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3. Out of the four cell types, which one of them does not belong to the immune system?
A) Basophils
B) Endothelial cells
C) Mast cells
D) Red blood cells
6. Danny went on a hiking trip with his friends when he accidentally slipped and fell. He suffered a deep cut at his elbow. Swelling, pain, heat
and redness were observed. A few days later, pus started to form at the wounded site.
True or False
1) The Fc receptor on mast cells binds IgE very strongly and cross-linking by antigen leads to mast cell activation and initiation of an
inflammatory reaction
2) IgE coating mast cells is a second line of defense of external body surfaces when they have been penetrated by an infectious agent
3) The first immunoglobulin heavy chain class to be expressed on the surface of a newly produced B-cell is IgD
A231 Immunology
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a. In the case of Antigen A, which antibody class is dominant in Response X and Y respectively?
Response X – IgM; Response Y – IgG\
On the other hand, Antigen B is exposed to the immune system for the first time; hence, there is an absence of memory B cells in order
for an increased production of antibodies.
COMPLEMENT SYSTEM:
• Fc and complement receptors on phagocytes trigger the uptake and degradation of antibody-coated bacteria (Taken from Janeway, 2001 -
http://www.ncbi.nlm.nih.gov/bookshelf/br.fcgi?book=imm&part=A1233&rendertype=figure&id=A1238)
Antibody coating also induces activation of the complement system and the binding of complement components to the bacterial surface.
These can interact with complement receptors (for example CR1) on the phagocyte. Fc receptors and complement receptors synergize in
inducing phagocytosis.
Bacteria coated with IgG antibody and complement are therefore more readily ingested than those coated with IgG alone. Binding of Fc and
complement receptors signals the phagocyte to increase the rate of phagocytosis, fuse lysosomes with phagosomes, and increase its
bactericidal activity.
• http://pathmicro.med.sc.edu/2006-immpdf/Complement06.pdf
Antigen interaction B-cell receptor (membrane Ig) binds antigen T-cell receptor binds antigen on MHC
Nature of antigens Protein, polysaccharide, lipid Peptide
Binding soluble antigens Yes No
Epitopes recognized Accessible Internal linear peptides produced by antigen
processing (proteolytic degradation)
Antibiotics are medicines that eliminate/ kill bacteria. Antibodies are proteins generated by the cells in the body to help eliminate pathogens
(e.g. bacteria, viruses etc).
Immunisation - process of rendering a subject immune, or of becoming immune. Immunization can occur naturally as your own body creates
immunities to fight off a disease, or through the administration of a vaccine.
Vaccination - is the use of vaccines to help prevent certain diseases. The vaccine is a suspension (in liquid form given orally or by injection)
consisting of weakened or dead pathogens.
Practice Questions
1. All the following are characteristics of both MHC class I and class II molecules except:
B MHC class I molecules are expressed on nearly all nucleated cells, but the constitutive expression of MHC class II molecules is more
limited (B cells, dendritic cells, and thymic epithelial cells). MHC class II expression can be induced on other cell types (such as
macrophages, endothelial cells, and human T cells) by cytokines.
2. A patient is admitted with multiple bacterial infections and is found to have a complete absence of C3. Which complement-mediated
function would remain intact in such a patient?
A) lysis of bacteria
B) opsonization of bacteria
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E All these functions are mediated by complement components that come after C3 and in its absence cannot be activated.
E All are correct. Immunodeficiency disorders may result from defects in the development of bone marrow stem cells into lymphocytes
and other cells that participate in the immune response. They can also result from defects in phagocyte functions, which are important in
phagocytosis and presentation of antigen. Immunodeficiency disorders may also result from defects in complement function — an
absence or malfunction of one or more of the complement components, their activators, or regulators.
A) bind b2-microglobulin.
B) prevent peptide binding to MHC molecules.
C) are part of the proteasome.
D) transport peptides into the endoplasmic reticulum for binding to MHC class I.
E) transport peptides into the endoplasmic reticulum for binding to MHC class II.
D The products of the TAP-1 and -2 genes selectively transport peptides 8—9 amino acids in length from the cytoplasm into the ER
where they bind to MHC class I molecules.
5. Which of the following is incorrect concerning the processing of an antigen, such as a bacterial protein, in the acid compartments of the
cell?
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D CD8+ T cells are generally not activated by processing in acid compartments; exogenous antigen processing in acid compartments
results in the generation of peptides, some of which can displace the CLIP fragment of the invariant chain from the MHC class II binding
groove. The peptide—MHC class II complexes move to the cell surface and can interact with a CD4+ T cell with the appropriate
receptor.
6. Which component(s) of complement could be missing and still leave the remainder of the complement system capable of activation by
the alternative pathway?
D C3 is required for the alternative pathway of complement activation, while C1, C2, or C4 are not required.
A) anaphylatoxin production.
B) depression of factor B.
C) production of chemotactic factors.
D) activation of C
E) activation of C5.
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• T cell receptor (TCR) is unable to discern between extracellular/endogenous and intracellular/exogenous pathogens, and yet the
immune system needs to involve CD4+ T cells and CD8+ in reactions against the respective antigens (derived from the pathogens).
• To sove this problem, the peptides generated from extracellular proteins and the peptides generated from intracellular proteins are
segregated in the cell and trafficked to different classes of MHC molecules. These different classes in turn involve different T-cells:
CD4+ T cells – MHC Class II; CD8+ - MHC Class I.
