Toll-Like Receptor 3-4 Expression Decrease in BALB c Diabetic Mouse Models | Rifa’i | Proceedings of The Annual International Conference, Syiah Kuala University - Life Sciences & Engineering Chapter 5995 12378 1 SM
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Toll- Lik e Re ce pt or 3 - 4 Ex pr e ssion D e cr e a se in BALB/ c
D ia be t ic M ou se M ode ls
1*
Muhaim in Rifa’i, 1 Aris Soewondo, and
1
M. Sasm it o Dj at i
1
Depart m ent of Biology, Facult y of Mat hem at ics and Nat ural Sciences, Brawij aya Universit y,
Malang 65145, I ndonesia
* Corresponding Aut hor: [email protected]
Abst ra ct
I n t his st udy , we observ ed t he effect of propolis ex t ract on im m unological funct ion in diabet ic
m ouse m odels wit h t he aim of highlight ing t he dy nam ics of im m unological st at us in t y pe- 2
diabet es. I n t his st udy we t est ed t he abilit y of propolis t o norm alize hom eost asis. Here we
showed t hat propolis im prov e hom eost asis by slight ly incr easing t he lev el of TLR ex pression.
The result s of t his st udy differ from prev ious findings which report ed t hat TLR ex pression
increased in diabet es m ellit us and m ost t reat m ent is int ended t o suppress t he ex pression of
TLR. I n t his st udy we found t hat TLR- 3 and TLR- 4 ex pression decreased in m ouse m odels of
STZ induced diabet es m ellit us. Furt herm ore, we found t hat adm inist rat ion of propolis showed
an increase of red blood cell precur sor s ( TER- 119) and im pr ov e t he rat io of CD4: CD8 dependent
m anner.
Ke y w ords: Diabet es m ellit us, propolis, im un om odulat or, Toll- lik e recept or
I nt r oduct ion
Current ly about 348 m illion people worldwide suffer from diabetes m ellit us ( DM) . DM is t he m ost
deadly diseases and has penet rated all count ries. I t is current ly known, 4.6% of t he t otal I ndonesian
populat ion suffer from diabet es m ellit us, and this will increase to 7.8% in 2030. During t his t im e, t he
t reat m ent of various diseases including diabet es really dependent on synt het ic drugs. On the ot her
hand, I ndonesia has abundant of nat ural resources wit h t he potent ial t o cure various diseases.
Propolis is one of I ndonesia's pride t radit ional m edicine which is often claim ed by t he com m unit y wit h
abilit y t o cure diabetes m ellit us. But unfortunat ely t here have not enough research been done on t his
exist ing claim s. On t he ot her hand, pat ients with diabet es m ellit us want to get t he right m edicine to
im prove t he qualit y of t heir life. I n general, t he severit y of diabetes is t riggered by inflam m at ion
fact ors known as proinflam m atory cyt okines. Adm inist rat ion of propolis to diabet ic m ice results in a
significant decrease of blood sugar levels and increases t he num ber of naïve T cells. When propolis
was given t o norm al or diabet ic m ice, it can increase t he num ber of regulat ory T cells. I n m ouse
m odels of diabetes m ellit us t he developm ent of effect or cells can be suppressed after propolis
adm inist rat ion. I n individual wit h diabetes m ellit us propolis t ends t o suppress I FN- γ but rather
increased TLR product ion.
M a t e r ia ls a nd M e t hods
Pr ocedur e
Preparat ions of Et hanol Ext ract of Propolis.
Propolis ext ract preaparat ion consisted of t hree phases including drying, ext ract ing, and evaporat ing.
The drying process began by washing the sam ple, cut t ing it int o sm all pieces, and put t ing t hem in t he
oven with 40- 60º C degree. Sam ples were dried and crushed by a blender. As m uch as 100 gram s of
dry sam ples were weighed and put in 1 L Erlenm eyer glass, soaked wit h ethanol t o the volum e of 1 L
before t he ext ract ion process. Sam ple in et hanol was st irred for ± 30 m inutes and allowed t o st and
overnight t o sett le. Supernatant was rem oved and filt ered. The ext ract of propolis was dried by using
an evaporat or and weighed. Propolis powder was t hen dissolved in dist illed wat er for oral t reatm ent .
Type 2 diabet es m ice m odels were t hen t reated wit h t he et hanol ext ract of propolis for 7 weeks. Mice
were m aintained in pat hogen free facilit y. Mice were fed wit h pellets BR1 by 5- 10 gram s per day and
drink was provided ad libit um .
