DEPARTMENT OF CLINICAL LABORATORY SCIENCE
CLINICAL IMMUNOLOGY- WRTG
4 Credit Hours
Marguerite E. Neita, Ph.D., MT (ASCP)
Welcome to Clinical Immunology! This course is designed to meet the requirements of the National Accrediting Agency for Clinical Laboratory Sciences (NAACLS) for basic immunology and clinical immunology (including serological techniques). The course will provide you with a basic knowledge of the principles of immunology, the application of these principles to diagnostic immunology, and exposure to current developments in the field.
This class syllabus is a valuable resource; it provides useful information; acts as a study guide and will familiarize you with the pace of the course. Read it carefully and have it available for use at all times! For your convenience the syllabus is available on Blackboard 5.0
It is your responsibility to become familiar with all aspects of this document. Please review the course outline and objectives for each lecture, note the dates and times of examinations, and plan your study schedule accordingly. The course is designed to motivate students to become actively involved in the learning process. This should enable the student to improve their comprehension, analyze and synthesize information, apply the information to clinical situations, and perform better on examinations (Metzler and Palau 1996).
A: Active Learning Strategies
Listed below are some recommendations which should help you to be successful in this class:
· Prior to attending class students should review and outline the assigned reading with special attention to:
- Chapter title
- Learning objectives
- Chapter introduction
- Italicized terms and abbreviations
- Headings and subheadings
- Illustrations, charts, tables, graphs
- End of chapter summaries
- End of chapter questions
· Apply the stages of active learning/critical thinking listed below (this will help you to comprehend and retain information).
1. Think about what you already know about the topic you are studying
2. Decide what you need to learn (what you don't know); these are your learning issues
3. Formulate preview questions, based on your learning issues, which will allow you to interpret, apply and evaluate the information you have read
4. Devise a means to alert yourself when you do not understand information you read or hear in class
5. Devise a method for improving your comprehension
Punctuality and regular class attendance are required. You are to sign the daily attendance sheet at the beginning of each class period. The attendance sheets will be closed ten minutes after class begins. There will be no make-up laboratory sessions, unless the student presents a documented excuse (medical certificate etc.). A grade of zero (0%) will be earned for each missed laboratory session. Assignment due dates and examination dates published in the syllabus are NOT subject to change.
There will be two in class examinations and a final comprehensive examination. These exams will include multiple choice, essay, and short answer questions. Students who miss an exam will be allowed to take the exam at a later date if prior arrangements are made with the professor.
B: Writing Across The Curriculum (WAC)
Clinical Immunology 709 is a writing intensive course which fulfills the third writing requirement for the Department of Clinical Laboratory Sciences.
Prerequisites: All students enrolled in this course must have earned a passing grade in ENGL-003.
Objectives: The purpose of a writing intensive course is two-fold. It is designed to improve the students’ writing ability, especially their professional writing (learning to write), and to improve the students critical, analytical, and problem solving skills (writing to learn).
The writing components of Clinical Immunology will consist of pre-writing strategies (Journals, outlines, summaries), laboratory notebooks, and a formal case report on an assigned immunological disorder. In these exercises there will be various stages of drafting and review (including peer-review). Writing assignments will be evaluated based on organization, clarity of expression, grammar and spelling, as well as a clear understanding of the immunological concepts taught in the class. Be sure to read the statement on plagiarism included on page 7 of this syllabus.
WAC Components for CLLS-709.
A. Journals (15 %):
Each student is required to keep a bound notebook (loose-leaf papers will NOT be accepted) in which entries will be made in journal format. This will enable you to be as creative as you like but your ideas must be organized in a coherent fashion and must be based on the course material.
· It is suggested that each entry in the Journal notebook be a minimum of one (1) handwritten page, with a maximum of two (2) pages. Occasionally journal entries will be made in class or submitted by e-mail.
· There will be regularly assigned topics which will include summaries of class concepts, analogies of your choice to describe a particular concept to a specified audience (lay person, school children, and scientific community) and abstracts.
· You may make relevant comments on class discussions, your assessment of the strengths and weaknesses of the class presentations, and comments as well as questions on the subject matter.
· You are required to reserve the first three pages of your journal notebook for a glossary of new Immunology vocabulary terms, definitions, and phrases.
· All entries must be dated and given a title. Entries that are not legible, dated, and titled will not receive a grade unless revised and resubmitted when the notebooks are next collected.
· Journals will be collected on Monday and will be returned to you on Wednesday of each week.
· If you are reading this e-mail me now at firstname.lastname@example.org for an additional 3 points on your first exam (This offer is valid until 10.00 am August 25, 2004).
