|Year : 2018 | Volume
| Issue : 2 | Page : 63-67
Plasma complement 4, tumor necrosis factor-alpha, and interleukin 4 in patients co-infected with Plasmodium confection and filarial worm
Mathew Folaranmi Olaniyan1, Kehinde Oluwanifemi Ojo2, Mufutau M Azeez2, Temitayo Afolabi2
1 Department of Medical Laboratory Science, Edo University, Iyamho, Edo State, Nigeria
2 Department of Medical Laboratory Science, Achievers University, Owo, Nigeria
|Date of Submission||21-Oct-2018|
|Date of Acceptance||08-Nov-2018|
|Date of Web Publication||12-Dec-2018|
Prof. Mathew Folaranmi Olaniyan
Department of Medical Laboratory Science, Edo University, Iyamho, Edo State
Source of Support: None, Conflict of Interest: None
Study Background: Tumor necrosis factor-alpha (TNF-α) is a proinflammatory cytokine which worsen disease, interleukin-4 (IL-4) is an anti-inflammatory cytokine that enhance healing process while complement 4 mediates inflammation. Plasmodiasis and lymphatic filariasis can generate immune responses involving cytokines and complements.
Aim and Objective: This work was designed to determine plasma level of TNF-α, complement 4 (C4), and IL-4 in Plasmodium co-infection with filariasis.
Materials and Methods: The test individuals recruited for this study include microfilaria monoinfected patients (30), Plasmodium monoinfected patients (50), and microfilaria and Plasmodium coinfected patients (30) aged 32–73 years. Microfilaria and Plasmodium noninfected individuals (50) were studied as control. All individuals were seronegative to HIV, hepatitis C virus (HCV), and HBsAg. TNF-α, IL-4, C4, HIV, HCV, and HBsAg were determined in each of the patients by immunochemical technique using enzyme-linked immunosorbent assay method. Microfilaria was identify in each of the subjects by identifying microfilariae on Giemsa stained, thin, and thick blood film smears, using the “gold standard” known as finger prick test while identification of Plasmodium was carried out using thick and thin film technique using Giemsa and Leishman staining.
Results: The result obtained showed a significant increase in the plasma value of TNF-α, IL-4 and a significant decrease in plasma value of C4 in patients who were co-infected with both Plasmodium and microfilaria compared with Plasmodium monoinfected, microfilaria monoinfected patients, and also compared with the controls (P < 0.05).
Conclusion: This work has been used to reveal a significant increase in plasma value of TNF-α, IL-4, and significant decrease in plasma value of C4 in microfilaria and Plasmodium coinfection. Routine laboratory evaluation of these parameters in microfilaria and Plasmodium coinfection will provide useful direction for the management of the disease condition.
Keywords: Complement 4, confection, filarial worm, interleukin-4, Plasmodium, tumor necrosis factor-alpha
|How to cite this article:|
Olaniyan MF, Ojo KO, Azeez MM, Afolabi T. Plasma complement 4, tumor necrosis factor-alpha, and interleukin 4 in patients co-infected with Plasmodium confection and filarial worm. Imam J Appl Sci 2018;3:63-7
|How to cite this URL:|
Olaniyan MF, Ojo KO, Azeez MM, Afolabi T. Plasma complement 4, tumor necrosis factor-alpha, and interleukin 4 in patients co-infected with Plasmodium confection and filarial worm. Imam J Appl Sci [serial online] 2018 [cited 2019 Jun 16];3:63-7. Available from: http://www.e-ijas.org/text.asp?2018/3/2/63/247315
| Introduction|| |
Plasmodiasis and lymphatic filariasis are some of the largest public health problem with respect to vector-borne diseases in the world., These diseases are co-endemic in sub-Saharan Africa as the parasitic agents of plasmodiasis and lymphatic filariasis are transmitted by the same vector which is Anopheles spp. mosquito. Plasmodiasis and lymphatic filariasis can co-exist in both vectors and hosts. Co-infection of Plasmodium and lymphatic microfilaria in the host may increase susceptibility and disease severity which can alter immunological cytokines. Cytokines are proteins produced by the immune system. They regulate immunity and can act as inflammatory and anti-inflammatory cytokines. They can stimulate the production of antibodies and acute phase reaction in the presence of an infectious agent.Plasmodium infection can induce Th1 response causing increase in type 1 cytokines including tumor necrosis factor-alpha (TNF-α), resulting in an extreme inflammatory responses. Lymphatic filariasis can also cause Th1 and Th2 responses. The initial response which is Th1 biased induces inflammation and can also protect against incoming larvae. Subsequently, Th2 responses are stimulated, especially cytokines interleukin-4 (IL-4), that induce strong antibody responses to reduce the infection. The infection can increase Th2 levels to decrease Th1 response. Increasing our knowledge on how co-infection influence the control of infectious diseases is very important as evidence of meaningful pathogens-host interactions becomes well established.
