Understanding Mechanisms of Actions for Vaccine

Adjuvants Critical for Designing Effective Vaccines:

Faculty of Biological and Physical Sciences, Tom Mboya University College, Kenya

*Correspondence: Moses Wanyonyi Sichangi, Email: [email protected]/ [email protected]. Tel:

Summary
INTRODUCTION
Vaccine adjuvants enhance immunogenicity of antigens through one or a combination of
mechanisms that include; improved antigen delivery to the innate immune system or by
providing signals that activate the innate immune system. Activation may lead to induction of
cytokines and chemokines, recruitment of immune cells to the site of vaccine inoculation or
trafficking of innate immune cells to draining lymph nodes. These events culminate in activation
of adaptive immunity.

OBJECTIVE
To identify the Current status of knowledge on action modes for vaccine adjuvants,
modes under investigation and future directions in this important area of biomedical research.
influence on molecular and physical interactions between vaccine components and innate
immune cells that affect the degree of immune responses given first priority.

METHODOLOGY
Published studies in English language on vaccines and adjuvants were identified by key
words from Comprehensive searches with no formal assessments for risk of biases. The type of
publications included basic research using experimental animals and clinical research in human. A
recent study using recombinant hemmaglutinin (rH5) protein of highly pathogenic avian influenza
(HPAI) virus as antigen demonstrated a quicker antibody production. IL -17 and IFN-γ when
adjuvant combinations of CpG and nanoemulsion were used in comparison to nanoemulsion alone
[25]. Equally, potential vaccine adjuvants including pathogen associated molecular patterns (PAMPs)
derived from microbes with their synthetic analogs like cytosine and guanine (CpG)
oligodeoxynucleotide that targeted toll-like receptors (TLRs) and mast cell activating compound such
as compound 48/80 (C48/80) [23, 24]. Combination of cationic peptide HH2 with CpG induced IgG1
(Th2) and IgG2a (Th1) type antibodies in experimental animals [30]
Other vaccine adjuvants developed for human use included Monophosphoryl lipid A
(MPLA) and MF59 (oil in water emulsion). MPLA is a detoxified form of bacterial cell wall lipid
A from Salmonella Minnesota R595 combined with alum to augment immunogenicity of subunit
vaccines. When mediated through activation of TLR4 induced Th1 type immune responses [16].
Combined with alum for hepatitis B virus (FENDrix) Human Papilloma Virus (Cervarix) [15]
and then combined with a water-soluble triterpene glucoside as adjuvants for malaria vaccine
trials in human [17]. Combination of a mucopolysaccharide chitosan as a mucosal vaccine with
Norwalk norovirus demonstrated induction of antigen-specific antibody.

18 African Journal of Health Sciences Volume 33, Issue No.3, May- June, 2020
 Cytomegalovirus Glycoprotein B antigen when adjuvanted with MF59 induced a higher
antibody titer using a lower antigen dose compared to antibody titer induced by a higher dose of
the same antigen [32] Inactivated hepatitis A antigen induced a significantly higher
seroconversion rate at two weeks after the first injection with 100 U of antigen compared to 50 or
25 U of the antigen [31]. Intranasal meningococcal subtype B vaccine induced highly bactericidal
immunity with day 0, 7, 28 and 56 schedules than same vaccine given on days 0, 28 and 56 [34].
Induction of dsDNA released when bound by adjuvants formed a complex and trans-located into
the endosome to activate TLR9 cascading to MyD88 adaptor molecule [29, 51] immunity [43].
RESULTS (DATA SYNTHESIS)
This literature review has shown that, several strategies exist that can be applied in order to
maximize immune activation and improve vaccine efficacy. Such strategies include combination of
adjuvants that activate different pathways of the innate immunity. Combination of adjuvants
produced synergistic effect in immune responses and pathogen clearance, but individual adjuvants
more often provided a response that was narrow in its effect, being either Th1 or Th2 biased.
CONCLUSION
Selection of the right adjuvant for a vaccine antigen requires knowledge on the mode of
action of the adjuvant. Different adjuvants and different routes of vaccine administration could
generate various types of immune responses. The route of vaccine administration might influence
the type of cells in the innate immunity activated by an adjuvant. Antibodies with high avidity
strongly bound to antigenic determinants on the pathogen inducing destructive processes against
the pathogen. Intranasal adjuvants can be suitable for mass vaccination against respiratory
infections such as influenza and CORONA viruses ( SARS Cov-2 (COVID-19)). The ratio
between the antigen and adjuvant in a vaccine would influence the structure of the final complex
formed and its biological activities.
RECOMMENDATIONS
To effectively design an adjuvant within a vaccine formulation, first understand its
mechanisms of activity in order to develop a potent, effective and safe vaccine. Induce sufficiently
mature (high avidity) antibodies by a vaccine to avoid lack of protection.
Key words: Vaccine adjuvants, mechanisms of actions of vaccines, mechanisms of actions of
adjuvants, vaccines design and development.
[Afr. J. Health Sci. 2020 33(3) : 19 - 29 ]

