Student Number: Seat Number:

RHODES UNIVERSITY

FACULTY OF PHARMACY

EXAMINATION: JUNE 2022

BIOCHEMISTRY, MICROBIOLOGY, AND IMMUNOLOGY PC 221

Examiners: Professor R. Tandlich MARKS: 100

Moderator: Dr N. Mutingwende DURATION: 3 HOURS
PASS: 50

INSTRUCTIONS

1) The use of the Concise English Dictionary is NOT allowed.

2) ANSWER ALL QUESTIONS.

3) Start each new full question on a SEPARATE PAGE.

4) FILL in the question numbers on the FRONT COVER of your answer book.

5) Do NOT use red ink anywhere on your script.

6) ALL answers, including drawings, must be written in INK.

Work written in PENCIL WILL NOT be marked.

7) Write your student number and seat number on each page of the answer book.

8) PLEASE HAND IN THE QUESTION PAPER.

PLEASE DO NOT TURN OVER THIS PAGE UNTIL TOLD TO DO

SO.

Biochemistry, Microbiology, and Immunology 2 – June2022 Page 1 of 4

 QUESTION 1
Outline the factors that contribute to the pathogenicity of animal viruses.

 Kill immune cells.

 Cytopathic effects: - The visible effects of viral infection on host cell. Some
effects will kill the cell, and some will just change the cells.
 Viruses stop DNA, RNA and/or protein synthesis e.g., Herpes virus block
mitosis.
 Lysosomal autolysis of host cells e.g., Influenza: bronchiolar epithelium.
 Production of inclusion bodies (visible viral parts inside the cell) can identify a
particular virus e.g., Rabies virus: Negri bodies.
 Syncytium formation (neighboring cells fuse together) e.g., Varicella Zoster
virus.
 Change in cell function e.g., Measles, production of interferons by host cell
(triggers host immune response), induce antigenic changes on host cell surface
(triggers destruction of infected cell by host immune response).
(5)

QUESTION 2
Briefly discuss the steps of endospore formation and describe the importance of endospores in
determining sterilization conditions.

(7)
The stages of endospore formation are as follows:
- Activation
- Germination
- Outgrowth
- Vegetative growth
- Sporulation
- Lysis
Endospores are resistant to heat, desiccation, chemicals, and radiation. They enable bacteria
to lie dormant for long periods. Endospores are important in determining sterilization
conditions because they are resistant to many of the physical and chemical agents used for
sterilization.

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 QUESTION 3
3.1 Describe the various viral classification systems.
(8)
 Based on their shape
- Icosahedral
- Helical
- Enveloped
- Complex

3.2 Which of the classification systems referred to in 3.1 is the most clinically significant?
Justify your answer.
(2)

QUESTION 4
4.1 Describe the differences between the following terms:
i. Virulence and pathogenicity.
Virulence is the capacity of a pathogen to cause disease. Pathogenicity is the
capacity of a microbe to cause damage in a host.
ii. Invasiveness and toxigenesis.
Invasiveness is the propensity of an introduced species to invade a recipient
ecosystem. Toxigenesis is the ability to produce toxins.
(4)
4.2 Describe the cell extensions seen in eukaryotes and explain how they differ from cell
extensions of prokaryotes. Provide a detailed discussion about why these extensions
are a major factor in bacterial virulence.
Eukaryotic cells have a variety of extensions that are used for different purposes. These
extensions include cilia, flagella, and microvilli. Cilia and flagella are used for movement, while
microvilli are used for absorption¹. In contrast, prokaryotes do not have these extensions.
Instead, they have pili and fimbriae. Pili are used for attachment to surfaces and other cells,
while fimbriae are used for attachment to surfaces².
Bacterial virulence is often associated with the presence of pili and fimbriae. These structures
allow bacteria to attach to host cells and tissues, which is often the first step in the infection
process⁶. Once attached, bacteria can then use other virulence factors to invade host cells and
tissues⁶.
(6)

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 4.3 With reference to 4.1 and 4.2, discuss the factors that influence the degree of
pathogenicity and the progression of an infection or disease.
(6)

Factors that Influence the degree of Pathogenicity and the Progression of Infection and Disease
 Host factors: Age, sex, ethnicity, nutrition (diet), hormonal status; personal hygiene and
immune status; Underlying disease or medical condition; Antibiotic or drug usage;
Presence of foreign object (e.g., splinter, catheter, sutures, etc.); Innate differences
between hosts
 Microbial factors: Bacterial virulence factors; Inoculum size (dosage)
 External factors (e.g., crowding; seasonal variations; hygiene, sanitation and public
health; food processing, storage and preparation; etc.

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 QUESTION 5
A new pharmaceutical company, specializing in the production of antibiotics, is looking for a
pharmacist with a microbiology background to take up an advisory role. You applied for this
position and have been called for an interview with the company's CEO. The company is
trying to develop a new antibiotic that will overcome the antibiotic resistance that bacteria X
has developed to antibiotic Y.

The CEO poses questions to you relating to the development of this new antibiotic. Provide
your answer to each of the following interview questions:

i. What factors can affect the selection and use of antibiotic X?

