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研究生:文詩妮
研究生(外文):Shy-Ni, Wen
論文名稱:經PMA,Ionomycin和IL-4活化過之NaiveCD8+T細胞其功能特性的維持
論文名稱(外文):Functional Fate of Naive CD8+ T cells activated by PMA + Ionomycin + IL-4
指導教授:孔祥智孔祥智引用關係
指導教授(外文):John T. Kung
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:免疫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:1999
畢業學年度:87
語文別:中文
論文頁數:46
中文關鍵詞:第二型T細胞分化CD4-8- T細胞
外文關鍵詞:type 2 T cellspolarizationCD4-8- T cells
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近年來,許多研究報告指出CD8+ T細胞可依其分泌之細胞激素 ( cytokine ) 類型而分群 : 第一型T細胞又稱Tc1 T細胞主要分泌IL-2,IFN-和TNF- ; 及第二型T細胞又稱Tc2 T細胞主要分泌IL-4,IL-5和IL-10。而在初次活化培養時外加IL-4可迫使T細胞往第二型T細胞方向發展。
在活體中,已分化至第一型或第二型之CD4+和CD8+ T細胞其分化狀態之持續已有文獻報導。文獻中指出不論是在CD4+或CD8+ T細胞中,第一型和第二型T細胞在活體中存活期間不會改變其細胞激素分泌類型,亦即分化結果不可逆。
PMA和ionomycin皆為T細胞之活化原。綜合PMA,ionomycin及IL-4之作用可使從未被活化之CD8+ T細胞降低其細胞表面CD8分子之表現,往第二型T細胞之分化發展並且呈現有效的細胞毒殺能力。
此篇論文主要之目的是為了探討CD8+ T細胞在被PMA + ionomycin + IL-4於活體外活化後,其功能上之特性及其特性之穩定度。在此特別討論三功能特性之穩定性及可逆性 : IL-4分泌能力,CD8分子表現之失去及細胞毒殺能力。
結果中指出,被PMA + ionomycin + IL-4於活體外活化後,B10 T細胞可維持IL-4分泌能力,但2C轉殖基因T細胞無法維持其IL-4 分泌能力。B10及2C轉殖基因T細胞於失去細胞表面CD8分子表現後,不會再度回復其表現,則此為不可逆現象。被活化之B10 T細胞具有良好的細胞毒殺能力,且此能力於活體中可被維持下去。

According to distinct cytokine secreting profile, CD8+ T cells can be defined as two populations : type 1 or Tc1 T cells that secret IL-2, IFN- and TNF- ; type 2 or Tc2 T cells that secret IL-4, IL-5 and IL-10. The presence of IL-4 in primary activation culture will drive CD8+ T cells to differentiate into Tc2 phenotype.
In vivo persistence of type 1 and type 2 polarization have been documented in CD4+ and CD8+ T cells. The reports demonstrated that both in CD4+ and CD8+ T cells, neither type 1 or type 2 T cells would change their cytokine secreting profile after surviving in vivo.
PMA and ionomycin are mitogens for T cells. The combine actions of PMA + ionomycin + IL-4 can induce naive CD8+ T cells to down-regulate surface CD8 expression, undergo Tc2 differentiation and express efficient cytotoxic activity.
The main purpose of this study is to identify the functional fate of naive CD8+ T cells activated in vitro by PMA + ionomycin + IL-4. Specifically, stability and potential reversibility with respect to IL-4 producing ability, loss of CD8 expression and ctyotoxic activity were examined.
The results indicated that conferred IL-4 producing ability of B10 T cells can persist in vivo but 2C tg T cells can not retain this functional characteristic. Down-regulation of CD8 expression is irreversible both in B10 and 2C tg T cells. Activated B10 T cells are potent cytotoxic effectors, and their potentials of regaining cytotoxic activity can exist in vivo.