Introduction
Vaccination is the most successful way of After vaccination, those antigens first activates
prevention against human infectious diseases [1, 2]. the host innate immune system, whose products then
The main goal was to induce pathogen-specific activate the adaptive immune system [5 -7]. Effective
immune responses using inactivated pathogen activation of innate immune cells was required for
molecules that generate protective immunity against generation of optimum adaptive immune responses.
future infections by similar virulent pathogens [3]. Vaccine antigens that poorly activate the innate
Vaccine antigens may exist in multiple forms such as; immune system could lead to less effective immunity.
- live-attenuated microorganisms,
- inactivated pathogenic microorganisms, Studies have shown that weaker immune
- purified components of microbial pathogens, responses to vaccine antigens were results of original
- polysaccharide-carrier protein conjugates poor immunogenicity most common with purified and
- or recombinant proteins of pathogenic subunit vaccines.
microorganisms [4].

African Journal of Health Sciences Volume 33, Issue No. 3, May - June 2020 19
 Incorporating relevant adjuvants in the vaccine in combination with a water-soluble triterpene glucoside
formulation could enhance the immunogenicity of as adjuvants for malaria vaccine trials in human [17].
vaccine antigens [3, 8]. Although vaccines formulated
using whole pathogens were highly immunogenic, Further studies on MPLA involved
ineffective inactivation of the pathogens was combination with a mucopolysaccharide chitosan as a
potentially infective. Consequently, most recent mucosal vaccine with Norwalk norovirus
vaccines were being produced as recombinant subunit demonstrating induction of antigen-specific antibody
proteins of the pathogens [9]. For example, hepatitis B responses [18]. MF59 (an oil in water emulsion) had
virus (HBV) and human papilloma virus (HPV) been used to improve the immunogenicity of influenza
vaccines were subunit vaccines. vaccine and demonstrated induction of protective
immunity against influenza virus in humans [19].
Although subunit vaccines had high purity, they
were often poorly immunogenic. Hence, vaccine One of the significant observation about
adjuvants were included in subunit vaccine formulations vaccination outcomes was that, different adjuvants and
to enhance immunogenicity and subsequent efficacy different routes of vaccine administration could
[3]. Different vaccines (antigens or antigen-adjuvant generate different types of immune responses [9, 20].
combinations) activated specific pathways of the innate The immune responses may differ in a number of
immunity, generated varying quantities and profiles of aspects including Th1 versus Th2 type immune profiles
immune mediators that determined the quality of the or antibody responses versus T cell mediated responses
adaptive immune response. or a combination of these with variations in magnitudes
or ratios of responses.
Understanding the modes (mechanisms) of
actions of vaccines and accompanying adjuvants was The knowledge about a vaccine was sensitive
therefore crucial in designing effective and safe because certain types of infections (pathogens) could
vaccines. The fact that, vaccines and their adjuvants require induction of specific type of immune responses
in order to be cleared [21]. That might require unique
target the innate immunity, it was prudent to identify
forms of adjuvant activity which activate specific
target cells and specific receptors (pathways) activated
pathways of innate immunity. The route of vaccine
by each vaccine antigen-adjuvant combination in order
administration might influence the type of cells in the
to know the expected immunomudulatory molecules.
innate immunity activated by an adjuvant. Different
Vaccine adjuvants applied sites of vaccine inoculation might differ in the types
and distribution of first-line immunity cells such as
in human vaccines mast cells, dendritic cells and macrophages.
Aluminum compounds (alum) was the earliest For instant, some antigen presenting cells could
and the most widely used adjuvant in human vaccines be abundant at mucosal sites while others would be
[10]. Alum haS been used in numerous vaccines including
intradermal or subcutaneous affecting their
diphtheria-tetanus-pertussis, human papilloma virus and
accessibility to vaccine antigens and adjuvants.
hepatitis B vaccines [11].
Recent developments have identified a variety
Other vaccine adjuvants developed for human of potential vaccine adjuvants for possible future
use include monophosphoryl lipid A (MPLA) [12-14[ and application in human vaccines. These include pathogen
MF59 (oil in water emulsion). MPLA was a detoxified associated molecular patterns (PAMPs) derived from
form of bacterial cell wall lipid A from Salmonella microbes and their synthetic analogs such as cytosine
Minnesota R595. MPLA adjuvant had been applied in and guanine (CpG) oligodeoxynucleotide that target
combination with alum to augment immunogenicity of toll-like receptors (TLRs) [22] and mast cell activating
subunit vaccines. The adjuvant activity of MPLA was compound such as compound 48/80 (C48/80) [23, 24].
mediated through activation of TLR4 and was seen to
induce Th1 type immune responses [16]. MPLA was used Objectives
in combination with alum for hepatitis B virus (FENDrix) The objective of this review was to identify the
human papilloma virus (Cervarix), [15] and current strategies that can be applied in understanding

