O papel modulador do gene AIRE - capes

More documents

Recommendations

Info

227coincided (CBA/J) with expression of the RAG-1 and RAG-2 genes.In agreement with previous observations [17, 18] we observedthat the overall level of RAG-2 expression was lowerthan that of RAG-1 in Balb-c and C57Bl/6 mice, whereas theinverse situation was observed in CBA/J mice.Germline transcripts of TCRVβ were detected before theTCR β locus rearrangements [1, 19–22] and our results withthe CBA/J strain agree these observations. However, TCRVβwas one among the lesser transcribed genes.MHC-I and MHC-II displayed different expression patternsamong strains and there was a suggestion that theircontribution to the termination of TCR rearrangements isunlikely [23].The clustering of IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-6, IFγand IL-2Rβ genes diverged among strains; however, weobserved a common tendency of their expression after (Balbc)and before RAG-1 and RAG-2 genes (C57Bl/6 and CBA/J strains).We were able to quantify the expression levels of all thesegenes implicated in T-cell proliferation, V(D)J recombinationor T-cell clonal deletion, such as MMTV(SW) retroviralsuperantigen, during thymus ontogeny (14–17 days p.c.) innewborn pups and adults. The onset of the TCRVβ8.1-Dβ2.1rearrangement in fetal thymus was correlated with a previousexpression of germline TCRVβ8.1 in the CBA/J strain(15–16 days p.c.). The most evident feature demonstrated wasthe kinetics of MMTV(SW) mRNA overexpression and parallelreduction of the TCRVβ8.1-Dβ2.1 rearrangement in the17 days p.c. thymus DNA of the of the CBA/J strain (Fig. 1),suggesting a susceptibility of this strain to the expression ofMMTV(SW) superantigen in combination with its geneticbackground, including MHC H-2 k , upon reduction of T-cellclones bearing the rearranged Vβ8.1 DNA segment, whilecoinciding with overexpression of IL-7 gene that plays a rolein T-cell proliferation.Expression of superantigen proteins from endogenousMMTV proviruses carried by mouse germline usually resultsin the deletion of self-reactive T-cells during negative selectionin the thymus and the elimination of T-cells required forinfection by specific milk-borne MMTVs [24, 25].Our data contribute to establishing a direct correlationbetween the different expression profiling, including cytokines,and backgrounds of the strains with evolution of TCRVβ8.1-Dβ2.1 rearrangements, of particular interest for characterizationof CBA/J mice, since the expression of these genes arenot typically studied in this strain.However, due to the complexity of this phenomenon involvingthe participation of a range of known and unknowngenes, our model-system with inbred mouse strains openperspectives for further studies on large-scale gene expressionprofiling using DNA-arrays.AcknowledgementsA.R.E., C.M.J. and R.S.C. are predoctoral fellows supportedby FAPESP (00/01705-8, 96/07847-1 and 00/09994-9, respectively),C.M. is a fellow from the Ludwig Institute forCancer Research (São Paulo) in the Human Cancer TranscriptSequencing Project. We thank the INSERM-FAPESP