[CANCER RESEARCH 40, 2588-2595, July 1980]0008-5472/80/0040-OOOOS02.00

Transplantability of Human Lymphoid Cell Line, Lymphoma, and Leukemiain Splenectomized and/or Irradiated Nude Mice1

Shaw Watanabe2, Yukio Shimosato, Masahito Kuroki, Yuichi Sato, and Takashi Nakajima

Pathology Division. National Cancer Center Research Institute, 5-1-1. Tsukiji. Chuo-ku, Tokyo 104, Japan

ABSTRACT

The effects of splenectomy and/or whole-body irradiation of

nude mice before xenotransplantation of lymphoid cell lines,lymphoma, and leukemia were studied, Transplantation afterwhole-body irradiation resulted in the increased "take" rate of

three cultured cell lines (two of T-cell-derived acute lympho-cytic leukemia and one of B-cell derived acute lymphocyticleukemia) and in the tumorous growth of Burkitt-derived Raji

and spontaneously transformed lymphoblastoid cell lines. Withsplenectomy plus irradiation as a pretreatment, tumorousgrowth occurred in four other cell lines which were not trans-plantable after irradiation only (two cell lines of Epstein-Barrvirus-transformed cord blood cells and one each of null acutelymphocytic leukemia and nodular iymphoma-derived cell

lines).Direct transplantation of leukemia and lymphoma cells into

the pretreated mice was successful in 7 of 24 cases (29%). B-cell-derived diffuse large lymphoid lymphoma was transplant-

able in three of seven cases (43%). However, lymphoma andleukemia of peripheral T-cell origin was difficult to transplanteven with pretreatment, and only one pleomorphic T-cell lymphoma grew to a significant size (2 cm). One tumor each of B-cell-derived diffuse large lymphoid lymphoma and T-cell diffuse

lymphoblastic lymphoma became transplantable.

INTRODUCTION

Transplantation of human neoplasms into athymic nude micehas been used extensively and successfully in studies of manyoncological and treatment problems. However, it is recognizedthat leukemia and lymphoma are the most difficult tumors totransplant into nude mice (3, 5, 6, 9, 12, 17, 18, 21, 22, 25,29). The rich surface glycoprotein of the hematopoietic cellswas thought to provoke a thymus-independent immune system.

Higher activity of natural killer cells in nude mice also plays arole in the rejection of the transplanted lymphoma cells (1, 2,10, 15). Xenotransplantation i.e.,3 treatment with immunosup-

pressive drugs, administration of antilymphocyte serum, and/or whole-body irradiation have been used to increase theacceptance rate of these tumors (5-8, 23). Pretreatment withwhole-body irradiation successfully induced leukemia from human T-ALL cells in nude mice in our previous study (25). These

data suggest that reduction of the ability of the host to mount

' This work was supported by the Princess Takamatsu Cancer Research Fund

and by Grants-in-Aid for Cancer Research from the Ministry of Health and Welfare(No. 53-5) and the Ministry of Education, Science, and Culture (No. 301086).

2 To whom requests for reprints should be addressed.3 The abbreviations used are: i.e., ¡ntracranial; T-, T-cell-derived; ALL, acute

lymphocytic leukemia; B, B-cell-derived; null, null cell-derived; EBV, Epstein-Barrvirus; DLL, diffuse large lymphoid type; DLB, diffuse lymphoblastic type; AML,acute myeloid leukemia; CML, chronic myeloid leukemia.

Received September 4, 1979; accepted April 14, 1980.

an immune response increases the transplantability of tumorsinto nude mice.

Lymphoma and leukemia in experimental animals are verydifferent from those in humans. The experimental model ofhuman lymphoma in nude mice may contribute to the study ofthe host-lymphoma relationship and to an accurate analysis of

the activities of neoplastic cells in the in vivo state, i.e., response to drugs, mode of infiltration, or leukemic change.

This report deals with the effects of splenectomy, whole-

body irradiation, and their combination on the transplantabilityof cultured human lymphoid cell lines and of fresh specimensof leukemia and lymphoma in nude mice. The characteristicsof successfully transplanted tumors are reported, and the effects of pretreatment are discussed.

