advantages and limitations as compared with the previous algorithm.
10. Discuss the principles and clinical uses of conventional immunoassays and rapid tests for HIV infection.
11. Describe the principle of the Western blot test for HIV antibody, interpretation of the results, and limitations of the test.
12. Discuss reasons for false-positive and false-negative results in HIV antibody testing.
13. Explain the principle and clinical utility of flow cytometric methods for CD4 T-cell enumeration.
14. Discuss the role of qualitative nucleic acid tests in the detection and diagnosis of HIV.
15. Describe the molecular techniques performed for HIV viral load testing and drug-resistance testing and their clinical utility.
16. Select appropriate methods for HIV testing of infants and children younger than 18 months of age.