Since you are expected to keep abreast of current developments in the field of immunology, each student is required to submit summaries for three (3) current, full length articles in immunology from peer reviewed journals (written & electronic). Copies of the original articles are to be submitted with the summary, which must be written in language a lay person can understand (Appendix A). This will constitute 30% of the JOURNAL grade. These summaries are due on the following dates:
Late summaries will not be accepted without prior discussion with, and approval of the professor.
JOURNALS will be graded from a holistic perspective. Your writing will be evaluated on the basis of a general impression of the content, arrangement and style. Grades will be as follows:
A (15 points) outstanding
B (10 points) good
C (7.5 points) adequate
F (2.0 points) unsatisfactory
Entries which receive a Grade of F may be rewritten and resubmitted with the next journal entry for an upgrade (maximum upgrade is a B).
Laboratory Notebooks (15 %):
You will be required to keep a laboratory notebook. Sections 1-3 below must be done as a pre-lab write-up before each laboratory session. Entries for sections 4-5 below are to be made during the laboratory exercise and should include any calculations made, comments on technical errors, observations, and results.
Details of the Laboratory Write-up:
The record of the laboratory procedures must be completed in a clear, concise and accurate manner. Students should minimize the use of personal pronouns (I, we). All entries should be made in the passive voice.
E.g. incorrect: I pipetted 1 ml of reagent into each tube
Correct: 1 ml of reagent was pipetted into each tube
Do not use white-out or erasers to remove errors from your notebook, instead draw a line through the error and make your correction next to it. The detail in the laboratory write-up should be sufficient to allow a fellow student to use your notebook as a guide. Laboratory notebooks will be evaluated based on completeness, adherence to the format, organization and understanding of the exercise. (If you are reading this, e-mail me now for 3 Free Points!)
The outline for each laboratory write-up is as follows:
1. Introduction and Theory. This section will include a three- or four- sentence statement of the objective of the laboratory exercise. A brief discussion of the theory and principles of the methods used should follow this. A description of the techniques, instrumentation and chemical or biochemical reactions should be included.
2. Materials and Supplies. A complete list of all materials, reagents and equipment is to be provided. The sources and concentrations of all chemicals and solutions should be listed. Instrumentation is to be listed with reference to company name and model number.
3. Procedure. This section is to contain a flow chart to describe the procedure. The procedural details and any observations are then recorded while the laboratory procedure is performed. You should include all observations, such as color changes, that occur.
4. Results and Calculations. Results are to be recorded directly in your notebook, not on scraps of paper or paper towels. Results may be recorded in a tabular form where appropriate. Calculations and graphs where necessary, must be included.
5. Conclusion and Discussion. This section should contain a conclusion based on the laboratory results and a three- or four-sentence discussion of their clinical significance. You should compare your results with known normal values for the procedure. Outline any problems encountered during the procedure and comment on their effect on the results with recommendations for avoiding these in future laboratory exercises. Each laboratory session will have an accompanying grading sheet which must be completed and turned in at the end of the class ( Appendix B) .
Case - Report (10%):
Each student will be assigned the case study of a patient with an immunological disorder. You will analyze and discuss the case and the laboratory diagnosis of the disease. You will prepare a written case report on the assigned disorder.
The case report will include the following sections:
- an abstract (150 word or less)
- the introduction
- review of the literature
- laboratory results and analysis
- discussion of the results
To assist you in completing this assignment the time line developed for completion and submission shown below, will be strictly followed:
Assignment of Case Aug. 30
Annotated Bibliography (30%) Oct. 08
Outline (5%) Oct. 22
First Draft (10%) Nov. 17 *two (2) copies)
Peer Review due (5%) Nov. 22 ** reviewer will be graded
Final Paper (50%) Dec. 01
NO CHANGES WILL BE MADE TO THE ABOVE TIMELINE
The annotated bibliography (Appendix C) will consist of the citation and a brief summary of the material to be used in the final paper. To demonstrate adequate research, there should be a minimum of ten references, retrieved from various sources including books, journal articles and reputable electronic sources. The bibliography must be submitted on the 3.5 floppy diskette. Please read the section on research writing in A Writers Manual, by Hatcher.
Required Format (Appendix D - case report samples)
Abstract (150 words or less)
Body: (5 - 10 pages)
Introduction (recap of written content)
Text (with headings)
Back Matter: (one to five pages)
Case - Report continued:
The final paper must be typewritten using a size 11 font, double-spaced, with one-inch margins on each side. The reference format may follow the style in any journal of laboratory medicine, medicine or immunology (e.g. Immunology Today, Clinical and Diagnostic Immunology, Clinical Laboratory Science). You must submit with the first draft a copy of the journal style you will be using and all of the articles cited in the paper. The audience for this assignment will be your peers in the scientific community (Students who follow this procedure correctly will earn 5 points on their final exam).
Please read the statement on plagiarism, found on page 7 of this syllabus, and the section in the text A Writer’s Reference assigned for this class. .
