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Food Science and Human Wellness 3 (2014) 1–8

In vitro modulation of TH1 and TH2 cytokine expression by edible tuber ofDioscorea alata and study of correlation patterns of the cytokine expression

Priyankar Dey, Tapas Kumar Chaudhuri ∗Cellular Immunology Laboratory, Department of Zoology, University of North Bengal, Siliguri 734013, West Bengal, India

Received 20 November 2013; received in revised form 3 January 2014; accepted 17 January 2014

bstract

The underground tuber of Dioscorea alata is a palatable food and is also claimed to have therapeutic effects. Various groups of researchersemonstrated different activities of this tuber such as antioxidant, hepatoprotective, anti-ulcer and anti-diabetic activities. Therefore, the aim of theresent study was to evaluate the immunomodulatory potentialities of 70% hydro-methanolic extract of D. alata underground tubers by investigatingifferent parameters like the expression of interferon gamma (IFN-�), interleukin (IL)-2, IL-4 and IL-10, in addition to its effect on the proliferation

f murine splenic T-lymphocytes in vitro. Results demonstrated that D. alata can actively polarize the TH0 lymphocyte population towards thexpression of TH1 immune response by up-regulating the IFN-� and IL-2 expression and down-regulating the IL-4 and IL-10 expression. The tuberxtract also proved to possess mitogenic activity as evidenced by the proliferation of lymphocytes in vitro.

2014 Beijing Academy of Food Sciences. Production and hosting by Elsevier B.V. All rights reserved.

eywords: Anti-inflammatory; Cytokine; Dioscorea (yam); Immunomodulation; Lymphocyte; MTT; Proliferation

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. Introduction

CD4+ T-helper cells differentiate from the precursor TH0ells to either TH1 or TH2 cells. These cells differ in theirytokine expression patterns which primarily governs their func-ional nature. In general, TH1 cytokine response leads towardsell-mediated and inflammatory immune response whereasH2 cytokine response mediates humoral immune response.ytokines are the major immunoregulators of the body. Inter-

eron gamma (IFN-�) is a TH1 cytokine which primarily displaysntiviral, anti-tumour and counter-autoimmune capacities [1].ctivation and increase in antigen presentation by macrophages,

ugmentation in leukocyte migration, up-regulation of MHC

∗ Corresponding author at: Cellular Immunology Laboratory, Department ofoology, University of North Bengal, Siliguri 734013, West Bengal, India.

el.: +91 8927884784/9434377127.

E-mail addresses: dr tkc [email protected],[email protected] (T.K. Chaudhuri).eer review under responsibility of Beijing Academy of Food Sciences.

Production and hosting by ElsevierELSEVIER

213-4530 © 2014 Beijing Academy of Food Sciences. Production andosting by Elsevier B.V. All rights reserved.ttp://dx.doi.org/10.1016/j.fshw.2014.01.001

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xpression and co-stimulatory signals are other salient featuresf IFN-�. Patients under clinical studies with recombinant IFN-�xhibit a vast spectrum of activities which include enhancementf antibody-dependent cell-mediated cytotoxicity and increasen oxidative metabolism of macrophages [2]. IFN-� expres-ion on tumour growth induces CD8+ cytotoxic T-cells andhus results in potent tumour immunity against neuroblastoma,reast cancer and in methyl cholanthrene-mediated tumour mod-ls [3]. Currently IFN-� therapy is active in the treatment ofhronic granulomatous disease, osteoporosis, idiopathic pul-onary fibrosis and ovarian cancer [4]. Another TH1 cytokine