agreementwhich has permitted cooperation between the Brazilianand French groups. G.A.S.P. received grants from FAPESP(98/09789-4, 98/05584-9, 99/12135-9 and 00/12495-4).References1. Junta CM, Passos GAS: Emergence of TCRα/β V(D)J recombinationand transcription during ontogeny of inbred mouse strains. Mol CellBiochem 187: 67–72, 19982. Macedo C, Junta CM, Passos GAS: Onset of T-cell receptor Vβ8.1 andDβ2.1 V(D)J recombination during thymus development of inbredmouse strains. Immunol Lett 69: 371–373, 19993. Muegge K,Vila MP, Durum SK: Interleukin-7: A cofactor for V(D)Jrearrangement of the T-cell receptor gene. Science 261: 93–95, 19934. Sollof RS, Wang TG, Dempsey D, Jennings SR, Wolcott RM, ChervenakR: Interleukin 7 induces TCR gene rearrangement in adult marrowresidentmurine precursor T cells. Mol Immunol 34: 453–462, 19975. Candeias S, Muegge K, Durum SK: IL-7 receptor and V(D)J recombination:Trophic vs. mechanistic actions. Immunity 6: 501–508, 19976. Haks MC, Oosterwegel MA, Blom B, Spits H, Kruisbeek AM: Cellfatedecisions in early T-cell development regulation by cytokinereceptors and pre-TCR. Sem Immunol 11: 23–37, 19997. Di Santo JP, Rodewald HR: In vivo roles of receptor tyrosine kinasesand cytokine receptors in early thymocyte development. Current OpinImmunol 10: 196-207, 19988. Laky K, Lefrançois L, von Freeden-Jeffry U, Murray R, PuddingtonL: The role of IL-7 in thymic and extrathymic development of TCRgamma-delta cells. J Immunol 161: 707–713, 19989. Schlissel MS, Durum SD, Muegge K: The interleukin 7 receptor isrequired for T-cell receptor gamma locus accessibility to V(D)J recombinase.J Exp Med 191: 1045–1050, 200010. Weissman IL: Developmental switches in the immune system. Cell 76:207–218, 199411. Mombaerts P, Iacomini J, Johnson RS, Herrup K, Tonegawa S, PapaioannouVE: RAG-1 deficient mice have no mature B and T lymphocytes.Cell 68: 869–877, 199212. Hempel WM, Stanhope-Baker P, Mathieu N, Huang F, Schlissel MS,Ferrier P: Enhancer control of V(D)J recombination at the TCR betalocus: Differential effects on DNA cleavage and joining. Genes Dev12: 2305–2317, 199813. Eisen MB, Spellman PT, Brown PO, Botstein D: Cluster analysis anddisplay of genome-wide expression patterns. Proc Natl Acad Sci USA95: 14863–14868, 199814. Eisen M: Gene cluster treeview. Stanford University (http://genomewww.stanford.edu/serum),200115. Rugh R: The Mouse: Its Reproduction and Development. Burgess,Minneapolis, 196816. Rothenberg EV, Telfer JC, Anderson MK: Transcriptional regulationof lymphocyte lineage commitment. BioEssays 21: 726–742, 199917. Wilson A, Held W, MacDonald HR: Two waves of recombinase expressionin developing thymocytes. J Exp Med 179: 1335–1360, 1994
