|Year : 2014 | Volume
| Issue : 4 | Page : 258-262
Th1/Th2 profile in patients suffering with osteoarthritis and rheumatoid arthritis: An analytical observational study
Yatendra Singh1, Shadab Ahmad Khan2, Mohammad Owais3, Mazhar Abbas4, Anjum Parvez2, Athar Kamal2
1 Department of Medicine, Government Medical College, Haldwani, Uttarakhand, India
2 Department of Medicine, Jawaharlal Nehru Medical College, Aligarh, Uttar Pradesh, India
3 Department of Biotechnology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
4 Department of Orthopaedics, Jawaharlal Nehru Medical College, Aligarh, Uttar Pradesh, India
|Date of Web Publication||16-Oct-2014|
Dr. Yatendra Singh
Room No. 32, Sr Hostel, Government Medical College, Haldwani - 263 129, Uttarakhand
Source of Support: None, Conflict of Interest: None
Aims: To compare the cytokine (Th1/Th2) profile in serum as well as in synovial fluid of patients suffering with osteoarthritis (OA) and rheumatoid arthritis (RA). Settings and Design: Hospital-based analytical observational study. Materials and Methods: The present study comprised of 70 patients of arthritis, out of which, 40 patients were patients suffering from RA and 30 patients suffering from OA. Patients fulfilling the revised ARA Criteria for the Classification of Rheumatoid Arthritis are diagnosed as a case of RA and recruited for this study. Patients fulfilling the clinical and radiological features of osteoarthritis included in the study. Cytokine assay estimated using Western Blot (Immuno-blot transfer). Statistical Analysis Used: Analysis was performed using SPSS version 16.0 Statistical package for windows (SPSS, Chicago, IL). Results: Arthritis is commonly seen in females than males. Proinflammatory cytokines (IL-1, IL-2 and IFN-γ), anti-inflammatory cytokines (IL-4 and IL-10), regulatory cytokine (TGF-β) of blood as well as serum levels were raised in RA patients as compared with OA patients (P < 0.01). Tumor necrosis factor (TNF-α), an proinflammatory cytokine, levels were also raised significantly in OA. Conclusions: Our study shows that both Th1 and Th2 cells derived inflammatory markers as well as levels of IL-1 and TNF-α (both blood as well as synovial fluid) were significantly raised in RA as compared with OA patients. Thus, high levels of these substances have been found in inflammatory arthropathies, in particular in those characterized by a more aggressive and destructive outcome, such as RA.
Keywords: Cytokine, osteoarthritis, rheumatoid arthritis, Th1/Th2
|How to cite this article:|
Singh Y, Khan SA, Owais M, Abbas M, Parvez A, Kamal A. Th1/Th2 profile in patients suffering with osteoarthritis and rheumatoid arthritis: An analytical observational study. CHRISMED J Health Res 2014;1:258-62
|How to cite this URL:|
Singh Y, Khan SA, Owais M, Abbas M, Parvez A, Kamal A. Th1/Th2 profile in patients suffering with osteoarthritis and rheumatoid arthritis: An analytical observational study. CHRISMED J Health Res [serial online] 2014 [cited 2020 Feb 17];1:258-62. Available from: http://www.cjhr.org/text.asp?2014/1/4/258/142998
| Introduction|| |
Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by joint swelling, joint tenderness, and destruction of synovial joints, leading to severe disability and premature mortality. ,, Classically, osteoarthritis (OA), unlike RA, is defined as an inherently noninflammatory disorder of movable joints characterized by deterioration of articular cartilage and the formation of new bone at the joint surfaces and margins. 
Cytokines are protein messengers that convey information between and within the cells via specific cell surface receptor molecules. The release of specific cytokines into the systemic circulation has been observed in a variety of inflammatory diseases including RA. Their concentration levels usually reflect disease severity and prognosis. Cytokines are also divided into proinflammatory cytokines [tumour necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin (IL)-1, IL-2, IL-6, IL-8, IL-12 and IL-18], anti-inflammatory cytokines (IL-4, IL-10) and regulatory cytokines [transforming growth factor beta (TGF-β)]. According to classification of cytokines based on standard practice used in study of RA, IL-10 and TGF-β are classified as regulatory cytokine., In RA, the balance between pro-inflammatory and anti-inflammatory cytokines determines the degree and extent of inflammation, and thus can lead to different clinical effects.