nterleukin (IL-2) also displays a broad array of activities whichnclude body’s innate response against microbial infections,self’ and ‘non-self’ recognition and proliferation of antigen-pecific CD8+ and CD4+ T-cells. Impairment of IL-2 signallingue to the absence of IL-2 or IL-2 receptors results in generalizednflammatory syndrome and often cause fatal autoimmune col-tis [5]. At present, IL-2 immunotherapy has been approved forhe treatment of asthma, renal transplantation, multiple sclerosis,soriasis and ulcerative uveitis [4]. Pro-inflammatory cytokinesFN-� and IL-2 are the principal cytokines to mediate naturaliller (NK) cell activity and increases the number of CD4+ cellsn HIV-infected patients [6]. The anti-inflammatory cytokinesL-4 and IL-10 belong to TH2 sub-set. IL-4 mediates allergic

nflammation and may induce class switching to form � iso-ype immunoglobulin (Ig) E secreted by B-lymphocytes and beesponsible for allergic reactions [7]. High concentration of IL-4

http://crossmark.crossref.org/dialog/?doi=10.1016/j.fshw.2014.01.001&domain=pdfhttp://www.sciencedirect.com/science/journal/aip/22134530dx.doi.org/10.1016/j.fshw.2014.01.001mailto:[email protected]:[email protected]/10.1016/j.fshw.2014.01.001

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P. Dey, T.K. Chaudhuri / Food Scie

n the serum and bronchoalveolar lavage due to dust-mite aller-ens is the hallmark of atopic individuals [8]. Increased IL-4xpression is seen in cases of asthma, various allergies, hepatitis

infection, cutaneous T cell lymphoma, malaria and leish-aniasis. On the other hand, the immunosuppressive cytokine

L-10 down-regulates the cell-mediated immune response bynhibiting prostaglandin E production and suppression of vari-us pro-inflammatory cytokines like IL-2, TNF-�, IL-1, IL-6,nd IL-8 [9,10]. In various B cell lymphoproliferative diseases,L-10 may function as growth factor for malignant B cells11] and elevated levels of IL-10 was found to be associatedith the systemic infection, leprosy, HIV infection, rheumatoid

rthritis, multiple sclerosis, lung cancers, melanomas, gliomas,eukaemia and lymphomas [5]. Both IL-4 and IL-10 polarizehe TH0 lymphocyte differentiation towards TH2 response andoth are natural antagonist to the expression and functionalityf IFN-�. Immunomodulation of the T-cells by nutritive supple-ents and herbal medicines towards either TH1 or TH2 pathway

hiefly determines the pattern of the immune response.The underground tuber of Dioscorea alata L. is a dietary

upplement in south-eastern Asia and Africa [12]. It is also useds traditional medicine in China [13]. Antioxidant [14], antidi-betic [15], antiosteoporotic [13], hepatoprotective [16] andnti-ulcer [17] activities of D. alata were reported previously.he effect of D. alata tuber mucilage on splenocyte-mediatedytotoxicity, stimulation of antibody production and the phago-ytic capacity of monocyte and granulocytes were studied byhang et al. [18]. Previous report suggests that 70% methano-

ic extract of D. alata underground tuber possesses stimulatoryffect on various activities of murine macrophage, suppressionf lipopolysaccharide stimulated nitric oxide level and stimu-ation of humoral immune response [19]. Moreover, the samextract was also chemically characterized and found to containigh phenolic and flavonoid content which contributed to itsntioxidant capacity [20].

Therefore, the objective of the present study was to furthervaluate the effect of the 70% hydromethanolic extract of D.lata underground tuber for modulating TH1 (IL-2 and IFN-�)nd TH2 (IL-4 and IL-10) cytokine expression and its effectn the proliferation pattern of murine splenic T-lymphocytes.n addition, the correlation patterns of the cytokine expressionsnd the cell proliferation were also studied.

. Materials and methods

.1. Chemicals and reagents

All the cell culture reagents including RPMI-1640 (AL-62S), concanavalin A (con A), foetal bovine serumFBS), EZcountTM 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-etrazolium bromide (MTT) Cell Assay Kit etc. were procuredrom HiMedia Laboratories Pvt. Ltd. (Mumbai, India), unless

therwise indicated. Nystatin was obtained from Sigma AldrichUSA). Mouse IL-2, IFN-�, IL-4 and IL-10 ELISA kits wererocured from RayBio (Georgia, United States).

