中文摘要
去氫表雄固酮(DHEA)為腎上腺皮質網狀層分泌的主要雄性激素之一，在血清中則大部份以硫化物(DHEAS)存在，參與哺乳類體內多種生理反應。近年來的文獻指出，DHEA具有調節免疫反應的功能，並發現DHEA可作為老年人或年老小白鼠注射疫苗時的佐劑。然而，DHEA是否可增強幼年期動物之專一性抗體反應仍有待研究，且注射DHEA後，相關細胞激素的變化亦不明確。因此本實驗目的為研究DHEA在幼年小白鼠接受大腸桿菌(E.coli O78:H11)、三合一疫苗(DPT)或百日咳毒素(PT)的免疫注射時，是否可增強其專一性抗體反應，並進一步探討其與細胞激素的關係。實驗結果顯示，DHEA對於大腸桿菌或百日咳毒素引起之專一性抗體反應均可增強抗體反應；然而抗三合一疫苗抗體實驗中，顯示DHEA對三合一疫苗不具佐劑效應。感染過E.coli或以PT致敏的老鼠施打DHEA後，DHEA對E.coli或PT激發脾臟淋巴球製造IFN-g、IL-2與IL-4、IL-10等細胞激素的作用皆有加乘效應，可見DHEA的免疫增強功能可能是透過對細胞激素的調節。本研究不但證實DHEA可促進專一性抗體反應，且發現此免疫調節功能與細胞激素有關。

Abstract
Dehydroepiandrosterone (DHEA) is one of the major androgens secreted by adrenal cortex and play a multifunctional role in regulating physiological system in mammals. DHEA is predominantly converted to DHEA-sulfate (DHEAS) in serum . Recent reports demonstrated that DHEA is a potent immunomodulator. In addition, recent data showed that DHEA might act as an effective vaccine adjuvant in aged human and mice. However, the immunostimulatory effect on the children and young animal upon immunization remain to be studied. Furthermore, the profile of cytokine associated with DHEA-mediated regulatory effect remained to be defined. The major purpose of this study is to investigate the possible immunostimulatory effect of DHEA on the young mice immunized with Escherichia coli (E.coli O78:H11), combined diphtheria, tetanus, acellular pertussis vaccine (DPT vaccine) or pertussis toxin (PT), and to investigate the profile of cytokine associated with DHEA administration. Results indicated that DHEA has an augmented effect on the young mice immunized with Escherichia coli (E.coli O78:H11) or pertussis toxin (PT). However, DHEA did not show adjuvant effect on DPT vaccine immunized mice. In the subsequent study, the mice were infected with E. coli or challenged with PT, followed by subcutaneous injection with DHEA. Result indicated that DHEA synergistically enhanced E. coli or PT induced IFN-g, IL-2 and IL-4, IL-10 cytokine productions. Therefore, the immunoregulatory effect of DHEA might closely relate with its ability to modulate cytokine production. This study not only confirmed the ability of DHEA to regulate specific antibody response, but also demonstrated the role of DHEA in regulation of cytokine production.

參考文獻
Al-Janadi M., Al-Ball S., Al-Dalaan S., Rsziuddin S. (1993) Cytokine profile in systematic lupus erythromatosus, rheumatoid arthritis and other rheumatic disease .J.Clin.Immunol. 13 : 58-67.
Araghi-Niknam M., Zhang Z., Jiang S., Call O., Eskelson CD., Watson RR. (1997) Cytokine dysregulation and increased oxidation is prevented by dehydroepiandrosterone in mice infected with murine leukemia retrovirus. Exp. Biol. Med. 216 : 386-391.
Araneo BA., Dowell T., Woods ML., Daynes RA., Ajudd M., Evans T. (l995) DHEAS as an effective vaccine adjuvant in elderly humans. Ann. NY Acad. Sci. 774 : 232-248.
