IMMUNOHISTOCHEMICAL STUDIES OF ADENOVIRAS 12 T ANTIGEN IN CELLS The Institute of Medical Science, The University of TOKYO and Central Reseach Laborato
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1 IMMUNOHISTOCHEMICAL STUDIES OF ADENOVIRAS 12 T ANTIGEN IN CELLS The Institute of Medical Science, The University of TOKYO and Central Reseach Laboratoraries, Sankyo CO., Ltd. Hiroshi Misawa
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4 (1) (6) (11) c c0.1m Veronal buffer ph 7.5 (2) c c0.2m Tris buffert+0.01m Mgcl2 ph 7.5 (3) c c0.2m Tris buffer ph 7.5 (4) (5) (10) c cpbs ph 7.5 (7) (14) (17) c c0.005m Phosphate buffer ph 7.0 (8) (9) (12) c c0.1m Acetate buffer ph 5.6 (13) c c 0.1M Acetate buffer ph 5.0 (15) (16) c c0.1m Acetate buffer ph 4.8
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6 Summary In this paper, an immunohistochemical study on the tumor antigen (T) induced by adenovirus type 12 infection to hamster embryo cells (HaE) in vitro was explored by the use of immunofluores cence technique. The infected cells were subjected to observe any antigenic alteration of T antigen stained with fluorescein isothiocyanate conjugated anti-t antigen hamster antibody after treatment of those cells with the following various reagents. The antigenicity of T antigen was somewhat deteriorated by treatment with both proteolytic and carbohydrate-digesting enzymes and completely abolished with sodium periodate, phenol, deoxy cholate, and sulfuric acid. However, it resisted to treatment with formaldehyde and organic sol vents such as ethyl-ether, carbontetrachloride chloroform, while some hydrophilic organic sol vents, for instance, acetone, methyl- and ethyl alcohol acted somewhat deterioratively. Interestingly, preabsorption of the conjugate with lactose resulted in inhibiting the positive fluores cence in T antigen From these results, an assumption was made that adeno-12 induced T antigen in HaE would mainly consist of a protein which contains a part of polysuccharide like lactose as an immuno determinant group. 1) Andersen, B. R., Abele, D. C. and Vannier, W. E. (1968): Effects ot mild periodate oxi dation on antibodies. J. Immunol. 97, ) Black, P. H., Rowe, W, P., Turner, H. C., and Huebner R. J. (1963): A specific com plement-fixing antigen present in SV4o tum ors and tarnsformed cells. Proc. Natl Acad, Sci., 50, ) Gilead, Z., and Ginsberg, H. (1965): Chara-
7 cterization of a tumor like antigen in type 12 and type 18 adenovirus infected cells, J. Bact. 90, ) Habel, K. (1965): Specific complement-fixing antigens in polyoma tumors and transformed cells. Virology 25, ) Hollinshead, A. C., Bunnag, B. and Alford, T. C. (1967): Relationship between the subu nits and T antigens of adenovirus type 12. Nature 215, ) Huebner, R. J., Rowe. W. P., Turner, H. C. and Lane, W. T. (1963): Specific adeno virus complement-fixing antigens in virus free hamster and rat tumors. Proc. Natl. Acad. Sci., 50, ) Pope, J. H., and Rowe, W. P. (1964): Im munofluorescent studies of adenovirus 12 tumor and cells transformed or infected by adenovirus. J. Exp. Med. 120, ) Rapp, F., Kitahara, T., Butel, J. S. and Melnick, J. L. (1964): Synthesis of SV40 tu mor antigen during replication of simian Pa pova virus (SV40). Proc. Natl. Acad. Sci. 52, ) Shimojo, H., Sawamura, K., Wada, R. and Yoshikawa, E. (1966): The nature of tumor antigen of adenovirus type 12 and its forma tion in cultured cells after infection. Jap. J. Med. Sci. Biol. 19, ) Shimojo, H., Yamamoto, H. and Abe, C. (1967): Differentiation of adenovirus antigens in cultured cells with immunofluorescent ana lysis. Virology. 31, ) Tavitian, A., Peries, J., Chuat. J. and Boi von. M. (1967): Estimation of the molecular weight of adenovirus 12 tumor CF antigen by rate-zonal centrifugation. Virology 31, ) Trentin, J. J., Yabe, Y. and Taylar, G. (1962): The quest for human Cancer virus. Science 137,
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