I nduct ion of Type 2 diabetes m ellit us wit h STZ.
St rept ozot onin ( STZ) dissolved in 0: 01 M cit rate buffer, pH 4.5 and always prepared in a fresh
condit ion. STZ was int raperit oneally inj ected t o m ice in t he age of 5 days ( 100 m g/ kg BW). Five weeks
aft er STZ inject ion, diabet ic condit ion is confirm ed by m easuring t he glucose levels using a glucom eter
( GlucoCare Ult im a) .
270
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Results a nd D iscussion
I n t his study we showed t hat TLR- 3 expression in diabet ic m ice decreased 6 t im es lower t han norm al.
The suppression of TLR- 3 expression is suspect ed to affect t he qualit y of hom eost asis because TLR- 3
norm ally funct ions as an act ivat or of innate im m unit y. The low expression of TLR- 3 in t urn m ake
difficult y for elim inat ing infect ious agents in pat ient wit h diabetes m ellit us. So t hat in m any cases
people wit h diabet es m ellit us oft en wounds and difficult t o be cured. However, t he increase of TLR- 3
expression far beyond norm al will bring bad im pacts and also aggravat e diabet ic pat ient healt h
condit ions.
Figur e 1 . I n m ice m odels of diabetes m ellit us induced by STZ, expression level of TLR- 3 decreased
Left panel showed TLR- 3 expression on CD4 cells in norm al m ice. Right panel is an expression of TLR3 on CD4 T cells in m ouse m odels of DM. Cells were isolated from spleen and analyzed by flow
cyt om et ry. CD4 posit ive cells expressing TLR- 3 ( CD4 + TLR- 3 + ) were shown in each panel.
Observat ions on CD4 T cells clearly showed t hat the expression of TLR- 3 decreased in m ouse m odel of
diabetes. I n t his experim ent we cannot explain t he sit e effect of TLR decline on T lym phocyt es.
However, t here is a lit t le clue that t he decline of TLR expression m ay correlate wit h t he failure of
pat ient in elim inat ing foreign ant igen specially bact eria and virus. TLR- 3 has been known to be
associat ed with t he em ergence of signal t ransduct ion related t o virus elim inat ion. The decline of TLR- 3
expression in DM will lead t o t he em ergence of disorders associated wit h TLR- 3 funct ion.
Figur e 2 . Adm inist rat ion of propolis in m ice m odel of diabet es increase t he expression of TLR- 3 on
CD4 cells approaching norm al condit ions
From left panel to right is propolis t reat m ent at a dose of 50, 100, and 200 m g/ kg BW recpect ively.
The cells were isolat ed from spleen and analyzed by flow cyt om et ry. CD4 posit ive cells expressing
TLR- 3 ( CD4 + TLR- 3 + ) were shown in each panel.
Conclusions
TLR- 3 expression in diabet ic m ouse m odels decrease on CD4 T cells. Propolis can norm alize
hom eost asis by slight ly increasing t he level of TLR expression. I n general, diabet ic m ice m odels t hat
had received propolis develop int o healt hy signed by t he decline in glucose levels and also physiologic
im provem ents. The rat io of T lym phocyt es seem t o be norm alized after consum ing propolis for t wo
weeks. I nt erest ingly, propolis has abilit y t o increase the precursor of red blood cells.
Ack now ledgem e nts
We t hank Niluh W for careful reading and crit ical com m ents on t his art icle, and Bam bang P, Konit a,
and Yuyun for experim ent al help.
Re fe re n ce s
Arora, S., S. K. Oj ha, dan D. Vohora. 2009. Charact erizat ion of St rept ozot ocin I nduced Diabet es Mellit u s I n Swiss
Albino Mice. Global Journal Of Pharm acology . Vo. 3, No. 2. Pp. 81- 84.
Cet in, E., S. Silici, N. Cet in, dan B.K. Guclu. 2010. Effect s Of Diet s Cont aining Different Con cent rat ion of Propolis
On Hem at ological And I m m unological Variables I n Lay ing Hens. Poult ry Science No. 89. Pp. 1703 - 1708 .
271
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
D. Sawick a, H. Car, M. H. Borawsk a, and J. Nik linsk i, “ The ant icancer act iv it y of propolis,” Folia Hist ochem ica et
Cy t obiologica, v ol. 50, pp. 25–37 , 2012.