The complement system is an innate immune system that increases the potential of antibodies and phagocytic cells to remove pathogens and damaged cells from the body of an organism. Complement triggers phagocytosis (opsonization of antigens), inflammation, and membrane attack (rupturing cell wall of pathogens).
Little attention has been paid to the study of immunochemical status of the co-infection of Plasmodium with filarial worm compared to co-infections of HIV and Viral Hepatitis with other infectious agents hence the justification for this research work. This work was designed to determine the plasma level of TNF-α, IL-4, and complement 4 (C4) in patients with Plasmodium co-infection with a filarial worm.
| Materials and Methods|| |
The work was carried out at the Baptist Medical Centre, Saki West Local Government Area located at the Northern part of Oyo state in Nigeria. Its headquarters is Saki town. It has an area of 2014 km2 and a population of 278,002 at the 2006 census. Saki hosts 2nd Mechanized Division of Nigerian Army, Baptist Medical Centre, Muslim Hospital, School of Medical Laboratory Technology, Baptist School of Nursing and Midwifery, Saki, School of Basic Midwifery, Oyo State Hospital, The Oke-Ogun Polytechnic, and Oyo State Technical College, Saki is one of the largest cities in Oyo state.
A total of 160 individuals classified into controls who were neither infected with Plasmodium nor lymphatic filariasis (n = 50); Plasmodium-infected patients (n = 50); lymphatic filariasis patients (n = 30); and Plasmodium co-infected with lymphatic filariasis patients (n = 30) aged 32–73 years were recruited for the study from Saki-west local Government of Oyo State. Individuals who were seronegative to HBsAg, hepatitis C virus (HCV), and HIV were included in the study.
Tumor necrosis factor-alpha enzyme-linked immunosorbent assay
Plasma TNF-α was analyzed by enzyme-linked immunosorbent assay (ELISA) using Abcam's kit.
Test samples, control, and standard are added separately to microtiter wells coated with biotinylated monoclonal antibody specific TNF-α, incubated and washed followed by the addition of enzyme streptavidin-horseradish peroxidase, that binds the biotinylated antibody. The mixture was also incubated and washed. After this, 3, 3', 5, 5'-Tetramethylbenzidine substrate solution was added that acts on the bound enzyme for color formation. The intensity of the color generated is directly proportional to the concentration of TNF-α in the test samples, standard, and control.
Interleukin-4 enzyme-linked immunosorbent assay
IL-4 was determined using Abcam's IL-4 Human ELISA kit.
Test samples, control, and standard are added separately to microtiter wells coated with biotinylated monoclonal antibody specific IL-4, incubated and washed followed by the addition of horseradish peroxidase-conjugated streptavidin that binds the biotinylated antibody. The mixture was also incubated and washed. After this, 3, 3', 5, 5'-Tetramethylbenzidine substrate solution was added that acts on the bound enzyme for color formation. The intensity of the color generated is directly proportional to the concentration of IL-4 in the test samples, standard, and control. Stop Solution was therefore added, and the color changed from blue to yellow the intensity of which was measured at 450 nm.
Complement 4 enzyme-linked immunosorbent assay
C4 was determined by C4 human in vitro competitive ELISA using Abcam's kit.
Specific antibody to C4 was precoated onto microtiter wells. Samples of tests, standards, and control were added to the wells and subsequently biotinylated Complement C4. The mixture after incubation was washed with wash buffer. Streptavidin-Peroxidase was added to wash unbound conjugates. 3, 3', 5, 5'-Tetramethylbenzidine substrate was then added to make Streptavidin-Peroxidase enzymatic reaction visible. Streptavidin-Peroxidase catalyzed 3, 3', 5, 5'-Tetramethylbenzidine substrate to form a blue color that changes to yellow on the addition of acidic stop solution. The intensity of the yellow color is inversely proportional to the concentration of C4 in the samples.
Laboratory identification of Plasmodium
This was carried out using Geimsha-Thick film technique as described by Centers for Disease Control and Prevention.
Anti hepatitis C virus determination by enzyme-linked immunosorbent assay
This was determined using Anti-HCV Core Antigen-antibody (ab50288) Abcam kit.
Determination of HIV by enzyme-linked immunosorbent assay test technique
HIV test was carried out by enzyme immunoassay using Genscreen ultra HIV Ag-Ab Biorad Kit.
Determination of HBsAg enzyme-linked immunosorbent assay test technique
This was assayed using Diagnostic Automation/Cortez Diagnostics, INC kit by ELISA method.
Laboratory diagnosis of filarial worm
This was carried out using BinaxNow filariasis Immunochromatographic test kit.