20 African Journal of Health Sciences Volume 33, Issue No.3, May- June, 2020
the mechanisms of action for vaccine adjuvants with nanoemulsion were used compared to nanoemulsion
regard to: alone [25].
1. Molecular pathways and physical interactions of
the innate immune system. Combination of adjuvants was seen to produce
2. Vaccine and adjuvant doses synergistic effect in immune responses and pathogen
3. Vaccination regimens for each vaccine. clearance, but individual adjuvants more often
4. The current status of knowledge on mechanisms provided a response that was narrow in its effect, being
of actions for known adjuvants and those under either Th1 or Th2 biased. Cationic molecules such as
investigation. KLKLLLLLKLK predominantly induce Th2 type
5. Future directions in this important area of immune responses against co administered vaccine
antigens [26], while other classes of molecules such as
biomedical research.
CpG enhance antibody affinity maturation [27] and
Methodology provide Th1 type immune responses [22, 28].
Despite adjuvants of different molecular
Publications on vaccine and adjuvants research
properties exhibit distinct immune activation properties
and development were identified and downloaded from
several data sources that included PubMed (NLM), when combined, they provide superior immune responses.
PubMed Central, Library of congress, LISTA Studies have shown that, combination of CpG with alum
(EBSCO), Google Scholar, Science Direct and Web of enhanced affinity maturation of anti-hepatitis B vaccine
Science (TS) databases. Articles were searched using antibody responses [27]. Combination of CpG with
key words as title or subject without restrictions on cationic peptides has shown to form complexes that
types of publications that were presented in English facilitate delivery of antigen to APCs [29] and to induce
language. Since comprehensive searches were both Th1 and Th2 type antibody responses.
conducted on multiple databases as mentioned, no Use of adjuvants was reported to induce
formal assessments for risk of bias were conducted. diversified immune responses as depicted by
The type of publications included basic research using combination of cationic peptide HH2 with CpG that
experimental animals and clinical research in human. induced IgG1 (Th2) and IgG2a (Th1) type antibodies
in experimental animals [30]. Consequently, this nature
Literature Review of immune response ends up being more effective than
Strategies Shown to Improve a narrow type of immune response. Due to the different
modes of action by vaccine adjuvants, their effects
Vaccine Adjuvant Activities
could work in synergy to provide a more diversified
From the literature review there were several and effective immune response.
strategies that have been applied in the development of
effective vaccines in the context of adjuvants 2. Varying the Dose of Vaccine
incorporated in the vaccine. Some of the key strategies Antigens and the Interval between
are discussed below:
Vaccinations Influence Immune
1. Combination of Vaccine Adjuvants Responses.
that Activate Different Molecular Some vaccine studies reported that, increasing
Pathways Generate Effective antigen dose was one strategy used to reduce the
Immune Responses number of immunizations while maintaining a
Studies have shown that combination of desirable immune response. For instance;
adjuvants provide superior immune responses compared A study conducted in human subjects using
with individual adjuvants. A recent study using inactivated hepatitis A antigen induced a
recombinant hemmaglutinin (rH5) protein of highly significantly higher seroconversion rate at two
pathogenic avian influenza (HPAI) virus as antigen weeks after the first injection with 100 U of
demonstrated a quicker antibody production, IL-17 and antigen compared to 50 or 25 U of the antigen
IFN-γ when adjuvant combinations of CpG and [31].