T helper cells can be subdivided into two subpopulations defined broadly by the cytokines they produce. Th1 cells secrete IFN-γ, TNF-α and IL-2 whereas Th2 secrete IL-4, IL-13 and IL-10., The overproduction of cytokines and growth factors from the inflamed synovium may play a role in the pathophysiology of OA. The low-grade OA synovitis is itself cytokine-driven, although the levels of proinflammatory cytokines are lower than in rheumatoid arthritis (RA). In particular, TNF-α and IL -1 have been suggested as key players in OA pathogenesis, both in synovial inflammation and in activation of chondrocytes.,,
| Materials and Methods|| |
The present study conducted in department of medicine and orthopedics, Jawaharlal Nehru Medical college and hospital. Study comprised of 70 patients of arthritis, out of which, 40 (57.14%) patients were of RA and 30 (42.86%) patients were of OA. Patients fulfilling the Revised ARA Criteria for the Classification of Rheumatoid Arthritis , are diagnosed as a case of RA and recruited for study. Patients fulfilling the clinical and radiological features of OA included in the study, as diagnosis is made with reasonable certainty based on history and clinical examinations. , Patients having psoriatic arthritis, connective tissue disorder (like SLE),Gout, hemophilic arthritis, diabetes mellitus, ANA positive and on DMARDs are excluded from the study. Synovial fluid was aspirated from the knee joint under aseptic precaution and stored at (−70°C) for cytokine estimation. Blood was collected by venipuncture and serum was separated by centrifugation, stored at (−70°C) for cytokine estimation. Rheumatoid factor positivity estimated using (RHELAX RF) kit following the manufacturers' instructions. Study was approved by institutional ethical committee. Cytokine assay estimated using Western Blot ( Immuno-blot transfer). Minimum detection levels of cytokines in blood as well as in synovial fluid were IL-1 ≥ 30 pg/ml, IL-2 ≥ 30 pg/ml, IFN-γ ≥ 10 pg/ml, TNF-α ≥ 30 pg/ml, TGF-β ≥ 20 mg/ml, IL-4 ≥ 25pg/ml, IL-10 ≥ 10 pg/ml.
Analysis was performed using Statistical Package for the Social Sciences software (SPSS version 16.0, Chicago, IL). Continuous variables were expressed as mean ± standard deviation (Gaussian distribution) or range and frequency data was expressed as percentage. Unpaired 't' test for independent samples was used in comparing continuous data between two groups. All P values were two tailed and values of <0.05 were considered to indicate statistical significance. All confidence intervals were calculated at 95% level.
| Results|| |
Age and sex wise distribution
Out of 70 cases studied, 26 (37.14%) were males and 44 (62.86%) were females. Arthritis is commonly seen in females than males. Among the RA patients (40), there were 28 (70%) females and 12 (30%) males with 2.3:1 sex ratio (F: M) and in osteoarthritis, the sex ratio was 1.1:1 comprising of 16 (53.33%) females and 14 (46.67%) males. In RA, maximum patients were between 31-40 years of age group, whereas in OA maximum patients were in 51-60 years of age group [Figure 1]. Mean age of OA patients (53.75 years) was more than RA (42.87 years). In total, 40 patients were of RA, 24(60%) were of rheumatoid factor positive.
Comparison of cytokine profile in blood
On comparison of cytokine profile in blood, proinflammatory cytokines (IL-1, IL-2, IFN-γ), anti-inflammatory cytokine IL-4 and IL-10, regulatory cytokine TGF-β blood levels were raised in RA patients as compared with OA patients i.e. highly significant (P < 0.01). TNF-α (proinflammatory cytokine) levels were also raised in RA patients as compared with OA patients i.e. significant (P < 0.05) [Table 1] and [Figure 2]a-c.
