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*Corresponding author: E-mail: gideonbenjamin.y@gmail.com; 
 
 
 

Asian Journal of Immunology 
 
2(1): 64-71, 2019; Article no.AJI.53159 
 

 
 

 

 

Interleukin-12 (P70) Concentrations in Malaria 
Patients Attending Some Hospitals in Zaria,  

Kaduna State, Nigeria 
 

Gideon Yakusak Benjamin1*, B. E. Moses2, E. D. Jatau1 and C. M. Z. Whong1 
 

1
Department of Microbiology, Ahmadu Bello University, Zaria, Nigeria. 

2
Department of General Studies, School of Agricultural Technology, Nuhu Bamalli Polytechnic, Zaria, 

Nigeria. 
 

Authors’ contributions  
 

This work was carried out in collaboration among all authors. Author GYB designed the study, wrote 
the protocol and performed the laboratory experiments and statistical analysis. Author BEM read and 

contributed to literature searches. Authors EDJ and CMZW co-supervised the research. All authors 
read and approved the final manuscript.  

 
Article Information 

 
Editor(s): 

(1) Dr. Tania Mara Pinto Dabes Guimaraes, Associate Professor, Faculty of Pharmacy, Federal University of Minas Gerais, 
Belo Horizonte, Minas Gerais, Brazil. 

(2) Dr. Cynthia Aracely Alvizo Baez, Professor, Laboratory Immunology and Virology, Faculty of Biological Sciences, 
Autonomous University of Nuevo Leon, Mexico. 

(3) Dr. Darko Nozic, Professor, Higher Medical School in Belgrade, University of Belgrade, Serbia. 
Reviewers: 

(1) Muhammad Ghafoor Ali, Pakistan. 
(2) Fadia M. Attia, Suez Canal University, Egypt. 

Complete Peer review History: http://www.sdiarticle4.com/review-history/53159 

 
 
 
 

Received 15 October 2019  
Accepted 19 December 2019 
Published 25 December 2019 

 
 

ABSTRACT 
 

Background and Aim: Malaria is the most important parasitic disease of man, and it remains one 
of the major threats to public health and economic development in Africa. Interleukin-12 is a 
heterodimeric cytokine which has potent effects on innate and adaptive immunity.  This study was 
aimed at determining Interleukin-12 (p70) concentrations among malaria patients attending some 
hospitals in Zaria, Kaduna State.  
Methods: A cross sectional hospital based study was conducted on consenting participants in 
Zaria. Four hundred blood samples were collected, from which Giemsa-stained thick and thin blood 
films were prepared and examined for the presence of Plasmodium species by microscopy. 
Enzyme Linked Immunosorbent Assay (ELISA) was used to determine concentrations of 
interleukin-12 (p70) in malaria positive samples and control samples. 

Original Research Article 



 
 
 
 

Benjamin et al.; AJI, 2(1): 64-71, 2019; Article no.AJI.53159 
 

 

 
65 

 

Results: Males had higher malaria prevalence (37.2%) than females (24.7%). The difference was 
statistically significant (P = 0.01). Pregnant women had a prevalence of 17.8% which was lower 
than the 27.9% obtained in non-pregnant women (P = 0.07), and the highest malaria prevalence 
(20.0%) was found in pregnant women in their third trimester (P = 0.65). Interleukin-12 (p70) was 
present at a significantly (P = 0.00) higher level in the plasma of participants in the malaria positive 
group than in the control group (those who tested negative for malaria). 
Conclusion: Gender was significantly associated with malaria in this study. The prevalence of 
malaria was higher in males than females; males are therefore encouraged to take more 
precautions to prevent malaria. Despite the fact that the exact role of cytokines in malaria 
pathogenesis is unclear, we can infer from the findings of this study that more interleukin-12 (p70) 
is produced during malaria infection. 
 

 
Keywords: Interleukin-12(p70); plasmodium; prevalence; malaria; cytokine. 
 

