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Clinical Laboratory Immunology, 1st Edition Test Bank

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    Exam Name___________________________________ MULTIPLE CHOICE. Choose the one alternative that best completes the statement or answers the question. 1) Which of the following defines commensalism? 1) A) Biologic relationship between two organisms in which one benefits and the other is harmed. B) Biologic relationship between two organisms in which both are harmed. C) Biologic relationship between two organisms in which both benefit. D) Biologic relationship between two organisms in which one benefits and the other remains unharmed. Answer: D 2) Microbes that colonize a particular person for a long period of time are called 2) A) nonpathogenic microbial flora. B) usual microbial flora. C) transient microbial flora. D) resident microbial flora. Answer: D 3) Each body site below is paired correctly with an expected component of the normal microbial flora except 3) A) colon/enterobacteriaceae. B) vagina/lactobacilli. C) upper respiratory tract/viridians streptococci and diphtheroids. D) skin/ Staph epidermis and Propionibacterium. E) All of the above are paired correctly. Answer: E 4) Normal microbial skin flora is primarily concentrated on the 4) A) dry areas of skin with heavy hair. B) moist areas of skin near sweat glands. C) dry areas of skin with light hair. D) fingernails and toenails. Answer: B 5) What is the collective purpose of nasal hair, mucus secretions in the naspopharyngeal tract, phagocytic cells in the throat, bronchial lysozyme, sneezing, and coughing? 5) A) To keep organisms from entering the lower respiratory tract and causing infection. B) To keep normal flora in the oropharynx from entering the eustachian tube causing infection. C) To keep normal flora in the oropharynx from causing infection. D) To maintain a sterile environment in the oropharynx. Answer: A 6) Quantitatively, the most important defense against food - borne pathogens in the gastrointestinal tract is 6) A) peristaltic movement. B) ciliated cells. C) mucus cells. D) resident phagocytic cells. E) low pH and high enzymatic content of the stomach. Answer: E 1

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    7) A microorganism that typically does not cause disease in a normal host but that does cause disease if the host is compromised in some way is called 7) A) symbiotic. B) commensal. C) opportunistic. D) parasitic. E) pathogenic. Answer: C 8) Which of the following situation(s) represents an event that could result in an opportunistic infection? 8) A) Subject is stabbed with a knife. B) Subject has dental surgery. C) Subject has leukemia. D) Subject is undergoing radiation for cancer treatment. E) All of the above. Answer: E 9) A microorganism that consistently causes disease in normal people with competent immune systems is a 9) A) virulent pathogen. B) opportunistic pathogen. C) commensal pathogen. D) true pathogen. E) symbiotic pathogen. Answer: D 10) Which of the following are mechanisms by which microbes evade phagocytosis? 10) A) Produce collagenase B) Produce leukocidins C) Produce hyaluronidases D) Motility Answer: B 11) Which microbial product below helps organisms penetrate tissues? 11) A) Hemolysins B) Hyaluronidase C) Capsule D) Fimbrae E) Coagulase Answer: B 12) Which microbial product below helps Staphylococcus aureus create a protective fibrin wall around itself? 12) A) Fimbrae B) Capsule C) Coagulase D) Hyaluronidase E) Hemolysins Answer: C 2

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    13) The microbial structures that promote attachment of the microbe to tissues, making phagocytosis more difficult, are 13) A) Panton - Valentines. B) fimbrae. C) cell walls. D) capsules. E) flagella. Answer: B 14) Which organism below produces a protein called Protein A that binds to the Fc portion of IgG? 14) A) Chlamydia B) Hemophilus influenzae C) Streptococcus pyogenes D) Streptococcus pneumoniae E) Staphylococcus aureus Answer: E 15) Which organism below is easily phagocytized but is capable of surviving and multiplying intracellularly? 15) A) Staphylococcus aureus B) Streptococcus pyogenes C) Hemophilus influenzae D) Chlamydia E) Streptococcus pneumoniae Answer: D 16) All of the following organisms are virulent at least in part due to exotoxins they secrete except 16) A) Staphylococcus aureus B) Mycobacterium tuberculosis C) Streptococcus pyogenes D) Corynebacterium diphtheriae E) Clostridium botulinum Answer: B 17) Which of the following antimicrobial substance is(are) secreted by mucous membranes? 17) A) Lactoferrin B) Lysozyme C) Antibodies D) All of the above Answer: D 18) Phagocytes are most effective against 18) A) protozoan parasites. B) extracellular bacteria. C) intracellular viruses. D) Phagocytes are equally effective against all the microbes listed. E) intracellular bacteria. Answer: B 3

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    19) Which of the following can function as chemotactic agents? 19) A) Complement fragments B) Bacterial products C) Cytokines produced by white blood cells responding to an inflammatory site D) Mediators released from damaged tissue E) All of the above Answer: E 20) Release of inflammatory mediators causes increased vascular permeability. Which of the four hallmarks of inflammation does this cause? 20) A) Heat B) Erythema C) Pus D) Pain E) Edema Answer: E 21) When an infection occurs in a fetus or early in the life of the neonate, the immune response may be inadequate due to 21) A) the induction of tolerance. B) suppression of the immune system. C) interference from maternal T cells. D) antigenic variation. Answer: A 22) What do infections with HIV, cytomegalovirus, and Epstein - Barr virus all have in common? 22) A) Evasion of the immune system by antigenic variation. B) An induction of immune tolerance to other viruses. C) Poor production of antibody by the host. D) Immunosuppression of the host. Answer: D 23) Which organism below evades the adaptive immune response by antigenic variation? 23) A) Borrelia recurrentis B) Plasmodium vivax C) Brucella abortis D) Listeria monocytogenes E) Mycobacterium tuberculosis Answer: A 24) Which organism below evades the adaptive immune response by surviving inside host liver cells? 24) A) Mycobacterium tuberculosis B) Borrelia recurrentis C) Listeria monocytogenes D) Plasmodium vivax E) Brucella abortis Answer: D 25) An important feature of many pathogenic viruses is their resistance to 25) A) lysozyme. B) interferon. C) interleukin - 2. D) interleukin - 1. E) lactoferrin. Answer: B 4

