ARTICLE INFO

Article Type

Original Research

Authors

Abtahi Froushani   S.A. (* )
Esmaeili Gouvarchin Ghaleh   H. (1 )
Mansouri Mothlagh   B. (1 )
Babaei   M. (1 )






(* ) Microbiology Department, Veterinary Faculty, Urmia University, Urmia, Iran
(1 ) Microbiology Department, Veterinary Faculty, Urmia University, Urmia, Iran

Correspondence

Address: Microbiology Department, Veterinary Faculty, Nazlu Pardis, Kilometer 11th of Sarv Road, Urmia, Iran
Phone: +984432770508
Fax: +984432771926
meysamabtahi@hotmail.com

Article History

Received:  December  17, 2013
Accepted:  August 16, 2014
ePublished:  September 23, 2014

BRIEF TEXT


… [1-3] Estrogen (17 beta-Estradiol) is one of the sexual female hormones. Estrogen plays an important role in regulating the homeostasis of bone marrow activity, cardiovascular system, and central nervous system [4]. Through their many biological functions, steroid compounds, such as Estradiol, change expressions of a wide range of genes [1]. Steroid hormones, especially Estrogen, considerably increase in three last months of pregnancy [5, 6]. There are 17 beta-Estradiol receptors in the immune cells including monocytes/macrophages, dendritic cells, and T and B lymphocytes [7, 8].

Steroid hormones powerfully affect all levels of inherent and acquired immune responses [7]. Glucocorticoid hormones have been highly investigated, while the effects of sexual hormones, such as estrogen, on immune system are less likely to be discussed.

The aim of this study was to investigate probable moderating effects of estrogen on cellular and humoral immune responses following challenge with sheep red blood cells in rat model.

This is a case/control experimental study.

6-week age-range male albino rats from animal home of Faculty of Veterinary Medicine of Urmia University (Iran) were studied.

14 rats were studied.

The rats were randomly divided into two groups including control and treatment groups. 0.1ml SRBC 1×109 intra-peritoneal injection was done on the rats of treatment group at the beginning of experiment and one week later. From the beginning of the study, the rats of treatment group received 5mg/kg 17 beta-Estradiol through intra-peritoneal injection for 2 weeks [9]. As previous group, the rats of control group underwent challenges with SRBC anti-gene. From the beginning of immunization, 0.1ml PBS containing 2% DMSO intra-peritoneal injection was done on the rats with one day interval between every two injection. The rats having been anaesthetized 5 days after the last injection, blood was taken from their hearts. 48 hours before blood-taking, 0.1ml SRBC 1×109 was injected to the left feet of the animals. And, simultaneously, 0.1ml PBS was injected to the right feet of the animals. After 48 hours and before blood-taking, foot thickness of the rats was measured, using caliper (Mauser Dial Caliper; Germany) and Cell-mediated Immunity was computed by the following formula [10, 11]. Cell-mediated Response Index= (Swelling of the Right Foot-Swelling of the Left foot)/Swelling of the Right foot To assess inherent immune response, the severity of respiratory burst in phagocytic cells of the spleen was assessed, using NBT method. After blood-taking, spleen of the animal was removed under sterile conditions and weighted. Cell culture results were read at 540nm wave-length, using ELISA device [12, 13]. Assessing proliferation rate of the immune cells through MTT method, color intensity was determined at 490nm wavelength and Stimulation Index was computed by the following formula [12, 13]. Stimulation Index=OD Blank at the Presence of OD Peptide/OD Blank at the Absence of OD Peptide Data was compared using Mann-Whitney U test and SPSS 19 software. … [14-17]

The mean antibody titer (reverse of the last dilution of antibody with the potential to cause hem-agglutination) in control group was 39.67±7.66, while there was a significant increase (426.7±147.81) in treatment group. There was more inflammatory response (right foot swelling percentage) in treatment group (0.53±0.11) than control group (0.24±0.08). Results of NBT Revival Test (light absorption intensity in 540nm wavelength) showed that there was a significant decrease in the capability of respiratory burst of spleen monocytes/macrophages in treatment group (0.64±0.02) than control group (0.96±0.03). Results of MTT test (light absorption intensity in 490nm wavelength) showed that there was an increase in lymphocyte proliferation rate in treatment group (1.24±0.22) than control group (0.69±0.16). There was a significant increase in mean spleen weight of the rats of treatment group (28.62±1.76g) than control group (21.00±1.83g).