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nd Human Wellness 3 (2014) 1–8

.2. Preparation of plant extract

D. alata tubers were collected from Sonitpur district ofssam, India and authenticated by Botanical Survey of India,hillong, Meghalaya, India (accession number 78051). Seventyercent hydro-methanolic extract of underground tubers of D.lata was prepared according to a previously described method21]. In brief, the tubers were washed properly with double dis-illed water and dried in the shade at room temperature for 2eeks followed by grinding to powder. The powder (100 g)as mixed with 70% methanol (1000 mL) and kept overnight

t 37 ◦C in a shaking incubator (160 r/min). The mixture wasentrifuged at 5000 r/min for 15 min and the supernatant (liq-id phase) was collected. The remaining pellet was mixed with0% methanol (1000 mL) and kept in a shaking incubator asreviously described. The supernatant (liquid phase) was againollected after centrifugation and mixed with the supernatant ofrevious phase. The liquid was then filtered and the filtrate wasoncentrated under reduced pressure in a rotary evaporator. Theesultant was lyophilized and stored at −20 ◦C until further use.

.3. Preparation of murine splenocytes

Swiss albino mice were sacrificed under proper anaesthe-ia and the spleen was removed aseptically. The cells of thepleen were passed through the mesh of a tissue grinder and theuspension was prepared in RPMI-1640. The cell suspensionas washed thrice for 10 min using RPMI-1640 (2000 r/min)

nd then resuspended in 1 mol/L NH4Cl to eliminate red bloodells (RBC). After 10 min the cells were again centrifuged andesuspended in RPMI-1640.

All the experiments were approved by the ethical commit-ee of the Department of Zoology, University of North Bengaldate: 15/09/10) and in accordance with the legislation for therotection of animals used for scientific purposes.

.4. MTT cell proliferation assay

The cell proliferation assay was performed using EZcountTM

TT Cell Assay Kit according to the manufacturer’snstructions. Briefly, splenocyte suspension was made2 × 106 cells/mL) in RPMI-1640 (supplemented with 50 U/mLenicillin, 50 U/mL streptomycin, 50 U/mL nystatin and 10%BS) and the cells were seeded into different wells of 96 wellulture plates in presence of 5 �g/mL con A. To this, 50 �Lf different concentrations (0–80 �g/mL in RPMI-1640) of. alata extract was added. The plate was incubated for 24 hnder 5% CO2 and humidified atmosphere of 90% air at 37 ◦Cemperature. After the incubation 10 �L MTT (5 mg/mL)olution was added to each well. The plate was wrapped withluminium foil to avoid exposure to light and incubated for

h. Then 100 �L solubilizing reagent was added to each wellnd the absorbance was measured at 570 nm using Bio-RadMarkTM microplate absorbance reader.

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Fig. 1. Effect of D. alata on the proliferation of splenic lymphocytes. All dataare presented as the mean ± SD of six measurements. Murine splenic lympho-cytes (2 × 106 cells/mL) were cultured in vitro with con A (5 �g/mL) alongwith different doses of D. alata (0–80 �g/mL) extract. After 24 h of incubation,tP