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    SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 26) Discuss how the nature of a particular body site influences the normal microbial flora located there. 26) Answer: In order to colonize a particular body site, a microorganism must be able to evade the normal defense mechanisms and exploit the local conditions to survive. Thus, areas of the body with low oxygen, such as the gum crevices around the teeth, can support anaerobes that are resistant to antimicrobial substances such as lysozyme in the saliva. Secreted fatty acids on the skin are normally antimicrobial, but Propionibacterium species can metabolize them. Females of reproductive age secrete glycogen vaginally, allowing lactobacilli to flourish. Breast - fed babies have lower pH in the gastrointestinal tract, and acid - tolerant species such as Bifidobacterium colonize. 27) Discuss the benefits of normal microbial flora for the host. 27) Answer: Newborns normally existed in a sterile environment up until their birth, so the exposure to nonpathogenic organisms is the first stimulation to their immune systems and is a necessary step in the development of a fully functional immune system. The other advantage the normal flora provide is competition with pathogens for limited space and nutrients on the host, and antibiotics that kill off large numbers of normal flora actually make the host more susceptible to pathogens. 28) Differentiate virulence and pathogenicity. 28) Answer: If an organism is capable of causing a disease in a particular body site or situation, it is a potential pathogen. Thus, a normally nonpathogenic organism can be a pathogen in an compromised host. A very virulent pathogen is easily acquired and/or has very serious outcomes, including death by uncompromised people with normal immune systems. 29) List ways in which the skin resists the penetration of microorganisms. 29) Answer: 1. Physical barrier of keratinized skin 2. Bacteriocidal substances secreted by sebaceous glands 3. Normal flora — low pH, competition for nutrients and space 4. Constant shedding of outermost skin layer — reduces microbial load 30) List ways in which microorganisms evade phagocytosis. 30) Answer: 1. Production of protein that binds to Fc portion of IgG (e.g., Staph aureus ) 2. Possess capsule (e.g., Streptococcus pneumoniae ) 3. Adhere to host cells via fimbrae (e.g., Streptococcus pyogenes ) 4. Produce leukocidins and hemolysins (toxic to WBCs) 5. Produce coagulase to protect themselves with fibrin wall (e.g., Staph aureus ) TRUE/FALSE. Write 'T' if the statement is true and 'F' if the statement is false. 31) Normal microbial flora never cause disease in immunocompetent hosts. 31) Answer: True False 5

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    32) A Boy Scout goes camping in the woods for a week and doesn't take a shower. When he gets home, the microbes that he carried all week long are washed away in the shower, never to return. The microbes that the boy acquired in the woods and carried all week would be called transient microbial flora. 32) Answer: True False 33) A particular microorganism may be commensal in one body site but parasitic in another body site. 33) Answer: True False 34) A human could never harbor a pathogenic microorganism as resident flora. 34) Answer: True False 35) No microorganism is capable of directly penetrating keratinized, intact human skin. 35) Answer: True False 36) The movement of white blood cells through blood vessel walls is called chemotaxis. 36) Answer: True False 37) Erythema in an inflamed site is caused by release of inflammatory mediators that increase blood flow to the affected area. 37) Answer: True False 38) People with immune systems suppressed by HIV are more susceptible to viral, fungal, and protozoal infections. 38) Answer: True False 39) Some organisms evade the immune response by changing the antigens expressed on their surface during the course of an infection. 39) Answer: True False 40) Once an organism is engulfed by macrophage, it is always digested and killed. 40) Answer: True False SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 41) Microorganisms that are consistently associated with human disease are called ________. 41) Answer: pathogens 42) Antibiotic associated colitis has been associated with the organism ________. 42) Answer: Clostridium difficile 43) Microbes that have very low infective doses and/or that are very harmful to the host are very ________. 43) Answer: virulent 44) Microorganisms with capsules are better able to evade ________ by neutrophils and monocytes. 44) Answer: phagocytosis 6

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    45) The causative organism of botulism is particularly virulent because of the ________ it secretes. 45) Answer: exotoxin 7

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    Exam Name___________________________________ MULTIPLE CHOICE. Choose the one alternative that best completes the statement or answers the question. 1) The products of the major histocompatibility complex that are responsible for tissue type are called ________ because they were initially identified on white blood cells. 1) A) leukocyte thymic education antigens B) Gorer white antigens C) human leukocyte antigens D) Landsteiner leukocyte markers Answer: C 2) What role do HLA (MHC) antigens play in the thymus? 2) A) Selection of T cell clones that bind to foreign antigen B) Selection of T cell clones capable of binding to HLA (MHC) antigens C) Elimination of autoreactive T cell clones D) All of the above Answer: D 3) The proteins of the HLA system are said to belong to the ________ superfamily because most have a constant and variable region. 3) A) immunoglobulin B) complement C) immune receptor D) autoantigen Answer: A 4) In what manner are MHC genes expressed? 4) A) Activated B) Silent C) Dominant D) Codominant E) Recessive Answer: D 5) Which of the following is true regarding inheritance of MHC genes? 5) A) Alleles can be homozygous or heterozygous. B) Inheritance of MHC genes follows Mendelian genetics (Law of Segregation or Law of Independent Assortment). C) The rate of gene recombination is low, but there are some "hot spots" of increased recombination. D) All of the above. Answer: D 6) Which set of HLA gene(s) is considered the "classic" set of Class I MHC molecules? 6) A) HLA A, B, and C B) classic complement components C) HLA DP, DQ, and DR D) beta 2 microglobulin E) alpha/beta heterodimer Answer: A 1