Araneo BA., Woods ML., Daynes RA. (1993) Reversal of the immunosenescent phenotype by dehydroepiandrosterone : hormane treatment provides an adjuvant effect on the immunization of aged mice with recombinant hepatitis B surface antigen. J. Infect. Dis.167 : 830-840.
Ben-Nathan D., Lachmi B., Lustig S., Feuerstein G. (1991) Protection by dehydroepiandrosterone in mice infected with viral encephalitis. Arch. Virol. 120 : 263-271.
Ben-Nathan D., Lustig S., Kobiler D., Danenberg HD., Luou E., Feuerstein G. (1992) Dehydroepiandrosterone protects mice inoculated with West Nile virus and exposed to cold stress. J. Med. Virol. 38 : 159-166.
Blauer KL., Poth M., Rogers WM., Bernton EW. (1991) Dehydroepiandrosterone antagonizes the suppressive effects of dexamethasone on lymphocyte proliferation. Endocrinol. 129 : 3174-3179.
Carr DJ. (1998) Increased levels of IFN-gamma in the trigeminal ganglion correlate with protection against HSV-1-induced encephalitis following subcutaneous administration with androstenediol. J. Neurol. 89 : 160-167.
Cheng GF., Tseng J. (2000) Regulation of murine interleukin-10 producyion by dehydroepiandrosterone. J. Interferon Cytokine Res. 20 : 471-478.
Chopra U., Vohra H., Chhibber S., Ganguly NK., Sharma S. (1997) TH1 pattern of cytokine secretion by splenic cells from pyelonephritic mice after in-vitro stimulation with hap-65 of Escherichia. J. Med. Microbiol. 46 : 139-144.
Christal CB., John DC. (2001) Differential biological and adjuvant activities of cholera toxin and Escherichia coli heat-labile enterotoxin hybrids. Infect. Immun. 69 : 1528-1535.
Christina H., Bengt A., Agnes EW. (2000) Gram-positive bacteria are potent inducers of monocytic Interleukin-12 (IL-12) while Gram-negative bacteria preferentially stimulate IL-10 production. Infect. Immun. 68 : 3581-3586.
Coleman DL. (1988) Therapeutic effects of dehydroepiandrosterone (DHEA) and its metabolites in obesehyperglycemic mutant mice. Prog. Clin. Biol. Res. 265 : 161-175
Danel L., Souweine G., Monier JC., Saze S. (1983) Specific estrogen binding sites in human lymphoid cells and thymic cells. J. Steroid Biochem. 18 : 559-563.
Danenberg HD., Ben-Yehuda A., Zakay-Rones Z., Friedman G. (1995) Dehydroepiandrosterone enhances influenza immunization in aged mice. Ann. NY Acad. Sci. 774 : 297-299.
Danenberg HD., Alpert G., Lustig S., Ben-Nathan D. (1992) Dehydroepiandrosterone protects mice from endotoxin toxicity and reduces tumor necrosis factor production. Microbiol. Agent chemother. 36 : 2275-2279.
Daynes RA., Araneo BA.（1989）Contrasting effects of glucocorticoids on the capatity of T cells to produce the growth factors interleukin-2 and interleukin-4. Eur. J. Immunol. 19:2319-2325.
Daynes RA., Dudiay DJ., Araneo BA.（1990）Regulation of murine lymphokine production in vivo ：II. Dehydroepiandrosterone is a natural enhancer of interleukin 2 synthesis by helper T cells. Eur. J. Immunol. 20：793-802.
Delpedro AD., Barjavel MJ., Mamdouh Z., Bakouch O. (1998) Activation of human Monocytes by LPS and DHEA. J. Interferon. Cytokine. Res. 18 : 125-135.
Edinger M., Bessler WG., Kleine B. (1994) Synergistic stimulation of human B lymphocytes by anti-CD40 monoclonal antibodies and synthetic lipopeptide analogues from Escherichia coli lipoprotein. Immunol. 81 : 205-210.