Endhart i AT, Rifa'i M, Shi Z, Fuk uok a Y, Nak ahara. 2005. Cu t t ing edge: CD8 + CD122 + regulat ory T cells produce I L10 t o suppress I FN- gam m a produ ct ion and proliferat ion of CD8+ T cells. Journal of im m unology 175 ( 11) ,
7093- 709 7
Ganong, F.G. 2005. Endocrine funct ion s of t he pancreas & regulat ion of carbohy drat e m et abolism . I n: Rev iew of
Medical Phy siology . New York : McGraw- Hill Book Com pany . pp. 336- 355.
Kusum awardani, R. K. 2011. Det ect ion of I GF- 1 in seru m of t ipe- 2 diabet es m ellit us pat ient Biology Dept , Fak ult as
of Sciences, Brawij ay a Univ er sit y .
M Rifa’i, Y Kawam ot o, I Nak ashim a, H Suzuk i. 2004. Essent ial roles of CD8 + CD12 2 + regulat ory T cells in t he
m aint enance of T cell hom eost asis. The Journal of ex perim ent al m edicine 200 ( 9) , 11 23- 113 4
M Rifa’i, Z Shi, SY Zhang, YH Lee, H Shik u, K I sobe, H Suzuk i. 2008. CD8 + CD12 + regulat ory T cells recognize
act iv at ed T cells v ia conv ent ional MHC class I –αβTCR interaction and become I L- 10- pr oducing act iv e
regulat ory cells. I nt ernat ional im m unology 20 ( 7) , 937- 947
M Rifai. 2010. Andrograph olide am eliorat e rheum at oid art hrit is by prom ot ing t he dev elopm ent of regulat ory T cells.
Journal of Tropical Life Science 1 ( 1) , pp. 5- 8
M Rifa’i, N Widodo. 2014. Significance of propolis adm inist r at ion for hom eost asis of CD4 + CD25 + I m m unoregulat ory
T cells cont rolling hy pergly cem ia. SpringerPlus 3 ( 1) , 1- 8
Sy am sudin, Rit a, D. Marlet t a, dan Kusm ardi. 2008. I m m u nom odulat ory and in v iv o Ant iplasm odial Act iv it ies of
Propolis Ex t ract . Global Journal of Pharm acology . Vol. 2, no. 3. pp. 37- 40.
Sforcin, J. M. 2007. Propolis and I m m une Sy st em : A Rev iew . J. Et hnopharm acol. No. 113. pp. 1- 14
Say ed, E. M., O. M. Abo- Salem , H. A. Aly , dan A. M. Mansour. 2009. Pot ent ial Ant idiabet ic And Hypolipidem ic
Effect s Of Propolis Ex t ract I n St rept ozot ocin- I nduced Diabet ic Rat s. Pak . J. Pharm . Sci., Vol.22, No. 2.pp.168174
Sart ori, D.R.S., C. L. Kawak am i, C.L. Orsat t i, J.M., Sfor cin. 2 009. Propolis Effect On St rept ozot ocin- I nduced Diabet ic
Rat s. J Venom Anim Tox in in cl Tr op Dis. Vol. 15 . No. 1.pp.93- 102
YH Lee, Y I shida, M Rifa’i, Z Shi, K I sobe, H Suzuk i. 2008. Essent ial role of CD8 + CD1 22 + regulat ory T cells in t he
recov ery from ex perim ent al aut oim m une encephalom y elit is. The Journal of I m m un ology 180 ( 2) , 825- 832
YH Lee, M Rifa'i. 2011. CD4 + CD2 5 + FOXP3 + Regulat ory T Cells I n Allogeneic Hem at opoiet ic Cell Transplant at ion.