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    7) What is the best current technology for determining HLA types? 7) A) Flow cytometry detecting mixed lymphocyte cytotoxicity reactions. B) Flow cytometry using labeled antibodies to cell surface antigens. C) Molecular determination of which HLA alleles are in the DNA. D) Molecular determination of which HLA alleles are expressed by the RNA. Answer: C 8) All of the following cell types express Class I MHC antigens except 8) A) mature red blood cells. B) mature lymphocytes. C) brain cells. D) platelets. E) differentiated muscle cells. Answer: A 9) Why are red blood cell transfusions not rejected in the same way that transplanted tissues are? 9) A) MHC Class II molecules are unstable on red blood cells once they are removed from the body. B) Mature red blood cells do not express MHC Class II. C) Mature red blood cells do not express MHC Class I. D) Refrigerator storage of red blood cells inactivates both MHC Class II and I. E) MHC Class I molecules are unstable on red blood cells once they are removed from the body. Answer: C 10) Which of the following is TRUE of both MHC Class I and II receptors? 10) A) nonfunctional if alpha and beta chains are not properly associated. B) heterodimers composed of alpha and beta chains. C) alpha chain similar in structure to an immunoglobulin heavy chain and beta chain similar in structure to an immunoglobulin light chain. D) necessary to present foreign antigen to lymphocytes. E) All of the above are true. Answer: E 11) What is beta - 2 - microglobulin? 11) A) The beta chain of the Class II MHC molecule B) The alpha chain of the Class I MHC molecule C) The alpha chain of the Class II MHC molecule D) The beta chain of the Class I MHC molecule Answer: D 12) Which of the following cell types is not associated with MHC Class II expression? 12) A) Lymphocytes B) Macrophages C) Dendritic cells D) Muscle tissue E) Monocytes Answer: D 2

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    13) An important component of the immune response that Class III MHC genes code for is 13) A) immunoglobulins. B) complement components. C) lysozyme. D) interferon. E) myeloperoxidase. Answer: B 14) What antigen(s) are Class I and II MHC molecules capable of presenting to T cells? 14) A) Antigens endogeneous to the normal host cell B) Antigens digested from phagocytized material C) Antigens produced by viruses inside the host cell D) All of the above Answer: D 15) When foreign tissue is transplanted, the recipient's T cells react against 15) A) public epitopes. B) private epitopes. C) alloepitopes. D) All of the above. Answer: D 16) Panel reactive antibody (PRA) testing in organ transplantation is a system to test 16) A) organ donors for preexisting HLA - primed T cytotoxic cells. B) organ recipients for preexisting HLA - primed T cytotoxic cells. C) organ recipients for preexisting HLA antibodies. D) organ donors for preexisting HLA antibodies. Answer: C 17) All the following are contraindications to harvesting organs from a particular donor except 17) A) cardiac activity. B) presence of antibodies to human immunodeficiency virus. C) presence of metastatic carcinoma. D) neurologic activity. Answer: A 3

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    18) A donor organ has the following HLA and ABO types: A positive HLA type — A1, A3, B4, B5, DR 6, DR7 Observe the potential recipient information below. Based on UNOS criteria, which recipient should receive this organ? 18) A) Recipient 4 — 47 years old, never received an organ Current medical condition — poor A positive HLA type — A1, A3, B4, B10, DR 6, DR7 Panel reactive antibodies — positive; cross - match with donor tissue positive Time on waiting list — 15 years B) Recipient 2 — 5 years old, never received an organ Current medical condition — stable A positive HLA type — A2, A3, B4, B5, DR 6, DR9 Panel reactive antibodies — negative Time on waiting list — 1 year C) Recipient 1 — 65 years old, never received an organ Current medical condition — poor B positive HLA type — A1, A3, B4, B5, DR 6, DR7 Panel reactive antibodies — negative Time on waiting list — 5 years D) Recipient 3 — 29 years old, received an organ 2 years ago and has rejected it Current medical condition — poor O positive HLA type — A2, A3, B1, B5, DR 6, DR7 Panel reactive antibodies — negative Time on waiting list — 2 years Answer: B 19) Which of the following is(are) used to transplant progenitor cells (pluripotent stem cells)? 19) A) Growth factor stimulated peripheral blood B) Umbilical cord blood C) Bone marrow D) All of the above Answer: D 20) In order to transplant allogeneic progenitor (stem) cells, the recipient must 20) A) have a normal immune system. B) be completely immunodeficient. C) be on immunosuppressive therapy. D) be cytokine stimulated to promote engraftment. Answer: B 4

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    21) Which of the following patients is a candidate for autologous progenitor (stem) cell transplant?180 21) A) Breast cancer patient B) Multiple myeloma patient C) Acute myelogeneous leukemia patient D) Patient with severe combined immune deficiency disease (SCID) cause by genetic ADA deficiency E) Sickle cell anemia patient Answer: A 22) Which statement below best summarizes how information regarding a patient's HLA type is used with respect to diseases associated with specific HLA types? 22) A) Odds ratios can be calculated for certain HLA types and the diseases with which they are associated. This is used as supplemental information in diagnosis and counseling. B) HLA typing is done to exclude the presence of a disease and is an important part of diagnosis. C) HLA typing is done to confirm the presence of a disease and is an important part of diagnosis. D) HLA associations with diseases are so unreliable that they are never used in diagnosis. Answer: A 23) Patients with seronegative inflammatory joint problems could benefit from HLA typing to see if they are ________ positive. 23) A) A1 B) DQ1 C) A56 D) B11 E) B27 Answer: E 24) Certain HLA types have been associated with the progression of ________. 24) A) rheumatoid arthritis. B) systemic lupus erythematosus. C) herpes varicella zoster (chicken pox). D) AIDS. Answer: D 25) People with the HLA haplotype A1, B8, DR3 have an increased nonresponse rate to which vaccine? 25) A) Polio B) Hepatitis B virus (HBV) C) Measles/mumps/rubella (MMR) D) Influenza virus E) Diptheria/pertussis/tetanus (DPT) Answer: B SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 26) The cluster of genes found on the short arm of chromosome 6 that determine "tissue type" is called ________. 26) Answer: major histocompatibility complex (MHC) 27) The number of alleles for genes present in the major histocompatibility complex make this the most ________ set of human genes ever identified. 27) Answer: polymorphic 28) When T cells do not recognize an antigen presented by a cell in conjunction with MHC Class I or II molecules, this is called ________. 28) Answer: tolerance 5