Tutorial assistance is also available to students at The Writing Center. There tutors can diagnose your writing deficiencies and assist with solutions. Tutors will not edit your writing, but will assist with a variety of problems such as lack of organization and incorrect grammar.
Information and referrals to the support resources available to students at Howard University are available from the professor.
C: Computer Assisted Instruction (CAI)
Computer assisted instructional materials are available for use by all students as review materials, auto-tutorials, and as an adjunct to lecture material. In addition students may use simulated laboratory models to practice special laboratory techniques. Review of some computerized tutorials will be done as group projects and will be required before certain lectures. A list of the software available for this course can be found on page 12.
Your grade is computed as follows:
Journal 15 %
Laboratory Notebook 5 %
Examinations (I & II) 30 %
Case Report 10 %
Final Examinations 30 %
Examination 1 Friday, October 1, 2004
Examination 2 Friday, November 12, 2004
Final examination TBA (scheduled by students)
IN ACCORDANCE WITH THE AMERICANS WITH DISABILITIES ACT (ADA) 1990, IF YOU WANT TO BE IDENTIFIED AS A PERSON WITH DISABILITIES AND NEED TO HAVE SPECIAL ACCOMODATIONS MADE PLEASE SEE THE PROFESSOR BY MAKING AN APPOINTMENT.
STATEMENT OF PLAGIARISM
Plagiarism is the representation of another person’s word and ideas as your own. This misrepresentation is a breach of ethics that seriously compromises a person’s reputation. Professional careers have been ruined by revelations of plagiarism.
Researchers, therefore, must scrupulously acknowledge sources to give proper credit for borrowed materials. The following rules should be observed to make sure that the distinction between your own words and ideas and those of others is justly maintained. Of course, submitting a paper that is completely the work of another person is plagiarism in its most extreme form.
1. Words, phrases, and sentences of another person should be enclosed in quotation marks and cited in proper form.
2. Paraphrases and summaries of the ideas of ideas of others should be properly cited. These paraphrases and summaries should not represent merely the rearrangement of sentence elements but should be written in your own style.
3. Quotations, paraphrases, and summaries should be introduced with the name of the writer being cited.
4. Every item in the paper (i.e., all sources of others’ word and ideas) should appear in the bibliography in proper form. (If you are reading this e-mail mneita @ howard.edu for 2 free points! Offer valid until Saturday, September 4, 2004).
5. Citations should contain all the information required by standard conventions and specifically indicate the location of the material cited. Page numbers should be checked for accuracy before the paper is submitted; the reader must be able to find the source of the material quoted, paraphrased, or summarized. Forms for citations and bibliographies should conform to those indicated by the professor.
For a further discussion of plagiarism, read the pertinent sections in A Writer’s Reference, by Hacker.
If you plagiarize all or part of an assignment, you can expect severe penalties, ranging from failure in that assignment to being recommended for a hearing before a judiciary body of the University. For a list of penalties, which may be imposed, see The Student Code of Conduct and Judiciaries in the Student Reference Manual and Directory of Classes.
Students should be aware that any incident of plagiarism or other form of academic dishonesty in this class will be dealt with severely.
CLINICAL 1MMUNOLOGY - WRTG
OUTLINE AND READING ASSIGNMENTS:
TEXT. Hacker, Diana, Ed. A Writer’s Reference. 5th Ed. St. Martin’s Press, Boston, 1995.
Stevens, Christine Doresteyn. Clinical Immunology & Serology: A Laboratory Perspective. 2nd ed. F.A. Davis Co. Philadelphia, 2003
Chapter 1. Historical Perspectives and Introduction
Chapter 2. Natural Immunity
a. First Line of Defense innate / nonspecific immunity
b. Adaptive / specific immunity
c. Factors associated with immunologic disease
Chapter 3. Components of the Immune System
The cells and cellular activities of the immune system
