CHINHOYI UNIVERSITY OF TECHNOLOGY

SCHOOL OF AGRICULTURE

DEPARTMENT OF BIOTECHNOLOGY

Name: NOKUTHABA MITCHELLE SIYAFA

Registration: C18134182X

Programme: BSBIO

Course code and Title: CUBT ANIMAL BIOTECHNOLOGY

Lecturer: DR CHISANGO

Due date: 27 MAY 2020

Question: Describe any 3 options that are being employed for the

production of the Covid-19 vaccines highlighting

challenges being faced

Mark:

Comment:
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Vaccination consists of stimulating the immune system with an infectious agent, or components of
an infectious agent, modified in such a manner that no harm or disease is caused, but ensuring that
when the host is confronted with that infectious agent, the immune system can adequately
neutralize it before it causes any ill effect. For over a hundred years vaccination has been effected by
one of two approaches: either introducing specific antigens against which the immune system reacts
directly; or introducing live attenuated infectious agents that replicate within the host without
causing disease synthesize the antigens that subsequently prime the immune system.

A vaccine imitates the infection to give the immune system a preview of the disease. Vaccination
became a public health tool after Edward Jenner showed in 1796 that inoculation with the less
virulent cowpox could prevent smallpox. Vaccines prepare the immune system by generating
disease-fighting proteins called antibodies, which seek out and attack if the real infectious virus ever
shows up.

Traditional vaccines against viruses are either weakened versions of the whole virus that are unable
to cause disease; or they are made from signature viral proteins called antigens, that then spark an
immune response. An antigen in the new coronavirus SARS-CoV-2 is the crown-like spike (S) protein
through which the virus latches to the lung and respiratory cells. Currently there are over 110
vaccines being produced for the Covid-19 virus and amongst the leading/ promising eight are
adenoviral vector based vaccines, one of which is AD5-nCoV from Beijing, China, mRNA vaccines the
leading one by a biotechnology campany, Mordena as well as inactivated pathogens, the leading
campany being from Wuhan, China.

1. Adenoviral vector based vaccines.

There are currently more than 100 COVID-19 vaccine candidates in development globally, Ad5-nCoV
is the first to be evaluated in humans. It uses a weakened adenovirus, which infects human cells
readily but is incapable of causing disease, to deliver genetic material that codes for the SARS-CoV-2
spike protein to the cells. These cells then produce the spike protein and travel to the lymph nodes
where the immune system creates antibodies that will recognise that spike protein and fight off the
coronavirus.

Adenoviruses are one of the most genetically diverse DNA viruses and cause non-life-threatening
infections in the ocular, respiratory, or gastrointestinal epithelium of a diverse range of hosts.

Adenoviruses activate several innate immune signaling pathways that result in the secretion of a
number of proinflammatory cytokines. These proinflammatory cytokines pave the way for effective
immune cell stimulation and result in the induction of robust adaptive humoral and cellular immune
responses. To resolve infections with intracellular pathogens such as viruses, CD8+ cytotoxic T
lymphocyte (CTL) responses are critical. Transgene antigens carried by adenoviral vectors are
presented to T cells via MHC class I molecules, and therefore, they induce efficient and robust CTL
responses. The CTLs efficiently recognize and kill virus-infected cells, intracellular pathogens, and
cancerous cells. These properties make adenoviral vectors promising as vaccine vectors. A number of
human clinical trials have been conducted for adenoviral vector-based vaccines against different
infectious diseases including Ebola virus, Zika virus, influenza viruses, HIV, Mycobacterium
tuberculosis and now, for Covid-19 virus. Adenoviruses are highly immunogenic and can induce both
robust innate and adaptive immune responses in mammalian hosts and they have a large genome
size, making the manipulation of the genetic DNA much more convenient. Adenoviruses do not
integrate the viral genomic DNA into the hosts' genome, which reduces the risk of insertion
mutagenesis.

Scientists have revealed the first COVID-19 vaccine candidate, adenovirus type 5 vectored COVID-19
(Ad5-nCoV), to be tested in humans is safe, well-tolerated and able to generate an immune
response. The trials in China, Beijing, where the vaccine is being developed demonstrates that a
single dose of the new adenovirus type 5 vectored COVID-19 (Ad5-nCoV) vaccine produces virus-
specific antibodies and T cells in 14 days, making it a potential candidate for further investigation
(Professor Wei Chen, Beijing Institute of Biotechnology, 2020.)