Despite the fact that the rats foot swelling increased due to 17 beta-Estradiol, showing macrophage and T cells integration in a classical DTH reaction, respiratory burst capacity of macrophage cells was considerably reduced by 17 beta-Estradiol. Following challenge with tuberculosis, there are faster and more expanded granuloma reactions in castrated female rats than healthy ones. In addition, there is higher death-rate in the castrated female rats than healthy ones [1]. Due to estrogen, there is a significant reduction in pro-inflammatory cytokines production, such as TNF-α, IL1-β, and IL-6, from the neutrophils and macrophages [18, 19]. It seems that the above effects are due to estrogen through messaging inhibition via NFκB [7, 19, 20]. More TGF-ß surface and tissue growth factors are produced by macrophages treated by estrogen [21, 22]. There was a reduction in the respiratory burst capacity in the phagocytic population of mono-nucleus cells of the spleen. There is total reduction in the capacity of the cell immune reactions in the pregnant rats or rats treated by estrogen following challenges with Flu viruses [17, 23]. 17 beta-Estradiol affects Fas/FasL gene expressions. There is a reduction in Fas ligand expression on the surface of T lymphocyte cells, followed by an increase in life duration of the cells as they are in the presence of the Estradiol compounds [20, 24]. There was an increase in the delayed hypersensitivity reaction and proliferation of the spleen lymphocytes. This might due to a reduction in Fas ligand expression and an increase in the life duration of T lymphocytes cells activated by 17 beta-Estradiol. … [25, 26] IgG antibody production and the life duration of the spleen B cells increase due to 17 beta-Estradiol [27]. There was a significant increase in the antibody titer in treatment group than control group. This might due to the effects of 17-beta-Estradiol on B lymphocyte cells and enhancement of humoral immunity.

In the experimental models of tissue-specific auto-immune disease, the effects of 17 beta-Estradiol should be investigated by the method of the present study. Probable polarizing effects of 17 beta-Estradiol on T lymphocytes should be studied.

Non-declared

It seems that 17 beta-Estradiol can be noticed as the moderating compound of immune system.

The researchers appreciate Immunology Laboratory and Animal Home technicians (Faculty of Veterinary Medicine; Urmia University).