a. Granulocytic cells
b. The Monocytes and macrophages – Functions in host defense
- Inflammatory Response
c. Lymphocytes and plasma cells Development and differentiation
d. Organization and function of the lymphoid tissues
-primary lymphoid tissues
-secondary lymphoid tissues
e. Lymphoid and non-lymphoid surface markers
f. T-cells and B-cells
- Accessory cells
g. Cell products - the cytokines
h .Cell receptors and adhesion molecules
Chapter 6/7. Soluble Mediators of the Immune Response
- Acute phase reactants (APRs)
Chapter 4. The Major Histocompatibility Complex (MHC) [Not in the text- Lecture Notes]*
- Tissue distribution
- Structure and function
- Inheritance of HLA
- Detection of HLA
- Clinical importance- Association with disease, tissue transplantation & paternity
Chapter 4/ 5. Mechanisms of the Specific Immune Response
- The nature of antigens and adjuvants
- Antibody structure and function
- Antigen recognition and processing
- Regulatory mechanisms
Immunologic Techniques (Serology)
Chapter 8. Safety and Basic Techniques
Overview of Antigen - Antibody Interactions
Chapter9. Precipitation & Immunoelectrophoresis
- Types of reactions
- Applications of the technique
- Phases of agglutination
- Types of agglutination reactions
- Additional reagents
- Enhancement media
- Anti-human globulin (AHG) procedures
Chapter 11. Labeled immunoassays
- Enzyme labels
- Radioactive labels
- Immunofluorescent Assays
Chapter 10. Automated Procedures
- Lymphocyte subset analysis
- Lymphocyte phenotyping in HIV infection and leukemias
Other immunological techniques
- Complement assays
Chapter 11. Molecular Techniques
Examination 1: Friday, October 1, 2004
A. Immune Disorders - Theory and Diagnostic Procedures
Chapter 13 Hypersensitivity Reactions – Types 1, II, III, IV
Chapters 14. Factors influencing the development of autoimmunity
The spectrum of autoimmunity
Organ non -specific diseases systemic)
- Systemic lupus erythematosus (SLE)
- Rheumatoid arthritis
- Sj`gren's syndrome
Organ-specific Autoimmune Diseases
Endocrine gland disorders - autoimmune thyroiditis
Pancreas - IDDM
Gastrointestinal disorders – pernicious anemia
Immune Deficiency Disorders
Chapter 16. Accessory Cell Dysfunction
Primary Immune Deficiency Disorders
- Humoral immune deficiency - Bruton's agammaglobulinemia
- Selective IgA deficiency
- Cellular immune deficiency - DiGeorge Syndrome
- Combined humoral and cellular immune deficiency - SCID
- Wiskott-Aldrich syndrome
Secondary Immune Deficiency Disorders
Chapter 15 Monoclonal gammopathy - Multiple myeloma
- Waldenstrom's macroglobulinemia
- Other monoclonal disorders
Examination 2: Friday, November 12
B. Infectious Disease Serology: Chapters 13 - 22.
Chapter 19 The immune response in infections
Spirochetal infections: Syphilis and Lyme disease (chapter 15 & 16)
- Etiologic agents
- Stages of the disease
- Direct detection methods
- Serologic diagnosis
- Treatment and prevention
Chapter 20. Streptococcal infection and serology
- Streptococcal infection and sequelae
- Laboratory diagnosis
Chapters 21. Viral infections and Serology
- Epstein-Barr virus infection
- Viral hepatitis
Chapter 22 Acquired Immunodeficiency Syndrome - HIV infection
Chapters 17 Transplant Immunology
- Types of grafts
- Graft acceptance and rejection
- Tissue typing
Chapter 18 Tumor Immunology
- predisposing genes
- role of oncogenes
- tumor markers
- cancer treatment
Final Examination - TBA
Department of Clinical Laboratory Science
Instructional Material for Clinical Immunology – CLLS 700
Software for Computer Assisted Instruction (CAI)
Clinical Immunolgy-wrtg (CLLS 709)
(Software can be viewed in the Clinical Laboratory Science Computer Room or in Dr. Neita’s Office
60 2. Electrophoresis Tutor
70 3. ANA-tutor
4. Immunology Interactive CD-Rom
5. MARIAä 1.0
Part I - Introduction to Immunology
By the end of the semester the student will be able, with 85% accuracy, to:
1.0 Discuss the chronological development of immunology from its origins as a technology for the
diagnosis of infectious diseases to the development of immunology as a science.
2.0 Show an appreciation for the contributions of various scientists to the field of immunology e.g. (Jenner, Metchnikoff, Pasteur, Yalow, etc.)