MATERIALS AND METHODS

Animals. Male athymic nude mice, 3 weeks old, with aBALB/c genetic background, were supplied by the CentralInstitute for Experimental Animals, Kawasaki, Japan, wheremice were bred and maintained in plastic film isolators underspecific-pathogen-free conditions. They were kept in a plasticfilm isolator or in a laminar flow rack (Sanki Co., Tokyo, Japan)in our laboratory and were given sterilized pellets (CL-2; Clea

Japan, Tokyo, Japan) and tap water ad libitum throughout theexperiment.

Splenectomy and Irradiation. Splenectomy was performedat 8 to 10 weeks of age through a 5-mm-long incision in the

left upper flank. After the hilar vessels were tied and cut off,the spleen was removed. One or 2 stitches were enough toclose the wound, which usually healed within 1 week withoutany treatment. Three days before transplantation, whole-bodyirradiation (400 rads) was given with a 6-MeV machine at a rate

of 320 rads/min to mice placed in a paper box covered with apaper filter. Most animals used for transplantation were 12 to15 weeks old.

Leukemia and Lymphoma Cells Transplanted. The following cell lines (1 to 5 x 107 cells) were used for transplantation:

(a) Molt-4F strain (16) of human T-ALL origin with tetraploidkaryotype; (b) Ichikawa strain (25) of human T-ALL origin withhypotetraploid karyotype; (c) Takahashi strain, childhood B-

ALL origin with pseudotetraploid karyotype; (d) Raji strain (20),a Burkitt's Iymphoma-derived cell line with a mode of diploid

karyotype; (e) NH-AR strain (27), follicular lymphoma of large

lymphoid type origin containing 47 chromosomes with a markerchromosome; (O KOP strain," baby null-ALL origin with

45,XX,-13,14p+ karyotype; (g) Shibata strain from a childhood null-ALL; and (h) EBV-LC and EBV-L6 strains, umbilicalcord blood-derived lymphoblastoid cell lines transformed by

* Manuscript in preparation.

2588 CANCER RESEARCH VOL. 40

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

Transplantability of Lymphoid Cells in Nude Mice

infection with EBV strain B95-8, with normal diploid karyotypes

(26).Direct transplantation of biopsied tissue was attempted from

malignant lymphoma cases, consisting of 7 of DLL of B-cellorigin, 4 of pleomorphic T-cell lymphoma (28), one of DLLwithout any detectable marker, one of childhood DLB with T-cell marker, 2 follicular lymphomas, and one Hodgkin's lym

phoma, nodular sclerosis type. Diagnosis was based on theclassification of Dorfman (4). Transplantation of neoplastic cellsfrom the peripheral blood of patients was carried out in 1 caseof Sezary's syndrome, 1 case of ALL, 3 cases of AML, 1 case

of CML, and 2 cases of acute monocytic leukemia. Two or 3mice were given s.c. injections of 5 x 107 cells from these

cases.Histology and Cytology. Nude mice were autopsied at the

time of tumor transfer or sacrificed 6 months after transplantation. Various organs were fixed in 4% formaldehyde solutionfor routine histological examination. Tumor imprints werestained by May-Giemsa solution. Tumors were also fixed forelectron microscopy in 2.5% glutaraldehyde in 0.1 M phosphate buffer with 5% sucrose and postfixed in 1% osmiumtetroxide in the same buffer. Blocks were embedded in epoxyresin, and ultrathin sections were made after selection of theblocks by examining 1-/im-thick méthylèneblue-stained sec

tions. They were observed in a Hitachi 11E electron microscope after double staining with uranyl acetate and lead citrate.

Chromosome Analysis. A cell suspension was made fromthe tumor in Roswell Park Memorial Institute Medium 1640,and cells were expanded with 0.075 M KCI, fixed 4 times withmethyl alcohohacetic acid (3:1) solution, and dropped ontocold glass slides. The slides were quickly flame dried andstained with Giemsa solution. The standard G and/or Q bandtechnique was also used for further characterization. Chromosomes were counted, and karyotypes on photomicrographswere analyzed.