According to the report released from Beijing Institute of Technology in Beijing, China, the trial
assessed the safety and ability to generate an immune response of different dosages of the new
Ad5-nCoV vaccine in 108 healthy adults between the ages of 18 and 60 years who did not have
SARS-CoV-2 infection. Volunteers were enrolled from one site in Wuhan, China, and assigned to
receive either a single intramuscular injection of the new Ad5 vaccine at a low dose (5 × 1010 viral
particles/0·5ml, 36 adults), middle dose (1×1011 viral particles/1.0ml, 36 adults) or high dose (1.5 x
1011 viral particles/1.5ml, 36 adults). The researchers tested the volunteers’ blood at regular
intervals following vaccination to see whether the vaccine stimulated both a humoral (antibody) and
(T) cell-mediated response. The ideal vaccine would generate both to combat SARS-CoV-2. The paper
published in the Lancet reveals the Ad5-nCoV vaccine was well tolerated at all doses with no serious
adverse events reported within 28 days of vaccination. Most adverse events were mild or moderate
and included mild pain at the injection site (54 percent), fever (46 percent), fatigue (44 percent),
headache (39 percent) and muscle pain (17 percent). After 14 days, all doses of Ad5-nCoV triggered
production of SARS-CoV-2 binding antibodies and some participants had detectable neutralising
antibodies (28 percent of low-dose group; medium dose 31 percent; high dose 42 percent). Most
participants had a four-fold increase in binding antibodies after 28 days, at which point half of the
low- and medium-dose groups and 75 percent of those in the high-dose group showed neutralising
antibodies against SARS-CoV-2.

Ad5-nCoV also stimulated a rapid T cell response in the majority of volunteers, with levels peaking at
14 days after vaccination (low-dose group 83.3 percent; medium 97.2 percent; high-dose group 97.2
percent at 14 days). Further analyses showed that 28 days after vaccination, the majority of
recipients showed either a positive T cell response or had detectable neutralising antibodies against
SARS-CoV-2 (Professor Feng-Cai Zhu, 2020.)

A randomised, double-blinded, placebo-controlled phase 2 trial of the Ad5-nCoV vaccine has been
initiated in 500 health adults in Wuhan. It aims to determine whether there are any adverse events
up to six months after vaccination. Thee protocol states 250 volunteers will be given a middle dose,
125 given a low dose and 125 given a placebo as a control. This trial will also include participants
over 60 years old, an important target population for a potential COVID-19 vaccine.

2. mRNA vaccine. (Moderna's mRNA 1273 vaccine

The mRNA vaccines are hailed as the alternative approach to the traditional vaccines. The RNA-
based vaccine work by carrying the molecular instructions to make the protein which is known as the
mRNA sequence. This sequence instructs the human cells to build viral proteins to fight the virus. As
a result, the body’s immune system gears up to fight the virus.
The vaccine candidate mRNA-1273 which is being developed by Moderna for Covid-19, carries the
mRNA strand of the spike protein of the novel coronavirus, which will then be used by the human
cells to produce the antigen required to fight the virus, just by using the genetic information of a
specific protein.

One of the biggest advantages of using RNA-based vaccines is its rapid manufacturing rate which can
prove to be a game-changer, especially in the face of current COVID-19 pandemic. They are also
usually safer to use as they do not carry weakened viruses in them.

As of now, Moderna’s mRNA-1273 vaccine has become the first vaccine candidate to develop
protective antibodies against the virus and stimulated the immune system to fight it. The findings
come from a small early trial conducted in the month of March where 8 people out of the 45
subjects showed positive immune system response and developed protective antibodies against the
virus. All the 8 people received 100 microgram dose of the vaccine and developed minor side effects
including redness, chills and soreness where the injection was given. The symptoms were said to
subside within a day.

Basically, an RNA vaccine works like this; rather than injecting a pathogen’s antigen into the body,
the body is instead given the genetic code needed to produce that antigen itself. When the antigens
appear on the outside of the body cells, the body’s immune system attacks them—and learns how to
defeat future intruders in the process. The body is essentially turned into its own vaccine
manufacturing unit. Because RNA vaccines let the body do most of the work, they don’t require
much material. That makes them much faster to manufacture.

Vaccines work by training the body to recognise and respond to the proteins produced by disease-
causing organisms, such as a virus or bacteria. Traditional vaccines are made up of small or
inactivated doses of the whole disease-causing organism, or the proteins that it produces, which are
introduced into the body to provoke the immune system into mounting a response.

mRNA vaccines, in contrast, trick the body into producing some of the viral proteins itself. They work
by using mRNA, or messenger RNA, which is the molecule that essentially puts DNA instructions into
action. Inside a cell, mRNA is used as a template to build a protein. ‘An mRNA is basically like a pre-
form of a protein and its (sequence encodes) what the protein is basically made of later on,’( Prof.
Bekeredjian-Ding, 2020)

To produce an mRNA vaccine, scientists produce a synthetic version of the mRNA that a virus uses to
build its infectious proteins. This mRNA is delivered into the human body, whose cells read it as
instructions to build that viral protein, and therefore create some of the virus’s molecules
themselves. These proteins are solitary, so they do not assemble to form a virus. The immune
system then detects these viral proteins and starts to produce a defensive response to them.