African Journal of Health Sciences Volume 33, Issue No. 3, May - June 2020 21
 However, other studies have demonstrated that effective and safe vaccine. Aluminum compounds were
this effect is dependent on antigen type. For example; the longest serving vaccine adjuvants [35]. Although
Cytomegalovirus Glycoprotein B antigen when aluminum-based adjuvants have been used for decades
adjuvanted with MF59 (oil in water emulsion) with human vaccines, their mechanisms of action have
induced a higher antibody titer using a lower not been fully elucidated [11]. It has been
antigen dose compared to antibody titer induced demonstrated that aluminum compounds forms a depot
by a higher dose of the same antigen [32]. at the site of vaccine inoculation that then releases the
antigen in small doses to stimulate the immune cells
Those observations demonstrated that, for each [36-38]. Recent studies suggest that although alum
antigen type, an optimum dose should be determined forms antigen depots, depot formation is not required
through a dose-response study. That was more so with for its adjuvant activity [36].
adjuvant included in the vaccine because antigen-
adjuvant interactions would lead to formation of Studies have also shown that alum and other
secondary complexes that would differ in their capacity pore forming adjuvant molecules activate the
to be delivered to activate the immune cells. inflammasome in macrophages and dendritic cells as a
possible intermediate in its adjuvant activity
The ratio between the antigen and adjuvant in [39]. Inflammasomes are multiprotein complexes in
the vaccine would also influence the structure of the dendritic cells and macrophages activated by danger
final complex formed and its biological activities. signals that lead to the recruitment and activation of
Besides the vaccine dose, studies have shown that, caspase 1. Caspase 1 activation then leads to
intervals between the primary and booster processing of pro-inflammatory cytokines such as IL-
immunizations may influence the magnitude of both 1β and IL-18 that participate in immune responses
antibody and cellular immune responses (33). against insulting pathogens [40]. Although activation
of the inflammasome was considered a possible
This observation suggested that a critical time mediator of adjuvant activity by alum [41], similar to
existed after priming the immune system at which later the depot formation other studies showed that
immunizations effectively activated immune cells to inflammasome activation of this pathway by alum was
achieve maximum responses. The interval seemed to not required for antigen specific immunity [39].
influence the frequency of antigen-specific memory B
and T cells. Comparably MF59, oil in water emulsion
Practically, intranasal meningococcal subtype B adjuvant was seen to activate the inflammasome in
vaccine induced highly bactericidal immunity vitro studies. However, that activity was not required
with a day 0, 7, 28 and 56 schedules than same for in vivo adjuvant activity because there was no
vaccine given on days 0, 28 and 56 (34). difference between wild type mice and inflammasome
deficient mice [42]. The activities of alum and MF59
The results suggested that the inclusion of a suggested that adjuvant molecules could activate some
shorter interval between the primary and first booster molecular pathways or cause changes in the behavior
enhanced the production of immune factors with higher of the antigen in the host but those changes might not
bactericidal activities. It is important to note that the be immunologically important. More importantly, it is
optimum time intervals required between the primary well documented that those adjuvants augment antigen
and booster inoculations may vary with different activation of the innate immune cells lead to enhanced
antigen types or types of adjuvants in the vaccine and immune responses.
the respective antigen dose.
Toll-like receptors (TLRs) were identified
Current Understanding of Modes among PRRs as important targets for mediating the
activity of some vaccine adjuvants.
of Actions for Vaccine Adjuvants
Immunization of vaccine antigens combined with
To effectively design an adjuvant within a TLR agonist monophosphoryl lipid A (MPLA)
vaccine formulation, there was a need to understand its specifically activates TLR4 to stimulate innate
mechanisms of activity in order to develop a potent, immunity [43]. MPLA has been used