B. Basic Principles
1. Demonstrate knowledge of the concepts of Non-specific Immunity
.1 Explain the concepts of susceptibility and non-susceptibility
.2 List and identify the functions of the epithelial barriers involved in non-specific immunity
.3 Explain the inflammatory response, with emphasis on the vascular and cellular responses
.4 Identify chemokines and discuss their role in the inflammatory response
.5 Describe the process of phagocytosis, the cells, and the mechanisms involved
.6 Identify and discuss the role of the nonspecific soluble (humoral) mediators of natural immunity such as:
- Acute phase reactants (CRP, haptoglobin, fibrinogen)
.7 Explain the role played by chemokines and cell adhesion molecules in innate immunity
2. Identify the components of the complement system and its activation during the immune response
.1 List the protein components of the complement system
.2 Classify them according to their activity
-membrane attack unit
.3 Compare the classical, alternate, and MBL pathways of complement activation
.4 Outline the order of reaction of each component in the classical and alternative pathways
.5 Describe the conditions necessary for the activation of each pathway
.6 Identify the accessory factors needed for the activation of complement
.7 Describe the formation and biological effects of the split products of complement activation
.8 Identify the enzymes activated during C' activation and amplification
.9 Identify and describe the activity of the control mechanisms of the complement system
.10 Discuss with examples the consequences of deficiencies in components of the complement system
.11 Identify the other factors that mediate and control nonspecific resistance to disease including but not limited to the following:
(a) Species indifference
(b) Antagonism of indigenous flora
(c) Nutritional factors
(e) hormonal influences
3. Explain the principles that define Specific Immunity (adaptive immunity)
.1 List and discuss the characteristics that define the specific immune response:
(a) Recognition / discrimination
.2 Outline the development, maturation and commitment of the components of the immune response
.3 Demonstrate knowledge of the structures of the sites of development:
(a) Primary lymphoid tissue
(b) Secondary lymphoid tissue
.4 Describe the acquisition of the specific immune response in terms of active and passive immunity with examples of each of the following:
-naturally acquired active immunity
-artificially acquired active immunity
-naturally acquired passive immunity
-artificially acquired passive immunity
4. Describe and identify the components of the specific immune response. Relate the structures and products of each component to its role in immune function
.1 Differentiate between the different lymphoid cells involved in the immune response
.2 Describe the structure and immune function of:
-T-cells and the T-cell sub-population
.3 Describe the developmental pathway and acquisition of specific markers for T-cells, B-cells and NK-
.4 Discuss the use of (antigen specific receptors markers for the identification and evaluation of cell
.5 List the common T-cell and B-cell mitogens and state their utility in determination of cell numbers and
.6 List and identify the characteristics of the nonspecific soluble mediators of the immune response, e.g.,
cytokines ( monokines and lymphokines)
.7 Discuss the interactions between cytokines and cellular mediators of the immune response
.8 Discuss the role of cell adhesion molecules (CAM) in lymphocyte trafficking
.9 Diagram and label the basic antibody unit
.10 Describe the structure, occurrence, and physiochemical properties of the five immunoglobulin classes;
IgG, IgM, IgA, IgD and IgE and their subclasses
.11 Describe the organization and expression of the immunoglobulin genes
.12 Describe the gene rearrangements that give rise to antibody heavy (H) and light (L) chains
.13 Define alleleic exclusion
.14 Describe isotype switching
.15 Describe the production of monoclonal antibodies, and discuss their application to the practice of clinical and laboratory medicine
5. Define antigens and discuss the physical and chemical characteristics that affect their induction of an immune response.
.1 Define and differentiate between antigens, immunogens, haptens
.2 Define paratopes, idiotopes, and epitopes
.3 Outline the characteristics required for immunogenicity and antigenicity
.4 Define adjuvant and discuss its relationship to the development of an immune response
.5 Distinguish between T-dependent and T-independent antigens
.6 Define the following terms used to describe antigens: -allogeneic -autogeneic , syngeneic and
.7 Define heterophile antigens and their use in diagnostic procedures
6. Discuss the factors involved in triggering the immune response
.1 Outline the steps involved in processing of T-dependent and T-independent antigens
.2 Discuss the presentation of the antigen to the immune cells
.3 Identify and explain the cytosolic and endocytic pathways of antigen processing
.4 Outline the stages involved in T-cell activation
.5 Discuss the production of signals necessary for activation of T-cells
.6 Outline B-cell activation and the response of the B-cells to T cell produced lymphokines (e.g.
Interleukin -4; Interleukin-6)
.7 Define super antigens and discuss their roles in disease pathology
7. Outline the general features of the Major Histocompatibility Complex (MHC)
.1 List and define the Class I and Class II and Class III of the MHC
.2 Differentiate between Class I and Class II MHC antigens based on:
(a) Tissue distribution
.3 List examples of diseases associated with the presence of specific HLA antigens
Part 2 - Immunodiagnostics
By the end of the semester the student will, with 85% accuracy:
A. Antigen - Antibody interactions
1. Explain the mechanisms of antigen antibody interactions and explain the principles involved in the practical applications of those mechanisms.
.1 Explain the primary interactions between antibody and antigen
-discuss the forces involved
-list and explain the factors which effect Ag:Ab interactions
-define and explain affinity and avidity
.2 Discuss the secondary Ag: Ab interactions with reference to a) type of antigen b) type of antibody c) formation of complexes
.3 Explain the practical applications of the primary and secondary Ag:Ab interactions by discussing the principles and applications of the following immunologic techniques
-Precipitation techniques - including RID, CIE, IEP, IFE, etc.
-Agglutinations, and agglutinations inhibition (including hemagglutinations)
-Reactions involving complement
-Neutralization of toxins and viruses
-Fluorescent antibody reactions
-Enzyme linked immunoassay (ELISA)
-Enzyme inhibition assays (EIA)
2. Tests for cellular immunity - Explain the principles of test procedures used to enumerate
and evaluate the function of immune cells.