Fischer JE., Johnson TR., Peebles RS., Graham BS. (1999) Vaccination with pertussis toxin alters the antibody response to simultaneous respiratory syncytial virus challenge. J. Infect. Dis. 180 : 714-719.
Garg M., Bondada S. (1993) Reversal of age-associated decline in immune response to Pnu-immune vaccine by supplementation with the steroid hormone dehydroepiandrosterone. Infect. Immun. 61 : 2238-2241.
Girouard L., Mandrekar P., Catalano D., Szabo G. (1998) Regulation of monocyte interleukin-12 production by acute alcohol : a role for inhibition by interleukin-10. Alcohol Clin. Exp. Res. 22 : 211-216.
Gordon GB., Bush DE., Weisman HF. (1988) Reduction of atherosclerosis by administration of dehydroepiandrosterone. J. Clin. Invest. 36 : 712-720.
Hadely ME. (1988) Adrenal steroids: In endocrinology prentice-hall international. Inc. New Jersey. 349-383.
Harlow E., Lane D. (1988) Making the assay quantitative. Antibodies. 566.
Hessle C., Hanson LA., Wold AE. (2000) Interleukin-10 produced by the innate immune system masks in vitro evidence of acquired T-cell immunity to E.coli. Scan. J. Immunol. 52 : 13-20.
Hidvegi T., Feher GK., Heher T., Koo E., Fust G. (1984). Inhibition of complement activation by an adrenal androgen:Dehydroepiandrosterone. Complement. 1 : 201-206.
Inserra P., Zhang Z., Ardfstani SK., Araghi-Niknam M., Liang B., Jiang S., Shaw D., Molitor M., Elliott K., Watson RR. (1998) Modulation of cytokine production by dehydroepiandrosterone (DHEA) plus melatonin (MLT) supplementation of old mice. Exp. Biol. Med. 218 : 76-82.
Jacobson MA., Fusaro RE., Galmarini M., Lang W.（1991）Decreased serum dehydroepisterone is associated with an increases progression of human immunodeficiency virus infection in men with CD4 counts of 200-499. J. Inf. Dis. 164 : 864-868.
Kim HR., Ryu SYM., Kim HS., Choi BM., Lee EJ., Kim HM., Chung HT. (1995) Administration of dehydroepiandrosterone reverses the immune suppression induced by high dose antigen in mice. Immunol. Invest. 24 : 583-593.
Klapproth JM., Donnenberg MS., Abraham JM., James SP. (1996) Products of enteropathogenic E. coli inhibit lymphokine production by gastrointestinal lymphocytes. Amer. J. Physiol. 271 : G841-G848.
Kosecka U., Marshall JS., Crowe SE., Bienenstock J., Perdue MH. (1994) Pertussis toxin stimulates hypersensitivity and enhances nerve-mediated antigen uptake in rat intestine. Amer. J. Physiol. 267 : G745-G753.
Longcope C. (1996) Dehydroepiandrosterone metabolism. J. Endocrinol. 150 : S125-S127
Loria RM., Padgett DA., Huynh PN. (1996) Regulation of the immune response by dehydroepiandrosterone and its metabolites. J. Endocrinol. 150 Suppl : S209-S220.
Luciene ML., Asher M., Gordon D., Benjamin MC. (2000) Inhibition of T-cell response by Escherichia coli heat-labile enterotoxin-treated epithelial cells. Infect. Immun. 68 : 6891-6895.
MacEwen EG., Maki H., Kurzman ID. (1990) Dehydroepiandrosterone effects on cholesterol and lipoprotein. In the Biological role of dehydroepiandrosterone (DHEA). M.Kalimi and W.Regelson, Eds.pp299-315. Walter de Gruyter Berlin. New York.
Magistris MD., Romano M., Bartoloni A., Rappuoli R., Tagliabue A. (1989) Human T cell clones define S1 subunit as the most immunogenic moiety of pertussis toxin and determine its epitope map. J. Exp. Med. 169 : 1519-1532.