1.  INTRODUCTION 
 
Malaria is a life-threatening disease caused by 
parasites that are transmitted to people through 
the bites of infected female Anopheles 
mosquitoes. It is preventable and curable [1]. 
Malaria is the most important parasitic disease of 
man. Approximately 5% of the world’s population 
is infected. It remains one of the major threats to 
public health and economic development in 
Africa. It is estimated that three million deaths 
result from malaria throughout the world, with 
Africa having more than 90% of this burden [2]. 
About half of the world’s population is at risk of 
malaria [3]. In 2016, an estimated 445 000 
malaria deaths occurred worldwide from an 
estimated 216 million reported cases of malaria; 
90% of these cases were in the World Health 
Organization (WHO) African Region. Of the 91 
countries reporting indigenous malaria cases in 
2016, 15 countries – all in sub-Saharan Africa, 
except India – carried 80% of the global malaria 
burden [1]. In 2017, Plasmodium falciparum 
accounted for 99.7% of estimated malaria cases 
in the WHO African Region, as well as in the 
majority of cases in the WHO regions of South-
East Asia (62.8%), the Eastern Mediterranean 
(69%) and the Western Pacific (71.9%) [1]. 
 
According to the level of malaria transmission 
and immunity acquisition, vulnerable populations 
differ in endemic areas. In highly endemic 
settings, children under five years and pregnant 
women are the most affected, constituting the 
main target population of new malaria control 
strategies as recommended by the World Health 
Organization [4]. There are now a large number 
of regular prevalence surveys of childhood 
parasitemia [5], as most malaria deaths occur in 
children. However, the prevalence of parasitemia 
in adults remains of scientific interest, not only 
because clinical attacks in adults remain an 

important cause of death in adults [6,7], as well 
as of morbidity and health service use [8], but 
also because adults form a community reservoir 
of infection for children. With the current 
sustained implementation of malaria control and 
prevention strategies across most African 
countries and the consideration of elimination in 
some settings [9,10], the impact of this adult 
reservoir in these control strategies needs to be 
assessed. In pregnancy, there is a transient 
depression of cell-mediated immunity that allows 
fetal allograft retention which in the other hand 
interferes with resistance to various infectious 
diseases such as malaria [11]. On the top of 
host, pregnant women, immunossupression; 
studies showed that immunological interactions 
between protozoan and helminths infection can 
intensify the impact of parasitic infection when 
they co-exist [12]. In addition, as the 
epidemiology of malaria changes across Africa 
there are likely to be changes in the disease 
pattern with adults becoming susceptible to 
severe disease and this trend should be 
monitored. 
 
While there are many studies that associate 
patterns of cytokines to disease, results may be 
different depending on the cohort population. 
Thus, there is an association between elevations 
in certain cytokines and disease outcomes, but it 
is hard to generalize these associations to 
different patient populations. IL-12 acts on 
antigen stimulated CD4+ T cells, promoting the 
differentiation of T cells into the Th1 subset [13], 
which acts on macrophages not only to stimulate 
their microbicidal functions, but also to increase 
their production of IL-12. The elevated levels of 
IL-12 also modulate the macrophage activity, 
which is associated with the increased 
erythrocyte destruction, bone marrow 
dyserythropoiesis [14] and thrombocytopenia 
[15]. During the intraerythrocytic life cycle of 



 
 
 
 

Benjamin et al.; AJI, 2(1): 64-71, 2019; Article no.AJI.53159 
 

 

 
66 

 

Plasmodium falciparum, macrophages avidly 
phagocytize parasite specific products, leading to 
the impairment of macrophage functions [16] and 
cytokine production [17]. 
 

2. MATERIALS AND METHODS 
 
2.1 Study Area 
 
The study was carried out in four selected 
hospitals in Zaria Nigeria; Major Ibrahim B. 
Abdullahi Memorial Hospital Zaria, Hajiya Gambo 
Sawaba Hospital Kofan Gaya Zaria, Salama 
Hospital and St. Luke’s Hospital Wusasa Zaria. 
Zaria is a city found in Kaduna state, Nigeria. It is 
located at 11.11 latitude and 7.72 longitude and 
it is situated at elevation 644 meters above sea 
level. Zaria has a population of 975,153 making it 
the second largest city in Kaduna [18]. 
 