Abbreviation...................................................I
English Abstract.............................................III
中文摘要.......................................................V
Introduction...................................................1
Material and Method............................................6
Mice.........................................................6
Solution.....................................................6
Panning Plate................................................7
Staining and Sorting.........................................8
Purification of B10 / 2C naive CD8+ T cells .................9
Generation of P + I + IL-4 activation B10 / 2C T cells......10
Generation of B10 / 2C CD8- memory T cells..................11
Functional Assay............................................13
Preparation of B10 / B10A B cell blasts for stimulation...13
IL-2 / IL-4 Bioassay......................................15
IFN-gamma ELISA...........................................16
IL-4 Producing Ability....................................18
CD8 Expression............................................18
Cytotoxicity Activity.....................................19
Results.......................................................22
Discussion....................................................28
Figures.......................................................33
Reference.....................................................42

1. Zangemeister-Wittke, U., Kyewski, B. and Schirrmacher, V. 1989. Recruitment and activation of tumor-specific immune T cells in situ. J. Immunol. 143 : 379.
2. Fan, S. T. and Edgington, T. S. 1989. Sufficiency of the CD8+ T cells lineage to mount an effective tumoricidal response to syngeneic tumor-bearing novel class I MHC antigens. J. Immunol. 143 : 4287.
3. Yasukawa, M., Inatsuki, A. and Kobayashi, Y. 1989. Differential in vitro activation of CD4+CD8- and CD8+CD4- herpes simplex virus-specific human cytotoxic T cells. J. Immunol. 143 : 2051.
4. Mizuochi, T., Hugin, A. W., Morse III, H. C., Singer, A. and Buller, R. M. L. 1989. Role of lymphokine-secreting CD8+ T cells in cytotoxic T lymphocyte responses against vaccinia virus. J. Immunol. 142 : 270.
5. Binnendijk, R. S. V., Poelen, M. C. M., Vries, P. D., Voorma, H. O., Osterhaus, A. D. M. D., and Uytdehaag, F. G. C. M. 1989. Measles virus-specific human T cells clones. J. Immunol. 142 : 2847.
6. Fabio, S. D., Mbawuike, I. N., Kiyono, H., Fujihashi, K., Couch, R. B. and McGhee, J. R. 1994. Quantitation of human influenza virus-specific cytotoxic T lymphocytes : correlation of cytotoxicity and increased numbers of IFN- producing CD8+ T cells. Int. Immunol. 6 : 11.
7. Salgame P., Abrams, J. S., Clayberger, C., Goldstein, H., Convit, J., Modlin, R. L. and Bloom, B. R. 1991. Differing lymphokine profiles of functional subsets of human CD4 and CD8 T cells clones. Science 254 : 279.
8. Seder, R. A., Boulay, J. L., Finkelman, F., Barbier, S., Ben-Sasson, S. Z., Gros, G. L. and Paul, W. E. 1992. CD8+ T cells can be primed in vitro to produce IL-4. J. Immunol. 148 : 1652.
9. Croft, M., Carter, L., Swain, S. L. and Dutton, R. W. 1994. Generation of polarized antigen-specific CD8 effector population : reciprocal action of interleukin ( IL )-4 and IL-12 in promoting type 2 versus type 1 cytokine profiles. J. Exp. Med. 180 : 1715.
10. Sad, S., Marcotte, R. and Mosmann, T. R. 1995. Cytokine-induced differentiation of precursor mouse CD8+ T cells into cytotoxic CD8+ T cells secreting Th1 or Th2 cytokines. Immunity 2 : 271.
11. Sad, S. and Mosmann, T. R. 1996. The expanding universe of T-cell subsets : Th1, Th2 and more. Immunol. Today 17 : 138.
12. Li, L., Sad, S., Kagi, D. and Mosmann, T. R. 1997. CD8Tc1 and Tc2 cells secrete distinct cytokine pattern in vitro and in vivo but induce similar inflammatory reactions. J. Immunol. 158 : 4152.
