Sarhat E. R, Albarzanji Z. N. M, Pambuk C. I. A. Estimation of Some Interleukins in Cerebrospinal Fluid in Children with Meningitis. Biomed Pharmacol J 2019;12(4).

---

Manuscript received on :17-07-2019
Manuscript accepted on :29-11-2019
Published online on: 27-12-2019

---

Plagiarism Check: Yes
Reviewed by: Nour Rano 
Second Review by: ANDRE ANDRADE
Final Approval by: Dr Jihan Seid Hussein

---

How to Citeclose    |   Publication History close

 Views: (Visited 684 times, 1 visits today)    PDF Downloads: 417

Entedhar R. Sarhat¹ , Zubaidah N. M Albarzanji², Chateen I. Ali Pambuk¹

¹Department of Basic science, Dentistry College, Tikrit University, Tikrit

²Department of Basic science, Dentistry collage university of Kirkuk

Corresponding Author E-mail: entedharr@tu.edu.iq¹

DOI : https://dx.doi.org/10.13005/bpj/1850

Abstract

Meningitis is often associated with cerebral compromise which may be responsible for neurological squeal in nearly half of the survivors. Little is known about the mechanisms of CNS involvement in bacterial meningitis. the current study was to analyze the clinical significance of cerebrospinal fluid (CSF) concentrations of interleukin (IL)- 6, IL-1, IL-8, IL-10, tumor necrosis factor-alpha (TNF-α), and C-reactive protein in children with meningitis (n=35) and compared with control subjects (n=28). Serum total protein,  interleukine-6, IL-1 , IL-8, IL-10 , and, CRP, and TNF-α levels were found to be significantly high in study group, whereas serum glucose IL-10 levels were significantly low comparing with control group. Positive correlation was observed between IL-6with glucose , CRP  , and IL-8 , also between IL-10 with protein  , TNF with IL-8  ,and IL-1 with CRP  . whereas there was negative correlation  between IL-6 with protein  , TNF ,  IL-1 , IL-10 , also between IL-1 with IL-8  ,between  IL-10  with, and TNF ,  IL-10 with  glucose .  More studies performed in suitable models of meningitis are required in order to establish the routine use of inflammatory markers in the diagnosis of infectious diseases of the central nervous system

Keywords

Interleukin; cerebrospinal fluids; meningitis

Introduction

Central nervous system(CNS) cytokine production may lead to protection or damage of neural cells, depending on the specific cytokine, timing of its production, and the amount produced. Production of pro-inflammatory cytokines by microglia can induce neuronal damage or death,  but the effects of pro-inflammatory cytokines may be modulated by production of anti-inflammatory cytokines, or by other proinflammatory cytokines. Disruption of BBB in meningitis cytokines and chemokines could cross the BBB in an area of breakdown and affect brain tissue directly, and stimulate cytokine and chemokine production by microglial cells and astrocytes⁽¹⁾. Cytokines are small polypeptides with a wide range of inflammatory ,metabolic and immunomodulatory properties^(2,3).There is differ­ent types of cytokines including interleukin. Studies have indicated the pathological role of interleukins in both pe­ripheral and CNS diseases such as shock, meningitis and head injury⁽⁴⁾.

The present study aimed at analyzing cytokine profiles(IL-6 , IL-1 , IL-10, IL-8),TNF-α, and CRP among patients with meningitis in comparison with healthy controls.

Material and Methods

A prospective hospital based study was done during a period of one year ( from 1st.March / 2017 to 1st.April 2018). All patients from one month to 7.5 years admitted to the Salahalddin Teaching Hospital in Tikrit, and Pediatric Hospital in Kirkuk, Iraq with a presumptive diagnosis of meningitis were included in the study.

The fluid was withdrawn by lumbar puncture, using a spinal needle No.20).The patient lies on a hard bench, taking the lateral reclined position and the needle is gently placed above or beneath the forth lumber vertebra. The amount of withdrawn C.S.F is not fixed, but usually in the range of (1-3) ml. The CSF was examined grossly for appearance and color .Freshly collected specimens were stored at 4C°.Turbid specimens were centrifuged at 3000 rpm for 10 minutes before storage. The biochemical parameters such as IL-6 , IL-1 , IL-10, IL-8 , CRP, and TNF were measured by using the commercial enzyme-linked immunosorbent assay (ELISA) kits.

The C.S.F samples were divided into two groups. Normal control group , this group comprises 26 healthy individuals, and meningitis group which they were classified into four groups  (N=  26 patients)

Statistical analysis

The statistical package for social science (SPSS version 21 for Windows® Microsoft, USA) was used for data analysis. Comparison between patients and controls was made using the t-test for quantitative data and χ2 -test for qualitative data. P < 0.05 was considered statistically significant.

Results

Table 1: Demographic characteristic of study population.