2.2 Sample Size  
  
The sample size was determined using a 
prevalence of 23.45% [19] and the following 
formula as described by Naing, et al. [20]:  
 

n =  
        

   
 

n= number of samples 
 

p=prevalence rate of previous study = 23.45% = 
0.2345  
 

z=standard normal distribution at 95% 
confidence limit = 1.96 
 

d=absolute desired precision of 5% = 0.05 
z=1.96  
 

n=1.96
2 
*0.2345(1-0.2345)   

                    0.05
2 

 

n=275 samples  
 
Four hundred (400) blood samples were however 
collected for this study. 
 

2.3 Administration of Structured 
Questionnaire  

 
A structured questionnaire was used to collect 
data from consenting participants. 
 

2.4 Sample Collection 
 
A total of 400 blood samples were collected from 
Major Ibrahim B. Abdullahi Memorial Hospital 

Zaria (former Limi Hospital), Hajiya Gambo 
Suwaba Hospital Kofan Gaya Zaria, Salama 
Hospital and St. Luke’s Hospital Wusasa Zaria 
(100 samples from each hospital). Venipuncture 
technique was used for blood sample collection. 
A soft tubing tourniquet was fastened to the 
upper arm of the patients to enable the index 
finger to feel a suitable vein. The puncture site 
was then cleansed with Methylated spirit 
(methanol) and venipuncture was made with the 
aid of a 21 G needle attached to a 5 ml syringe. 
When sufficient blood (3 ml) was collected, the 
tourniquet was then released and the needle 
removed immediately while the blood was 
transferred into an EDTA bottle [21]. 
 

2.5 Determination of Malaria Parasitemia 
 

The malaria parasitemia was determined using a 
grading scheme of + =1-10 parasites, ++ = 11-20 
parasites, +++ = more than 20 parasites per 
microscopic field was used to establish the levels 
of parasitemia [19]. 
 

2.6 Determination of Interleukin-12 (p70) 
Concentrations 

 
The concentrations of interleukin-12 (p70) were 
determined in eighty eight (88) serum samples, 
using Boster’s interleukin-12 ELISA kit 
purchased from Boster Biological Technology 
Co. Ltd. (Fremont, CA USA). The samples were 
divided into two groups; the malaria positive 
group (59) and the control group; without 
parasitemia (29). The interleukin-12 
concentrations were determined according to 
manufacturer’s instruction. 
 

2.7 Statistical Analysis 
 

The data obtained were analyzed using 
Statistical Package for Social Sciences (SPSS 
Inc., Chicago, IL,USA). Chi square and odds 
ratio were used to check for association. P≤0.05 
was considered significant. 
 

3. RESULTS 
 

Table 1 shows malaria prevalence in relation to 
gender. Out of the 113 males examined; 42 
(37.2%) were positive, while out of the 287 
females screened; 71 (24.7%) were positive. 
Therefore males had a higher prevalence 
(37.2%) than females (24.7%). The P value and 
Odds ratio showed significant statistical 
association (P = 0.01, Odds ratio=1.800, 
Confidence interval= 1.129-2.869, Chi square = 
6.180). 



 
 
 
 

Benjamin et al.; AJI, 2(1): 64-71, 2019; Article no.AJI.53159 
 

 

 
67 

 

The age related prevalence of malaria among the 
study population is shown in Table 2. The 
prevalence rate in the table reveals that the 
highest prevalence of 50.0% was found in the 
age group 66 years and above, followed by the 
age group 56-60 years with 44.4% prevalence, 
51-55 years with 40.0% prevalence, 11-15 years 
with 37.5% prevalence, 41-45 years with 33.3% 
prevalence and 16-20 years with 32.9% 
prevalence. The lowest prevalence was in the 
age group 46-50 years (0.00%). The age group 
31-35 years also had a low prevalence of 13.3%. 
Statistically, there was no significant difference 
among the age groups (χ

2
=11.620, P 

value=0.56). 
 
Table 3 shows the prevalence of malaria in 
relation to pregnancy status. Pregnant women 
had a prevalence of 17.8% which was lower than 
the 27.9% obtained in non-pregnant women. The 
difference was not statistically significant 
(P>0.05). 
 