Non-declared

Non-declared

Non-declared


CITIATION LINKS

[1]Bini EI, Espinosa DM, Castillo BM, Payan JB, Colucci D, Cruz AF, et al. The influence of sex steroid hormones in the immunopathology of experimental pulmonary tuberculosis. PLoS One. 2014;9(4):e93831.
[2]Eidinger D, Garrett TJ. Studies of the regulatory effects of the sex hormones on antibody formation and stem cell differentiation. J Exp Med. 1972;136(5):1098-116.
[3]Cutolo M, Seriolo B, Villaggio B, Pizzorni C, Craviotto C, Sulli A. Androgens and estrogens modulate the immune and inflammatory responses in rheumatoid arthritis. Ann N Y Acad Sci. 2002;966:131-42.
[4]Socas-Rodríguez B, Hernández-Borges J, Asensio-Ramos M, Herrera-Herrera AV, Palenzuela JA, Rodríguez-Delgado MA. Determination of estrogens in environmental water samples using 1,3-dipentylimidazolium hexafluorophosphate ionic liquid as extraction solvent in dispersive liquid-liquid microextraction. Electrophoresis. 2014;doi: 10.1002/elps.201400024
[5]Bukovsky A, Cekanova M, Caudle MR, Wimalasena J, Foster JS, Henley DC, et al. Expression and localization of estrogen receptor-alpha protein in normal and abnormal term placentae and stimulation of trophoblast differentiation by estradiol. Reprod Biol Endocrinol. 2003;1:13.
[6]Edelstam G, Karlsson C, Westgren M, Löwbeer C, Swahn ML. Human chorionic gonadatropin (hCG) during third trimester pregnancy. Scand J Clin Lab Invest. 2007;67(5):519-25.
[7]Nadkarni S, McArthur S. Oestrogen and immunomodulation: New mechanisms that impact on peripheral and central immunity. Curr Opin Pharmacol. 2013;13(4):576-81.
[8]Bouman A, Heineman MJ, Faas MM. Sex hormones and the immune response in humans. Hum Reprod Update. 2005;11(4):411-23.
[9]Shahabi N, Khaksari M. Effect of 17 beta-estradiol on wound healing in overiectomized rats. Koomesh. 2002;3(1-2):1-10. [Persian]
[10]Zimecki M, Wieczorek Z. Differential patterns of cyclosporine A-induced inhibition of humoral and cellular immune responses to sheep erythrocytes in mice. Pol J Pharmacol. 2001;53(5):495-500.
[11]Hassan ZM, Ebtekar M. Immunological consequence of sulfur mustard exposure. Immunol Lett. 2002;83(3):151-2. [Persian]
[12]Abtahi Froushani SM , Delirezh N, Hobbenaghi R , Mosayebi Gh. Therapeutic effects of all-trans retinoic acid on experimental autoimmune encephalomyelitis and its role in T-helper lymphocyte responses. Tehran Univ Med J. 2012;69(11):710-7.
[13]Abtahi Froushani SM, Delirezh N, Hobbenaghi R, Mosayebi G. Synergistic effects of atorvastatin and all-trans retinoic acid in ameliorating animal model of multiple sclerosis. Immunol Invest. 2014;43(1):54-68.
[14]Haq A, Lobo PI, Al-Tufail M, Rama NR, Al-Sedairy ST. Immunomodulatory effect of Nigella sativa proteins fractionated by ion exchange chromatography. Int J Immunopharmacol. 1999;21(4):283-95.
[15]Jones RE, Kaler L, Murphy S, Offner H. Tissue-dependent expression of estrogen receptor β in 17β-estradiol-mediated attenuation of autoimmune CNS inflammation. Open Autoimmun J. 2010;2:197-204.
[16]Offner H, Polanczyk M. A potential role for estrogen in experimental autoimmune encephalomyelitis and multiple sclerosis. Ann N Y Acad Sci. 2006;1089:343-72.
[17]Escribese MM, Kraus T, Rhee E, Fernandez-Sesma A, López CB, Moran TM. Estrogen inhibits dendritic cell maturation to RNA viruses. Blood. 2008;112(12):4574-84.
[18]Toyoda Y, Miyashita T, Endo S, Tsuneyama K, Fukami T, Nakajima M, Yokoi T. Estradiol and progesterone modulate halothane-induced liver injury in mice. Toxicol Lett. 2011;204(1):17-24.
[19]Murphy AJ, Guyre PM, Pioli PA. Estradiol suppresses NF-kappa B activation through coordinated regulation of let-7a and miR-125b in primary human macrophages. J Immunol. 2010;184(9):5029-37.
[20]Singh NP, Singh UP, Nagarkatti PS, Nagarkatti M. Prenatal exposure of mice to diethylstilbestrol disrupts T-cell differentiation by regulating Fas/Fas ligand expression through estrogen receptor element and nuclear factor-κB motifs. J Pharmacol Exp Ther. 2012;343(2):351-61.
[21]Ashcroft GS, Dodsworth J, van Boxtel E, Tarnuzzer RW, Horan MA, Schultz GS, et al. Estrogen accelerates cutaneous wound healing associated with an increase in TGF-beta1 levels. Nat Med. 1997;3(11):1209-15.
[22]Morales DE, McGowan KA, Grant DS, Maheshwari S, Bhartiya D, Cid MC, et al. Estrogen promotes angiogenic activity in human umbilical vein endothelial cells in vitro and in a murine model. Circulation. 1995;91(3):755-63.
[23]Pazos MA, Kraus TA, Muñoz-Fontela C, Moran TM. Estrogen mediates innate and adaptive immune alterations to influenza infection in pregnant mice. PLoS One. 2012;7(7):e40502.
[24]Zhang J, Chen X, Zhang S, Zhou G, Xia X, Lu L. Effects of transdermal estrogen therapy on expressions of estrogen receptors and T-lymphocyte apoptosis in surgically menopausal women. Cell Mol Immunol. 2009;6(4):277-83.
[25]Matalka KZ. The effect of estradiol, but not progesterone, on the production of cytokines in stimulated whole blood, is concentration-dependent. Neuro Endocrinol Lett. 2003;24(3-4):185-91.
[26]Xiong YH, Yuan Z, He L. Effects of estrogen on CD4(+) CD25(+) regulatory T cell in peripheral blood during pregnancy. Asian Pac J Trop Med. 2013;6(9):748-52.
[27]Fu Y, Li L, Liu X, Ma C, Zhang J, Jiao Y, You L, Chen ZJ, Zhao Y. Estrogen promotes B cell activation in vitro through down-regulating CD80 molecule expression. Gynecol Endocrinol. 2011;27(8)593-6.