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    29) Transplants are most successful if the recipient is both ________ and ________ compatible with the donor's tissue 29) Answer: HLA , ABO 30) The HLA types DR2 and DQ1 are associated with ________. 30) Answer: narcolepsy TRUE/FALSE. Write 'T' if the statement is true and 'F' if the statement is false. 31) One of the reasons that compatible organs for transplantation are so difficult to find is that there is a relatively high rate of recombination in the MHC genes. 31) Answer: True False 32) The probability that two siblings from the same parents have identical HLA types is 25%. 32) Answer: True False 33) MHC Class I expression in a particular cell type remains constant through that cell's life cycle once it matures. 33) Answer: True False 34) MHC pseudogenes express alpha and beta MHC receptor chains that are similar to heavy and light immunoglobulin chains. 34) Answer: True False 35) Organs for transplant in the United States are allocated based on which person on the waiting list is the sickest. 35) Answer: True False 36) The final decision as to whether a person receives a particular organ for transplant in the United States is made by UNOS (United Network of Organ Sharing). 36) Answer: True False 37) The best donor for progenitor (stem) cell transplantation if the recipient himself. 37) Answer: True False 38) The greatest immediate risk after allogeneic progenitor (stem) cell transplant is graft versus host disease. 38) Answer: True False 39) There is no benefit from graft versus host disease to any patient population. 39) Answer: True False 40) To date, the strongest association of an HLA type with a disease is the association of 5 - CREG with Hodgkin's disease. 40) Answer: True False 6

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    SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 41) Explain why as a general rule a child's HLA type is only 50% identical to his father's and 50% identical to his mother's. 41) Answer: Each parent has two sets of MHC genes, and because of the polymorphism of the MHC system, these gene sets would virtually never be identical. Each child gets one of those two sets from each parent. Because the genes are closely linked, they are usually inherited intact * by classic Mendelian genetics (Law of Segregation or Law of Independent Assortment). The inherited alleles are expressed codominantly, meaning that both the mother's and the father's genes are expressed and to equal degrees. The following example shows that any possible child would only have 50% of a parent's HLA type. Mother's genotype = a and b Father's genotype = c and d Possible children = a and c, a and d, b and c, b and d * The recombination rate of the HLA genes is only about 2%. 42) Explain linkage disequilibrium and how it relates to predicting the frequency of particular HLA alleles. 42) Answer: Example: An organ transplant candidate inherited MHC gene set A from his mother and MHC gene set B from his father. An organ must be found that has both A and B MHC genes. If the frequency of A in the general population is 1% and the frequency of B is 1% than the predicted probability of finding the two together is frequency of A x frequency of B = 0.01 x 0.01 = 0.0001 or 1 in 10,000 In many cases, the prediction illustrated above is wrong, and in certain populations the incidence of particular gene combinations is higher than predicted. This is called linkage disequilibrium. Since many examples of linkage disequilibrium seem to be related to ethnicity and/or nationality, knowledge of the gene pool in various ethnic populations can be useful to more easily identify compatible donors. 43) Describe the composition of the Class I MHC molecule on a cell's surface. 43) Answer: The Class I MHC molecule is composed of an alpha chain and a beta chain complex. The alpha chain spans the cell membrane with an external domain, a transmembrane domain, and an internal domain. The alpha chain is similar in structure to a heavy chain immunoglobulin molecule. The beta chain, also called beta - 2 - microglobulin, is similar in structure to a light chain immunoglobulin molecule, but its structure is constant and highly conserved. The external domain of the Class I molecule is used to present foreign antigen to T cytotoxic cells. 7

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    44) Discuss the nomenclature of the HLA alleles. 44) Answer: Initially, the HLA A and B genes were numbered sequentially as they were discovered (e.g., HLA B2). Newly discovered genes were given the provisional designation "w" until it was certain that the discoveries were genuine (e.g., HLA Bw2). To prevent confusion between the HLA complement genes and the HLA C genes, the "w" designation was permanently given to all HLA C genes. As typing methods became more sophisticated, the specificities of different alleles become more defined, and epitopes shared by different alleles were identified. Some previously identified allele epitopes were characterized as "splits" when two or more individual epitopes were found to be components of the original epitopes. This means that related epitopes may not be numbered consecutively and that ever more sophisticated nomenclature for the alleles is developing. 45) Discuss the optimal circumstances to approach a family about organ donation. 45) Answer: The facility should first verify that the prospective donor meets all the criteria necessary for donation. The family should be notified that the donor will not survive, and they should be given some time to assimilate that information. A health care provider who has previously interacted successfully with the family should gather the family in a private room away from the bedside to discuss the possibility of organ donation. 8

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    Exam Name___________________________________ MULTIPLE CHOICE. Choose the one alternative that best completes the statement or answers the question. 1) The regulatory body for HLA testing for transplant programs is 1) A) College of American Pathologists. B) Foundation for the Accreditation of Cellular Therapy. C) American Society of Histocompatibility and Immunogenetics. D) United Network for Organ Sharing. Answer: C 2) The complement dependent cytotoxicity method of HLA testing can be used for 2) A) cross - matching recipient plasma with donor cells. B) HLA typing of Class I antigens. C) identifying recipient HLA antibodies. D) HLA typing of Class II antigens. E) All of the above. Answer: E 3) Which of the following is (are) TRUE regarding acceptable specimens for HLA typing by serological methods? 3) A) Lithium heparin is the anticoagulant of choice for HLA typing. B) Peripheral blood, spleen tissue, and lymph node tissue can all be used for HLA typing. C) Specimens should be stored at 2 – 6° C after collection. D) Specimens preserved in sodium heparin or acid citrate dextrose are acceptable for up to one week after collection. E) All of the above. Answer: B 4) Can sodium oxalate and EDTA be used for HLA typing, antibody identification, and cross - matching by conventional serologic methods? 4) A) Yes, as long as the specimen is kept at room temperature. B) Yes, as long as the specimen is kept at 2 – 6° C. C) No, because EDTA and oxalate chelate Mg ++ and Ca ++ . D) No, because EDTA and oxalate are toxic to white blood cells. Answer: C 5) The Ficoll Hypaque technique isolates mononuclear cells by 5) A) cell surface marker affinity. B) cellular adherence. C) cell membrane solubility. D) cell density. Answer: D 6) How can monocytes be removed from lymphocyte preparations? 6) A) Washing Ficoll - Hypaque density separation with 5% ethanol B) Specific monocyte destruction with monoclonal antibodies C) Adding iron filings for monocyte phagocytosis D) A and B E) B and C Answer: E 1