.1 Describe the tests for evaluation, enumeration, and analysis of
i. Lymphocyte proliferation assays
ii. Analysis of serum antibodies
i. T-cell subsets by fluorescent assays
ii T-cell function
-DTH skin testing
-Lymphocyte proliferation assays
-Mixed lymphocyte cultures
3. Tests for Acute-phase reactants
.1 Discuss the current and potential uses of C-reactive protein determination in evaluating the clinical course of:
(a) Bacterial and viral infections
(b) Rheumatic diseases & SLE
(c) Myocardial infarctions
(d) Burn injuries
(e) Surgical recovery
(f) Renal transplantation
.2 Explain the principles of the laboratory tests for C - reactive protein (CRP)
.3 Evaluate the status of an individual based on the levels of CRP in the blood
B. Laboratory Practice
By the end of the semester the student will, with 90% accuracy, be able to:
1. Specimen preparation and safety precautions
.1 Determine the appropriate method for collection, separation and inactivation of tests samples
.2 Discuss the reasons for heat inactivating or not inactivating test sera
.3 Identify the correct temperature and time for the inactivation of serum samples
.4 Outline the procedure for labeling, handling and disposing of potentially infectious material
.5 Define titer, and explain the significance of the use of acute and convalescent samples in determination of past, or current infections
.6 Prepare accurate serial dilutions, and determine antibody titers
.7 Specify the correct handling procedure for various laboratory specimens
.8 Initiate the use of safety procedures and personal protective equipment in the laboratory
2. For each laboratory session the student will:
.1 Attend the laboratory punctually and in appropriate safety attire
.2 Organize the work area to facilitate efficient manipulation of laboratory equipment (racks, tubes, etc.)
.3 Prepare samples as outlined in the laboratory procedure using universal precautions
.4 Select and perform the appropriate controls and instrument calibrations
.5 Perform the laboratory procedure with 90% accuracy
.6 Demonstrate an understanding of the principle of each test procedure
.7 Analyze the results obtained, with reference to the test controls
.8 Maintain a clean work area during the laboratory session.
.9 Dispose of waste appropriately and clean-up laboratory area after the session exercising acceptable safety precautions.
.10 Maintain a neat laboratory notebook for the recording of results laboratory exercises
.11 Discuss and explain the results obtained for each laboratory procedure and document the information in the laboratory notebook
.12 Correlate and evaluate the results based on in the case studies attached to the laboratory exercise
.13 Discuss and justify the confidentiality of patient results in the clinical laboratory.
.14 Review and adhere to the safety regulations outlined on page 19 of the syllabus
Working in any laboratory can expose you to many hazardous and infections agents. It is important that you are aware of the practices and procedures that are instituted for your protection. The safety precautions outlined below are important and should be adhered to in every laboratory session.
1. Experiments and procedures are to be carried out only with the supervision of the instructor.
2. If you have any questions about the procedure, equipment, materials or reagents, STOP what you are doing and ASK the instructors for assistance
3. All book bags, text books and clothing should be left outside of the laboratory area. Place these items in your locker! The only items on the bench top should be your lab notebook and a pen or pencil
4. Your lab coat should be worn at all times in the laboratory, long hair should be tied back and gloves worn for all procedures. If you have a sensitivity to latex gloves speak to the instructor immediately
5. After putting on your lab coat, clean the bench top by wiping with disinfectant and wash your hands.
6. Eating, drinking, smoking, taking medication, applying cosmetics, licking labels, storing food or any activity which involves putting any item in your mouth is STRICTLY PROHIBITED.
7. MOUTH PIPETTING IS STRICTLY FORBIDDEN. Avoid techniques that may cause the formation of aerosols; remove stoppers carefully, do not squirt pipette or syringe contents etc.
8. Gloves and other contaminated materials must be carefully discarded in the receptacles provided. DO NOT DISCARD ANY USED MATERIALS IN THE TRASH! This is a violation of OSHA requirements. Do not re-cap syringes or lay them on the lab bench. Place all sharps in the sharps container.
9. Notify the instructor immediately of any spills, cuts, needle sticks or other laboratory accident.
10. In the event of a spill of potentially infectious material follow this procedure.
A). Remove your lab coat if it has been contaminated
B) Cover the spill with paper towels and flood with 10% Clorox solution.
