Suppressive Effect of Garcinia kola on the Humoral Immune Response of Mice to Hepatitis B Virus Subunit Vaccine

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*Corresponding author: E-mail: damian.odimegwu@unn.edu.ng, nonsodimegwu@yahoo.co.uk;
International Journal of TROPICAL DISEASE
& Health
30(1): 1-13, 2018; Article no.IJTDH.40868
ISSN: 2278–1005, NLM ID: 101632866
Suppressive Effect of Garcinia kola on the Humoral
Immune Response of Mice to Hepatitis B Virus
Subunit Vaccine
Ifeoma A. Onah
1
, Ebele B. Onuigbo
1
and Damian C. Odimegwu
1*
1
Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences,
University of Nigeria, Nsukka, Enugu State, Nigeria.
Authors’ contributions
This work was carried out in collaboration between all authors. Author DCO conceptualized the study.
All authors contributed in the design of the study. Authors DCO and IAO wrote the protocol, performed
the experiments, statistical analysis and wrote the manuscript. All authors managed the analyses of
the study, literature searches, read and approved the final manuscript.
Article Information
DOI: 10.9734/IJTDH/2018/40868
Editor(s):
(1)
Arthur V. M. Kwena, Professor, Department of Medical Biochemistry, Moi University, Kenya.
Reviewers:
(1)
Oti Baba Victor, Nasarawa State University, Nigeria.
(2)
Sylvester Yao Lokpo, School of Allied Health Sciences, University of Health and Allied Sciences, Ghana.
Complete Peer review History:
http://www.sciencedomain.org/review-history/24065
Received 30
th
January 2018
Accepted 5
th
April 2018
Published 9
th
April 2018
ABSTRACT
Aim:
Due to the health benefits of most medicinal plants, humans have resorted to their frequent
and sometimes daily consumption, thus there is need to investigate the effect of medicinal plants
consumption while on vaccination.
Study Design: This study was designed to investigate the biological interaction between Garcinia
kola (GK) seed extract and Hepatitis B virus Surface Antigen.
Place and Duration of Study: The research was carried out in the Department of Pharmaceutical
Microbiology and Biotechnology, University of Nigeria Nsukka, within six (6) months.
Methodology: Fresh GK seeds were obtained, identified, dried, pulverized and stored in an air-tight
container until extraction. Cold maceration technique was used for extraction. Methanol was the
solvent used. Locke's method of acute toxicity testing was used to ascertain the toxicity of the
extract. Afterwards, the experimental animals were grouped and vaccinated accordingly. After
vaccination, sera collected from the animals were used for immunogenicity studies while the whole
Original Research Article

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2
blood was used for total white blood cell count. During the study period, the experimental animals
were monitored frequently and weighed.
Results: The percentage yield after extraction was 16.7%. The extract was non-toxic up to 5000
mg/kg. The vaccination induced antibody responses (IgM, IgG1 and IgA) in all the groups but the
response in the hepatitis B vaccine group was significantly higher than that of the hepatitis B
vaccine/GK extract combination group (P<0.05), suggesting an inhibitory/ suppressive effect of G.
kola on immune response to the hepatitis B surface antigen. The total white blood cell count equally
revealed a suppressive effect of the extract on the hepatitis B virus surface antigen. The periodic
weight monitoring reveals similar growth pattern across all other groups except the hepatitis B
vaccine/GK combination group that seems not to be growing rapidly.
Conclusion: The outcome of this present study shows that at ≥250 mg/kg body weight GK seed
extract demonstrates a suppressive effect on the immunogenic responses to hepatitis B surface
antigen. Therefore, cautious consumption or total abstinence from GK is adviced in subjects
receiving hepatitis B vaccination.
Keywords: Vaccine; hepatitis B virus; Garcinia kola; antibody responses; immunogenicity.
1. INTRODUCTION
Microorganism surrounding our environment
causes numerous diseases, most of which have
no cure, making preventive measures the
effective means of managing such diseases.