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    7) Class II HLA antigens are best determined from 7) A) monocytes. B) T lymphocytes. C) B lymphocytes. D) neutrophils. Answer: C 8) In a complement dependent cytotoxicity assay used for HLA antigen typing, which dye can be used to identify cells that have been lysed by corresponding antibody? 8) A) Ethidium bromide B) Fluorescein isothiocyanate C) Acridinium chloride D) Carbonylmethyl fluorescein diacetate Answer: A 9) How can transplant recipients acquire HLA antibodies? 9) A) Blood transfusion B) Prior organ transplant C) Pregnancy D) All of the above Answer: D 10) Panel reactive antibody testing for preexisting HLA antibodies is not done on recipient's ________ 10) A) liver. B) progenitor (stem) cell. C) heart. D) kidney. Answer: D 11) In which situation below is transplant contraindicated? 11) A) Heart transplant recipient with negative T cell cross - match and positive B cell cross - match with donor cells. B) Kidney transplant recipient with positive T cell cross - match and negative B cell cross - match with donor cells. C) Liver transplant recipient with negative T cell cross - match and positive B cell cross - match with donor cells. D) All of the above. Answer: B 12) An advantage of performing HLA cross - matches using flow cytometry is that 12) A) flow cytometers are considerably less expensive than fluorescent microscopes. B) Both T and B cell cross - matches can be performed in one tube. C) the completed cross - match tubes can be stored for about a week until analysis is convenient. D) viable cells are not required. Answer: C 2

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    13) Observe the flow cytometry cross - match protocol in the table and interpret it. FITC is fluoroscein isothiocyanate which fluoresces green, and PE stands for phycoerythrin which fluoresces red. Tube 1 Tube 2 Step 1 Donor cells + recipient serum Donor cells + recipient serum Step 2 Incubate, wash Incubate, wash Step 3 Add FITC - anti - IgG Add FITC - anti - IgG Step 4 Incubate, wash Incubate, wash Step 5 Add PE - anti CD3 Add PE - anti CD19 Step 6 Incubate, wash Incubate, wash Step 7 flow cytometry: No fluorescence detected flow cytometry: green & red fluorescence detected 13) A) This is a positive B cell cross - match. B) This procedure must be repeated because it was performed incorrectly. C) This procedure must be repeated because no fluorescence was detected in tube 1. D) This procedure must be repeated because both fluorochromes were detected in tube 2. E) This is a positive T cell cross - match. Answer: A 14) What is an important advantage of HLA testing by molecular methods? 14) A) Molecular methods are inexpensive. B) Many alleles thought to be different based on serological methods have actually been shown by molecular methods to be the same. C) There is more lot - to - lot variability in antisera than there is in molecular oligonucleotide primers. D) Although cells used for typing must still be viable, fewer cells are required to get an accurate typing. Answer: C 15) If a unrelated donor is being considered for progenitor cell transplant, which molecular HLA typing method below is required before final transplant can occur? 15) A) Low - resolution PCR sequence specific oligonucleotide primer typing B) High - resolution PCR sequence specific oligonucleotide primer typing C) Sequence - based typing D) High - resolution PCR sequence specific probe typing E) Low - resolution PCR sequence specific probe typing Answer: C 16) Which of the following is not acceptable for specimens to be used for HLA typing? 16) A) ACD anticoagulant B) Refrigerator storage up to 14 days C) Room temperature storage up to five days D) EDTA anticoagulant E) Heparin anticoagulant Answer: E 3

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    17) When DNA has been extracted from a sample for HLA molecular typing 17) A) it must be amplified immediately to insure accuracy of the process. B) it can be stored one month at – 20° C. C) it can be stored in the refrigerator no more than 24 hours prior to amplification. D) it can be stored indefinitely at –70 ° C. Answer: D 18) A high resolution HLA sequence - specific primer PCR reaction was done in a 96 well microtiter plate. Each well contains primers to a different HLA allele and the same primers to a sequence of DNA to be used as an internal control. A negative control consists of distilled water left in an open tube during the specimen processing. After PCR, the contents of each well were electrophoresed and visualized on gel. Which finding automatically indicates a potentially invalid reaction that requires repeat analysis? 18) A) In the well that an HLA - A gene amplified, the control sequence also amplified (i.e., two bands were visible). B) Two different alleles for the HLA - A locus were amplified. C) An allele determined by PCR does not match the allele determined by classic serology. D) No bands were visualized in the negative control. E) An allele identified by the high resolution procedure does not match the allele identified by the low - resolution procedure. Answer: E 19) Under the Clinical Laboratory Improvement Amendments (CLIA) regulations, HLA testing is considered 19) A) low complexity. B) high complexity. C) waived. D) moderate complexity. Answer: B 20) For CLIA compliance, HLA testing labs can be inspected by 20) A) College of American Pathologists (CAP). B) American Association of Blood Banks (AABB). C) Centers for Medicare and Medicaid Services. D) American Society of Histocompatibility and Immunogenetics (ASHI). E) All of the above. Answer: E 21) HLA labs that support transplant programs can be accredited by 21) A) Centers for Medicare and Medicaid Services. B) American Society of Histocompatibility and Immunogenetics (ASHI). C) College of American Pathologists (CAP). D) American Association of Blood Banks (AABB). E) All of the above Answer: B 22) On - site laboratory inspection for accreditation of HLA testing laboratories must occur a minimum of every 22) A) 2 years. B) 1 year. C) 5 years. D) 6 months. Answer: A 4