|Figure 2: (a) Comparision of blood cytokines levels in rheumatoid arthritis and osteoarthritis patients (b) Western blot analysis showing levels of various cytokines in blood of RA Patients. (N1 - Control, R1 and R2- TNF-β, R3- IL-4, R4- IL-10) (c) Western blot analysis showing levels of various cytokines in serum of OA Patients. (N1– Control, R1- TNF-β, R2 and R4- IL-10, R3 -IL-4)|
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|Table 1: Comparison of cytokine levels in blood of patients suffering with rheumatoid arthritis and osteoarthritis|
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Comparison of cytokine profile in synovial fluid
On comparison of cytokine profile in synovial fluid, proinflammatory cytokines (IL-1, IL-2, IFN-γ, TNF-α), anti-inflammatory cytokines (IL-4 and IL-10), regulatory cytokine TGF-β of synovial fluid levels were raised in RA patients as compared with osteoarthritis patients i.e. highly significant (P < 0.01) [Table 2] and [Figure 3]a-c.
|Figure 3: (a) Comparision of synovial fl uid cytokines levels in rheumatoid arthritis and osteoarthritis patients (b) Western blot analysis showing levels of various cytokines in synovial fl uid of RA Patients (N1, N2- Control, R1 -IL-1,R2- IL-2,N3- TNF-α, R3- IFN-β) (c) Western blot analysis showing levels of various cytokines in synovial fl uid of OA patients (N1 and N2 – control, R1- IL-1, R2- TNF-α, R3 -IL-2)|
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|Table 2: Comparison of cytokine levels in synovial fluid of patients suffering with rheumatoid arthritis and osteoarthritis|
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| Discussion|| |
Arthritis in general is commonly seen in females than males. This is consistent with information provided by USA National Center for Biotechnology Information 2010. In RA, the sex ratio (M: F) was 1: 2.3 whereas in OA, the sex ratio was 1: 1.1. Most of the studies (Gabriel et al, Symmons et al, Alamanos and Drosos) show that females are two-three times more likely to develop RA compared with males.,, The Framingham Knee Osteoarthritis study suggests that knee osteoarthritis increases in prevalence throughout the elderly years, more so in women than in Men. Guillemin et al, Shichikawa et al, Alamanos and Drosos, and most epidemiological studies suggest an age of disease onset during or after the fifth decade of life.,, Our study have almost similar results as shown by these epidemiological studies mentioned.
On comparing blood cytokine profile in both group, proinflammatory cytokines (IL-1, IL-2, IFN-γ), anti-inflammatory cytokines (IL-4 and IL-10), regulatory cytokine (TGF-β) blood levels were raised in RA patients as compared to osteoarthritis patients i.e. statistically highly significant (P < 0.01). TNF-α (proinflammatory cytokine) levels were also raised in RA patients as compared to osteoarthritis patients i.e. statistically significant (P < 0.05). Paramalingam et al, studied in vivo pro and anti-inflammatory cytokines in normal and patients with RA and observed that the pro-inflammatory cytokines were significantly elevated in RA patients as compared to control while anti-inflammatory cytokines were not elevated.
Fong et al, observed the cytokine concentrations in the synovial fluid and plasma of RA patients and compared it with other arthritic lesions like osteoarthritis. His findings suggest that plasma cytokines levels may reflect synovial levels. In his study, synovial fluid and plasma concentrations of IL-1 alpha, IL-2, TNF-alpha, interferon-alpha (IF-alpha) and interferon-gamma (IF-gamma) were measured. There were no differences in the IFN-alpha, IFN-gamma or TNF-alpha levels between groups but IL-1and IL-2 were significantly raised in RA as compared to other arthritic lesions. There was difference in observation regarding cytokine levels on comparing with other arthritic lesions. Petrovic-Rackov et al, studied the cytokines in RA and OA patients. There results have shown that IL-18, IL-15, IL-12 and TNF-alpha levels in Serum (S) and synovial fluid (SF) of patients with RA were significantly higher than the levels obtain from patients with OA (P < 0.01). Observation regarding TNF-α was almost similar in our study.
In OA patients, mean TNF-α levels (115.7 pg/ml in SF and 80.34 pg/ml in blood) were markedly raised although less than RA patients (minimum detection limit 30 pg/ml). Our finding supported by the fact that TNF-α and IL-1 have been suggested as key players in OA pathogenesis. Future directions in the research and treatment of osteoarthritis will be based on the emerging picture of pathophysiological events by cytokines that modulate the initiation and progression of OA.