Vaccination exercise is one of the best known
and effective means of preventing infectious
diseases having been shown to reduce the rate
of chronic infection in many countries [1].
Hepatitis-B virus (HBV) infection is the major
cause of the development of liver cirrhosis and
hepatocellular carcinoma [2]. Approximately 2
billion people worldwide are infected with the
virus [3] with the highest infection in sub-Saharan
Africa and East Asia [4]. Majority of individuals
who get infected in highly endemic areas acquire
the infection either perinatally or in early
childhood, whereas in low prevalence areas the
infection is acquired primarily in adulthood [5].
Hepatitis B virus belongs to the Hepadnaviridae
family, possess a 3.2 kb partially double-
stranded DNA, is an enveloped virus, and infects
hepatocytes by interacting with sodium-
taurocholate co-transporting polypeptide present
on the surface of these cells [6,7]. Treatment of
hepatitis B virus infection could be either
palliative or preventive. Currently, the preventive
approach involves vaccination with the
recombinant hepatitis B virus vaccine based on
the recombinant Hepatitis-B surface antigen
(rHBsAg) [8,9].
Garcinia kola (GK) is highly valued in Africa thus
highly consumed; it is used socially as
refreshment and medicinally for the treatment of
abdominal pain, cough, laryngitis, liver disease,
infection and erectile problems [10]. The seeds
are also considered a poison anti-dote. The
seeds have been proven to possess numerous
physiological and pharmacological effects which
include hepatoprotective, antiinflammatory,
antioxidant, antifertility, haematological and
anticancer effects [11]. The seeds are mostly
eaten by the elderly because of their belief that it
could prolong life. Chinedu et al. [12] in his report
encourages the daily consumption of GK as it
induces hypoglycaemic effect. Phytochemical
screening of the seed revealed the presence of
flavonoids, bioflavonoids, xanthones, triterpenes,
cycloartenol, benzophenones, phenol, alkaloids,
saponin and tannin [13]. GK, locally called bitter
kola and known variously as “Orogbo” (Yoruba),
“Namijin gworo” (Hausa) and “Aki ilu” (Igbo) is an
angiosperm, belonging to the family of Guttiferae.
It is found in moist forest and is widely distributed
throughout west and central Africa.
Booster doses of the HBV vaccine are required
to achieve a lasting immune response against
the virus [9,14]. This trend seems to suggest the
inherent relative weakness of the vaccine
immunogen or the possibility that factors that
tend to suppress the immune response to HBV
vaccine exist. Is it possible that frequent
consumption of Garcinia kola (as evaluated in
our study) negatively impact the HBV vaccine
response? Our current findings reported in this
paper attempt to answer this question. Therefore,
our study evaluated the effect of Garcinia kola
seed extract on HBV vaccine response.
2. MATERIALS AND METHODS
2.1 Collection, Identification and
Extraction of Plant Material
GK seeds were purchased from Ogige market in
Nsukka area of Enugu State, Nigeria and

Onah et al.; IJTDH, 30(1): 1-13, 2018; Article no.IJTDH.40868
3
authenticated by a botanist from the Department
of Botany, University of Nigeria, Nsukka having
the voucher number UNH NO 55 (University of
Nigeria Herbarium number 55). These seeds
were air-dried under shade (for 21 days) to avoid
sunlight thereby conserving the volatile
constituent in the plant materials, then pulverized
and the powder obtained was stored in an air
tight container until extraction. Methanolic extract
of the pulverized seeds was obtained using cold
maceration method [15]. Exactly 300 g of the
pulverized G. kola was macerated with 2.5 litres
of methanol (Sigma-Aldrich) using a glass jar.
The preparation was sealed properly to avoid
evaporation. This was shaken vigorously after 24
hours. At 48 hours, the extract was filtered using
a muslin cloth and Whatman No. 1 filter paper.
The filtrate obtained was evaporated to dryness
at room temperature for 1 week [16].