Journal of Tropical Life Science 1 ( 2) , 69- 75
Z Shi, Y Ok uno, M Rifa'i, AT Endhart i, K Ak ane, K I sobe, H Suzuk i . 2009. Hum an CD8 + CXCR3 + T cells hav e t he
sam e funct ion as m urine CD8 + CD122 + Treg
European j ournal of im m unology 39 ( 8) , 21 06- 21 19
Z Shi, M Rifa’i, YH Lee, H Shik u, K I sobe, H Su zuk i. 200 8. I m port an ce of CD8 0/ CD86 –CD2 8 int eract ions in t he
recognit ion of t arget cells by CD8 + CD122 + regulat ory T cells. I m m unology 124 ( 1) , 1 21- 128
272
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Toll- Lik e Re ce pt or 3 - 4 Ex pr e ssion D e cr e a se in BALB/ c
D ia be t ic M ou se M ode ls
1*
Muhaim in Rifa’i, 1 Aris Soewondo, and
1
M. Sasm it o Dj at i
1
Depart m ent of Biology, Facult y of Mat hem at ics and Nat ural Sciences, Brawij aya Universit y,
Malang 65145, I ndonesia
* Corresponding Aut hor: [email protected]
Abst ra ct
I n t his st udy , we observ ed t he effect of propolis ex t ract on im m unological funct ion in diabet ic
m ouse m odels wit h t he aim of highlight ing t he dy nam ics of im m unological st at us in t y pe- 2
diabet es. I n t his st udy we t est ed t he abilit y of propolis t o norm alize hom eost asis. Here we
showed t hat propolis im prov e hom eost asis by slight ly incr easing t he lev el of TLR ex pression.
The result s of t his st udy differ from prev ious findings which report ed t hat TLR ex pression
increased in diabet es m ellit us and m ost t reat m ent is int ended t o suppress t he ex pression of
TLR. I n t his st udy we found t hat TLR- 3 and TLR- 4 ex pression decreased in m ouse m odels of
STZ induced diabet es m ellit us. Furt herm ore, we found t hat adm inist rat ion of propolis showed
an increase of red blood cell precur sor s ( TER- 119) and im pr ov e t he rat io of CD4: CD8 dependent
m anner.
Ke y w ords: Diabet es m ellit us, propolis, im un om odulat or, Toll- lik e recept or
I nt r oduct ion
Current ly about 348 m illion people worldwide suffer from diabetes m ellit us ( DM) . DM is t he m ost
deadly diseases and has penet rated all count ries. I t is current ly known, 4.6% of t he t otal I ndonesian
populat ion suffer from diabet es m ellit us, and this will increase to 7.8% in 2030. During t his t im e, t he
t reat m ent of various diseases including diabet es really dependent on synt het ic drugs. On the ot her
hand, I ndonesia has abundant of nat ural resources wit h t he potent ial t o cure various diseases.
Propolis is one of I ndonesia's pride t radit ional m edicine which is often claim ed by t he com m unit y wit h
abilit y t o cure diabetes m ellit us. But unfortunat ely t here have not enough research been done on t his
exist ing claim s. On t he ot her hand, pat ients with diabet es m ellit us want to get t he right m edicine to
im prove t he qualit y of t heir life. I n general, t he severit y of diabetes is t riggered by inflam m at ion
fact ors known as proinflam m atory cyt okines. Adm inist rat ion of propolis to diabet ic m ice results in a
significant decrease of blood sugar levels and increases t he num ber of naïve T cells. When propolis
was given t o norm al or diabet ic m ice, it can increase t he num ber of regulat ory T cells. I n m ouse
m odels of diabetes m ellit us t he developm ent of effect or cells can be suppressed after propolis
adm inist rat ion. I n individual wit h diabetes m ellit us propolis t ends t o suppress I FN- γ but rather
increased TLR product ion.
M a t e r ia ls a nd M e t hods
Pr ocedur e
Preparat ions of Et hanol Ext ract of Propolis.
Propolis ext ract preaparat ion consisted of t hree phases including drying, ext ract ing, and evaporat ing.
The drying process began by washing the sam ple, cut t ing it int o sm all pieces, and put t ing t hem in t he
oven with 40- 60º C degree. Sam ples were dried and crushed by a blender. As m uch as 100 gram s of
dry sam ples were weighed and put in 1 L Erlenm eyer glass, soaked wit h ethanol t o the volum e of 1 L
before t he ext ract ion process. Sam ple in et hanol was st irred for ± 30 m inutes and allowed t o st and
overnight t o sett le. Supernatant was rem oved and filt ered. The ext ract of propolis was dried by using
an evaporat or and weighed. Propolis powder was t hen dissolved in dist illed wat er for oral t reatm ent .
Type 2 diabet es m ice m odels were t hen t reated wit h t he et hanol ext ract of propolis for 7 weeks. Mice
were m aintained in pat hogen free facilit y. Mice were fed wit h pellets BR1 by 5- 10 gram s per day and
drink was provided ad libit um .