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    23) Personnel who work in HLA laboratories earn certification from the 23) A) National Credentialing Agency for Laboratory Personnel. B) American Society of Clinical Pathologists. C) American Board of Histocompatibility and Immunogenetics. D) American Association of Blood Banks. E) All of the above. Answer: C 24) Which process below includes all the controls utilized to make sure that a particular laboratory analysis is performing properly? 24) A) Continuous quality improvement (CQI) B) Quality assurance (QA) C) Quality control (QC) D) Total quality management (TQM) Answer: C 25) For quality control purposes, which reagent below must be tested against the antibody screening panel to insure that it lacks HLA antibodies? 25) A) Trypan blue B) Complement C) Protein supplement D) Antiglobulin Answer: C SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 26) For HLA typing, the cell of choice is the ________. 26) Answer: lymphocyte 27) HLA typing is acceptable on lymphocyte preparations in which > 95% of the cells can ingest the dye ________ ________. 27) Answer: trypan blue 28) The specimen required from a transplant recipient to do panel reactive HLA antibody testing and crossmatching with donor cells is ________. 28) Answer: serum 29) Gels for HLA typing by PCR should be interpreted by at least ________ (how many?) people. 29) Answer: 2 30) After progenitor cell transplant, if STR DNA regions from both donor and recipient are detected, this is called a genetic ________. 30) Answer: chimera TRUE/FALSE. Write 'T' if the statement is true and 'F' if the statement is false. 31) If a transplant is going to fail due to immune attack, it will be because of recipient production of antibodies or cytotoxic T cells. 31) Answer: True False 32) Traditional HLA typing always requires viable cells. 32) Answer: True False 5

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    33) A lymphocyte preparations in which < 95% of the cells have ingested trypan blue must be discarded and not used for HLA typing. 33) Answer: True False 34) If ethidium bromide is used as the supravital dye in a complement dependent cytotoxicity assay, then cells injured by HLA antibody will appear orange. 34) Answer: True False 35) An advantage of flow cytometry cross - matching is that B cells and T cells do not have to be separated in lymphocyte preparations prior to analysis. 35) Answer: True False 36) Although flow cytometry donor:recipient cross - matching is not as sensitive as the traditional complement dependent cytotoxicity assays, it is widely used because it is automated and significantly reduces turnaround time. 36) Answer: True False 37) When DNA is extracted from a sample for HLA typing, the purity of the sample is best determined by the ratio of the sample's absorbance at 260 and 280 nm. 37) Answer: True False 38) PCR sequence specific oligonucleotide probe methods are usually preferred for large volumes of specimens over PCR sequence specific primer methods. 38) Answer: True False 39) It is still normal a year after progenitor cell transplant to perform STR analysis and find both donor and recipient DNA in peripheral blood white cells. 39) Answer: True False 40) One of the most frequent reasons for death after a progenitor cell transplant is graft versus host disease. 40) Answer: True False SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 41) List three ways that T and B cells can be separated from each other using mononuclear cell preparations. 41) Answer: 1. T cells possess the CD2 receptor that binds to sheep RBCs. When sheep RBCs are added to a mononuclear preparation, T cells will bind to them, and the heavier cell clusters can be removed from B cells by Ficoll - Hypaque density gradients. Lysis of the sheep RBCs liberates the T cells. 2. Aliquots can be mixed with magnetic particles coated with monoclonal antibodies to T and B cells. For example, when anti – T cell particles are added, T cells bind to the particles and are held to the side of the test tube with a magnet while B cells are decanted off. 3. Nylon wool non - specifically bind B cells and monocytes, allowing T cells to be washed through and collected. A cold wash then removes B cells. 6

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    42) List the steps in a complement dependent cytotoxicity assay used for HLA antigen typing. 42) Answer: A complement dependent cytotoxicity assay proceeds as follows: 1. Aliquots of cells are mixed with different HLA antibodies and incubated. 2. Complement is added. 3. Supravital dye is added. Injured cells cannot exclude ethidium bromide and will be orange while intact cells are capable of retaining carbonylmethyl fluorescein diacetate and appear green. 4. Formalin is added to fix cells. 5. Lysis is determined by the presence of dye in cells. 43) How can the sensitivity of the cross - match between the recipient's serum and the donor's cells be enhanced? 43) Answer: After the initial serum:cell incubation in a cross - match, the cells can be washed (Amos modification) to remove any unbound antibodies that have fixed complement. Antihuman globulin can then be added to enlarge any formed immune complexes. Fresh complement is then added, and the larger immune complexes should activate it efficiently. Detection of Class I antibodies in the presence of Class II antibodies can be enhanced by first absorbing the recipient's serum with donor platelets that express Class I but not Class II antigens. 44) Describe the technique of the mixed lymphocyte culture reaction (MLC). 44) Answer: When live lymphocytes recognize foreign Class II HLA antigen on other cells, they stimulate proliferation of several white cell lines to destroy the foreign cells. For an MLC, two mononuclear cell populations are prepared, one from the transplant recipient and one from the donor. The cells from the donor are treated with an inhibitor of DNA synthesis, so that while they might stimulate a reaction from the recipient's cells, they are not capable of mounting their own response to the recipient's cells. The two cell preparations are mixed in a culture medium that contains tritiated thymidine. If the recipient's cells do not recognize any HLA antigens in the donor's cells as foreign, no proliferation occurs, and the cells do not take up the tritiated thymidine to any significant degree. If the recipient's cells do recognize any HLA antigens in the donor's cells as foreign, proliferation occurs and the cells take up the tritiated thymidine aggressively. 45) What is a short tandem repeat (STR), and how can it be used to determine progenitor cell engraftment? 45) Answer: Throughout each chromosome are areas of repeated sequences that are not translated as part of any known gene. Variable number tandem repeats (VNTRs) are repeats of 9 – 80 base pairs while short tandem repeats (STRs) are repeats of 2 – 7 base pairs. Since the repeated sequences are not translated, mutations in these areas are highly tolerated, leading to substantial differences between individuals regarding the actual number of base pairs repeated in each of these regions. These differences can be used as markers to identify the individual who is the source of a piece of DNA. Prior to progenitor cell transplant, the STR regions of donor and recipient DNA are tested until an "informative locus" is found. An informative locus is one in which the donor and recipient STRs differ enough that their DNA can be differentiated. Several loci typically need testing as progenitor cell transplants are often done with related donors. Once an appropriate locus is determined, STR analysis of white cell DNA in the recipient can be done to determine the degree to which the donor's cells did or did not successfully engraft. 7