RESULTS

Transplantability of Cultured Cells and Fresh Materials.The transplantability of various lymphoid cell lines in untreated,irradiated, and/or splenectomized mice is summarized in Table1. Irradiation increased transplantability in the cell lines withaneuploid chromosomes in the tetraploid range (Molt 4F, Ichi-kawa, and Takahashi). Latent periods for these tumors were 5to 7 days, and the tumors reached about 3 cm in diameter after4 weeks. Transplantation of cells in the diploid range (Raji, NH-AR, KOP, Shibata, EBV-LC, and EBV-L6) seemed to be moredifficult, although their latent periods were also 5 to 7 days andthey reached their maximum size in 4 to 5 weeks. The NH-AR

strain became transplantable in nude mice pretreated withsplenectomy and irradiation. Serial transfer, however, has beenunsuccessful thus far. The maximum tumor size was 2 cm indiameter after the initial transplantation; it decreased to 0.8 cmin diameter after the first passage, and no nodular growth wasobserved in the second transfer. The KOP strain also grew inthe splenectomized and irradiated nude mice, and serial transplantation was possible in irradiated nude mice but not inunirradiated ones (Fig. 1a). EBV-transformed cell lines were

also tumorigenic in the conditioned mice, although the tumorregressed spontaneously after reaching a diameter of 2.5 cmand transplantation was unsuccessful (Fig. 2a).

The effects of pretreatment and the route of transplantationwere analyzed statistically by using the data on transplantationof cells in the diploid range (Table 2). Injection s.c. yielded thehighest takes by irradiation (p < 0.01 ). Injection i.p. producedi.p. tumors in cell lines with high chromosome abnormality inthe tetraploid range, whereas transplantation by that routeseemed difficult with cells in the diploid range (Raji, KOP).Injection i.v. caused leukemia in only the Ichikawa strain.

Tumors from 3 of 7 cases of B-DLL grew to 3 cm in diameter,

and one of them became serially transplantable (Table 3,

Table 1Transplantabi/ity of cultured human lymphoid cell lines

TransplantabilityCell

lineMolt

4FIchikawaTakahashiRajiNH-ARKOPShibataEBV-LCEBV-L6OriginALLALLALLBurkittFollicular

lymphomaALLSpontaneouslymphoblastoidEBV-transformedcellsSame

as abovePheno-typeTTBBBNullBBBChromosomeTetraploidHypotetraploid

(mode88.35%)Hypotetraploid44-87

(mode46.65%)47,XY,

+145,XX,-13Normal

diploidNormal

diploidNormal

diploidRoute

oftrans-planta

tions.c.i.p.I.V.s.c.I.p.I.V.s.c.i.p.I.V.s.c.l.p.I.V.s.c.s.c.i.p.i.V.s.c.8.C.s.c.Untreated1/2"0/23/3All0/121/30/20/20/60/30/60/20/20/20/60/60/2Irradiated2/21/20/3All12/123/33/40/26/61/60/30/60/20/20/24/60/6Sple

nectomy+irradia

tion6/66/64/46/66/64/4

' Number of cases producing tumors/number of cases transplanted

JULY 1980 2589

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

S. Watanabe et al.

Tumor 126-79). All T-cell lymphoma and Sézarycells wererejected, except for one T-DLL (pleomorphic) and one T-DLB

lymphoma from a pleural effusion (Figs. 3 and 4). One of 3AML transplants and one CML transplant grew to 1.5 cm indiameter, but the transplant of the first passage was rejected.Other lymphoma and leukemia transplants failed to grow evenin conditioned nude mice. The total acceptance rate of theinitial transplantation was 29%.

Characterization of Transplanted Tumors. The characteristics of the Ichikawa cells were described previously (25).Takahashi cells (B-ALL) transplanted i.p. resulted in tumorous

proliferation in the abdominal cavity, producing no ascitic fluid.The histology of these 2 and of the Molt 4F tumor was that ofDLL, although occasional giant cells were intermingled with theother cells. Raji and NH-AR cells formed cutaneous nodules

with a histology like that of DLL lymphoma without definitenodular growth. Neoplastic cells had large vesicular nuclei andlarge nucleoli. Medium-sized lymphoid cells were occasionally

intermingled with these cells. Ultrastructurally, a moderateamount of cytoplasm was filled with polyribosomes and contained an occasional long, lamellar, rough endoplasmic retic-ulum. There were short cytoplasmic projections, but the laby-

Table 2Transportability of cell lines with karyotypes in diploid range

Difference between s.c. and i.p. in irradiated groups is significant by x2 lest

(p < 0.01). Difference between untreated, irradiated, and splenectomy plusirradiation treatments in s.c. groups is also significant by \2 'est (p < 0.01).