22 African Journal of Health Sciences Volume 33, Issue No.3, May- June, 2020
 in combination with alum adjuvant (AS04) for MyD88 pathway. The danger signal on epithelial cells
human papilloma virus vaccine [15]. may easily be generated by pore forming molecules
used as adjuvants in intranasal or oral vaccines that
This adjuvant system Was approved for use in make contact with epithelial cells lining mucosal
humans in Europe and USA. model studies displayed surfaces. Release of IL-33 forms another possible
the activation signals generated through TLRs involved mechanism of adjuvant activity through non-TLR
recruitment and activation of adaptor molecules ligands that activate MyD88 [46].
including MyD88 (44).
MyD88 was found downstream of TLRs, It is postulated that when released by adjuvant
except TLR3, and were critical in transmitting or antigen activated epithelial cells, IL-33 activates its
the activation signals that lead to generation of target cells, CD25+CD44+ intraepithelial innate
cytokines. Cytokines mediated adjuvant lymphoid cells (ILCs) that express the IL-33 receptor
activities on adaptive immune responses ST2/T1. Other target cells for IL -33 include
[45].Stimulation of MyD88 adaptor protein macrophages that were reported to produce IL-5 when
commonly originate from engagement of TLRs activated by IL-33 [47].
by vaccine adjuvants or microbes or their Early activation (within hours) of the innate
components. Activation of MyD88 then signals immune system has been demonstrated by studies to
the induction of pro-inflammatory cytokines by induce release of serum cytokines such as IL-5, G-
innate immune cells that modulate other CSF, IL -6 and KC [48] that drive subsequent events
immune responses. leading to adaptive response. These cytokines can
therefore be used as biomarkers of effective activation
Early (within hours) cytokine release had also
of the innate immunity by adjuvants in vaccines.
been demonstrated by studies with MF59 adjuvant that
was dependent on MyD88 activation in vivo [42]. That Studies conducted to understand modes of
could be a non-TLR activation of MyD88. Dependency activation of innate immunity reported that, double
on MyD88 was demonstrated by significant decrease in stranded DNA (dsDNA) could activate certain
the production of IL-5 and G-CSF in MyD88 knockout cytosolic DNA sensors such as stimulator of interferon
mice, with a significant reduction in serum antigen- genes (STING) within innate immune cells including
specific IgG antibody titers. DCs [49].
Therefore, understanding the mechanisms of STING was seen to drive the interferon response
adjuvant activity was important for deciphering their factor 3 (IRF3) activation and subsequent generation of
contributions in the induction of immune responses. IFN cytokines. Damaged cells could release double
Apart from MyD88, Mc Lachlan and co-workers have stranded DNA as a danger signal which might be the case
exhibited mast cells activation can enhance induction with pore forming adjuvants or vaccine molecules. As a
of immune responses [23]. The immune activation was possible pathway of adjuvant mechanism activity, dsDNA
mediated by release of preformed cytokines within the released after administration of adjuvants that cause local
mast cells including TNF that were released upon mast cellular injury activate intracellular STING to activate
cell degranulation. This observation had identified cytokine genes through IRF3.
mast cell activators as potential vaccine adjuvants, Host dsDNA released from damaged cells also
identifying a unique target cell of the innate immunity had the potential to activate innate immune cells
for vaccine design. including dendritic cells in either TLR9-dependent or
Although mast cells were reported to play a TLR9-independent pathways. Generation of dsDNA
role in undesired allergic reactions, studies have was a potential mechanism through which localized
demonstrated that mast cells activation can be induced death-inducing adjuvants could activate MyD88-
in a safe manner to provide beneficial immunological dependent activity, besides the IL-33 pathway.
functions. Further suggestions that, IL-33 cytokine
released by epithelial cells as a result of cell injury The TLR9-independent activity by host dsDNA
(danger signal) can induce Th2 type cellular immune might account for MyD88-independent activity by some
responses by its activation of innate immunity through adjuvants and that could involve the activation of