The results of table(2)indicate the mean serum of Total protein,  interleukine-6, IL-1 , IL-8, and, CRP, and TNF-α , IL-10 levels was higher in patients with (63.05 ± 8.08 g/l), (0.487 ± 0.184 ng/mL),( 11.25 ± 1.10 pg/ml), (1.12 ±2.80 ng/mL), (5.532 ±  0.566 mg/dl) and (57±142 pg/ml), (70  ± 176) respectively than in controls (25.80 ±4.30 g/l), (0.1635±0.0340 ng/mL  ),( 6.450 ± 0.887 pg/ml), (0.135 ± 0.339 ng/mL ) (1.940 ± 0.227 mg/dl ), and (5.8± 14.5 pg/ml) , (0.1635±0.0340), and (24.3 ± 60.7) respectively. Also the results show that there is a significant decrease in serum Glucose level in patients(33.1± 82.8 mg/dl)) comparing with control group (120 ± 300 mg/dl) .

Table 2: Biochemical parameters of patients and the controls.

Correlation within Parameters

The results revealed that there was positive correlation  between IL-6with glucose (0.178),  CRP  (0.131), and IL-8 (0.155), also between IL-10 with protein  (0.265), TNF with IL-8  (0.272),and IL-1 with CRP  (0.106).  whereas there was negative correlation  between IL-6 with protein  (-0.049), TNF (-0.221),  IL-1 (-0.209), IL-10 (-0.036), also between IL-1 with IL-8  (-0.176),between  IL-10  with(-0.110), and TNF (-0.123).between  IL-10 with  glucose (-0.402), CRP ( -0.065).

Table 3: Correlation within Parameters

Discussion

Interleukin-6 (IL-6) is a multifunctional inflammatory cytokine that plays an important role in the response to environmental stress and has been implicated in the pathogenesis of many chronic diseases.  Interleukin-6 is a proinflammatory cytokine involved in the inflammatory response in the CNS. It is produced within CNS, i.a. by astrocytes and glial cells as a response to initiated or ongoing infection. Its production is also affected by other cytokines, i.a. TNF and IL-1. IL-6 has mainly proinflammatory properties. It persuades the synthesis of acute-phase proteins and contributes to BBB. IL-6 may act as an anti-inflammatory cytokine. It is validated by leukocyte transmission from blood to CSF^(5-7).The level of IL-6 is higher in patients with meningitis  than control group. Our findings are in the line with^(8,9),whereas disagreement with the results obtained by Pinto et al. ⁽¹⁰⁾.

Interleukin-10 is an acid-labile anti-inflammatory cytokine produced by monocytes, macrophages, B and T lymphocytes, brain cells such as neurons and microglia that capable of inhibiting proinflammatory responses^(11,12).The level of IL-10 is higher in patients with meningitis ,These findings are in harmony with preceding reports^(13,14).who stated that initially high IL-10 concentration gradient across the blood-brain barrier, comparably low amounts of IL-10 were detected in serum in the acute phase of meningitis, suggesting an initial compartmentalized release of IL-10 in the subarachnoid space in patients presenting with meningitis without persistent circulatory impairment.

Interleukin-8 is a pro-inflammatory cytokine which  acts as a chemoattractant for neutrophils to the site of inflammation⁽¹⁴⁾.Serum level of IL-8 was found to be significantly elevated in patients with meningitis, Because IL-8 contributes to leucocyte extravasation during infectious meningitis of viral and bacterial origin. The role of this molecule has been implicated in the blood–brain barrier disruption and meningeal infiltration by immune cells⁽¹⁵⁾.Similar observation was supported by ^(13,16).

Tumor necrosis factor alpha (TNF-a) and interleukin-1 (IL-1) a proinflammatory cytokine  are present in the CSF which are produced in the subarachnoid space by different cells, e.g., leukocytes, astrocytes, and microglia., Therefore, TNF-a contributes to the accumulation of leukocytes in the CSF, brain edema , blood-brain barrier damage ,and damage of cells within the CNS⁽¹⁷⁾.TNF may leak from CSF to serum during meningitis. However, in spite of the marked leakage of protein, and the high concentration gradient of TNF across the blood-brain barrier, the negligible amounts of bioactive TNF leak from CSF to serum during meningitis. By leakage from the subarachnoid space to the systemic circulation there is a dilution factor of 10-20.

Interleukin-1 (IL-1) produced within tissues and has multifunctional cytokine contributes to local inflammatory reactions, primarily complicated in the regulation of inflammatory processes, it mediates stimulation of fibroblast growth, and differentiation of B and T cells^(18,20).