22818. Turka LA, Schatz DG, Clettinger MA, Chun JJM, Gorka C, Lee K, McCormack WT, Thompson CB: Thymocyte expression of RAG-1 andRAG-2: Termination by T-cell receptor cross-linking. Science 253:778–781, 199119. Mallick CA, Dudley EC, Viney JL, Owen MJ, Hayday AC: Rearrangementand diversity of T-cell receptor beta chain genes in thymocytes:A critical role for the beta chain. Cell 73: 513–519, 199320. Godfrey DI, Kennedy J, Mombaerts P, Tonegawa S, Zlotnik A: Onsetof TCRβ gene rearrangement and role of TCRβ expression duringCD3-, CD4-, CD8- thymocyte differentiation. J Immunol 152: 4783–4791, 199421. Chung S, Strominger JL: Regulation of T-cell antigen receptor (TCR)α-chain expression by TCRβ chain transcripts. Proc Natl Acad Sci USA92: 3712–3716, 199522. Guidos CJ, Williams CJ, Wu GE, Paige CJ, Danska JS: Development ofCD4+ CD8+ thymocytes in RAG-deficient mice through a T-cell receptorβ chain-independent pathway. J Exp Med 181: 1187–1195, 199523. Robey E, Fowlkes BJ: Selective events in T-cell development. Ann RevImmunol 12: 675–705, 199424. Acha-Orbea H, Shakhov AN, Scarpellino L, Kolb E, Muller V, VessazshawA, Fuchs R, Blochlinger K, Rollini P, Billotte J, Sarafidou M, MacDonaldHR, H. Diggelmann H: Clonal deletion of V-beta 14-bearing T-cellsin mice transgenic for mammary tumour virus. Nature 350: 207–211,199125. Barnett A, Mustafa F, Wrona TJ, Lozano M, Dudley JP: Expression ofmouse mammary tumor virus superantigen mRNA in the thymus correlateswith kinetics of self-reactive T-cell loss. J Virology 73: 6634–6645, 199926. Montgomery RA, Dallman MJ: Analysis of cytokine gene expressionduring fetal thymic ontogeny using polymerase chain reaction.J Immunol 147: 554–560, 199127. Hikida M, Mori M, Kawabata T, Takai T, Ohmori H: Characterizationof B-cells expressing recombination activating genes in germinalcenters of immunized mouse lymph nodes. J Immunol 158:2509–2512, 199728. Le Bon A, Desaymard C, Papiernik M: Neonatal impaired response toviral superantigen encoded by MMTV(SW) and Mtv-7. Int Immunol7: 1897–1903, 1995
Page 1 and 2:
Universidade de São PauloFaculdade
Page 3 and 4:
Macedo, ClaudiaO papel modulador do
Page 5 and 6:
Dedico Especialmente este TrabalhoM
Page 7 and 8:
AgradecimentosAgradeço do fundo do
Page 9 and 10:
ÍndiceRESUMO .....................
Page 11 and 12:
INDICE DE FIGURASFigura 1. Desenvol
Page 15 and 16:
Promiscuous gene expression in medu
Page 17 and 18:
INTRODUÇÃO1. INTRODUÇÃO1.1. A m
Page 19 and 20:
INTRODUÇÃORecentemente surgiram e
Page 21:
INTRODUÇÃOmigram para a região s
Page 24 and 25:
INTRODUÇÃOCD4 + CD8 + , e Ccl21 (
Page 26 and 27:
INTRODUÇÃOos timócitos duplo-pos
Page 28 and 29:
INTRODUÇÃOgrande diversidade nas
Page 30 and 31:
INTRODUÇÃOAs células mTECs são
Page 32 and 33:
INTRODUÇÃODerbinski (2004). Os ti
Page 34 and 35:
INTRODUÇÃOinvertebrados multicelu
Page 36 and 37:
INTRODUÇÃOCoraçãoLínguaEstôma
Page 38 and 39:
INTRODUÇÃO1.3. O papel do gene Au
Page 40 and 41:
INTRODUÇÃOdefinido pela atividade
Page 42 and 43:
INTRODUÇÃOpara abordarmos nossa p
Page 44 and 45:
INTRODUÇÃOesses modelos levam em
Page 46 and 47:
Hipóteses
Page 48 and 49:
Objetivos
Page 50 and 51:
Material e Métodos
Page 52 and 53:
MATERIAL E MÉTODOSFigura 6. Padrã
Page 54 and 55:
MATERIAL E MÉTODOScom estes RNAs f
Page 56 and 57:
MATERIAL E MÉTODOSdos complexos de
Page 58 and 59:
MATERIAL E MÉTODOSAo RNA total rec
Page 60 and 61:
4.5. PCRs semi-quantitativas para o
Page 62 and 63:
MATERIAL E MÉTODOSminutos a 4 °C.