African Journal of Health Sciences Volume 33, Issue No. 3, May - June 2020 23
cytosolic sensors of dsDNA such as STING. Although [53]. This showed lack of inflammasome–dependent
the specific mediator that binds dsDNA to signal IRF3 adjuvant activity in vivo. The striking differences
was not very clear, mitochondrial antiviral signaling between WT and caspase 1-/- mice was the
protein (MAVS) [50] and stimulator of interferon significantly lower neutrophil infiltration at the site of
genes (STING) had been proposed [49]. IRF3 could inoculation in the absence of caspase 1 [53].
induce interferon response genes that induce
In addition to molecular activation, cationic
transcription and release type I interferon.
molecules have been shown to delay antigen clearance
That hypothesis showed there were two from the site of inoculation thereby enhancing antigen
possible pathways by which damage associated uptake, processing and presentation. This theory was
adjuvants could activate MyD88; demonstrated by studies with cationic poly-L-arginine
1. By induction of IL-33 and subsequent that showed increased retention and absorption of
activation of ST2/T1 receptors dextran through the nasal epithelium in rats [54].
2. By induction of dsDNA release that when Similar studies using cationic nanogels, chitosan and
bound by adjuvants forms a complex and trans- other nanoconjugate molecules had demonstrated
locate into the endosome to activate TLR9 prolonged antigen residence in the nasal cavity leading
cascading to MyD88 adaptor molecule [29, 51]. to enhanced immune responses in mice [55].
immunity [43].
It was reported that, when the vaccine antigen
Synthetic cationic peptides inclusive of
was retained for a longer time at the inoculation site, there
KLKL5KLK and innate defense regulator (HH2)
was an increase in the induction of antigen-specific
peptides had demonstrated effective adjuvant activities
immune response. Some vaccine adjuvants function by
in mice [26, 29]. In addition to their cationic nature,
enhancing antibody avidity for the vaccine antigen.
these peptides possess non-specific cell penetrating
Studies disclosed that, failure to induce sufficiently
properties that might contribute to cell lysis. They were
mature (high avidity) antibodies by a vaccine can lead to
active on many cell types and could have had diverse
lack of protection [56]. Antibodies with high avidity
models for adjuvant activity.
strongly bind to antigenic determinants on the pathogen
Other studies concluded that, inflammasome inducing destructive processes against the pathogen.
activation was a possibility to link innate and adaptive
On the contrary weakly binding (low avidity)
immunity induced by adjuvants through activation of
antibodies might lack the capacity to induce
intracytoplasmic PRRs such as NALP3 [39, 41]. The
neutralization of the pathogen. Antibody avidity could be
specific events that lead to inflammasome activation
used to determine pathogen-neutralizing ability of
remain unclear. It was postulated that events that lead to
antibodies. Despite in vitro pathogen, neutralization was
pore formation and potassium efflux, generation of
used as a correlate of protection by an immune response
Reactive Oxygen Species (ROS), lysosomal damage and
in certain circumstances of pathogen antigenic epitopes
release of cathepsin B could activate the inflammasome.
which might not have been be available in sufficient
The fact cationic peptides, penetrated cells amounts on mature virions in vitro to allow
leading to pore formation, they could induce generation neutralization. This may lead to lack of in vitro pathogen
of ROS [52] and activate the inflammasome. The same neutralization despite the same antibodies having
mechanism of pore formation in cells of innate immunity sufficient in vivo pathogen clearance activity.
might apply to aluminum compounds that were assumed
Another school of thought was that, in vivo,
to activate the inflammasome. That theory was further
protection could be enhanced by antibody opsonization of
supported by studies using a cationic peptide melittin that
the pathogen for clearance by phagocytic cells and T cell
demonstrated in vitro activation of inflammasome in mice
immune responses. That could make the antibodies with
Bone Marrow-Derived Macrophages (BMDM).
poor pathogen neutralizing capacity in vitro more
In vivo adjuvant activity, similar antigen- effective in vivo [57]. From the forgoing literature, it was
specific antibodies in wild-type (WT) and caspase 1-/- evident that mechanism studies in vaccine adjuvants was
(inflammasome-deficient) mice was demonstrated an active area of current and future investigations.