13. Halverson, D. C., Schwartz, G. N., Carter, C., Gress, R. E. and Fowler, D. H. 1997. In vitro generation of allospecific human CD8+ T cells of Tc1 and Tc2 phenotype. Blood. 90 : 2089.
14. Sad, S., Krishnan, L., Bleackley, R. C., Kagi, D., Hengartner, H. and Mosmann, T. R. 1997. Cytotoxicity and weak CD40 ligand expression of CD8+ type 2 cytotoxic T cells restricts their potential B cell helper activity. Eur. J. Immunol. 27 : 914.
15. Cerwenka, A., Carter, L. L., Reome, J. B., Swain, S. L. and Dutton, R. W. 1998. In vivo persistence of CD8 polarized T cell subsets producing type 1 or type 2 cytokines. J. Immunol. 161 : 97.
16. Maggi, E., Giudizi, G. M., Biagiotti, R., Annunziato, F., Manetti, R., Piccinni, M. P., Parronchi, P., Sampognaro, S., Giannarini, L., Zuccati, G. and Romagnani, S. 1994. Th2-like CD8+ T cells showing B cell helper function and reduced cytolytic activity in human immunodeficiency virus type 1 infection. J. Exp. Med. 180 : 489.
17. Erard, F., Wild, M. T., Garcia-Sanz, J. A. and Le Gros, G. 1993. Switch of CD8 T cells to noncytolytic CD8-CD4- cells that make TH2 cytokone and help B cells. Science 260 : 1802.
18. Maggi, E., Manetti, R., Annunziato, F., Cosmi, L., Giudizi, M. G., Biagiotti, R., Galli, G., Zuccati, G. and Romagnani, S. 1997. Functional characterization and modulation of cytokine production by CD8+ T cells from human immunodeficiency virus-infected individuals. Blood 89 : 3672.
19. Le Gros, G. and Erard, F. 1994. Non-cytotoxic, IL-4, Il-5, IL-10 producing CD8+ T cells : their activation and effector functions. Curr. Opin. Immunol. 6 : 453.
20. Cronin II, D. C., Stack, R. and Fitch, F. W. 1995. IL-4 producing CD8 T cell clones can provide B cell help. J. Immunol. 154 : 3118.
21. Zamoyska, R. 1998. CD4 and CD8 : modulators of T-cell receptor recognition of antigen and of immune response ? Curr. Opin. Immunol. 10 : 82.
22. Straus, D. S. and Weiss, A. 1992. Genetic evidence for the involvement of the lck tyrosine kinase in signal transduction through the T cell antigen receptor. Cell 70 : 585.
23. Veillette, A., Bookman, M. A., Horak, E. M. and Bolen, J. B. 1988. The CD4 and CD8 T cell surface antigens are associated with the internal membrane tyrosine-protein kinase p56lck. Cell 55 : 301.
24. Renard, V. Romero, P., Vivier, E., Malissen, B. and Luescher, I. F. 1996. CD8 increase CD8 coreceptor function and participation in TCR-Ligand binding. J. Exp. Med. 184 : 2439.
25. Garcia, K. C., Scott, C. A., Brunmark, A., Carbone, F. R., Peterson, P. A., Wilson, I. A. and Teyton, L. 1996. CD8 enhances formation of stable T-cell receptor/MHC class I molecule complex. Nature 384 : 577.
26. Jameson, S. C. and Bevan, M. J. 1995. T cell receptor antagonists and partial agonists. Immunity 2 : 1.
27. Renard, V., Delon, J., Luescher, I. F. Malissen, B., Vivier, E. and Trautmann, A. 1996. The CD8 polypeptide is required for the recognition of an altered peptide ligand as an agonist. Eur. J. Immunol. 26 : 2999.
28. Kwan-Lim, G. e., Ong, T., Aosai, F., Stauss, H. and Zamoyska, R. 1993. Is CD8 dependence a true reflection of TCR affinity for antigen ? Int. Immunol. 5 : 1219.