2.2 Test Animal
Young female Swiss albino mice (7 - 9 weeks
old) purchased from Faculty of Veterinary
Medicine, University of Nigeria Nsukka and kept
under standard pathogen-free conditions in an
animal facility of the Department of
Pharmacology and Toxicology of the University
of Nigeria, Nsukka was used. These animals
were fed with standard feed and water ad libitum
throughout the study period. A total of 39 animals
were used. The use and care of laboratory
animals in the study were in accordance with
ethical guidelines of the Faculty of
Pharmaceutical Sciences, University of Nigeria,
Nsukka.
2.3 Test Vaccine
Hepatitis B vaccine from Berna Biotech Korea
Corporation known as Hepavax-Gene®
(Hepatitis B virus vaccine, recombinant) was
used.
2.4 Acute Toxicity Study
Lorke’s method for acute toxicity study, as
modified by Bulus et al. [17] was employed for
the acute toxicity study. The study was carried
out in two stages:
Stage 1: Twelve (12) mice were used in this
stage, these were divided into four groups (A, B,
C and D) of 3 mice each. Group A, B, C were
given 10, 100 and 1000 mg/kg BW of the
methanolic GK seed extract respectively, Group
D (control group) received distilled water (DW)
(10 ml/kg BW). All these were given in a single
dose via parenteral route after 4 days of
acclimatization.
Stage 2: This stage was carried out based on the
result of the first stage. Here further specific
doses of 1600, 2900 and 5000 mg/kg BW of the
extract were administered to 3 mice (one mouse
per dose) to further determine the correct LD
50
value.
All animals were observed frequently on the day
of treatment and surviving animals were
monitored daily for 2 weeks for delayed signs of
toxicity. At the end of 14 days, all surviving mice
were sacrificed and the vital organs isolated. The
weights of these organs were taken and the
mean organ-body weight ratios calculated and
compared with those of the control group. The
body weight changes in the mice were also
recorded.
2.5 Vaccination
Twenty-four (24) mice randomly divided into four
groups (groups A - D) of six mice each was used.
These were immunized on days 0, 28 and 56 as
follows; Group A received 0.1 ml of distilled water
only (control); Group B received 0.1 ml (2 µg) of
Hepatitis B vaccine only; Group C received 0.1ml
GK (250 mg/kg BW) only; Group D received 0.1
ml (2 µg) Hepatitis B vaccine + 0.1 ml GK (250
mg/kg BW). The vaccine was administered
intramuscularly while the GK was given
intraperitoneally.
2.6 Immunogenicity Studies
The sera were harvested by eye puncture at the
retro-orbital plexus and analyzed for IgG1, IgA
and IgM rHBsAg specific antibody levels using
conventional ELISA method as described by
Ternette et al. [18]. ELISA plate (Brandplates
(immunograde), Wertheim, Germany) were
coated with hepatitis B vaccine antigen at a dose
of 2 µg/ml (0.2 µg / 100 µl / well) overnight at 4
º
C
in coating carbonate-bicarbonate buffer (pH 9.5).
The plates were blocked with Phosphate
Buffered Saline (PBS) containing 0.05% Tween
80 (PBS-T) and 5% fat-free milk for 1 hour at
room temperature. The plate was washed three
times with PBS-T. Mice sera diluted in PBS-T
with 2% fat-free milk were added to each well
and incubated for 1 hour at room temperature.
After washing three times with PBS-T the IgG1,
IgA and IgM subclass specific antibodies
conjugated with horseradish peroxidase were

Onah et al.; IJTDH, 30(1): 1-13, 2018; Article no.IJTDH.40868
4
added to the different wells and incubated for 1
hour at room temperature. The plates were
washed as done previously and 3, 3’5, 5’-
Tetramethylbenzidine (TMB) substrate
solution (Sigma Aldrich, Taufkirchen, Germany)
was added. The reaction was stopped
with 2 M H
2
SO
4
and absorbance at 450 nm
was measured using an ELISA reader (GM
2000).
At the end of the 3-times vaccination schedule,
the mice were sacrificed, livers excised,
examined, weighed and the liver homogenate
was equally screened for the presence of these
antibodies.