May M., Holmes E., Rogers W., Poth M. (1990) Protection from glucocorticoid induced thymic involution by dehydroepiandrosterone. Life Sci. 46 : 1627-1631.
McLachlan JA., Srekin CD., Bakouche O. （1996）Dehydroepiandrosterone modulation of lipopolysaccharide-stimulated monocyte cytotoxicity. J. Immunol. 156 : 328-335.
Meikle AW., Dorchuck RW., Araneo BA., Stringham JD., Evans TG., Spruance SL., Daynes RA.（1992）The presence of a dehydroepiandrosterone-specific receptor binding complex in murine T cells. J. Steroid Biochem. Mol. Biol. 42 : 293-304.
Mendenhall CL., Finkelman F., Means RT Jr., Sherman KE., Nguyen VT., Grossman CE., Morris SC., Rouster S., Roselle GA. (1999) Cytokine response to BCG infection in alcohol-fed mice. Alcohol. 19 : 57-63.
Moynihan JA., Callahan TA., Kelley SP., Campbell LM. (1998) Adrenal hormone modulation of type 1 and type 2 cytokine production by spleen cells: dexamethasone and dehydroepiandrosterone suppress interleukin-2, interleukin-4 and interferon-g production in vitro. Cell Immunol. 184 : 58-64.
Mu HH., Sewell WA. (1993) Enhancement of interleukin-4 production by pertussis toxin. Infect. Immun. 61 : 2834-2840.
Na HR., Zhu X., Strwart GL., Seelig LL Jr. (1997) Ethanol consumption suppresses cell-mediated inflammatory responses and increases T-helper type 2 cytokine secretion in Trichinella spiralis-infected rats. Alcohol Clin. Exp. Res. 21 : 1179-1185.
Nestler JE., Clore JN., Blackard WG. (1992) Dehydroepiandrosterone : the missing link between hyperinsulinemia and atherosclerosis ? FASEB 6 : 3073-3075
Ockenfels HM., Keim-Maas C., Nussbaum G., Goos M. (1996) Ethanol enhances the IFN-gamma,TGF-alpha and IL-6 secretion in psoriatic co-cultures. Bri. J. Dermatol. 135 : 746-751.
Orskov F., Orskov I. (1992) Escherichia coli serotyping and disease in man and animals. Canad. J. Microbiol. 38 : 699-704.
Padgett DA., Loria RM. (1994) In vitro potentiation of lymphocyte activation by dehydroepiandrostrone, androstenediol, and androstenetriol. J. Immunol. 153 : 1544-1552.
Padgett DA., Sheridan JF., Loria RM. (1995) Steroid hormone regulation of a polyclonal Th2 immune response. Ann. NY Acad. Sci. 774 : 323-325.
Parker LN. (1989) Adrenal androgens in clinical medicine. 118-184. Academic press. San Diego.
Rasmussen KR., Martin EG., Healey MC. (1993) Effects of DHEA in immunosuppressed rats infected with Cryptosporidium parvum. J. Parasitol. 79 : 364-370.
Risdon G., Kumar V., Bennett M. (1991) Differential effects of dehydroepiandrosterone (DHEA) on murine lymphopoiesis and myelopoiesis. Exp. Hematol. 19 : 128-31.
Robel P., Bauliue EE. (1995) Dehydroepiandrosterone (DHEA) is a neuroactive neurosteroid. Ann. NY Acad. Sci. 774 : 82-110.
Rom WN., Harkin T. (1991). Dehydroepiandrosterone inhibits the spontaneous release of superoxide radical by alveolar macrophage in vitro in asbestosis. Environ. Res. 55 : 145-156.
Rowe J., Macaubas C., Monger TM., Holt BJ., Harvey J., Poolman JT., Sly PD., Holt PG. (2000) Antigen-specific responses to diphtheria-tetanus-acellular pertussis vaccine in human infants are initially Th2 polarized. Infect. Immun. 68 : 3873-3877.