Page 64 and 65:
MATERIAL E MÉTODOS(Merck), soluç
Page 66 and 67:
MATERIAL E MÉTODOSEnsaios de co-hi
Page 68 and 69:
MATERIAL E MÉTODOSRNA TotalANELAME
Page 70 and 71:
MATERIAL E MÉTODOSa) Preparação
Page 72 and 73:
MATERIAL E MÉTODOScentrifugação
Page 74 and 75:
MATERIAL E MÉTODOScanais (Cy3 e Cy
Page 76 and 77:
MATERIAL E MÉTODOSSpotfinderQuanti
Page 78 and 79:
MATERIAL E MÉTODOSOnde Xp(i) e Xc(
Page 80 and 81:
MATERIAL E MÉTODOSO programa SAM p
Page 82 and 83:
MATERIAL E MÉTODOSextrair automati
Page 84 and 85:
DELINEAMENTO EXPERIMENTAL5. DELINEA
Page 86 and 87:
RESULTADOS6. RESULTADOS6.1. Inibiç
Page 88 and 89:
RESULTADOSPCR semiquantitativa gene
Page 90 and 91:
RESULTADOS6.3. RT-PCRs semi-quantit
Page 92 and 93:
RESULTADOS11.00%6,50%AdequadaNão A
Page 94 and 95:
RESULTADOS6.6. Quantificação e no
Page 96 and 97:
RESULTADOSPara um melhor entendimen
Page 98 and 99:
RESULTADOSFigura 25. Agrupamento hi
Page 100 and 101:
RESULTADOSFigura 26. Gráfico ilust
Page 102 and 103:
RESULTADOS6.8. Influência do silen
Page 104 and 105:
Discussão
Page 106 and 107:
DISCUSSÃOA tolerância do repertó
Page 108 and 109:
DISCUSSÃOCom relação à técnica
Page 110 and 111:
DISCUSSÃOgene AIRE (do inglês aut
Page 112 and 113:
DISCUSSÃOdesse modo a tolerância
Page 114 and 115:
DISCUSSÃOPML (leucemia promielocit
Page 116 and 117:
DISCUSSÃOBoehm et al. (2003) afirm
Page 118 and 119:
DISCUSSÃOprograma Genenetwork (Wu
Page 120 and 121:
Conclusões
Page 122 and 123:
Referências Bibliográficas
Page 124 and 125:
REFERÊNCIAS BIBLIOGRÁFICASBarthlo
Page 126 and 127:
REFERÊNCIAS BIBLIOGRÁFICASDissert
Page 128 and 129:
REFERÊNCIAS BIBLIOGRÁFICASGotter
Page 130 and 131:
REFERÊNCIAS BIBLIOGRÁFICASMatsumo
Page 132 and 133:
REFERÊNCIAS BIBLIOGRÁFICASby mito
Page 134 and 135:
REFERÊNCIAS BIBLIOGRÁFICASRossi S
Page 136 and 137:
REFERÊNCIAS BIBLIOGRÁFICASof thym
Page 138 and 139:
Anexo I123
Page 140 and 141:
ANEXO IEST IMAGE:582370 Expressed s
Page 142 and 143:
ANEXO IDhps IMAGE:583332 Deoxyhypus
Page 144 and 145:
ANEXO IMapkapk2 IMAGE:572979 MAP ki
Page 146 and 147:
ANEXO IGm608 IMAGE:583350 Gene mode
Page 148 and 149:
ANEXO ICcdc72 IMAGE:640628 Coiled-c
Page 150 and 151:
ANEXO ITranscribed locusTranscribed
Page 152 and 153:
Page 154 and 155:
ANEXO ITranscribed locusTranscribed
Page 156 and 157:
ANEXO Irepair deficiency, complemen
Page 158 and 159:
ANEXO Imember 1EST IMAGE:583824 Exp
Page 160 and 161:
ANEXO ITranscribed locus IMAGE:5830
Page 162 and 163:
ANEXO ITfdp1 IMAGE:640119 Transcrib
Page 164 and 165:
Page 166 and 167:
ANEXO I2410022M11Rik IMAGE:640727 R
Page 168 and 169:
ANEXO Icontaining 1Kif14 IMAGE:5836
Page 170 and 171:
ANEXO Imember 3In multiple clusters
Page 172 and 173:
Page 174 and 175:
ANEXO IAnkrd10 IMAGE:583740 Ankyrin
Page 176 and 177:
Page 178 and 179:
ANEXO IVcam1 IMAGE:576563 Vascular