2.7 Determination of Total White Blood
Count (WBC)
Using the method outlined by Verma [19] 0.38 ml
(380 µl) of diluting fluid (10% glacial Acetic acid
and 1 drop of Gentian violet) was measured and
dispensed into a small tube and 0.02 ml (20 µl)
volume of well-mixed EDTA anticoagulated blood
was added. The Neubauer Counting Chamber
(Sigma-Aldrich, Germany) was assembled. The
diluted blood was re-mixed using a micropipette
and one of the grids of the chamber was filled
with the sample. The rulings of the chamber and
white cells were focused carefully using the 10x
objective until they appeared as small black dots.
One area/chamber was counted and the total
white blood cell is calculated using the formula
below:

1(ℎ)× 0.1(ℎℎℎ ) × 20()
2.8 Periodic Body Weight Assessment
The mice used for the study where weighed
periodically using a digital sensitive weighing
balance (Sartorius, Germany) and their weight
recorded accordingly.
2.9 Statistical Analysis
The data obtained were expressed as a mean ±
standard error of the mean (Mean ± SEM) being
consistent with the Tukey’s test. One way
analysis of variance (ANOVA) followed by pair
wise multiple comparisons according to Tukey’s
post hoc test was used to test for significance.
P˂0.05 was considered significant.
Graphpad Prism (version 6.0) was used for the
analysis.
3. RESULTS AND DISCUSSION
3.1 Physical Properties of the Garcinia
kola Seed Extract
Polar solvents have been shown to be effective
in extracting organic and inorganic materials from
plants [20]. Methanol, the polar solvent used
gave a percentage yield of 16.7 as shown in
Table 1 below.
Table 1. Some physical properties of the
methanolic extract of
Garcinia kola
seed
Physical
properties
Garcinia kola
seed
extract
Weight extracted 300 g
Weight of extract 50.1 g
Percentage Yield 16.7%
Description A brown sticky extract that
contains oil
3.2 Acute Toxicity
The acute toxicity test was carried out on the
extract to determine possible toxicity/ safety
margin [21] and it revealed a mean lethal dose
(LD
50
) greater than 5000 mg/kg as there was no
mortality recorded after 24 hours of treatment
(Table 2). Furthermore, no death was recorded in
all the groups throughout the 14 days monitoring
period (Table 2). Kennedy et al. [22] reported
that substances with LD
50
values higher than
5000 mg/kg are regarded as being safe or
practically non-toxic, thus the extract is
practically non-toxic. The LD
50
gotten is in line
with the findings of Komolafe et al. [23]. At the
end of the 14 days monitoring period the percent
body weight change and mean organ-body
weight ratio was determined. Increase in body
weight was noticed in all the groups. The percent
body weight change for GK doses of 10, 100,
1000, 1600, 2900 and 5000 mg/kg were 35.36 ±
3.07, 28.96 ± 5.18, 29.80 ± 4.51, 22.70, 41.92
and 28.49 % respectively and the distilled water
(DW) control group had a body weight change of
17.04 ± 0.93% (Fig. 1). Macroscopic examination
of the internal organs showed no sign of
congestion, inflammation or change of colour.
The mean organ-body weight ratio revealed
there was no significant difference in the weight
of the various organs when compared with the
control groups as indicated in Fig. 2 suggesting
the extracts have no effect on normal growth.
The usefulness of weighing organs in toxicity
studies includes their sensitivity to predict

Onah et al.; IJTDH, 30(1): 1-13, 2018; Article no.IJTDH.40868
5
toxicity, enzymes induction, physiologic
perturbation and acute injury. It correlates well
with histopathological changes [24]. The organ
weight was expressed as a percentage of the
body weight (organ-body weight ratio) rather than
as absolute weight so as to take into
consideration differences in the organ weight that
may solely be attributable to differences in the
body weight of the respective mice. The result of
the study revealed that the heart, spleen,
kidneys, liver and lungs did not show clinical sign
of toxicity throughout the treatment since there
was no significant (P<0.05) reduction or increase
in the organ-body weight ratio of the treated
animals. The body weight change monitored to
serve as a sensitive indication of the general
health status of the experimental animals [25].