Synovial fluid cytokines
On comparing synovial fluid cytokine profile in both groups, proinflammatory cytokines (IL-1, IL-2, IFN-γ, TNF-α), anti-inflammatory cytokine IL-4 and IL-10, regulatory cytokine TGF-β synovial fluid levels were raised in RA patients as compared to osteoarthritis patients i.e. statistically highly significant (P < 0.01). Our finding of high IL-1 and TNF-α levels in synovial fluid of RA patients correlated with study conducted by Lettesjo et al. This is in accordance with work done by Dinarello et al, where it was shown that TNF-α stimulates IL-1 production and that TNF-α and IL-1 act synergistically. Gerald Partsch et al, investigated the concentrations of T cell derived cytokines in the synovial fluids of patients with psoriatic arthritis (PsA) in comparison with RA and OA. Results shows that Th1 type cytokines (IL2, TNF β, and INFγ) and Th2 type cytokines (IL4, IL10) were present in higher concentration in RA patients while in OA patients synovial fluid generally lacked any detectable T cell cytokines altogether. Schlaak et al, studied different cytokine profiles in the synovial fluid of patients with osteoarthritis, RA and seronegative spondylarthropathies. Their data shows that compared to patients with OA, all cytokines (IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-8, IL-10, IL-12, TNF-alpha, TGF-beta 1 and IFN-gamma) were found in higher levels in patients with RA.
Steiner et al studied cytokine production by synovial T cells in RA and comparison with OA and revealed the facts that almost 60% of RA sera contained at least one of the cytokines investigated, although in low concentrations, whereas cytokines were generally not detectable in sera from OA patients. In contrast, cytokines were found in virtually all SF. Majority of SF from RA patients contained IFN-gamma (median level 17 pg/ml) in addition to the monokines IL-6 (4700 pg/ml) and TNF-alpha (157 pg/ml). Cytokine expression was also revealed in study particularly IL-2 and IFN-gamma, and to a lesser extent also IL-4.
Ridderstad et al, studied cytokines in RA and proposed that macrophage-derived cytokines such as IL-1, TNF-alpha, have usually been detected in large quantities, whereas T cell produced cytokines (IL-2, IL-4, IFN-gamma) are absent or present in small quantities. Our observation was similar regarding IL-1, TNF-alpha levels but not consistent with fact that T cell produced cytokines (IL-2, IL-4, IFN-gamma) are absent or present in small quantities in this study. Our study shows that both Th1 and Th2 cells derived as well as IL-1 and TNF- alpha levels (both blood as well as synovial fluid) were significantly raised in RA as compared with osteoarthritis patients. Thus, high levels of these substances have been found in inflammatory arthropathies, in particular in those characterized by a more aggressive and destructive outcome, such as RA.
| Conclusion|| |
In keeping with their role, the determination of cytokines in synovial fluid may be proposed for clinical purposes, including both diagnostic as well as prognostic assessments. Furthermore, as some of these cytokines may reflect disease activity, their determination may also be useful in the evaluation of therapy. In this study, we have tried to examine the role of a variety cytokines in the pathogenesis of rheumatoid arthritis (RA) and osteoarthritis (OA), which might be useful for future possible applications in the treatment of these diseases. Recent data on the cytokine profile in RA and OA implies that alternative treatment strategies should be considered.
| References|| |
Wolfe F. The natural history of rheumatoid arthritis. J Rheumatol Suppl 1996;44:13-22.
Mitchell DM, Spitz PW, Young DY, Bloch DA, McShane DJ, Fries JF. Survival, prognosis, and causes of death in rheumatoid arthritis. Arthritis Rheum 1986;29:706-14.
Isomaki H. Long-term outcome of rheumatoid arthritis. Scand J Rheumatol Suppl 1992;95:3-8.
Hough AJ. Pathology of osteoarthritis. In: McCarty DJ, Koopman WJ, editors. Arthritis and Allied Conditions. Philadelphia and London: Lea and Febiger; 1993. p. 1699-723.
Vervoordeldonk MJ, Tak PP. Cytokines in rheumatoid arthritis. Curr Rheumatol Rep 2002;4:208-17.
Feldmann M, Brennan FM, Maini RN. Role of cytokines in rheumatoid arthritis. Annu Rev Immunol 1996;14:397-440.
Mosmann TR, Sad S. The expanding universe of T-cell subsets: Th1, Th2 and more. Immunol Today 1996;17:138-46.
Romagnani S. The Th1/Th2 paradigm. Immunol Today 1997;18:263-6.