Table 4 shows the distribution of malaria 
according to pregnancy trimester. Pregnant 
women in their third trimester had the highest 
prevalence (20.0%), followed by those in their 

second trimester (19.6%) and those in their first 
trimester (10.5%). The difference was not 
statistically significant (P>0.05). 
 
Table 5 shows the mean concentrations of              
IL-12 (P70) among malaria positive individuals 
and control group. The malaria positive group 
had a mean concentration of 28.31 pg/ml               
which was high compared to the 19.23 pg/ml 
mean concentration of the control group               
(those without malaria infection). The                
statistical difference was not significant  
(P=0.06). 
 
Fig. 1 shows the mean concentrations of IL-12 
(p70) in relation to malaria parasitemia. The ++ 
parasitemia group had the highest mean 
concentration of 39.39 pg/ml. The + parasitemia 
group had 22.59 pg/ml mean concentration of IL-
12 (p70) which was lower than the 27.67 pg/ml 
mean IL-12 (p70) concentration found in the +++ 
parasitemia group. The mean IL-12 (p70) 
concentration in each of the three parasitemia 
groups (+,++,+++) was more than the mean IL-
12 (p70) concentration in the control group 
(19.23 pg/ml). The difference was statistically 
significant (df=3, P value=0.00). 

 
Table 1. Malaria prevalence in relation to gender 

 

Gender No. examined No. positive % Prevalence  χ
2
 P value OR 95%CI 

Male 113 42  37.2 6.180 0.01* 1.800 1.129-2.869 

Female 287 71 24.7     

Total 400 113 28.3     
Key: No=Number, OR=Odds ratio, CI=Confidence interval, *=Significant 

 
Table 2. Age-related prevalence of malaria among the study population 

 

Age(years) No. examined No. positive % prevalence χ
2
 P value 

0-5 43 11 25.6 11.620 0.56 

6-10 28 9 32.1   

11-15 24 9 37.5   

16-20 70 23 32.9   

21-25 74 20 27.0   

26-30 61 17 27.9   

31-35 30 4 13.3   

36-40 26 7 26.9   

41-45 18 6 33.3   

46-50 8 0 0.00   

51-55 5 2 40.0   

56-60 9 4 44.4   

61-65 2 0 00.0   

66> 2 1 50.0   

Total 400 113 28.3   
Key: No=Number, χ

2
=Chi square 



 
 
 
 

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68 

 

Table 3. Malaria prevalence in relation to pregnancy 
 

Pregnant No. examined No. positive  % Prevalence χ
2
 P value 

Yes  90 16 17.8 3.412 0.07 
No 197 55 27.9   
Total 287 71 24.7   

Key: No=number χ
2
= Chi square 

 

Table 4. Distribution of malaria according to pregnancy trimester 
 

Trimester No. examined No. positive % prevalence χ2 P value 

First 19 2 10.5 0.867 0.65 
Second 56 11 19.6   
Third 15 3 20.0   
Total 90 16 17.8   

Key: No=number χ
2
= Chi square 

 
Table 5. Mean concentrations of IL-12 (P70) among malaria positive individuals and control 

group 
 

Malaria No. Mean(pg/ml) SE P value 

Positive 59 28.31 2.003 0.06 
CG 29 19.23 2.125  
Total 88 25.32 1.576  

Key: CG= control group, SE=Standard error, No.= number of samples 
 

 
 

Fig. 1. Mean concentrations of IL-12 (p70) in relation to malaria parasitemia 
(P value<0.05) 

KEY: CG= control group, A= +, B=++, C=+++, (+=1-10, ++=11-20, +++=20> parasites per microscopic field) 

 

4. DISCUSSION 
 
In the current study, we found a high prevalence 
of malaria in males than females. The difference 
observed was statistically significant (p<0.05). 
This agrees with the findings of Muntaka and 
Opoku-Okrah [22] who reported the percentage 
of males with malaria to be higher than females; 
19.4% and 10.7% for males and females 
respectively. Reza and Taghi [23] also reported a 
similar finding. Our finding is in contrast to that of 