24 African Journal of Health Sciences Volume 33, Issue No.3, May- June, 2020
 engineered towards the targets while deleting the
Conclusions and Recommendations
nonessential ones to minimize toxicity effects. In
Vaccines (with or without adjuvants) should be summary, this review has identified the following key
safe to the host, and also effective in their functions. The facts:
safety and efficacy of vaccines can be predicted by 1. Evidence accrued that, combination of
deciphering their mechanisms of actions. The knowledge
adjuvants with different modes of action was a
about the modes of actions for vaccines and any
better strategy to induce a more effective
combined adjuvants is useful for designing and
immune response than use of a single adjuvant.
developing effective vaccines against particular
pathogens. Such knowledge can aid in directing an 2. Vaccine regimens with fewer doses were more
immune response towards a desired type of protective desirable because they ensured high compliance
immunity to effectively eliminate a particular pathogen. than multiple doses. However, the antigen type,
antigen dose and adjuvant activity could
It was reported that, different infectious agents
influence the number of immunizations and the
could require different profiles of immunomodulatory
intervals between vaccine administrations.
molecules for their clearance that might require
Optimum doses for each antigen-adjuvant
specific adjuvant activity [21]. In contrast;
combination could be determined
the activity of cholera toxin (CT) from Vibrio
independently through dose-response studies.
cholera and heat labile toxin (LT) from
Escherichia coli as adjuvants in experimental 3. Studies have demonstrated that, activities of
animals predominantly induced Th2 type T cell vaccine adjuvants are dependent on a variety of
immune responses with characteristic CD4+ T molecular pathways including; MyD88 activation
cells that secrete IL-4, IL-5, IL-6 and IL-10 [58]. mast cell activation or inflammasome activation
This knowledge was useful for the fact that, (macrophages and dendritic cells). A new class of
the right choice of an adjuvant can lead to generation adjuvants prolonged antigen retention at the site
of immunological mediators that will clear a particular of inoculation to enhance activation of the
infection. immune system. Surprisingly, the mechanism of
adjuvant activity for the oldest aluminum
Conclusively, understanding how a vaccine
adjuvants is still under intense investigations. It
(antigen-adjuvant complex) interacts with the immune
was clear that an adjuvant molecule might
system can aid in retaining immunogenic components in
activate multiple pathways but only one or few
the vaccine while deleting toxic ones to ensure safety. were relevant to their immunological functions.
Although studies had shown that CT and LT 4. The recent status of knowledge on mechanisms of
were effective vaccine adjuvants, they were highly action for vaccine adjuvants suggested future
toxic to the host animals [59, 60]. It was therefore studies to be focused on identifying more safe
necessary to retain potent genes in cholera toxin while vaccine adjuvants for human use. Previousily
deleting the virulent ones in order to retain desired
there were very few options on adjuvants
adjuvant activity but also increase safety [61, 62]. A
approved for human use that included aluminum
similar challenge applied to the potential use of mast
compounds MF59 and monophosphoryl lipid A.
cell activating c48/80 adjuvant which was a polymer of
many molecules. There was need to isolate the specific The limitation with those adjuvants was that,
monomers that activate mast cells to provide adjuvant they had been approved for use with injectable
activity and removal of nonessential molecules. vaccines. There was very limited development
In order to separate essential components from of adjuvants that could be safely applied with
redundant deleterious components in most adjuvants one intranasal vaccines. Intranasal vaccines were
must perform model studies. Mechanism studies will help desired because they were pain - free and
to identify the target cells and receptors for each relatively easy to administer. They require less
component on the larger molecules that are important for technical training as opposed to injectable
effective immune activation so that vaccines are vaccines. Injectable vaccines had more risks

African Journal of Health Sciences Volume 33, Issue No. 3, May - June 2020 25
 associated with use of needles such as safe Babiuk LA, Mutwiri G, et al. Strategies to link
disposal of needles and possible transmission of innate and adaptive immunity when designing
blood borne pathogens. vaccine adjuvants. Vet Immunol Immunopathol.
Intranasal adjuvants can be suitable for 2009 Mar 15;128(1-3):184-91.
mass vaccination campaigns especially against
respiratory infections such as influenza and 10. McKee AS, Munks MW, MacLeod MK,
CORONA viruses (e.g. SARS Cov-2 (COVID- Fleenor CJ, Van Rooijen N, Kappler JW, et al.
19)). Therefore, mast cell activating molecules Alum induces innate immune responses through
such as c48/80, mastoparan peptides and macrophage and mast cell sensors, but these
mucoadhesive molecules such as chitosan can sensors are not required for alum to act as an
be explored for use with intranasal vaccines. adjuvant for specific immunity. Journal of
immunology (Baltimore, Md : 1950). 2009 Oct
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