The percentage body weight change (Fig. 1)
revealed all animal demonstrated an increase in
body weight during the period. This increase in
weight might be as a result of the extract having
an appetite-stimulating effect, which resulted to
increase in food intake and consequently
increase in weight. The increase in body weight
might also be as a result of natural growth since
the mice were young animals.
Table 2. Acute toxicity effect of methanolic extract of
Garcinia kola
seed administered
intraperitoneally to swiss albino mice
Experiment
Dose (mg/kg BW)
No. of mortality
after 24 hours
No. of mortality
after 14 days
Survival rate
Stage 1
10
100
1000
-
-
-
-
-
-
100%
100%
100%
Control
10 ml/kg - - 100%
Stage 2
1600
2900
5000
-
-
-
-
-
-
100%
100%
100%
Fig. 1. Percentage body weight change in mice treated with
Garcinia kola
during
acute toxicity test
Mice (n=3 for DW, 10, 100, 1000 mg/kg and n=1 for 1600, 2900 and 5000 mg/kg dose) were weighed at the end
of the acute toxicity study and the body weight change illustrated.
DW = Distilled water, GK = Garcinia kola

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Table 3. Effect of the intraperitoneal administration of methanolic extract of
Garcinia kola
seed on percent organ-body weight ratios of mice
following acute toxicity study
Organs Control
(DW)
10 mg/kg bw 100 mg/kg bw 1000 mg/kg bw 1600 mg/kg bw 2900 mg/kg bw 5000 mg/kg bw
Kidneys 1.378 ± 0.054 1.175 ± 0.016 1.304 ± 0.025 1.253 ± 0.020 1.240 1.168 1.316
Liver 6.426 ± 0.063 6.418 ± 0.148 6.370 ± 0.111 6.640 ± 0.061 6.284 6.502 6.117
Lungs 0.829 ± 0.055 0.810 ± 0.095 0.681 ± 0.011 0.731 ± 0.006 0.656 0.703 0.703
Spleen 0.778 ± 0.052 0.684 ± 0.064 0.708 ± 0.083 0.675 ± 0.035 0.646 0.703 0.699
Heart 0.464 ± 0.034 0.407 ± 0.003 0.405 ± 0.009 0.463 ± 0.016 0.477 0.494 0.410

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3.3 Effect of the Co-administration of
Vaccine and GK Extract on the
Humoral Immune Response
The immunogenicity studies revealed HBsAg –
specific serum antibody response. Antibodies
revealed were the IgG1, IgA and IgM. Following
vaccination, the antibody response (total
immunoglobulin i.e IgA, IgG1 and IgM) increased
from first vaccination through second vaccination
and declined after the third vaccination for all the
groups (Fig. 3) except the DW groups which did
not decline after the third vaccination. The
antibody titre (OD) trend as observed were 0.052
± 0.001 to 0.064 ± 0.007, then 0.075 ± 0.004 for
the distilled water (DW) treated group; 0.137 ±
0.009 to 0.337 ± 0.018, then 0.182 ± 0.011 for
hepatitis B vaccine (HV) group; 0.104 ± 0.003 to
0.281 ± 0.010, then 0.175 ± 0.007 for GK (GK)
treated group; 0.113 ± 0.009 to 0.267 ± 0.012,
then 0.148 ± 0.010 for the hepatitis B
vaccine and the GK (HV + GK) combination. The
antibody response seen in the HV + GK
group was less than that seen in the HV
positive control group in all the three vaccinations
with the decrease observed after second
and third vaccination in the HV + GK group
been significant (P˂0.05), this decrease was
more evident in the IgA specific responses
(Fig. 6).