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.5. Estimation of cytokines

Mouse splenocytes were cultured with varying concentra-ions (0–80 �g/mL) of D. alata tuber extract as previouslyescribed (Section 2.4) for 48 h. After incubation, suspensionsrom each well were centrifuged (2850 × g) for 5 min and theulture supernatants were collected to estimate the concentra-ions of IL-2, IFN-�, IL-4 and IL-10 using RayBio ELISA kitsccording to the manufacturer’s instructions. Briefly, 100 �Lulture supernatant from different experimental groups weredded to the anti-mouse cytokine (IL-2, IFN-�, IL-4, IL-10)oated wells of the 96 well ELISA plate. The plate was cov-red and incubated at room temperature for 2.5 h. The solutionere then discarded and the wells were washed 4 times with

ssay buffer (300 �L). Next, 100 �L biotinylated anti-mouseytokine (IL-2, IFN-�, IL-4, IL-10) antibody was added to eachell and the plate was incubated at room temperature with con-

inuous but gentle shaking. After 1 h, the wells were washedith assay buffer as previous. Then 100 �L of horseradish per-xidase (HRP)–streptavidin conjugate solution was added andhe plate was incubated for 45 min at room temperature withentle shaking. Next, the wells were washed as previous and00 �L 3,3,5,5′-tetramethylbenzidine (TMB) one-step substrateeagent was added followed by 30 min incubation at room tem-erature in dark. Then 50 �L of 0.2 mol/L sulphuric acid wasdded to each well to stop the reaction. Absorbance was immedi-tely measured at 450 nm using a Bio-Rad iMarkTM microplatebsorbance reader. Standards were run in parallel to the samples.

.6. Statistical analysis

All data are presented as the mean ± SD of six measurements.he comparisons between the control group and the test groupsere performed by one-way analysis of variance (ANOVA) withunnett’s test using KyPlot version 2.0 beta 15 (32 bit). P < 0.05as considered significant. The linear correlation analysis waserformed by Microsoft Excel 2010. Principal component anal-sis (PCA) based on the correlation matrix was performed inrder to analyse the pattern and inter-relations of up- and down-egulation of the cytokines using IBM SPSS statistics version0.0.

. Results

.1. Cell proliferation assay

Fig. 1 displays the results of the splenic lymphocyte prolif-ration using plant extract. Increased metabolic activity of theells in a dose-dependent manner were found as evidenced fromhe cell proliferation. At 80 �g/mL dose, the percentage of celliability was (178.46 ± 6.08)%, much higher than that of theontrol.

.2. Estimation of cytokines

The results after 48 h of incubation displayed significantP < 0.05) dose-dependent up-regulation of IL-2 and IFN-�

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he lymphocyte viability was measured by MTT method. **, P < 0.01; ***; < 0.0001 compared with 0 �g/mL.

nd down-regulation of IL-4 and IL-10 (Fig. 2), comparedo the control (0 �g/mL). At 80 �g/mL, the concentrationf IL-2 ((28.7 ± 2.24) pg/mL) was significantly (P < 0.001)igher than that of the control ((9.94 ± 1.56) pg/mL), account-ng for (195.06 ± 65.75)% increase in the IL-2 secretionFig. 2A). IFN-� concentration increased (100.22 ± 23.48)%ompared to control group (Fig. 2A). At 80 �g/mL dose,he IFN-� concentration was (356.00 ± 12.52) pg/mL, whichas significantly higher (P < 0.001) than the 0 �g/mL group

(179.04 ± 16.09) pg/mL).D. alata tuber extract considerably down-regulated IL-

secretion to 50.06 ± 10.76%. At the dose of 80 �g/mLFig. 2C), the concentration of IL-4 in the culture supernatantas (1.35 ± 0.20) pg/mL and this was significantly (P < 0.001)

ower than that of the control group ((3.22 ± 0.20) pg/mL).he secretion of IL-10 was down-regulated by the plantxtract from 2206.33 ± 184.69 pg/mL in the control group to1101 ± 132.71) pg/mL at the highest dose (Fig. 2D). Com-ared to the control, the IL-10 concentration has significantlyP < 0.01) decreased at 80 �g/mL dose, which accounted for aotal of (49.71 ± 8.79)% decrease in IL-10 secretion.

The IFN-�/IL-4 ratio, a major determinant of immunomod-

latory activity, was also found to be elevated from 0 �g/mL to0 �g/mL. At 0 �g/mL the ratio was 56.02 ± 4.91 whereas at0 �g/mL the ratio was increased up to 268.86 ± 51.05.