TransplantabilityRoute

oftransplanta

tions.c.

¡.p.i.v.TotalUntreated0/2830/50/20/43Irradiated10/26

Õ38.5)6

1/8(12.5)0/511/40Splenec

tomy + irradiation32/32

(100)NDCNO32/32Total42/86

(48.8)1/13(7.7)0/743/106

Number of cases producing tumor/number of cases transplanted.0 Numbers in parentheses, percentage.c ND, not done.

rinthine structure of dendritic reticulum cells with desmosomeswas not observed. The chromosome pattern of NH-AR cells

was mosaic 47/48, in which the characteristic marker chromosome of the cell line remained.

The KOP strain (null-ALL) revealed a distinct growth pattern.

The neoplastic cells showed diffuse infiltrative growth in theskin, invading into the deep s.c. tissue and extending to thebasal layer of the epidermis (Fig. 1b). Individual cells wereregular in size with many mitotic figures (Fig. 1c). Vascularinvasion was common (Fig. 1oO.Nucleoli were distinct althoughnot very large. These features are those of DLB lymphoma.Ultrastructural features of KOP cells included poorly developedorganelles in the scant cytoplasm (Fig. 1e). The chromosomepattern of the cutaneous tumor was the same as that of thecultured cell line (Fig. 5).

Nodules which developed s.c. after transplantation of EBV-

transformed lymphoblastoid cell lines revealed diffuse proliferation of large lymphoid cells and did not show the starry-skypattern of the Burkitt's lymphoma (Fig. 2b). Ultrastructurally,

the tumor was composed of large lymphoid cells and plasmacells of varying degrees of maturity. No viral particles werefound by routine electron microscopy. The chromosome pattern was normal diploid, as in the established clone in the cellculture without a 14q+ chromosome (Fig. 6).

Three B-DLL lymphomas and one pleomorphic T-cell lym

phoma transplanted directly from the patients into the s.c.tissue of nude mice retained the same morphology as those ofthe biopsied specimens (Fig. 3). One B-DLL (Tumor 126-79)

became serially transplantable; its karyotype was in the diploidrange. Metastasis to the regional axillary lymph node wasobserved in the mouse receiving pleomorphic T-cell lymphoma,although the third transfer failed. One transfer of T-DLB cells

from pleural effusion yielded a transplantable tumor (Tumor78-79) and retained its original histology (Fig. 4). The kary

otype was normal diploid. Tumorous growth from AML wascomposed of immature myeloid cells; that from CML was composed of myeloid cells in various stages of maturity and ofmonocytic cells. Ultrastructurally, both tumors were seen tocontain typical azure granules. Both failed in the second passage.

Table 3Transplantabi/ity of human lymphoma and leukemia

Two or 3 animals were used for each transplantation.

Transplantability

DiagnosisMalignant

lymphomaDLLDLL

(pleomorphic)DLLDLBFollicularHodgkin

NSCALLSézaryAMLAMoLCMLTotalMarkerBTNullTBTTMyeloidMonocyticMyeloidNo.

ofcases7411211132124Irradiated1/10/10/10/10/11/5Splenectomy1/4a0/30/11/10/10/12/11Splenec

tomy +irradiation2/30/10/10/10/11/20/11/14/11Mouse

GroupNo. for serial

transplantation7-6-78,

71-79,126-79"102-7978-79"7-11-7895-797/24

(29%)a Number of cases producing tumors/number of cases transplanted.'' Serially transplantable cell lines obtained.c NS, nodular sclerosis type; AMoL, acute monocytic leukemia.

2590 CANCER RESEARCH VOL. 40

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

Transplantability of Lymphoid Cells in Nude Mice

DISCUSSION

Direct transplantation of human hematopoietic and lymphoidmalignant cells from patients to nude mice has been triedextensively, but only a few successes have been reported (5,12, 21). The presence of natural killer cells and of a T-inde-

pendent immune system was thought to act in the rejectionmechanism. Sublethal irradiation, which can reduce the immunity of animals, has been used for heterotransplantation ofhematopoietic cell lines by Toolan (24), Imamura et al. (11),and Prehn and Outzen (19). In this experiment, suppression ofthe immune system with splenectomy and/or irradiation yieldedthe increased tumor acceptance. Most cultured cell lines became transplantable in the conditioned mice. The difficulty indirect transplantation of hematopoietic and lymphoid tumorsfrom patients has not yet been resolved. There was initialacceptance in 7 of 24 cases (29%), and serially transplantabletumor lines were obtained in 2 of them. Thus far, the importanceof the pretreatment seems to be to increase the probability oftumor outgrowth from a primary tumor.