Fig. 2. Percent organ-body weight ratio of mice after
Garcinia kola
acute toxicity
study
Mice (n=3 for DW, 10, 100, 1000 mg/kg and n=1 for 1600, 2900 and 5000 mg/kg dose) were sacrificed and vital
organs isolated and weighed. The mean organ-body weight ratio was calculated and illustrated.
DW = Distilled water, GK = Garcinia kola, * Significant at P˂0.05.
Fig. 3. Total HBsAg – specific serum antibody response
Mice (n=6) were vaccinated 3 times, sera were collected after each vaccination and examined for the presence of
HBsAg specific antibodies, this was summed up and presented as total HBsAg – Specific serum antibody
DW = Distilled water, HV= Hepatitis B vaccine, GK= Garcinia kola, * Significant at P˂0.05.

Onah et al.; IJTDH, 30(1): 1-13, 2018; Article no.IJTDH.40868
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Fig. 4. HBsAg – specific serum IgM response
Mice (n=6) were vaccinated 3 times, sera were collected after each vaccination and examined for the presence of
HBsAg specific IgM.
DW = Distilled water, HV= Hepatitis B vaccine, GK= Garcinia kola, * Significant at P˂0.05.
Fig. 5. HBsAg – specific serum IgG1 response
Mice (n=6) were vaccinated 3 times, sera were collected after each vaccination and examined for the presence of
HBsAg specific IgG1.
DW = Distilled water, HV= Hepatitis B vaccine, GK= Garcinia kola, * Significant at P˂0.05

Onah et al.; IJTDH, 30(1): 1-13, 2018; Article no.IJTDH.40868
9
Fig. 6. HBsAg – specific serum IgA response
Mice (n=6) were vaccinated 3 times, sera were collected after each vaccination and examined for the presence of
HBsAg specific IgA.
DW = Distilled water, HV= Hepatitis B vaccine, GK= Garcinia kola, * Significant at P˂0.05
Vaccination is employed in the prevention or
elimination of disease because it is possible to
induce long-term protective immunity. This long-
term protection is conferred by the maintenance
of antigen-specific immune effectors and/or by
the induction of immune memory cells that may
be sufficiently efficient and rapidly reactivated
into immune effectors in case of re-exposure to
the pathogen. Vaccine-induced immune effectors
are essentially antibodies. Antibodies are the
main specific and robust defence against
pathogens [26]. These antibodies were produced
in HV and the HV + GK groups (Fig. 3) with the
response as seen in the HV group been higher. It
appears GK had a suppressive effect on HV
immune response as the response seen in the
HV + GK group was significantly (P<0.05) lower
than that in the HV group (Fig. 3).
Specifically, IgM, IgG1 and IgA titres were
maintained at low level in the HV + GK group
(Figs. 5, 6 and 7) compared to the HV group. IgA
titre, in particular, was significantly (P<0.05) low
in the HV + GK group. The low antibody titre
observed in this HV + GK group could be
attributed to the presence of GK in the group as
the group which received only the hepatitis B
vaccine was high. IgM, which is the first antibody
to appear in response to an initial exposure to an
antigen was suppressed in the HV + GK group
(Fig. 4), and this shows that the vaccine-extract
combination is no longer immunogenic. Earlier
research proved that HV is immunogenic [27,28].
Thus it is the presence of GK in our HV + GK
group that made it less immunogenic, thus
suggesting that GK could inhibit early immune
responses to hepatitis B virus surface antigen.
IgG1, the naturally most abundant of all
immunoglobulins was equally suppressed. The
IgG1 class acts against bacteria and viruses by
opsonizing and neutralizing them, thus
vaccination with the combination of HV
and GK could decrease the rate of opsonisation
of hepatitis B virus for recognition and
phagocytosis by neutrophils and macrophages.
IgA is principally found in saliva, respiratory,
mucosal, intestinal and genital tract secretions
[29] and hepatitis B virus is transmitted via this
infected body fluid and blood [12,30]. Any
vaccination that gives rise to abundant IgA in
blood and body fluids will help prevent the
transmission of the disease from one person to
another. In the presence of GK, however, this
may not be achieved.