4 P. Dey, T.K. Chaudhuri / Food Science and Human Wellness 3 (2014) 1–8

Fig. 2. Effect of D. alata on the expression pattern of various cytokines. All data are presented as the mean ± SD of six measurements. Murine splenic lymphocytes( 6 erent c thod.i > 0.05

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2 × 10 cells/mL) were cultured in vitro with con A (5 �g/mL) along with diffytokines (pg/mL) were performed from the culture supernatants by ELISA men IL-4 secretion; and (D) decrease in IL-10 secretion. N.S.: non-significant (P

.3. Correlation study

The correlation analysis of the results revealed that IFN- was highly correlated with IL-2 (R2 = 0.9496) and IL-4R2 = 0.9053). Similarly, IL-4 and IL-2 was also found to be

ighly correlated (R2 = 0.9122).

Linear negative correlation resided between IL-10 and IFN- (R2 = 0.8073), IL-2 (R2 = 0.8456) and IL-4 (R2 = 0.6278) as

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doses of D. alata (0–80 �g/mL) extract. After 48 h of incubation estimation of (A) Increase in IFN-� expression; (B) increase in IL-2 secretion; (C) decrease); *. P < 0.05; **. P < 0.01; ***. P < 0.001, compared with 0 �g/mL.

isplayed in Fig. 3. The correlation between the cell viabilitynd cytokine expression is demonstrated in Fig. 4.

Under given parameters, the variables were subjected to PCAnd the results of the loading plot (Fig. 5) completely matchedith the correlation matrix (Table 1). The loading of the first

nd second principal component (PC1 and PC2) accounted for4.19% and 4.29% of the variance, respectively. The loadinglot demonstrated that cell viability, IL-2 and IFN-� are heavily

P. Dey, T.K. Chaudhuri / Food Science and Human Wellness 3 (2014) 1–8 5

Fig. 3. Correlation between the expression patterns of various cytokines form splenic lymphocytes. Linear correlation analysis was performed by Microsoft Excel2010. (A) Positive correlation between IL-2 and IFN-�; (B) negative correlation between IL-10 and IFN-�; (C) negative correlation between IL-4 and IFN-�; (D)n -10 ad

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egative correlation between IL-4 and IL-2; (E) negative correlation between ILetermination unless otherwise stated.

oaded on PC1 with component score of 0.691, 0.767 and 0.748,espectively.

. Discussion

The complex interactions between the lymphoid, inflam-atory and hematopoietic cells are chiefly governed by the

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nd IL-2; and (F) positive correlation between IL-10 and IL-4. R2: coefficient of

ytokines which are generally divided into TH1 and TH2ub-sets. TH1 cells are the primary source of IL-2, IFN-�,ymphotoxins and other factors which characterize the cell-ediated immune responses [22]. IL-4, IL-5, and IL-10 are

roduced by the TH2 cells which not only mediate the humoralmmune responses, but also promote allergic reactions by induc-ng the function of IgE formation [7]. In the present study,

6 P. Dey, T.K. Chaudhuri / Food Science and Human Wellness 3 (2014) 1–8

Fig. 4. Correlation between splenic lymphocyte viability and cytokine expression. Linear correlation analysis was performed by Microsoft Excel 2010. (A) Positivecorrelation between IFN-� and cell viability; (B) positive correlation between IL-2 and cell viability; (C) negative correlation between IL-4 and cell viability; (D)negative correlation between IL-10 and cell viability.

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Table 1Correlation matrix of cell viability, IFN-�, IL-2, IL-14 and IL-10.