The route of transplantation also influenced the acceptancerate. Injection of neoplastic cells i.e., used by Epstein era/. (5),yielded a high transplantability rate. Schaad ef al. (21) succeeded in transplanting lymphoblastoid cell lines. Our preliminary work revealed that the number of collectable cells wasmuch higher from cutaneous tumors than from i.e. ones. Injection s.c. was more effective than were the i.p. or i.v. routes.The reason may be the better blood supply and the presenceof a connective tissue framework for fixation and proliferationof injected cells. The i.p. phagocytic system and systemicreticuloendothelial system would be activated promptly to provoke a systemic host immune reaction by i.p. or i.v. inoculationof cells, respectively.

Recently, congenially asplenic-athymic (Lasat) mice havebeen used for transplantation studies (8, 14). Gershwin ef al.(8) reported favorable results of heterotransplantation of tumors in nude mice treated with antilymphocytic serum and inLasat mice. Lozzio ef al. (13) reported the successful transplantation of the CML-derived K-562 strain to Lasat mice.

However, our preliminary transplantation of 3 fresh leukemiasinto Lasat mice failed. Further work on the comparison betweenLasat and pretreated mice for their ability to accept humanhematopoietic and lymphoid tumors is needed.

Nilsson ef al. (17) found that only old hematopoietic cell lineswith chromosome aneuploidy (hyperploidy) were transplantable. The same results were reported by Imamura ef al. (11 ) andZamecnic and Long (29). Diehl ef al. (3) reported that lymphoblastoid cell lines lacking a 14q+ marker chromosome failedto produce tumors in nude mice independent of diploidy oraneuploidy. They also failed to transplant lymphoblastoid celllines newly established from umbilical cord blood cells by EBVinfection. EBV-transformed lymphoblastoid cell lines with nor

mal karyotype, as well as the 2 lymphoma and leukemia celllines with near diploid chromosome pattern in this experiment,were able to proliferate to a significant size. However, of allthese tumors, only the KOP strain could be serially transplanted. It is not clear why the tumor that was once transplantedand grew to a significant size was rejected after passage evenin the conditioned mice. Failure of transfer or disturbed growthin the first or second transplantation could be attributed to theimpaired adaptation of the metabolic system of tumor cells to

the murine nutritional environment. Further investigation isneeded to resolve this program.

REFERENCES

1. Campanile, F.. Crino, L., Boumasser, E., Houchens, E., and Goldin, A.Radioresistant inhibition of lymphoma growth in congenially athymic (nude)mice. Cancer Res., 37: 394-398. 1977.

2. Cudkowicz, G. Rejection of bone marrow allografts by irradiated athymicnude mice. Proc. Am. Assoc. Cancer Res., 76 170, 1975.

3. Diehl, V.. Krause, P., Hellriegel. K. P., Busche, M., Schedel, I., and Laske-witz. E. Lymphoid cell lines: in vitro cell markers in correlation to tumori-genicity in nude mice. In: S. Thierfelder, H. Rodth, and E. Thiel (eds.),Hematology and Blood Transfusion, Vol. 20, pp. 289-296. Berlin: SpringerVerlag. 1977.

4. Dorfman, R. F. Pathology of the non-Hodgkin's lymphomas: new classifica

tions. Cancer Treat. Rep., 6). 945-952, 1977.

5. Epstein, A. L, Herman. M. M., Kim, H.. Dorfman, R. F., and Kaplan. H. S.Biology of the human malignant lymphomas. III. Intracranial hetero-trans-plantation in the nude, athymic mouse. Cancer (Phila.), 37: 2158-2176,1976.

6. Epstein, A. L., and Kaplan, H. S. Biology of the human malignant lymphomas.I. Establishment in continuous cell culture and hetero-transplantation ofdiffuse histiocytic lymphomas. Cancer (Phila.). 34: 1851-1872, 1974.

7. Gallie. B. L., Albert, D. M., Wong, J. J. Y.. Buyukmici. N., and Puliafito, C.A. Heterotransplantation of retinoblastoma into the athymic "nude" mouse.