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Although, Chinedu et al. [31] reported that the
daily consumption of GK guarantees the
longevity of human life due to its hypoglycaemic
effect. However, we found on vaccination, its
consumption may not be advised as it has been
shown to suppress the humoral immune
response to hepatitis B vaccine, although Nworu
et al. [16] had earlier reported the immune-
modulatory action of G. kola extract.
3.3 Effect of the Co-administration of
Hepatitis B Vaccine and GK Extract
on Total White Blood Cell Count of
Swiss Albino Mice
White blood cells form the basis for immune
responses to invading microbes and foreign
substances, some white blood cells function in
the innate system, whereas others are part of an
adaptive immune response. These white blood
cells are important components in the
surveillance and protection systems of host
defence [32]. The total white blood cell count
equally showed the HV + GK combination-
treated group has low white blood cell count
compared to the HV treated group.
3.4 Effect of the Co-administration of
Hepatitis B Vaccine and GK Extract
on Body Weight of Swiss Albino
Mice
The result from the periodic assessment of mice
body weight showed a progressive increase;
from day 1 to day 29 and then decreased down
to day 43, this was followed by a rapid increase
in body weight up to day 57 (Fig. 8). The Distilled
water group grew from 23.73 g ± 0.80 to 24.94 g
± 0.99 to 26.34 g ± 0.99 to 24.69 g ± 0.96 to
27.20 g ± 0.99 while the other groups grew as
follows 23.31 g ± 0.73 to 24.09 g ± 0.76 to 24.87
g ± 0.45 to 23.69 g ± 0.61 to 25.12 g ± 0.65 (HV
group); 23.65 g ± 0.86 to 25.49 g ± 0.96 to 26.42
g ± 1.01 to 26.10 g ± 1.22 to 27.96 g ± 0.56 (GK
group); 25.05 g ± 1.19 to 25.63 g ± 1.03 to 26.57
g ± 1.21 to 26.50 g ± 1.16 to 27.20 g ± 1.15 (HV
+ GK group); on days 1, 15, 29, 43 and 57
respectively (Fig. 8).
Animal body weight monitoring is important in
research studies, as it provides information as
regards the health status of the experimental
animal especially if infection or a disease state is
involved, physiological changes due to
administered immunogens could instigate
changes in body weight [33]. Periodic
body weight monitoring of the mice used in this
study revealed a similar growth pattern for all
groups (Fig. 8) except for the HV + GK group.
The animals showed a progressive increase in
body weight from day 1 up to day 29 for
all the groups except the HV + GK groups which
seems not to be growing, this was followed by a
sharp decrease up to day 43, the reason for this
sharp decrease is not known, it was however
quickly followed by a rapid increase in
body weight for all the groups (Fig. 8). The
increase in body weight seen can be attributed to
growth since the experimental animal was
young mice between the ages of 7 and 9 week,
thus are expected to grow progressively when
properly fed.
Fig. 7. Total WBC count after vaccination
DW= Distilled water, HV= Hepatitis B
Vaccine, GK= Garcinia kola, * Significant at P˂0.05.

Onah et al.; IJTDH, 30(1): 1-13, 2018; Article no.IJTDH.40868
11
Fig. 8. Mice body weight curve
Mice used for this study were monitored periodically by means of body weight measurement to ascertain their
health status.
DW = Distilled water, HV= Hepatitis B vaccine, GK= Garcinia kola.
4. CONCLUSION
The results of our study have shown that GK
suppresses the humoral immune response to
HBV vaccine. Although the mechanism of this
immune suppression is not known, it
nevertheless has far-reaching implications for the
design and administration of vaccination
schedules for HBV vaccines.
CONSENT
It is not applicable.
ETHICAL APPROVAL
As per international standard or university
standard, written approval of Ethics committee
has been collected and preserved by the
authors.
COMPETING INTERESTS
Authors have declared that no competing
interests exist.
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© 2018 Onah et al.; This is an Open Access article distributed under the terms of the Creative Commons Attribution License
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