Viability IFN-� IL-2 IL-4 IL-10

Viability 1.000IFN-� 0.976*** 1.000IL-2 0.973*** 0.974*** 1.000IL-4 −0.906** −0.951** −0.955*** 1.000IL-10 −0.918** −0.898** −0.919** 0.792* 1.000The matrix is based on the principal component analysis performed by SPSSstatistics version 20.0 software package. * correlation is significant at the 0.05lc

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ig. 5. Loading plot of PCA for cell viability, IFN-�, IL-2, IL-4 and IL-10. PC1
nd PC2 accounted for 94.19% and 4.29% variance, respectively. The expression

atterns of IFN-� and IL-2 are highly similar. Thus, they overlap each other inhe loading plot. PCA was performed using IBM SPSS statistics version 20.0.ydro-methanolic extract of D. alata underground tuber was

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evel (1-tailed); ** correlation is significant at the 0.01 level (1-tailed); ***.orrelation is significant at the 0.001 level (1-tailed).

ound to stimulate the splenic T-lymphocyte proliferation andodulate cytokine expressions in vitro.Spleen functions primarily against the antigens in the blood

tream. The outer red pulp of spleen is abundant in lymphocytes.on A, which is a typical T-lymphocyte mitogen, responsi-le for blastoid differentiation of the T-lymphocytes [23], mustave stimulated T-lymphocyte population out of the whole

plenocyte cells to secret the cytokines in our experiments asell. This is evidenced from the increase in the number of

ells. Previously, Liu et al. [24] have also demonstrated the

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itogenic effect of dioscorin, the storage protein of D. alatan the phytohemagglutinin induced cell proliferation. Besides0% methanolic extract of D. alata formerly showed the capac-ty to increase the splenic cellularity in murine model in vivo19]. Here, we have shown that the degree of stimulation ofplenocytes by D. alata extract at 80 �g/mL concentration alongith con A was 178.46 ± 6.08%, much higher than that of the

timulation of con A alone in group I (0 �g/mL). It may beypothesized that D. alata hydro-methanolic extract acted syn-rgistically with the external mitogenic stimulators to triggeritosis of the splenic cells. Therefore, D. alata possess potent

mmunogenicity towards the activation and proliferation of theplenic cells.

Profound increases in the TH1 cytokine (IL-2, IFN-�) levelsnd decrease in TH2 cytokine (IL-4 and IL-10) levels were evi-ent by the stimulation of D. alata. Similar findings [25,26] alsouggest the potentiality of various other plant materials to up-egulate the TH1 response. In the present study, dose-dependentncrease in the splenic T-lymphocyte was evident and the radiusf proliferation was much higher in case of con A along with thelant extract than con A alone induced stimulation. This couldossibly be due to the fact that the D. alata induced increasedxpression of IL-2, which is a natural T-lymphocyte stimulatingactor or the plant extract itself may posses mitogenic effect,erhaps, which have mimicked the function of IL-2 in the stim-lation of splenic T-lymphocytes.

IL-4 and IL-10 are known to suppress the expansion ofH1 subsets. Thus in the present study (Fig. 3), high negativeorrelation between IFN-� and IL-4 (R2 = 0.9053) and IL-10R2 = 0.8073) were evident and positive correlation between IL-

and IFN-� (R2 = 0.9496) were found, both of which are TH1ytokines. Considering the dose-dependent expression patternsf the four cytokines, the correlation matrix (Table 1) also dis-layed very high positive correlation (0.974) between IL-2 andFN-� and very high negative correlation of IL-4 with IFN-�−0.951) and IL-2 (−0.955).

Principal component analysis is a multivariate data reduc-ion technique which statistically allows visualizing the patternf inter-relations among the variables of the experiments [27].roups of data with correlated immunological parameters cane visualized in a simplified manner using this data reduc-ion technique [28]. Under univariate correlation analysis, biasay emerge due to antagonist or stimulatory relations of

ifferent cytokines. This bias is eliminated and the overallnter-correlation patterns are visualized using this technique.reviously, PCA analysis have been extensively used to anal-se the cytokine correlation patterns in different immunologicaltudies [29,30].