Invest. Ophthalmol. Vis. Sci.. 76: 256-259, 1977.8. Gershwin, M. E., Ikeda, R. M., Erickson, K., and Owens, R. Enhancement of

heterotransplanted human tumor graft survival in nude mice treated withantilymphocytic serum and in congenitally athymic-asplenic (Lasat) mice. J.Nati. Cancer Inst.. 67. 245-248, 1978.

9. Giovanella, B. C., Morgan, A. C.. Stehlin. J. S., Williams. L. J., and Mimford,B. C. Development of invasive tumors in "nude" thymusless mice injected

with human cells cultured from Burkitt lymphomas. Proc. Am. Assoc. CancerRes., 74. 20, 1973.

10. Greaves, M. F., Janossy, G., Roberts, M.. Rapson, N. T., Ellis, R. B..Chesells, J., Lister, T. A., and Catovsky. D. Membrane phenotyping: diagnosis, monitoring and classification of acute "lymphoid" leukemias. In: S.

Thierfelder, H. Rodth, and E. Thiel (eds.), Hematology and Blood Transfusion. Vol. 20, pp. 61-76 Berlin: Springer Verlag, 1977.

11. Imamura, T., Huang, C., Minowada, J., and Moore, G. E. Heterologoustransplantation of human hematopoietic cell line. Cancer (Phila.). 25. 1320-1331, 1970.

12. Kuga, N.. Yoshida. K., Seido, T., Oboshi. S., Koide, T.. Shimosato, Y., andNomura, T. Heterotransplantation of cultured human cancer cells and humancancer tissues into nude mice. Gann, 66. 547-570. 1975.

13. Lozzio, B. B., Machado, E. A., Lozzio, C. B., and Lair, S. Hereditary asplenic-athymic mice: transplantation of human myelogenous leukemic cells. J. Exp.Med.. 743. 225-231. 1976.

14. Lozzio. B. B.. and Wargon, L. B. Immune competence of hereditary asplenicmice. Immunology, 27. 167-178, 1974.

15. Martin, W. H., and Martin, S. E. Naturally occurring cytotoxic antitumorantibodies in sera of congenitally athymic (nude) mice. Nature (Lond.), 249.564-565, 1974.

16. Minowada, J., Ohmura, T.. and Moore, G. E. Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derivedlymphocytes. J. Nati. Cancer Inst., 49 891-895, 1972.

17. Nilsson, K., Giovanella, B. C.. Stehlin. J. S., and Klein, G. Tumorigenicity ofhuman hematopoietic cell lines in athymic nude mice. Int. J. Cancer, 79.337-344, 1977.

18. Povlsen, C. O., Fialkow, P. J., Klein. E., Klein. G., Rygaad, J., and Wiener,F. Growth and antigenic properties of a biopsy derived Burkitt's lymphoma

in thymusless (nude) mice. Int. J. Cancer, 77. 30-39, 1973.

19. Prehn, L. M., and Outzen. H. C. Primary tumor immunity in nude mice. Int.J. Cancer, 79. 688-691, 1979.

20. Pulfertaft, R. J. Cytology of Burkitfs tumor. Lancet, 7. 238-240. 1964.21. Schaad, M., Kirchner, H.. Fonatsh. C., and Diehl. V. Intracranial heterotrans

plantation of human hematopoietic cells in nude mice. Int. J. Cancer. 23:751-761, 1979.

22. Sordat, B., Tamaoki, N., and Povlsen, C. O. List of human tumors transplanted to nude mice. In: T. Nomura, N. Ohsawa, N. Tamaoki. and K.Fujiwara (eds.), Proceedings of the Second International Workshop on NudeMice. pp. 587-595. Tokyo: University of Tokyo Press. 1977.

23. Stanbridge, E. J., and Perkins, F. T. Tumorigenicity testing in ¡mmunosup-pressed mice: advantages and disadvantages. Dev. Biol. Stand.. 37: 211-217, 1977.

24. Toolan, H. W. Successful subcutaneous growth and transplantation of human tumors in X-irradiated laboratory animals. Proc. Soc. Exp. Biol. Med..77. 572-578, 1951.