C) Allow the Clorox to stay in contact with the spill for approx. 20 minutes
D) Do not touch broken glass with your hands
E) Use a dustpan to clean up the spill and discard the material in the biohazard bags
F) WASH YOUR HANDS
11. Remove your gloves and wash your hands any time you leave the laboratory, and at the end of the laboratory session
12. Before leaving the laboratory, clean your work area and disinfect with 10% Clorox solution. Remove your lab coat and then wash your hands. Lab coats should remain in the laboratory
Adapted from Immunology Investigations: A Laboratory Manual. Loreli Batina, 1999. STAR Publishing Co., California
Part 3 - Clinical Immunology
By the end of the semester the student will, with 90% accuracy:
A. Laboratory Determination of Infectious Diseases
l a. Syphilis -the disease
.1 Identify the etiologic agent of the disease (Treponema pallidum)
.2 distinguish between syphilis and the other human treponematoses
.3 Identify the three stages of syphilis and discuss the symptomatology, infectivity, and pathology of each stage
.4 Describe congenital syphilis
.5 Identify the types of antibodies developed during the course of the disease (non-treponemal or reagin and treponemal antibodies)
.6 Correlate the presence or absence of these antibodies with the various stages of the disease
.7 Discuss treatment of the disease and its correlation with the presence of antibodies
1b. Diagnostic Procedures
.1 Identify the non-treponemal antigen tests
.2 Determine the indications for use of each test
.3 Discuss the sensitivity, specificity, and incidence of false positives and false negative results for
(i) VDRL test procedure
(ii) RPR test procedure
.4 Discuss the principles of each test
.5 Identify the treponemal antigen tests (i) FTA-ABS (ii) IgMFTA (iii) TPI (historical)
.6 Determine the indications for use of treponemal antigen tests
.7 Discuss the sensitivity, specificity, and incidence of false positive and negative results
.8 Describe the principles of each test
.9 Given appropriate patient information specify the test procedures to be used and evaluate the results
.10 Perform the test procedures in a laboratory session
2. Infectious Mononucleosis
.1 Summarize the principles of the tests for heterophile antibodies in the diagnosis of Infectious Mononucleosis
(i) Paul-Bunnell presumptive test
(ii) The Davidsohn differential test
(iii) Rapid differential slide tests
.2 Identify other tests for detection of infectious mononucleosis
.3 Evaluate the sensitivity, specificity and usefulness of the test procedures
.4 Given appropriate information use the knowledge of heterophile antibodies and the characteristics of the disease to evaluate the patient's status
3. Cold Agglutinins and Streptococcus MG
.1 Define and explain the characteristics of cold agglutinins
.2 Associate the presence of cold agglutinins with the following disease states and conditions:
(a) Cold agglutinin syndrome
(b) Mycoplasma pneumoniae infections
(c) Other disease states
.3 Describe the handling of specimens for detection of cold agglutinins
.4 Discuss the tests for determination of cold agglutinating antibodies
.5 Describe the characteristics of streptococcus MG antibodies
.6 Describe the tests for determination of streptococcus MG antibodies
.7 Given the titer of a sample, evaluate the significance of the test results
.8 Correlate the results obtained in the test for streptococcus MG antibodies and the test for cold agglutinins
4. Streptococcal infections and related antibodies
.1 Identify the groups and serotypes of streptococci associated with the development of post-streptococcal sequelae
.2 Characterize the extracellular toxins implicated in the disease processes
(a) Streptolysin O
.3 Identify the antibodies produced in response to the extracellular toxins
(a) Antistreptolysin O
.4 Identify two important sequelae of streptococcal infections
(a) Acute glomerulonephritis (AGN)
(b) Rheumatic fever (RF)
.5 Associate the two types of streptococcal infections (pyoderma and pharyngitis) with the incidence of AGN and RF
.6 Describe the clinical symptoms of AGN and RF
.7 Identify the following laboratory procedures for determination of antibodies associated with AGN and RF
(a) Antistreptolysin O tube test (ASO)
(b) Antistreptolysin O latex slide text
©. Antihyaluronidase; mucin clot test (AHT)
(d) Antideoxyribonuclease B Test (Anti DNAse B)
(e) Streptozyme test
.8 Discuss and evaluate each of the tests listed above with reference to
(a) Principle of the test
(c) Preparation of reagents
(d) Selection of appropriate controls
(e) Limitations of the procedure
(f) Interpretation of the results
(g) Indications for use
.9 Given relevant case histories evaluate the test results and recommend additional courses of action necessary to complete a diagnosis
.10 Correlate the serologic test results with results from microbiology, hematology and clinical chemistry laboratories
5. Prenatal Screening - TORCH tests
For each of the following infections discuss the transmission and outline the clinical symptoms of the diseases. Identify the etiologic agent.
