Transformation &lt;i&gt;in vitro&lt;/i&gt; of the Embryonal Hamster Brain Cells by Human Adenovirus Type 12

Kusano, Toshihisa; Yamane, Isao

doi:10.1620/tjem.92.141

Cited by 10 publications

(3 citation statements)

References 8 publications

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“…Since the observation by McBride and Wiener (6) of the in vitro transformation of hamster kidney cells by human adenovirus type 12 (Adl2), several additional reports (2,3,5 Proc. 24:308, 1965) have appeared substantiating these findings and extending them to other cell types.…”

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Adenovirus Transformation of Hamster Embryo Cells

Casto

1968

J Virol

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Inoculation of hamster embryo cell cultures with human adenovirus type 12 (Adl2) or simian adenovirus (SA7) resulted in the formation of foci of morphologically transformed cells within 12 days. The rapid appearance of well-defined foci was dependent upon the transfer of cells into new plates, with sufficient dilution after virus adsorption, and was independent of virus dose. Dose-response studies showed linearity of focus formation with dilution of Adl2 or SA7. Results averaged from several experiments show plaque-forming unit to focus-forming unit ratios of approximately 1.8 X 108 for Adl2 and 2.6 x 105 for SA7. Other experiments showed that most of the adenovirus involved in transformation was adsorbed by 3 hr. Cell lines derived from SA7 transformed cells produced tumors within 19 days when inoculated intradermally into young adult hamsters. Such cell-induced tumors histologically resembled SA7 virus-induced hamster tumors. Formation of tumors with SA7 transformed cells was inhibited by prior immunization of test animals with SA7 or Adl2 virus.

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Adenovirus Transformation of Hamster Embryo Cells

Casto

1968

J Virol

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“…'-4 Primarily for this reason it has also been suspected of being involved in human neoplasia.5 In cell culture, adenoviruses have been shown to transform hamster, rat, and rabbit embryo cells. [6][7][8][9] The transformants are recognized by their altered morphology and by their ability to grow on top of one another. They form multilayered foci and can be readily distinguished from the background of untransformed cells.…”

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Human cell strains susceptible to focus formation by human adenovirus type 12.

Todaro¹,

Aaronson²

1968

Proc. Natl. Acad. Sci. U.S.A.

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Adenovirus type 12 has been shown to produce tumors when inoculated into newborn animals of various species.'-4 Primarily for this reason it has also been suspected of being involved in human neoplasia.5 In cell culture, adenoviruses have been shown to transform hamster, rat, and rabbit embryo cells.6-9 The transformants are recognized by their altered morphology and by their ability to grow on top of one another. They form multilayered foci and can be readily distinguished from the background of untransformed cells. The adenovirus type 12 tumors and transformed lines contain a virus-specific new intranuclear tumor (T) antigen'0 and virus-specific messenger RNA." The cells transformed in culture produce tumors when inoculated into the appropriate host, and these tumors continue to show the presence of virus-specific genetic and antigenic material.'The present experiments demonstrate that adenovirus type 12 is able to produce altered foci in certain "susceptible" human cell cultures. Discrete adenovirus-induced foci are first seen 10-15 days after infection. The system, using human diploid cells, provides a method of quantitating the focus-forming ability of at least one type of human adenovirus.Materials and Methods.-Cells: Human fibroblast strains were derived from embryo lung, newborn foreskin, and adult skin. The last were initiated from 4-mm skin-punch biopsies. The tissue was finely minced and inoculated into 20 cm2 plastic Petri dishes containing Dulbecco and Vogt's modification of Eagle's medium supplemented with 10% calf serum. After approximately 2 weeks there generally was sufficient growth to allow subculture. The cells were then passaged at a 1:2 to 1 :10 dilution every 4-5 days and were used in the assays before the tenth transfer generation. Skin fibroblast cultures have been obtained from normal individuals and from individuals with genetic diseases associated with an increased risk of tumor formation. In particular, cell strains have been initiated from individuals with Fanconi's anemia, an autosomal recessive disease associated with a high risk of leukemia.12 1" These strains have previously been shown to be particularly susceptible to transformation in culture by another oncogenic DNA virus, SV40.14 Virus: Adenovirus type 12 (Huie strain), titering 106 5 PFU/0.1 ml on human embryonic kidney (HEK) cells, was obtained from Dr. W. Rowe (NIH), and one pool was used in all experiments.Experimental: Previous studies with SV40 transformation of the mouse line 3T315and human diploid strains'4 have shown that the efficiency of transformation is increased if rapidly dividing cultures rather than stationary, nonproliferating cultures are infected, and also if the cells are allowed several divisions subsequent to infection before they become arrested by contact inhibition of cell division. A similar requirement for several cell divisions for the expression of transformation has been shown for adenovirus type 12 focus formation in hamster embryo cells.'6 For this reason rapidly dividing cultures were inocula...

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“…This method of the subcultivation was similar to that employed in the previous study. 2,3 Assay procedure of growth rate and nutritional requirement of HT-2 cells. In the previous paper, it was indicated that HT-2 cells as well as other cell lines transformed by adenovirus type 12 could luxuriantly grow in a 5% CO2-incubator with a small inoculum size of less than 104 cells per ml, but they could not propagate in the rubber-stoppered bottles at the same inoculation density.…”

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confidence: 99%