The proposal of the study was reviewed and approved by Ethical and Research Committee of Baptist Medical Centre, Saki, Nigeria before the commencement of this work. Informed consent was also obtained from each of the patient and controls.
The results obtained were analyzed statistically using SPSS 18.0 (IBM SPSS, Armonk, New York, United States) to evaluate the mean, standard deviation, student t-test, and P at 0.05 level of statistical significance.
| Results|| |
There was a significantly higher plasma level of IL-4 in patients co-infected with Plasmodium and filarial worm than the result obtained from the controls, Plasmodium monoinfected patients and filaria worm monoinfected patients with P > 0.05 [Table 1], [Table 2] and [Figure 1].
|Table 1: Mean and standard deviation of interleukin-4, complement 4, and tumor necrosis factor-alpha in patients and controls|
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|Table 2: Variations of interleukin-4, complement 4, and tumor necrosis factor-alpha in patients and controls|
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|Figure 1: Comparative description of interleukin-4, complement 4 and tumor necrosis factor-alpha obtained in patients and control groups|
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There was a significantly lower plasma level of C4 in patients co-infected with Plasmodium and filarial worm than the result obtained from the controls with P < 0.05 [Table 1], [Table 2] and [Figure 1]. There was a significantly higher plasma level of C4 level in Plasmodium monoinfected patients than the result obtained from patients co-infected with Plasmodium and filarial worm. There was a significantly lower plasma level of C4 level filarial worm monoinfected patient than the result obtained from the control with P < 0.05 [Table 1], [Table 2] and [Figure 1].
There was a significant higher plasma value of TNF-α in Plasmodium monoinfected patients compared to the result obtained from the control with P < 0.05 [Table 1], [Table 2] and [Figure 1]. There was a significantly higher plasma TNF-α in filaria worm and Plasmodium co-infected patients than the controls and filaria worm monoinfected patients with P < 0.05 [Table 1], [Table 2] and [Figure 1].
There was a significant higher plasma value of TNF-α in filaria worm monoinfected patients compared to the result obtained from the control. There was a significantly higher plasma TNF-α level in Plasmodium monoinfected patients than the result obtained from filarial worm and Plasmodium co-infected patients with P < 0.05 [Table 1], [Table 2] and [Figure 1].
| Discussion|| |
This work was used to determine plasma level of TNF-α, C4 and IL-4 in Plasmodium co-infection with filaria worm. The outcome of this work, therefore, includes the following; there was a significantly higher plasma IL-4 in patients co-infected with Plasmodium and filarial worm than the result obtained from the controls, Plasmodium monoinfected patients and filarial worm monoinfected patients. There was significantly lower plasma C4 in patients co-infected with Plasmodium and filarial worm than the result obtained from the controls, Plasmodium monoinfected patients and filaria worm monoinfected patinets.
There was also a significantly higher plasma TNF-α in filaria worm and Plasmodium co-infected patients than the controls and filaria worm monoinfected patients The findings of this work revealed a significant influence of co-infection of filarial worm and Plasmodium on the plasma level of IL-4, C4 and TNF-α compared with Plasmodium and filarial worm mono- and non-infected subjects.
These findings could be associated with the following facts that TNF-α is an inflammatory cytokine which according to Dinarello act to generate fever, acute phase reaction and can worsen a disease condition. Increase in this inflammatory agent could be triggered by mono and co-infection of filarial worm and Plasmodium.
In addition, TNF-α is synthesized by macrophages, lymphoid cells, mast cells, endothelial cells, cardiac myocytes, adipose tissue, fibroblasts, and neurons. It acts in conjunction with IL-6 on organs and systems. It stimulates attraction and migration of neutrophils and also promotes the expression of adhesion molecules. This might account for the increased plasma level of TNF-α. IL-4 stimulates activated B-cell, T-cell proliferation, differentiation of B cells and induction of B-cell class to switch to IgE. IgE is an antibody involved in parasitic infection,,,,, IL-4 also promotes inflammation and wound repair. The body's requirement for IL-4 for repair of damaged tissues/cells by Plasmodium/Filarial infections and the production of IgE antibody against Plasmodium and filarial infections causing increased synthesis and release might be responsible for the plasma increase.
Decreased C4 is an innate immune protein,, whose activation is triggered by pathogenic infections to lyse o invading pathogen. The reduction in the plasma level of C4 can due to excessive utilization to destroy Plasmodium and filarial worm.,,
| Conclusion|| |
This work has revealed a significant increase in plasma IL-4 and TNF-α as a result of filarial worm and Plasmodium co-infection including a significant decrease in plasma C4 as a result of filarial worm and Plasmodium co-infection.
A routine investigation of Plasmodium infection involving estimation of plasma IL-4, C4, and TNF-α is recommended for the management of malaria.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2]