-Rubella (German measles)
.2 Discuss the effect of intrauterine infection on the development of the fetus
.3 Evaluate the necessity for prenatal screening for these infections
.4 Discuss precautions necessary to prevent infections during pregnancy
.5 Determine the appropriate test procedure and evaluate the results obtained for detection of past or current infection
B. Immune Disorders
.1 Describe with examples in the in vivo effects of antigen: antibody or cell mediated reactions
.2 Describe the pathogenesis of the four types of hypersensitivity reactions
- Type I or IgE dependent hypersensitivity (allergy)
- Type II or cytotoxic hypersensitivity
- Type III hypersensitivity or immune complex reactions
- Type IV or delayed type hypersensitivity reactions
.3 Describe the tests used for diagnosis of hypersensitivity reactions (e.g. RAST, RIST)
2a. Autoimmune disorders
.1 List and briefly discuss the theories posited for the generation of autoimmune disorders:
-sequestered antigen release
-stem cell defects
-disruption of tolerance
.2 Describe the characteristics of autoimmune disorders
.3 Outline with examples the classification of organ specific and systemic disorders
.4 Identify the types of autoantibodies present and discuss their significance in the following autoimmune disorders
- Lupus erythematosus (SLE)
-Rheumatoid arthritis (RA)
-Auto immune thyroiditis
- Sjogren’s syndrome
.5 Describe sarcoidosis
.6 Discuss the imunological implications of the disease known as sarcoidosis
.7 Discuss the current theories regarding the etiology of the disease
2 b. Anti-nuclear antibodies in the diagnosis of auto-immune disorders
.1 Define antinuclear antibodies and discuss their association with autoimmune disorders in general and
with SLE specifically
.2 Describe and identify the different patterns of reaction obtained using immunofluorescent techniques
.3 Evaluate the fluorescent antinuclear antibody (FANA) procedure as a screening test for SLE
.4 State the principle of the Anti-DNA test
.5 Outline the procedures for detecting antibodies to dsDNA (nDNA)
.6 Compare the anti-DNA test with the FANA procedure for the diagnosis of SLE and for the evaluation of treatment
.7 Discuss other test procedures useful in the diagnosis of Systemic Lupus Erythematosus (SLE).
(complement components, anti-cardiolipin assays)
2c. Rheumatoid Factor in autoimmune diseases
.1 Explain the principles of the various Latex Agglutination tests for the detection of the rheumatoid factor
.2 Evaluate the sensitivity of the tests for use in the screening of patients for RA
.3 List and explain the significance of other tests used in the diagnosis of RA, e.g., CRP, complement
3 Immunodeficiency and Immunoproliferative disorders
3a. Immunodeficiency disorders
a. accessory cell dysfunction - qualitative or quantitative
b. primary B-cell deficiencies
c. primary T-cell deficiencies
d. combined B- and T-cell deficiencies
.2 Correlate the clinical manifestations of Immunodeficiency disorders with the development and function of cells of the immune system
.3 Discuss secondary or acquired Immunodeficiency syndrome with special reference to infection with the human Immunodeficiency virus (HIV) and AIDS
.4 Describe and discuss HIV/ AIDS with reference to:
a the etiologic agent
b basic structure of the virus
c effect on the immune system (T4 cell depletion)
d mode of transmission and disease prevention
e laboratory diagnosis
f opportunistic infections
g progress of the disease
h development of new drugs and vaccines
3b. Immunoproliferative disorders
(a) Monoclonal disorders
(b) Polyclonal disorders
.1 Describe specific immunoproliferative disorders including multiple myeloma, heavy chain disease, and Waldenstrom’s macroglobulinemia
.2 Characterize the laboratory findings as polyclonal or monoclonal based on serum protein electrophoresis and imunoelectrophoresis
4. Given the relevant clinical data the student will be able to choose the appropriate test procedures for evaluation of patient status.
5. Given specific test results, the student will be able to determine the patients' clinical condition and correlate the serology results with results from the microbiology, hematology and chemistry laboratories
C. Transplant Immunology
.1 Discuss the principles of tissues transplantation with reference to the genetic composition of donor and recipient and the immunologic and non-immunologic problems associated with transplantation
.2 Describe graft tissue according to the relationship between donor and recipient
.3 Relate the products of the MHC and ABO systems to the survival or rejection of grafted tissue
.4 Discuss chronic and acute rejection
.5 Describe the immunologic issues associated with tissue grafts from immunologically privileged and other sites with reference to the following
- Bone marrow
.6 Explain graft vs. host disease
.7 Outline the procedures for pre-transplant testing of donors and recipients
-choosing the donor
-mixed lymphocyte cultures
.8 Discuss the use of immunosuppression in the transplantation of tissues and name three immunosuppressive agents
D. Tumor Immunology
.1 Discuss the immune response of the body to tumor development
.2 Define the tumor associated antigens (TAA’s), tumor specific antigens (TSA’s) and oncofetal antigens
.3 Recall the tumor markers that are associated with diagnosis treatment and monitoring of cancer therapy
Writing Assignment 1:
1. Select two articles from different sources
2. Examine and NOTE differences in bibliography style
3. select a style for final term paper
1. Agglutination and Determination of serum antibody titers using serial dilutions
2. Agglutination - RPR Card Test
3. Streptozyme detection
4. Latex Agglutination procedures
5. Fluorescent antibody procedures
6. Enzyme immunoassays
7. Enzyme immunoassays