29. Lopes, M. F., Cunha, J. M. T., Bezerra, F. L., Gonzalez, M. S., Gomes, J. E. L., Silva, J. R. L. E., Garcia, E. S. and Reis, G. A. D. 1995. Trypanosoma cruzi : Both chemically induced and triatomine-derieved metacyclic trypomastigotes cause the same immunological disturbance in the infected mammalian host. Exp. Parasitol. 80 : 194.
30. Van-Dam, J. G., Damoiseaux, J. G., Van-der-Heijden, H. A., Grauls, G., Van-Breda-Vriesman, P. J. and Bruggeman, C. A. 1997. Infection with rat cytomegalovirus ( CMV ) in the immunocompromised host is associated with the appearance of a T cell population with reduced CD8 and T cell receptor ( TCR ) expression. Clin. Exp. Immunol. 110 : 349.
31. Perez, v. L., Lederer, J. A., Lichtman, A. h. and Abbas, A. K. 1995. Stability of Th1 and Th2 population. Int. Immunol. 7 : 869.
32. Sad. S. and Mosmann, T. R. 1995. Interleukin ( IL ) 4, in the absence stimulation, induces an anergy-like state in differentiated CD8+ TC1 cells : loss of IL-2 synthesis of other cytokines. J. Exp. Med. 182 : 1505.
33. Swain, S. L. 1994. Generation and in vivo persistence of polarized Th1 and Th2 memory cells. Immunity 1 : 543.
34. Brooks, E. G., Balk, S. P., Aupeix, K., Colonna, M., Strominger, J. L. and Groh-spies V. 1993. Human T-cell receptor ( TCR ) /+ CD4-CD8- T cells express oligoclonal TCRs, share junctional motifs across TCR V-gene families, and phenotypically resemble memory T cells. Proc. Natl. Acad. Sci. 90 : 11787.
35. Matsuzaki, G., Li, X. Y. Kadena, T., Song, F., Hiromatsu, K., Yoshida, H. and Nomoto, K. 1995. Early appearance of T cell receptor + CD4-CD8- T cells with a skewed variable region repertoire after infection with Listeria monocytogenes. Eur. J. Immunol. 25 : 1985.
36. Margolick, J. B., Carey, v., Munoz, A., Polk, B. f., Giorgi, J. V., Bauer, K. D., Kaslow, R. and Rinaldo, C. 1989. Development of antibodies to HIV-1 is associated with an increase in circulating CD3+CD4-CD8- lymphocytes. Clin. Immunol. Immunopatho. 51 : 348.
37. Zlotnik, A., Godfrey, D. L., fischer, M. and Suda, T. 1992. Cytokine production by mature and immature CD4-CD8- T cells. J. Immunol. 149 : 1211.
38. Takamoto, M., Kusama, Y., Takatsu, K. Nariuchi, H. and Sugane, K. 1995. Occurrence of interleukin-5 production by CD4-CD8- ( double-negative ) T cells in lung of both normal and congenitally athymic nude mice infected with Toxocara canis. Immunology 85 : 285.
39. Osborne, J. and Devaney, E. 1998. The L3 of Brugia induces a Th2-polarized response following activation of an IL-4-producing CD4-CD8-  T cell population. Int. Immunol. 10 : 1583.
40. Cho, B. K., Wang, C., Sugawa, S., Eisen, H. N. and Chen, J. 1999. Functional difference between memory and naive T cells. Proc. Natl. Acad. Sci. USA 96 : 2976.
41. Tanchot, C., Guillaume, S., Delon, J., Bourgeois, C., Trautmann, A. and Rocha, B. 1998. Modifications of CD8+ T cells function during in vivo memory or tolerance induction. Immunity 8 : 581.
42. Curtsinger, J. M.,Lins, D. C. and Mescher, M. F. 1998. CD8+ memory T cells ( CD44high, Ly-6C+ ) are more sensitive than naive cells ( CD44low, Ly-6C- ) to TCR/CD8 signaling in response to antigen. J. Immunol. 160 : 3236.

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