It is interesting to note the correlation patterns of splenic-lymphocyte proliferations with the levels of cytokine expres-ions. IFN-� and IL-2 increased expression pattern were foundo be similar to the PCA loading plot (Fig. 5) and thus, remainverlapped. The increase in cell viability, which correlated with

he TH1 cytokine release, was also placed in the same double-ositive quadrant in the loading plot. Consequently, under givenondition the D. alata-mediated modulation pattern of IL-2 andFN-� are very much similar, followed by nearly similar patterns

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nd Human Wellness 3 (2014) 1–8 7

f increase in cell viability. On the other hand, due to the sim-lar down-regulation in the expression of TH2 cytokines, IL-4nd IL-10 remained in the double-negative quadrant, but theirpartness in the loading plot (Fig. 5) was resulted due to the dif-erence in expression patterns of both the cytokines. Under theresent in vitro condition when linear correlation between anywo cytokine expression were considered (Fig. 3A), the highestorrelation (R2 = 0.9496) was found between IFN-� and IL-2.herefore, with the help of PCA coupled with correlation matrix

t is now feasible to analyse the regulation and correlation pat-erns considering multiple variables at a time in a single plot.he allocated quadrants of the four cytokines and viability inig. 5 are fixed for immunomodulation by D. alata extract underxact given conditions. Increase or decrease in variables underltered conditions may change the correlation patterns in theCA loading plot.

Recently, modulation of TH1 and TH2 balance has emerged as major determinant for the therapeutic activity of plant extracts.ased on the regulatory patterns of TH1/TH2 cytokine expres-

ion, immunomodulators are classified into TH1, TH2 or mixedH1/TH2 agents [31]. Moreover, TH1/TH2 balance has alsoeen implicated in anti-tumour immunity where IFN-� and IL-

producing TH1 cells-mediate cellular and tumour immunity,hereas IL-4 producing TH2 cells are responsible for suppres-

ion of cytolytic activity [32]. In the present study polarizationf the TH1 pathway was evident from the increase in the expres-ion of IFN-� and IL-2. Besides, the relative immunomodulatoryffect [31] of D. alata was higher for TH1 subset as evidentrom the IFN-�/IL-4 ratio, which is a commonly used index ofH1/TH2 immunity [33].

. Conclusion

The results from our experiments revealed the potentmmunomodulatory activity of the edible tubers of D. alata,onsumption of which may in turn stimulate the lymphocyteroliferation and navigate the immune response towards theH1 phenotype. We also hypothesize that the extract may pos-ess mitogenic activity and exert excellent pro-inflammatoryesponse upon consumption. The TH1/TH2 fate regulatory rolef D. alata was investigated through ELISA method by estimat-ng the expressed proteins in the culture medium. It is difficulto conclude from the present study whether D. alata possesshe potentiality to regulate the mRNA levels of the cytokinesr it simply regulates the protein expression patterns. There-ore, RT-PCR and FACS analysis are further required in additiono modulation study of cyclooxygenase, prostaglandin E2 anduclear factor-kappa � for in-depth analysis of the immunomod-latory and anti-inflammatory activities of D. alata.

onflict of interest

The authors declare no conflict of interest.

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cknowledgements

The financial assistance was provided by Departmentf Biotechnology, Government of India (sanction order no.T/09/NE/TBP/2010). The authors are thankful to Prof.nupam Chatterjee of the Department of Biotechnology &ioinformatics, North-Eastern Hill University for supplying thelant material and to Dr. Nripendranath Mandal, Division ofolecular Medicine, Bose Institute for his kind help in prepa-

ation of the plant extract.

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In vitro modulation of TH1 and TH2 cytokine expression by edible … · 2016. 12. 7. · Shillong,...

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Transcript of In vitro modulation of TH1 and TH2 cytokine expression by edible … · 2016. 12. 7. · Shillong,...