Otajevwo [24] who reported a higher infection 
rate in females than males. The high prevalence 
in males may be due to the fact that men are less 
likely to sleep under the insecticide treated bed 
nets than females [4]. According to Olapeju, et al. 
[25], ITN use tends to be higher among females 
than males especially in households without 
sufficient ITNs. In some societies, men have a 
greater occupational risk of contracting malaria 
than women if they work in mines, fields or 
forests at peak biting times. Leisure activities and 

0 

10 

20 

30 

40 

CG A B C 

19.23 
22.59 

39.39 

27.67 

M
ea

n
 c

o
n

ce
n

tr
a
ti

o
n

 

(p
g
/m

l)
 

Parasitemia 



 
 
 
 

Benjamin et al.; AJI, 2(1): 64-71, 2019; Article no.AJI.53159 
 

 

 
69 

 

sleeping arrangements may also be contributing 
factors, as men are more likely to sleep outdoors 
or be found outdoors during the active biting 
hours of Anopheles mosquito [26,27]. This can 
increase the human-vector contact, and 
consequently lead to Plasmodium infection [26]. 
 
Interleukin-12 (p70) was present at a significantly 
(p=0.000) higher level in the plasma of those in 
the malaria positive group than in the control 
group (those who tested negative for malaria). 
This is in agreement with the findings of Lyke, et 
al. [28]. However, Adrian, et al. [29] reported that 
concentration of IL-12 (p70) was significantly 
higher in the plasma of those with mild malaria 
than in the plasma of those with severe malaria. 
On comparing the different parasitemia groups 
and their mean interleukin-12 (p70) 
concentrations, the control group still had the 
lowest mean concentration of IL-12 (p70). The + 
and +++ parasitemic group had mean IL-12 (p70) 
concentrations which were less than the IL-12 
(p70) concentrations in the ++ parasitemia group. 
This is possible because low interleukin-12 (p70) 
activity has also been associated with severe 
Plasmodium falciparum malaria [30]. Our finding 
is however in contrast to that of Adrian, et al. [29] 
who reported that the acute-phase, pretreatment 
plasma IL-12 and alpha interferon (IFN-a) levels, 
as well as the acute-phase mitogen-stimulated 
whole-blood production capacity of IL-12, were 
significantly lower in children with severe rather 
than mild malaria.  It has been reported that early 
events in the cell-mediated immune response 
required for protection against malaria are 
initiated by the release of interleukin-12 (IL-12) 
from monocytes/macrophages, B cells, and 
perhaps other cell types [31,32]. In Plasmodium 
falciparum infection, IL-12 has immunoregulatory 
functions with effects on the immune response to 
the blood stage of disease, but also induces 
protection and reduces malarial anemia [33,34]. 
IL-12 has been shown to be involved in 
protective immunity against malaria by regulating 
gamma interferon. Pro-inflammatory cytokines 
like IL-12 are thought to be critical for controlling 
the erythrocytic and hepatic stages of 
Plasmodium infection [34] this may be one of the 
reasons why we had higher concentrations in the 
parasitemia group than the control group in this 
study.  
 

5. CONCLUSION 
 

This study found a significant association 
between the gender of participants and malaria. 
The prevalence of malaria was higher in males 

than females; males are therefore encouraged to 
take more precautions to prevent malaria. 
Although the concentrations of interleukin-12 (IL-
12 (p70) in this study differed with parasitemia, 
its concentrations reflect the role it plays in 
malaria infection. And despite the fact that the 
exact role of cytokines in malaria pathogenesis is 
unclear, the findings of this study suggest that 
more IL-12 (p70) cytokine is produced during 
malaria infection. 
 

CONSENT  
 

As per international standard, patient’s written 
consent has been collected and preserved by the 
author(s). 
 

ETHICAL APPROVAL 
 

The research protocol was read and approved by 
the Ethics and Research Committee (ERC) of 
Kaduna State Ministry of health, Nigeria. 

 
COMPETING INTERESTS 
 
Authors have declared that no competing 
interests exist. 
 

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© 2019 Benjamin et al.; This is an Open Access article distributed under the terms of the Creative Commons Attribution License 
(http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, 
provided the original work is properly cited. 

 
 

 
 

 

Peer-review history: 
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