(6)
 Antibiotic should possess activity against the widest possible range of organisms.
 Antibiotic should have a good oral absorption.
 Antibiotic should exhibit minimal toxicity and side effects for the human patient.
 Antibiotic should exhibit good penetration into tissues and cells.
 Antibiotic should not interact with other drugs.
 Antibiotic should not be prone to resistance development.
ii. What factors may influence bacteria X’s susceptibility or resistance to antibiotic Y?
(4)
 Concentration of antibiotic at the site of infection.
 Host defense.
 Specific infection
iii. What factors may lead to the misuse of antibiotic Y, as an active pharmaceutical
ingredient and in the context of the BMI course content?
(4)
 Treatment of non-responsive infections
 Therapy of fever of unknown origin
 Inappropriate reliance on chemth
 Lack of adequate bacteriological information

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 QUESTION 6
Some of the microbes that invade the immune system contain mannose residues on their
surface. These mannose residues trigger a cascade of events, leading to the opsonization,
destruction, and elimination of the microbial antigens from the body. Different active
proteases are involved during this process.

i. Give the name of the pathway that the immune system uses to protect the body from
these microbes. (2)

Classical pathway

ii. Describe the cascade of events that take place in this pathway i.e. from identification of
the microbe to its elimination from the body.

(3)
Classical pathway was named as it was discovered first and uses the plasma protein C1q
to detect antibody bound over the surface of microbes. Following the binding, C1q starts
the cascade which leads to lysis of the microbes.

Steps:

 C1 is a complex made of hexamer C1q and serine proteases C1r and C1s.
 C1q binds to Fc region of antibody. IgM is more efficient because of it’s high avidity
as it can bind to 3 C1q compared to IgG which can bind to only one.
 Activated C1s cleaves C4 and C4b either binds to antigen-antibody complex or to
the adjacent cell surface.
 C1s also cleaves C2 and the C4bC2a complex is the C3 convertase of classical
pathway.
 Once C3b is formed – it can either follow alternative pathway or bind to C4bC2a
(C3 convertase) to form C5 convertase.
 Similarly, MAC is formed.

Biochemistry, Microbiology, and Immunology 2 – June2022 Page 6 of

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iii. Discuss all the plasma proteins involved and their roles using an annotated diagram
ONLY.

(3)

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 QUESTION 7
Immunochemical methods of antigen quantification provide valuable research data in basic
and clinical sciences, or settings. Draw a schematic representation illustrating the principle of
the enzyme-linked immunosorbent assay.

(6)

QUESTION 8
8.1 Discuss the peptide-MHC interaction. (6)
There are some characteristic features of peptide-MHC interaction.
I. MHC class I and II molecules have a single peptide binding cleft that accommodates
one peptide at a time but can bind to different peptides.
II. The processed peptide that binds to MHC shares structural compatibility that
promotes their interaction.
III. MHC acquires the peptide over their cleft during the processing of the antigen inside
the cell.
IV. Only small populations of peptide loaded over the MHC molecules can elicit the
immune responses.
V. MHC molecules present both the self and non-self-peptide to the T cells. Remarkably
it is the T lymphocyte that decides to which the body should produce an immune
response. Majority of the MHC present in the body are loaded with the self-peptide
and T cells activated against the self-peptides are either killed or inactivated by the
host immune surveillance system. Hence, T cells normally do not respond to a self-
antigen.

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 8.2 Briefly describe the antigen processing by either Major Histocompatibility Complex
(MHC) class I or class II pathway. (6)
 The antigen containing endosomes are fused with the lysosome to form
endolysosome, the acidic pH of the endolysosome helps in the degradation of
proteins into smaller peptides.
 MHC class II molecules are synthesized in the endoplasmic reticulum and
transported to the endosomes with the help of invariant chain, which binds to the
peptide binding cleft of a newly synthesized MHC class II molecule.
 Class II molecules with bound invariant chain (CLIP) are transported to
endosomes and are degraded by proteolysis to release the invariant chain.
 The remaining part of CLIP is removed by HLA-DM present in the endosomes in
order to create space for peptide.
 Once CLIP is removed, the peptides are loaded over the MHC class II molecule.
 The MHC class II molecule bound to peptides is delivered to the surface for their
recognition by CD4+ T lymphocytes (humoral immune response).

8.3 Discuss CD40-CD40L interaction during T cell activation and differentiation. (6)
Naïve T-cells activated by peptide MHC complex causes the expression of CD 40 ligand (CD40L)
on T-cell. CD40L binds to CD40 present on dendritic cells or antigen presenting cells which lead
to expression of B7-1 and B7-2 molecules that binds to CD28 receptor present over the T-
lymphocyte. In addition, activated dendritic cell also secretes cytokines such as IL-12 which
further stimulates the proliferation and differentiation of T-lymphocytes.

 T cells recognize antigen causing expression of CD40 ligand on T cells

 CD40 ligand binds to CD40 on DC; leads to DC expression of B7; secretion of cytokines.
 Activated DCs stimulate T cell proliferation and differentiation.

QUESTION 9
Describe how an infection with Gram-negative bacteria triggers fever and why using anti-
prostaglandin medication temporarily reduces fever?
(8)
Gram-negative bacteria cause fever by releasing toxins that cause the body to produce more heat. Anti-
prostaglandin medications work by reducing the amount of toxins produced, which in turn reduces the amount
of heat the body produces¹.

Prostaglandins are hormone-like substances that are produced by the body in response to injury or infection.
They are responsible for causing inflammation and fever². Anti-prostaglandin medications such as aspirin and
ibuprofen can reduce fever by blocking the production of prostaglandins¹.

QUESTION 10
Explain how an antibody helps in the neutralization and opsonization of microorganisms.
(8)
The production of different form of antibodies requires class switching of the surface immunoglobulin present
over the B cells. Antibodies bind and prevent the pathogens, thus “neutralizing” the pathogens and block their
ability to infect host cells. IgA antibodies specifically act on the mucosal surface of the gastrointestinal and
respiratory tract and neutralize the invading pathogens. IgG antibodies coat pathogens and target them for
phagocytosis while IgM can activate the complement pathway.
Biochemistry, Microbiology, and Immunology 2 – June2022 Page 9 of
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 TOTAL MARKS: 100

END OF EXAMINATION PAPER

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