I nduct ion of Type 2 diabetes m ellit us wit h STZ.
St rept ozot onin ( STZ) dissolved in 0: 01 M cit rate buffer, pH 4.5 and always prepared in a fresh
condit ion. STZ was int raperit oneally inj ected t o m ice in t he age of 5 days ( 100 m g/ kg BW). Five weeks
aft er STZ inject ion, diabet ic condit ion is confirm ed by m easuring t he glucose levels using a glucom eter
( GlucoCare Ult im a) .
270
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Results a nd D iscussion
I n t his study we showed t hat TLR- 3 expression in diabet ic m ice decreased 6 t im es lower t han norm al.
The suppression of TLR- 3 expression is suspect ed to affect t he qualit y of hom eost asis because TLR- 3
norm ally funct ions as an act ivat or of innate im m unit y. The low expression of TLR- 3 in t urn m ake
difficult y for elim inat ing infect ious agents in pat ient wit h diabetes m ellit us. So t hat in m any cases
people wit h diabet es m ellit us oft en wounds and difficult t o be cured. However, t he increase of TLR- 3
expression far beyond norm al will bring bad im pacts and also aggravat e diabet ic pat ient healt h
condit ions.
Figur e 1 . I n m ice m odels of diabetes m ellit us induced by STZ, expression level of TLR- 3 decreased
Left panel showed TLR- 3 expression on CD4 cells in norm al m ice. Right panel is an expression of TLR3 on CD4 T cells in m ouse m odels of DM. Cells were isolated from spleen and analyzed by flow
cyt om et ry. CD4 posit ive cells expressing TLR- 3 ( CD4 + TLR- 3 + ) were shown in each panel.
Observat ions on CD4 T cells clearly showed t hat the expression of TLR- 3 decreased in m ouse m odel of
diabetes. I n t his experim ent we cannot explain t he sit e effect of TLR decline on T lym phocyt es.
However, t here is a lit t le clue that t he decline of TLR expression m ay correlate wit h t he failure of
pat ient in elim inat ing foreign ant igen specially bact eria and virus. TLR- 3 has been known to be
associat ed with t he em ergence of signal t ransduct ion related t o virus elim inat ion. The decline of TLR- 3
expression in DM will lead t o t he em ergence of disorders associated wit h TLR- 3 funct ion.
Figur e 2 . Adm inist rat ion of propolis in m ice m odel of diabet es increase t he expression of TLR- 3 on
CD4 cells approaching norm al condit ions
From left panel to right is propolis t reat m ent at a dose of 50, 100, and 200 m g/ kg BW recpect ively.
The cells were isolat ed from spleen and analyzed by flow cyt om et ry. CD4 posit ive cells expressing
TLR- 3 ( CD4 + TLR- 3 + ) were shown in each panel.
Conclusions
TLR- 3 expression in diabet ic m ouse m odels decrease on CD4 T cells. Propolis can norm alize
hom eost asis by slight ly increasing t he level of TLR expression. I n general, diabet ic m ice m odels t hat
had received propolis develop int o healt hy signed by t he decline in glucose levels and also physiologic
im provem ents. The rat io of T lym phocyt es seem t o be norm alized after consum ing propolis for t wo
weeks. I nt erest ingly, propolis has abilit y t o increase the precursor of red blood cells.
Ack now ledgem e nts
We t hank Niluh W for careful reading and crit ical com m ents on t his art icle, and Bam bang P, Konit a,
and Yuyun for experim ent al help.
Re fe re n ce s
Arora, S., S. K. Oj ha, dan D. Vohora. 2009. Charact erizat ion of St rept ozot ocin I nduced Diabet es Mellit u s I n Swiss
Albino Mice. Global Journal Of Pharm acology . Vo. 3, No. 2. Pp. 81- 84.
Cet in, E., S. Silici, N. Cet in, dan B.K. Guclu. 2010. Effect s Of Diet s Cont aining Different Con cent rat ion of Propolis
On Hem at ological And I m m unological Variables I n Lay ing Hens. Poult ry Science No. 89. Pp. 1703 - 1708 .
271
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
D. Sawick a, H. Car, M. H. Borawsk a, and J. Nik linsk i, “ The ant icancer act iv it y of propolis,” Folia Hist ochem ica et
Cy t obiologica, v ol. 50, pp. 25–37 , 2012.