9. After being injected with a vaccine for a microbe, both T and B-cell responses were observed. Identify the respective responses from
the graph and give an explanation for the responses.
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Response Y – B cells response. The vaccination induces prolonged serum antibody responses that can persist for months or years
after the vaccination.
10. The table below shows a comparison between two types of vaccines.
Pathogen A B
a. The antibodies against toxoid ‘B’ disappear from the blood circulation within a year. Yet why is it only necessary to vaccinate against B
every 10 years?
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b. Why is the vaccine against ‘B’ always effective, whereas the vaccine against ‘A’ does not offer protection on all occasions?
The toxoid is a stable molecule (does not mutate or change in structure). Hence antibodies and lymphocytes which recognise it will still
be effective later on. Pathogen A may mutate frequently. The previous year’s antibodies or the antibodies generated by the current
vaccine may not be effective against new strain of pathogen.
A231 Immunology: Revision Package (Problems 9 to 15) – with Answers to practice questions
Additional resources
• http://www.chem.uky.edu/courses/che554/Lectures/CHE554Lect9_WesternBlot_30March2010.pdf
• http://www.abdserotec.com/uploads/WesternBlottingBrochure.pdf
Transplantation:
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14
• Anaphylactic reactions:
15
http://www.ncbi.nlm.nih.gov/bookshelf/br.fcgi?book=imm&part=A1719&rendertype=figure&id=A1721
16
Taken from:
http://extension.entm.purdue.edu/publichealth/print/insects/stinging.html
17
o http://www.theintellectualdevotional.com/blog/2010/01/25/autoimmune-diseases-basically-an-epic-body-fail/
o http://www.telepathology.com/courses/immune/auto.htm
o http://www.ncbi.nlm.nih.gov/bookshelf/br.fcgi?book=imm&part=A1906#A1918
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1. Figure A and B below shows the Western Blot results detecting presence of complement components C1q and C4 present in the urine
sample of a patient suffering from bacterial infection. The human serum serves as a positive control for the study.
Figure A Figure B
C4
C1q
Human serum Urine sample
Human serum Urine sample
Based on the results shown above, it was postulated that the classical complement pathway has been activated in this patient. Explain
why. (5 marks)
• Protein bands detected in the urine sample are more intense compared to the positive control - increase in production of complement
component C1q and maybe C4.
• Increased level of antibodies binding to the pathogen (present in urine sample) C1q bind to the antibodies activation of the classical
complement cascade pathogen removal
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3. The graph below shows the survival percentages of two groups of liver graft patients. One group received blood transfusions from their
donors before transplantation surgery. The other group did not receive any prior blood transfusions from their donors.
a. Based on the above results, which group of patients received blood transfusions prior to transplantation surgery? Justify your answer. (5
marks)
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b. What are some of the possible pre-transplantation tests that can be conducted to increase the chances of graft survival? (3 marks)
HLA typing tests, ABO compatibility tests, DNA compatibility tests etc.
4. The presence or absence of Protein X (a 70 kDa protein) in patient’s serum is routinely established through the use of ELISA techniques
followed by a confirmatory test (if ELISA test is positive) using Western-blotting techniques.
a. Discuss why 2 tests are used to establish the presence of Protein X in patient’s serum. (Hint: Think of the overwhelming number of
patient’s samples to process in a clinical laboratory)
Any 4
Western blottings expts are slow and hence cannot process samples fast enough
Western blotting expts are able to pick up false positive ELISA results due to unspecific bindings of antibodies used.
Size of proteins reacting with antibodies are revealed in Western-blotting expts; this allow us to realize unspecific binding of
antibodies.
b. In the above 2 tests mentioned, ELISA and Western-blotting experiments, the same pair of antibodies were used.
Primary antibody is rabbit anti-Protein X IgG; while secondary antibody used was enzyme-conjugated horse anti-rabbit
IgG. To reduce economic cost and time of testings, Mr. Chin Kay Kiang decided to carry out both ELISA and Western
blotting experiments using only enzyme-conjugated primary antibody. The antibody used here were identical (apart being
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i. Assuming the conditions used for the new method and old method were the same, discuss why the new method fails to detect Protein X
from Sample B and D; but was able to detect Protein X from Sample A and C?
Any 3
ii. Looking at the results obtained for Sample E from Table 1, discuss why the new method was able to prevent a FALSE POSITIVE ELISA
results as compared to the old method.
Any 3
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Secondary antibody NOT used in new method, hence no false positive results.
5. In an attempt to reduce graft’s rejection, the lymphocytes of Michael (patient), was isolated and mixed individually with fixed lymphocytes of
potential donors, Mr. A, Mr. B and Mr. C, in culture vessels; to observe for lymphocyte proliferation after a period of time. Fixed lymphocytes
are lymphocytes that had been subjected to treatment and cannot proliferate. Hence any proliferation of lymphocytes observed can be
assumed to be due to that of Michael’s. Controls were also set up where the lymphocytes involved in the tests are placed in the culture
vessels. These 4 individuals have the same O blood group. The results are as shown below.
Michael’s lymphocyte with Mr. C’s fixed lymphocytes. Very Low Rate
a. Discuss if the results of this test is can be used to reduce the chances of graft’s rejection in Michael.
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b. Do you think there are any suitable donor(s) identified here? Justify your answer.
Mr. C fixed lymphocytes causes a very mild response from Michael’s lymphocytes (1mark).
Hence, Mr. C is a suitable donor. (1 mark)
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