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    Exam Name___________________________________ MULTIPLE CHOICE. Choose the one alternative that best completes the statement or answers the question. 1) The most primitive component of the immune response found even in some of the simplest forms of life is the ability to 1) A) distinguish self from non - self. B) produce phagocytes. C) mount a cellular immune response. D) produce antibodies. Answer: A 2) Immunology was originally a subdiscipline of 2) A) blood banking. B) transplantation. C) anatomic pathology. D) microbiology. Answer: D 3) Which of the following is (are) characteristics of nonspecific, innate immunity? 3) A) Only needs to recognize agent as "non - self" to react. B) Responds to all challenges equally. C) Has no "memory." It does not react differently when reexposed to the same agent. D) Reacts without prior exposure to the agent. E) All of the above. Answer: E 4) A child has chicken pox and recovers. Later, she is exposed to chicken pox and does not get sick, but when she is exposed to influenza she does get sick. This is an example of 4) A) innate immunity. B) vaccination. C) nonspecific immunity. D) adaptive immunity. Answer: D 5) A man opens up an old loaf of bread and inhales Penicillium spores from the organism growing inside. Inside the man's lungs, phagocytes engulf and digest the spores. The man never gets sick. This is an example of 5) A) vaccination. B) adaptive immunity. C) specific immunity. D) variolation. E) innate immunity. Answer: E 6) The definition of antigen is 6) A) the portion of the antibody that binds to the microorganism. B) an infectious, pathogenic microorganism. C) the portion of the white blood cell responsible for recognizing something as "non - self." D) the exact portion of a microorganism or chemical that the immune system reacts against. Answer: D 1

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    7) In the recognition phase of the immune response, the body must determine that 7) A) a microorganism is different that what is usually present in the normal flora. B) the antigen encountered is "non - self." C) a substance is capable of causing harm. D) a microorganism is capable of causing actual disease. Answer: B 8) The first description of inflammation came from ancient Rome and included all the components listed below except 8) A) swelling. B) pain. C) pus. D) heat. E) redness. Answer: C 9) In humans, after the recognition phase of the immune reaction, the response phase consists of 9) A) activation of the adaptive, specific immune components. B) activation of the innate, nonspecific immune components. C) both A and B. D) Cannot tell from this information. It depends on the immunizing agent as to whether A or B is used. Answer: C 10) The English physician who vaccinated a boy with cowpox to induce immunity to smallpox was 10) A) Robert Koch. B) Peter Medawar. C) Louis Pasteur. D) Edward Jenner. E) Celsus Galen. Answer: D 11) Simian immunodeficiency virus (SIV) causes a disease in monkeys similar to AIDS in humans caused by human immunodeficiency virus (HIV). In theory, why would SIV be a candidate organism to be used in a human AIDS vaccine? 11) A) SIV could stimulate a humoral immune response against HIV. B) SIV could stimulate a cellular immune reaction against HIV. C) Monkey viruses cause disease in humans. D) Monkey viruses cannot cause disease in humans. E) SIV could stimulate an immune response against SIV that cross - reacts with HIV. Answer: E 12) The person who won the Nobel prize in 1908 for his work in demonstrating phagocytosis by white blood cells is 12) A) Paul Erlich. B) Elie Metchnikoff. C) Edward Jenner. D) Robert Koch. E) Louis Pasteur. Answer: B 2

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    13) The first true vaccine given to a human containing the specific pathogen to which immunity was desired was 13) A) chicken fowl cholera. B) anthrax. C) tetanus. D) rabies. E) diphtheria. Answer: D 14) The first research identifying the activity of antibodies demonstrated that 14) A) the immunity could be transferred from one animal to another using the serum component of blood. B) the immunity was produced by B lymphocytes. C) the immunity could be transferred from one animal to another using the cellular component of blood. D) the immunity was produced by T lymphocytes. Answer: A 15) Hyperacute rejection of a solid organ transplant occurs if there is a mismatch of 15) A) HLA types. B) MHC types. C) ABO blood types. D) tissue types. Answer: C 16) The first successful human kidney transplant was in 1954. Why was it successful? 16) A) The recipient and the donor had identical tissue types. B) The recipient and the donor were identical twins. C) The recipient and the donor had identical ABO blood groups. D) All of the above. Answer: D 17) How were the cells responsible for cell - mediated immunity first identified as T lymphocytes? 17) A) Metchnikoff demonstrated phagocytosis of fungal spores by Daphnia blood cells. B) Ehrlich demonstrated it using cells isolated from the Bursa of Fabricius in chickens. C) Glick demonstrated it using cells isolated from chicken thymus. D) Landsteiner demonstrated it showing incompatibility between sera of people with different blood groups. Answer: C 18) In early studies of antibodies, which properties were recognized? 18) A) Ability to cause precipitation reactions. B) Ability to agglutinate particles. C) Antitoxin effects. D) All of the above. Answer: D 3

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    19) The selective/clonal selection theory of immune response ultimately proved to be correct, but the instructional theory, while incorrect, seemed logical because 19) A) if thousands of preformed antibodies were present to antigens that are never encountered, this is excessively wasteful and poor use of the body's resources. B) the immune system was shown to be capable of such a wide variety of substances that it seemed impossible that such a high number of different possibilities would be preformed without any prior exposure. C) if one gene produces one antibody, then it didn't seem that humans had enough DNA to produce thousands of possible antibodies. D) it seemed reasonable that a cell would be "instructed" to react to a specific antigen only when it was encountered. E) All of the above. Answer: E 20) Which breakthrough below is mostly responsible for supporting the clonal selection theory of the immune response? 20) A) Discovery of the T cell receptor B) Discovery of the genes encoding for different portions of antibody molecule C) Discovery of how monoclonal antibodies can be produced artificially D) Discovery of the chain structure of antibodies Answer: B 21) What is a monoclonal antibody? 21) A) An antibody produced by a mature, stimulated B cell. B) An antibody produced in response to an antigen with a repeating epitope. C) An antibody produced by a hybridoma cell. D) An antibody produced in response to an antigen with only one epitope. E) An antibody produced by an immature B cell. Answer: C 22) How can hybridoma cells be kept alive for extended periods of time? (Check as many as apply.) 22) A) They can't. Once removed from the host animal the cells die quickly. B) Growing them at refrigerator temperature. C) Placing them into a susceptible host where they will form a tumor. D) Using the appropriate cell culture techniques. E) Growing them in a field of ionizing radiation. Answer: C, D 23) Which of the cell types listed below needs to have antigen processed before it can be recognized? 23) A) B lymphocytes B) Neutrophils C) T lymphocytes D) Eosinophils E) Monocytes/macrophages Answer: C 4