Ryan M., McCarthy L., Rappuoli R., Mahon BP., Mills KHG. (1998) Pertussis toxin potentiates Th1 and Th2 responses to co-injected antigen: adjuvant action is associated with enhanced regulatory cytokine production and expression of the co-stimulatory molecules B7-1, B7-2 and CD28. Intl. Immunol. 10 : 651-662.
Sewell WA., De Moerloose PA., Hamilton JA., Schrader JW., Mackay IR., Vadas MA. (1987) Potentiation of delayed-type hypersensitivity by pertussigen or cyclophosphamide with release of different lymphokines. Immunol. 61:483
Starkenberg S., Waltenbaugh C. (2000) Alteration of cytokine levels and specific immune responses in alcohol consuming mice. Eur. Cytokine Netw. 11 Special issue ICS/ISICR:17.
Suzuki T., Suzuki N., Daynes RA., Engleman EG. (1991) Dehydroepiandrosterone enhances IL2 production and cytotoxic effector function of human T cells. Clin. Immunol. Immunopathol. 61 : 202-211.
Suzuki T., Suzuki N., Engelman EG., Mizushima Y., Sakane T. (1995). Low serum level of dehydroepiandrosterone production by lymphocytes in patients with systemic lupus erythematosus (SLE). Clin. Exp. Immunol.,99 : 251-255.
Tamura M., Nogimori K., Yajima M., Ase K., Ui M. (1983) A role of the B-oligomer moiety of islet-activating protein, pertussis toxin, in development of the biological effects on intact cells. J. Biol. Chem. 258 : 6756-6761.
Tseng J., Liou CJ. (1994) Dehydroepiandrosterone regulates immunoglobulin A production of murine B lymphocytes. Int. J. Immunotherapy. 10 : 95-101.
Van Vollenhoven RF., Engleman EG., McGuire JL. (1995) Dehydroepiandrosterone in systemic lupus erythematosus. Arthritis. Rheum. 38 : 1826-1831.
Waltenbaugh C., Starkenberg S., Munroe M. (2000) Alcohol consumption up-regulates murine Th2 cytokines and IgE levels. Eur. Cytokine Netw.11 Special issue ICS/ISICR:25.
Wang Y., Huang DS., Giger PT., Watson RR. (1994) Influence of chronic dietary ethanol on cytokine production by murine splenocytes and thymocytes. Alcohol. Clin. Exp. Res.18 : 64-70.
Weksler ME. (1993) Immune senescence and adrenal steroids:immune dysregulation and the action of dehydroepiandrosterone (DEHA) in old animals. Eur. J. Clin. Pharmacol. 45 suppl 1 : S21-S23 ; discussion S43-S44.
Wiedmeier SE., Araneo BA., Huang K., Daynes RA. (1991) Thymic modulation of IL-2 and IL-4 synthesis by peripheral T cells. Cell. Immunol. 135 : 501-518.
Wu M-F., H-L Chang and Tseng, J. (1998). Dehydroepiandrosterone induces the transforming growth factor b production by murine macrophages. Int. J. Tiss. Reac. XIX : 141
Yang BC., Lin CW., Cnen YC., Yu CK. (1998) Dehydroepiandrosterone suppress interleukin-2 , interleukin-4, and interferon-gamma production in vitro. Cell. Immunol. 184 : 58-64.
Yu CK., Yang BC., Lei HY., Chen YC., Liu YH., Chen CC., Liu CW. (1999) Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response. Clin. Exp. Allergy. 29 : 414-422.
Zepp F., Knuf M., Habermehl P., Schmitt HJ., Meyer C., Clemens R., Slaoui M. (1997) Cell-mediated immunity after pertussis vaccination and after natural infection. Dev. Biol. Standard. 89 : 307-314.
財團法人食品工業發展研究所 (1994) Escherichia coli serotype O78:H11.菌種目錄增訂版 Ⅱ: Bacteria 7.