Horton WE Jr, Udo I, Precht P, Balakir R, Hasty K. Cytokine inducible matrix metalloproteinase expression in immortalized rat chondrocytes is independent of nitric oxide stimulation. In Vitro
Cell Dev Biol Anim 1998;34:378-4.
Goldring MB. The role of cytokines as inflammatory mediators in osteoarthritis: Lessons from animal models. Connect Tissue Res 1999;40:1-11.
Martel-Pelletier J, Alaaeddine N, Pelletier JP. Cytokines and their inhibitors in the pathophysiology of osteoarthritis. Front Biosci 1999;4:D694-703.
Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, et al.
The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 1988;31:315-24.
Clegg DO, Ward JR. Diagnostic criteria in rheumatoid arthritis. Scand J Rheumatol Suppl 1987;65:3-11.
Altman R, Alarcon G, Appelrouth D, Bloch D, Borenstein D, Brandt K, et al.
The American College of Rheumatology criteria for the classification and reporting of osteoarthritis of the hip. Arthritis Rheum 1991;34:505-14.
Schlaak JF, Pfers I, Meyer Zum Buschenfelde KH, Märker-Hermann E. Different cytokine profiles in the synovial fluid of patients with osteoarthritis, rheumatoid arthritis and seronegative spondylarthropathies. Clin Exp Rheumatol 1996;14:155-62.
Gabriel SE, Crowson CS, O'Fallon WM. Comorbidity in arthritis. J Rheumatol 1999;26:2475-9.
Symmons DP. Epidemiology of rheumatoid arthritis: Determinants of onset, persistence and outcome. Best Pract Res Clin Rheumatol 2002;16:707-22.
Alamanos Y, Drosos AA. Epidemiology of adult rheumatoid arthritis. Autoimmun Rev 2005;4:130-6.
Felson DT. The epidemiology of knee osteoarthritis: Results from the Framingham Osteoarthritis Study. Semin Arthritis Rheum 1990;20 3 Suppl 1:42-50.
Guillemin F, Briancon S, Klein JM, Sauleau E, Pourel J. Low incidence of rheumatoid arthritis in France. Scand J Rheumatol 1994;23:264-8.
Shichikawa K, Inoue K, Hirota S, Maeda A, Ota H, Kimura M, et al
. Changes in the incidence and prevalence of rheumatoid arthritis in Kamitonda, Wakayama, Japan, 1965-1996. Ann Rheum Dis 1999;58:751-6.
Sivalingam SP, Thumboo J, Vasoo S, Thio ST, Tse C, Fong KY. In vivo
pro- and anti-inflammatory cytokines in normal and patients with rheumatoid arthritis. Ann Acad Med Singapore 2007;36:96-9.
Fong KY, Boey ML, Koh WH, Feng PH. Cytokine concentrations in the synovial fluid and plasma of rheumatoid arthritis patients: Correlation with bony erosions. Clin Exp Rheumatol 1994;12:55-8.
Petrovic-Rackov L, Pejnovic N. Clinical significance of IL-18, IL-15, IL-12 and TNF-alpha measurement in rheumatoid arthritis. Clin Rheumatol 2006;25:448-52.
Lettesjo H, Nordstorm E, Storm H, Nilsson B, Glinghammar B, Dahlstedt L, et al.
Synovial fluid cytokines in patients with Rheumatoid arthritis or arthritic lesions. Scand J Immunol 1998;48:286-92.
Dinarello CA, Cannon JG, Wolff SM, Bernheim HA, Beutler B, Cerami A, et al.
Tumor necrosis factor (cachectin) is in endogenous pyrogen and induces production of interleukin 1. J Exp Med 1986;163:1433-50.
Partsch G, Wagner E, Leeb BF, Bröll H, Dunky A, Smolen JS. T cell derived cytokines in psoriatic arthritis synovial fluids. Ann Rheum Dis 1998;57:691-3.
Steiner G, Tohidast-Akrad M, Witzmann G, Vesely M, Studnicka-Benke A, Gal A, et al.
Cytokine production by synovial T cells in rheumatoid arthritis. Rheumatology (Oxford) 1999;38:202-13.
Ridderstad A, Abedi-Valugerdi M, Möller E. Cytokines in rheumatoid arthritis. Ann Med 1991;23:219-23.
[Figure 1], [Figure 2], [Figure 3]
[Table 1], [Table 2]