Interleukin -1 contributes to disruption of the blood-brain barrier and CSF leukocyte recruitment and stimulates the production of other cytokines, such as IL-6 and TNF-α. The recruitment of polymorphonuclear leukocytes may induce damage not only to the bacteria but also to the brain⁽²¹⁾.Our analysis revealed statistically significant increase in the level of TNF-a and IL-1 in patients compared to the control group. This result is consistent with the other studies^(6,21,22),whereas disagree with⁽²³⁾.

C-reactive protein is produced by the monocytes of the tissue factor which initiates the coagulation process. C-reactive protein, together with fibrinogen, acts as a chemotactic factor. Fibrinogen is responsible for the adhesion of macrophages to the endothelial surface for their migration into the intima⁽²⁴⁾.Direct hepatic release of CRP in to plasma which then undergoes  ultra-filtration to form CSF meningeal irritation stimulate CRP production. Once CRP enters the  CSF it binds to the damaged tissues⁽²⁵⁾.So that the increased CRP concentrations in patients with meningitis may stimulate production by the monocytes of the tissue factor which initiates the coagulation process⁽²⁶⁾.The findings of the present study concurs with earlier studies^(27,28).Corral et al. found positive CSF CRP in 24/32 patients with culture-proved bacterial meningitis, while only 2/32 children with non-bacterial meningitis had CSF which was positive for CRP. According to Corral et al., this was a more sensitive test for differentiating bacterial from non-bacterial⁽²⁹⁾.

Conclusion

More studies performed in suitable models of meningitis are needed in order to establish the routine use of inflammatory markers in the diagnosis of infectious diseases of the central nervous system.

Financial and conflict interests disclosure

This work was not supported by any institution or company. The author has no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript. No conflict interests of any were reported during the course of this study. We have done the work completely by our self-funding.

Acknowledgments

Authors Acknowledgment is going to Kirkuk and Tikrit health  directorate that allowed us to conduct this research in Kirkuk and Tikrit hospitals,  in Tikrit Teaching Hospital , Kirkuk General Hospital and Kirkuk pediatric Hospital. Also, all private clinics and laboratories assisted our work . Our thanks and appreciation going to the staff, laboratory worker at Hospitals , private clinics and laboratories for their help during the enrollments of patients and sampling.