There’s a catch, though: its not yet known for sure yet if RNA is a viable platform for vaccines. Since
COVID would be the first RNA vaccine out of the gate, there is a need to prove both that the
platform itself works and that it creates immunity. Other outstanding questions include whether the
proteins that have been chosen for the vaccine are the right ones to prevent a coronavirus infection
in the body, how targeted the immune response is to this particular coronavirus, how long any
immunity would last, and whether it causes side-effects such as increased inflammatory responses
like redness and swelling or, in the worst case, aggravates disease.

3. Inactivated pathogen
The most traditional vaccine approach — one utilized over many decades — is to inject someone
with the inactivated virus. This stimulates the immune system to produce antibodies, while the virus
is either killed before injection or weakened sufficiently so that it cannot cause a serious infection.
Inactivated viruses are used against influenza, for example, and in the global effort to eradicate
polio.

Here once again the Chinese are in the lead. The Chinese company Sinovac, in partnership with a
number of leading medical research institutes in China, designed a vaccine by isolating SARS-CoV-2
samples from infected hospital patients and growing the virus in cell lines before inactivating it with
a chemical agent. It is called PiCoVacc (for “purified inactivated SARS-CoV-2 vaccine”).

An international team has a different approach, using a vaccine that is already widely deployed: the
BCG vaccine against tuberculosis. It has been shown to protect against other respiratory diseases,
too, so researchers are hoping it might be effective against COVID. (BCG is an inactivated bacterial
pathogen, not a virus.)

The Chinese team has made impressive progress with its inactivated viral COVID vaccine. In a paper
published in Science on May 6, the team reported that their candidate vaccine had “induced SARS-
CoV-2-specific neutralizing antibodies in mice, rats and non-human primates.” It also “provided
partial or complete protection in macaques” against deliberate infection with the virus. A Phase 1/2
clinical trial with 744 human participants is underway in China, with the first results predicted for
August.

Because BCG already has a decades-long history of safe use as a vaccine, trials to see whether it is
effective against COVID have gone straight to Phase 3. Trials are currently underway among 10,000
frontline health workers in Australia, run by Murdoch Children’s Research Institute, and in the
Netherlands among a further 1,500 health workers.

An inactivated vaccine (or killed vaccine) is a vaccine consisting of virus particles, bacteria, or other
pathogens that have been grown in culture and then lose disease producing capacity. In contrast,
live vaccines use pathogens that are still alive (but are almost always attenuated, that is, weakened).
Pathogens for inactivated vaccines are grown under controlled conditions and are killed as a means
to reduce infectivity (virulence) and thus prevent infection from the vaccine. The virus is killed using
a method such as heat or formaldehyde.

Inactivated vaccines are further classified depending on the method used to inactivate the virus.
Whole virus vaccines use the entire virus particle, fully destroyed using heat, chemicals, or radiation.
Split virus vaccines are produced by using a detergent to disrupt the virus. Subunit vaccines are
produced by purifying out the antigens that best stimulate the immune system to mount a response
to the virus, while removing other components necessary for the virus to replicate or survive or that
can cause adverse reactions. Because inactivated viruses tend to produce a weaker response
immune system than live viruses, immunologic adjuvants and multiple "booster" injections may be
required to provide an effective immune response against the inactivated pathogen.

The downside is that they’re time-consuming to make. There’s a ton of material in each dose of a
vaccine. Most of that material is biological, which means you have to grow it. That takes time,
unfortunately.
References

1. Petrovsky, Nikolai; Aguilar, Julio César (2004). "Vaccine adjuvants: Current state and future
trends". Immunology and Cell Biology. 82 (5): 488–496. doi:10.1111/j.0818-9641.2004.01272.x.
ISSN 0818-9641. PMID 15479434.

2. WHO Expert Committee on Biological Standardization (2016). "Influenza". World Health

Organization (WHO). Retrieved 16 May 2016

3. Advances in mRNA vaccines for infectious diseases, Cuiling Zhang1, Giulietta Maruggi 2, Hu
Shan1 and Junwei Li 1 College of Veterinary Medicine, Qingdao Agricultural University, Qingdao,
China, 2 GSK, Rockville, MD, United States, 2019

4. The COVID-19 vaccine development landscape, Tung Thanh Le, Zacharias Andreadakis, Arun
Kumar, 09 April 2020

5. COVID-19 vaccine candidates show progress amid challenges, Bruce Calson, The science advisory
board contributing writer, 23 April 2020