Endhart i AT, Rifa'i M, Shi Z, Fuk uok a Y, Nak ahara. 2005. Cu t t ing edge: CD8 + CD122 + regulat ory T cells produce I L10 t o suppress I FN- gam m a produ ct ion and proliferat ion of CD8+ T cells. Journal of im m unology 175 ( 11) ,
7093- 709 7
Ganong, F.G. 2005. Endocrine funct ion s of t he pancreas & regulat ion of carbohy drat e m et abolism . I n: Rev iew of
Medical Phy siology . New York : McGraw- Hill Book Com pany . pp. 336- 355.
Kusum awardani, R. K. 2011. Det ect ion of I GF- 1 in seru m of t ipe- 2 diabet es m ellit us pat ient Biology Dept , Fak ult as
of Sciences, Brawij ay a Univ er sit y .
M Rifa’i, Y Kawam ot o, I Nak ashim a, H Suzuk i. 2004. Essent ial roles of CD8 + CD12 2 + regulat ory T cells in t he
m aint enance of T cell hom eost asis. The Journal of ex perim ent al m edicine 200 ( 9) , 11 23- 113 4
M Rifa’i, Z Shi, SY Zhang, YH Lee, H Shik u, K I sobe, H Suzuk i. 2008. CD8 + CD12 + regulat ory T cells recognize
act iv at ed T cells v ia conv ent ional MHC class I –αβTCR interaction and become I L- 10- pr oducing act iv e
regulat ory cells. I nt ernat ional im m unology 20 ( 7) , 937- 947
M Rifai. 2010. Andrograph olide am eliorat e rheum at oid art hrit is by prom ot ing t he dev elopm ent of regulat ory T cells.
Journal of Tropical Life Science 1 ( 1) , pp. 5- 8
M Rifa’i, N Widodo. 2014. Significance of propolis adm inist r at ion for hom eost asis of CD4 + CD25 + I m m unoregulat ory
T cells cont rolling hy pergly cem ia. SpringerPlus 3 ( 1) , 1- 8
Sy am sudin, Rit a, D. Marlet t a, dan Kusm ardi. 2008. I m m u nom odulat ory and in v iv o Ant iplasm odial Act iv it ies of
Propolis Ex t ract . Global Journal of Pharm acology . Vol. 2, no. 3. pp. 37- 40.
Sforcin, J. M. 2007. Propolis and I m m une Sy st em : A Rev iew . J. Et hnopharm acol. No. 113. pp. 1- 14
Say ed, E. M., O. M. Abo- Salem , H. A. Aly , dan A. M. Mansour. 2009. Pot ent ial Ant idiabet ic And Hypolipidem ic
Effect s Of Propolis Ex t ract I n St rept ozot ocin- I nduced Diabet ic Rat s. Pak . J. Pharm . Sci., Vol.22, No. 2.pp.168174
Sart ori, D.R.S., C. L. Kawak am i, C.L. Orsat t i, J.M., Sfor cin. 2 009. Propolis Effect On St rept ozot ocin- I nduced Diabet ic
Rat s. J Venom Anim Tox in in cl Tr op Dis. Vol. 15 . No. 1.pp.93- 102
YH Lee, Y I shida, M Rifa’i, Z Shi, K I sobe, H Suzuk i. 2008. Essent ial role of CD8 + CD1 22 + regulat ory T cells in t he
recov ery from ex perim ent al aut oim m une encephalom y elit is. The Journal of I m m un ology 180 ( 2) , 825- 832
YH Lee, M Rifa'i. 2011. CD4 + CD2 5 + FOXP3 + Regulat ory T Cells I n Allogeneic Hem at opoiet ic Cell Transplant at ion.
Journal of Tropical Life Science 1 ( 2) , 69- 75
Z Shi, Y Ok uno, M Rifa'i, AT Endhart i, K Ak ane, K I sobe, H Suzuk i . 2009. Hum an CD8 + CXCR3 + T cells hav e t he
sam e funct ion as m urine CD8 + CD122 + Treg
European j ournal of im m unology 39 ( 8) , 21 06- 21 19
Z Shi, M Rifa’i, YH Lee, H Shik u, K I sobe, H Su zuk i. 200 8. I m port an ce of CD8 0/ CD86 –CD2 8 int eract ions in t he
recognit ion of t arget cells by CD8 + CD122 + regulat ory T cells. I m m unology 124 ( 1) , 1 21- 128
272