References

1. Ashok Kumar, Roopali Mittal, Hari ev Khanna, Sriparna Basu. Free Radical Injury and Blood-Brain Barrier Permeability in Hypoxic-Ischemic Encephalopathy Pediatrics . 2008, 122 (3) .722-727.
2. María Erta, Albert Quintana, and Juan Hidalgo. 2012.Interleukin-6, a Major Cytokine in the Central Nervous System. International Journal of Biological Sciences; 8(9):1254-1266.
3. Siham A.Wadee, Entedar R. Sarhat, Rajaa S. Najim. Effect of Moringa oleifera Extract on Serum Glucose and Interleukin-1, Interleukin-2 and Tumor Necrosis Factor α in Streptozotocin-Induced Diabetic Rats. Tikrit Medical Journal .2018; 24(1) :61 – 68.
4. Ziebell JM, Morganti-Kossmann MC. Involvement of pro- and an­ti-inflammatory cytokines and chemokines in the pathophysiol­ogy of traumatic brain injury. Neurotherapeutics. 2010;7(1):22–30
5. Sarhat E.R. Acute Myocardial Infarction: Melatonin, Apelin, and Visfatin as Predictors of Disease. Diyala Journal of Medicine. 13( 2), 2017; 11-17.
6. Hsieh CC, Lu JH, Chen SJ, Lan CC, Chow WC, Tang RB. Cerebrospinal fluid levels of interleukin-6 and interleukin-12 in children with meningitis. Childs Nerv Syst 2009; 25: 461-5.
7. Lucjan Kępa, Barbara Oczko-Grzesik, Anna Boroń-Kaczmarska. Cerbrospinal fluid interleukin-6 concentration in patients with purulent,bacterial meningitis- Own Observations. Przegl Epidemiol.2014; 68: 645 – 649
8. Dano ID, Sadou H, Issaka B, Oukem-Boyer OOM. Measurement of Interleukin-6 in Cerebrospinal Fluid for the Diagnosis of Bacterial Meningitis. Pak J Biol Sci.2016;19(4):185-190
9. Prasad, R., R. Kapoor, R. Srivastava, O.P. Mishra and T.B. Singh, 2014. Cerebrospinal fluid TNF-“, IL-6 and IL-8 in children with bacterial meningitis. Pediatr. Neurol., 50: 60-65.
10. Pinto, Jr. V.L.L., M.C. Rebelo, R.N. Gomes, E.F. de Assis, H.C. Castro-Faria-Neto and M.N. Boia, 2011. IL-6 and IL-8 in cerebrospinal fluid from patients with aseptic meningitis and bacterial meningitis: Their potential role as a marker for differential diagnosis. Braz. J. Infect. Dis., 15: 156-158.
11. Thompson CD, Zurko JC, Hanna BF, Hellenbrand DJ, Hanna A. The therapeutic role of interleukin-10 after spinal cord injury. J Neurotrauma 2013; 30: 1311–1324. pmid:23731227
12. Tatiana Barichello; Jaqueline S. Generoso; Allan Collodel; Ana Paula Moreira; Sérgio Monteiro de AlmeidaPathophysiology of acute meningitis caused by Streptococcus pneumoniae and adjunctive therapy approaches. Arq. Neuro-Psiquiatr. 70 (5 ). 2012;366-372
13. Francesco M Marincola. Interleukin-10,Medical Intelligence Unit. LandesBioscience Georgetown, Texas, U.S.A.2006:1.
14. Anne K. Lehmann, Alfred Halstensen, Steinar Sornes, Ola Rokke, and Anders Waage. High Levels of Interleukin 10 in Serum Are Associated with Fatality in Meningococcal Disease.Infection and Immunity.1995, 63, (6) .. 2109–2112
15. Sulik, A. Kroten, M. Wojtkowska& E. Oldak.Increased Levels of Cytokines in Cerebrospinal Fluid of Children with Aseptic Meningitis Caused by Mumps Virus and Echovirus 30 A. Scandinavian Journal of Immunology, 2014, 79, 68–72.
16. Yao R, Cao Y, Chen Y, Zeng Z. Diagnostic performance of interleukin-6 and interleukin-8 for bacterial meningitis: a meta-analysis. International Journal of Clinical and Experimental Medicine. 2015;8(5):7059-7068.
17. Carl Granert, Johan Raud, Anders Waage ,and Lars Lindquist. Effects of Polysaccharide Fucoidin on Cerebrospinal Fluid Interleukin-1 and Tumor Necrosis Factor Alpha in Pneumococcal Meningitis in the Rabbit. Infection and Immunity. 67(5)2071–2074.
18. Arai K, Lee F, Miyajima A, et al. “Cytokines: Coordinators of immune and inflammatory responses“, Annu Rev Biochem, 59, 783, (1990).
19. Lord PCW, Wilmoth LMG, Mizel SB, Mccall CE. “Expression of interleukin-1α and β genes by human blood polymorphonuclear leukocytes“, J Clin invest, 87, 1312, (1991).
20. Sarhat E.R. Evaluation of serum concentration Interleukins in Patients with Myocardial Infarction by ELISA teqnique. kirkuk university journal scientific studies;2018; 13(1), 43-51.
21. Mook-Kanamori, B.B., Geldhoff, M., van der Poll, T., van de Beek, D., 2011.Pathogenesis and pathophysiology of pneumococcal meningitis. Clin. Microbiol.Rev. 24, 557–591.
22. Luscinskas FW, Gerszten RE, Garcia-Zepeda EA, Lim YC, Yoshida M, et al. (2000) A. C-C and C-X-C chemokines trigger firm adhesion of monocytes to vascular endothelium under flow conditions. Ann N Y Acad Sci 902: 288–293.
23. Thwaites G E, Simmons C P, Quyen N T H, et al. Pathophysiology and Prognosis in Vietnamese Adults with Tuberculous Meningitis. J Infect Dis 2003;188:1105-15
24. Kumar A, Sivakanesan R. Cardiovascular Risk Factors in Normolipidemic Acute Myocardial Infarct Patients on Admission – Do Dietary Fruits and Vegetables Offer Any Benefits? Online J Health Allied Scs. 2010;9(3):3
25. Piyush S, Snehal V. Patel, Kruti Shah.2013. Resident.Role of CSF-CRP as a Bedside Diagnostic Test in Children with Meningitis. NHL Journal of Medical 2(1);54-56.
26. Sarhat E.R, Moayad MY Al-Anzy, Mutaz S. Ahmeid Thuraia R. Sarhat. Characteristic Abnormalities in Serum Biochemistry during Congestive heart failure. Tikrit Medical Journal – ;2018: 24 (1) :69 – 77 .
27. Kalpana K. Malla, Tejesh Malla, K. Seshagiri Rao, Sahisnuta Basnet, Ravi Shah. 2013.Is Cerebrospinal Fluid C-reactive Protein a Better Tool than Blood C-reactive Protein in Laboratory Diagnosis of Meningitis in Children?. Sultan Qaboos University Med J, 13, ( 1). 93-99
28. Palmer A, Carlin JB, Freihorst J, Gatchalian S, Muhe L, Mulholland K, et al. The use of CRP for diagnosing infections in young infants < 3 months of age in developing countries. Ann Trop Pediatr.2004;24(3):205–12.
29. Corral CJ, Pepple JM, Moxon R, Hughes WT.C-reactive protein in cerebrospinal fluid in children with meningitis. J Pediatr 1981; 99:365–9.