Page 180 and 181:
ANEXO ITegt IMAGE:639877 Testis enh
Page 182 and 183:
ANEXO I3Ehmt2Euchromatic histone ly
Page 184 and 185:
Page 186 and 187:
Page 188 and 189:
ANEXO I20 homolog (yeast)Ube1c IMAG
Page 190 and 191:
ANEXO IRasa3 IMAGE:582174 RAS p21 p
Page 192 and 193:
Page 194 and 195:
ANEXO I4930566A11Rik IMAGE:583433 R
Page 196 and 197:
Page 198 and 199:
Page 200 and 201:
Anexo II
Page 202 and 203:
ANEXO II1500002B03RikIMAGE:1330385
Page 204 and 205:
ANEXO IIEST IMAGE:640173 Expressed
Page 206 and 207:
ANEXO IICnot10 IMAGE:576406 CCR4-NO
Page 208 and 209:
ANEXO IIEST IMAGE:576142 Expressed
Page 210 and 211:
Anexo III
Page 212 and 213:
ANEXO III197Ifit1RelaIMAGE:575632Gr
Page 214 and 215:
ANEXO III199Nde1Nol5aIMAGE:640341Rp
Page 216 and 217:
Manuscrito da Tese
Page 218 and 219:
MANUSCRITOAbstractThe expression of
Page 220 and 221:
MANUSCRITOdominated the scenario in
Page 222 and 223:
MANUSCRITOserum autoantibodies, all
Page 224 and 225:
MANUSCRITOwere transfected with 10
Page 226 and 227:
MANUSCRITOwere analyzed using the M
Page 228 and 229:
MANUSCRITOpurified from ex vivo mou
Page 230 and 231:
MANUSCRITOinformation theory as ARA
Page 232 and 233:
MANUSCRITOKyewski B, Derbinski J, G
Page 234 and 235:
MANUSCRITOBruno R, Sabater L, Tolos
Page 236 and 237:
MANUSCRITOFigure 3. The gene expres
Page 238 and 239:
MANUSCRITOABFigure 5. Genetics netw
Page 240 and 241:
SÚMULACURRICULUM VITAE (Janeiro 20
Page 242 and 243:
ESTÁGIO NO EXTERIORSÚMULA• Labo
Page 244 and 245:
SÚMULAtranscript finishing initiat
Page 246 and 247:
SÚMULAlinhagens isogênicas de cam
Page 248 and 249:
SÚMULABOLSAS DE ESTUDO• Bolsa de
Page 250 and 251: T cell signaling gene networks duri
Page 252 and 253: IntroductionThe thymus is an import
Page 254 and 255: ontogeny using such technology has
Page 256 and 257: laboratory using a Generation III A
Page 258 and 259: cDNA microarray data analysisStatis
Page 260 and 261: Gene networksAs previously mentione
Page 262 and 263: cells, including human herpesvirus-
Page 264 and 265: gene was mostly indirectly positive
Page 266 and 267: lymphocytes and finally associates
Page 268 and 269: 4) Finally, Prrg2 was also identifi
Page 270 and 271: 14. Kishimoto H, Sprent J (2000) Th
Page 272 and 273: proliferation of a rat astrocyte ce
Page 274 and 275: IMMUNOLOGYORIGINAL ARTICLEOnset of
Page 276 and 277: Promiscuous gene expression in muri
Page 282 and 283: R.S. Cardoso et al. / Molecular Imm
Page 290 and 291: Molecular Immunology 42 (2005) 1043
Page 292 and 293: D.A.R. Magalhães et al. / Molecula
Page 294 and 295: D.A.R. Magalhães et al. / Molecula
Page 296 and 297: Molecular and Cellular Biochemistry
Page 298 and 299: 225Table 1. Target cDNA sequences a
show all

O papel modulador do gene AIRE - capes

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?