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    24) Which of the following is true regarding immunoglobulins? 24) A) Immunoglobulins exist preformed on the surface of B lymphocytes. B) They are the major mediator of specific humoral immunity. C) Immunoglobulins are also called antibodies. D) All of the above. Answer: D 25) T lymphocytes recognized foreign antigen and discriminate self from non - self using 25) A) the MHC complex. B) the T cell antigen receptor. C) ABO antigens. D) A and B. E) A and C. Answer: D SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 26) ________ are responsible for cell - mediated immunity while ________ are responsible for humoral immunity. 26) Answer: T lymphocytes/B lymphocytes or antibodies 27) The study of organ transplantation was vastly improved when the cause of different tissue types, the ________, was discovered by Snell, Dausett, and Benacerraf. 27) Answer: major histocompatibility complex 28) The person who discovered the ABO human blood groups is ________. 28) Answer: Karl Landsteiner. 29) Rejection of transplanted organs can be minimized if ________ testing is done to assess tissue compatibility between donors and recipients. 29) Answer: HLA or histocompatibility testing 30) ________ is when one's own tissues are attacked by one's own immune system. 30) Answer: Autoimmunity TRUE/FALSE. Write 'T' if the statement is true and 'F' if the statement is false. 31) The human immune system never responds against human tissues. 31) Answer: True False 32) The adaptive arm of the immune system is capable of memory. 32) Answer: True False 33) In terms of organism survival, cellular immunity is more important than humoral immunity. 33) Answer: True False 34) Antibodies can only be formed against microorganisms or products they produce, such as toxins. 34) Answer: True False 5

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    35) When serum is divided into fractions by an electrical field, antibodies migrate in the beta fraction. 35) Answer: True False 36) It is believed that the phagocytic cells observed by Metchnikoff were macrophages and polymorphonuclear neutrophils. 36) Answer: True False 37) Louis Pasteur accidentally discovered vaccination using attenuated microorganisms by injecting an old bacterial culture into chickens. 37) Answer: True False 38) Paul Erlich was the first to propose that white blood cells had some structure that could bind to antigens similar to a "lock and key." 38) Answer: True False 39) B lymphocytes can react against unprocessed antigen. 39) Answer: True False 40) T cells recognize antigens by virtue their corresponding receptors on their cell surfaces. 40) Answer: True False SHORT ANSWER. Write the word or phrase that best completes each statement or answers the question. 41) What was variolation? 41) Answer: Variolation was a crude vaccination attempt in which crusts from smallpox scabs were introduced into skin scratches or inhaled to try to induce immunity to smallpox. Although it worked in some cases, it was dangerous because if excessive, live smallpox virus was introduced, disease resulted. 42) Explain why Jenner was successful in making a boy immune from smallpox by vaccinating him with cowpox. 42) Answer: Cowpox is a milder disease than smallpox, but it still induces T cell response (cellular immunity) and B cell response (antibodies). The two viruses contain some similar antigens such that the specific immune response against cowpox cross - reacts with smallpox antigens and gives the host immunity to small pox. 43) Define attenuation and explain why attenuated organisms are useful for vaccines. 43) Answer: An organism that is old or otherwise weakened such that it cannot cause disease is said to be attenuated. It should still possess many or most of its antigens that could initiate an immune reaction. Therefore, if the weakened organism were injected into a host, the immune reaction should be specific against the appropriate antigens but the host should not get sick. 6

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    44) Briefly outline the clonal selection theory of adaptive immunity. 44) Answer: Immature uncommitted lymphocytes have the genetic material to respond to a large variety of antigens. During the maturation process, lymphocytes differentiate and become committed to a particular antigen. At all times, there are a small number of mature lymphocytes developed that are capable of responding to every possible antigen that the host can genetically react against. When a particular antigen is encountered, the lymphocyte clone(s) capable of responding to that particular antigen is (are) "selected" or stimulated by the antigen to multiply rapidly and activate an immune response. 45) Explain what a hybridoma cell is and what it does. 45) Answer: A hybridoma cell is a fusion of a malignant myeloma cell and an immunized lymphocyte. The malignant nature of the myeloma cell confers upon the hybridoma almost limitless life in cell culture, while the immunized lymphocyte confers the ability to produce an antibody of a particular type. The hybridoma cell, therefore, produces only one antibody type (monoclonal) in massive amounts artificially in cell culture. 7

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    Exam Name___________________________________ MULTIPLE CHOICE. Choose the one alternative that best completes the statement or answers the question. 1) How does normal microbial flora on the human body prevent infection? 1) A) Its presence stimulates lysozyme secretions. B) Its presence stimulates acute phase reaction proteins. C) Its secretions maintain a low pH inhibitory to most pathogens. D) Its presence reduces nutrients and space needed for pathogens to grow. E) All of the above. Answer: D 2) Each chemical below is paired correctly with its antimicrobial effect except 2) A) interferon/activation of cellular immunity and inhibition of viral replication. B) lactoferrin/binds complex carbohydrates (like lactose) so that organism can't utilize it. C) fibronectin/when bound to bacterial cells walls, promotes phagocytosis. D) transferrin/binds iron so that organisms can't utilize it. E) lysozyme/degrades bacterial cell walls. Answer: B 3) The granules in mature neutrophils contain all of the following except 3) A) transferrin. B) alkaline phosphatase. C) lysozyme. D) gelatinase. E) histamine. Answer: E 4) The process by which neutrophils leave blood vessels, squeezing between the endothelial cells to enter the tissues, is called 4) A) phagocytosis. B) margination. C) chemotaxis. D) diapedesis. E) opsonization. Answer: D 5) A clinical laboratory scientist observed a peripheral blood smear and noticed one to two pale blue inclusions in several neutrophils. Which of the following would be consistent with this observation? 5) A) C - reactive protein is increased. B) Several neutrophils also have vacuoles. C) Serum albumin and transferrin are decreased. D) The total white blood cell count is high with an absolute neutrophilia. E) All of the above. Answer: E 1

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