25. Watanabe, S., Shimosato. Y., Kameya, T.. Kuroki, M., Kitahara, T., Minato,K., and Shimoyama, M. Leukemic distribution of a human acute lymphocytic

JULY 1980 2591

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

S. Watanabe et al.

leukemia cell line (Ichikawa strain) in nude mice conditioned with whole- comparison with lymphoblastoid cell lines. Cancer (Phila)., in press, 1980.body irradiation. Cancer Res., 38. 3493-3498, 1978. 28. Watanabe, S., Tsutsumi, Y., Shimosato, Y., Shimoyama, M., Minato, K.. and

26. Watanabe, S.. Shimosato, Y., Kuroki. M., Hirohashi. S., Okada, H.. Suzuki. Nakao. N. Terminal deoxynucleotidyl transferase activity in leukemia andS., and Tsutsumi, Y. Tumorigenicity of Epstein-Barr virus transformed human lymphoma, with special reference to adult T cell related neoplasms. Actalymphocytes in conditioned nude mice. In: Proceedings of the Third Inter- Haematol. Jpn., 48: 15-25, 1980.

national Workshop on Nude Mice. Stuttgart: Gustav Fischer, in press. 1980. 29. Zamecnik. P. C.. and Long. J. C. Growth of cultured cells from patients with27. Watanabe. S., Shimosato. Y., Kuroki. M., Sato, Y., and Hasegawa. T. The Hodgkin's disease and transplantation into nude mice. Proc. Nati. Acad. Sci.

establishment of a cell line (NH-AR) from a human nodular lymphoma and a U. S. A., 74: 754-758, 1977.

Fig. 1. a, nude mouse into which KOP (null-ALL) strain was transplanted. Cutaneous infiltrative growth of the neoplastic cells is characteristic. In b. neoplasticcells diffusely proliferate just beneath the epidermis and invade s.c. tissue through muscular layer. H & E, x 90. In c, histologically neoplastic cells have fairly uniformnuclei with aggregated chromatin and small distinct nucleoli. Mitotic figures are numerous, and the histological criteria of diffuse lymphoblastic lymphoma aresatisfied. H & E, x 350. d, sample of vascular invasion, often seen. Periodic acid-Schiff. x 550. e, ultrastructure of KOP cells. Poorly developed cytoplasmicorganelles and abundant polysemes. Uranyl acetate and lead citrate, x 10,200.

Fig. 2. a. nude mouse into which EBV-transformed lymphocytes were transplanted. Tumorous growth is recognized, b, tumor composed of diffuse proliferation oflarge lymphoid cells with vesicular nuclei and conspicuous nucleoli. Starry-sky pattern is not seen. H & E. x 350.

Fig. 3. Transplanted tumor of pleomorphic T-cell lymphoma. Pleomorphism of the tumor is evident, composed of large cells similar to Reed-Sternberg cells andsmaller medium-sized cells. H & E, x 350.

Fig. 4. Newly established transplantable tumor of T-DLB lymphoma. Diffuse proliferation of regular-sized cells with round nuclei and small nucleoli. Numerousmitotic figures. H & E, x 350.

Fig. 5. Chromosome pattern of a tumor from the strain which developed in mice. 45.XX.-13.14p+.

Fig. 6. Chromosome pattern of a tumor formed after transplantation of EBV-transformed lymphocytes. Normal diploid with no 14q+ chromosome.

2592 CANCER RESEARCH VOL. 40

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

Transplantability of Lymphoid Cells in Nude Mice

|¿Õ e

JULY 1980 2593

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

S. Watanabe et al.

.'

i- ^ '

•'>-

«r*>«*•-^f." 4'- *î-«x* «<¿í¿»Al*««»<*•*««>*.*%:

s:'•x«V-.<*•••9* i

•v^TV •• % v a-- «t* - vT«A ^ '-' •v ' , * • »•*"" , % ' * KÎ>V^f^,i.fv^ <.*;' ''^*?£*-•--A1

; ^

¿"A^llk /*^ * **-.»•_•'-

f," • "•*•...

' t«.»«»

* •» .»^ • ^ «•^ ••*" * «^ ' ^r* ^^ 'fj * »

'*"'' ^ V* f '* ' *""* *r*v* ' '/ >^;fv^t^^ ^ V* A ' '

V*V; *' - •.'t •MV >•••-\ "' *•-'^ffiï^: '* "* •-

vvVfc' *, MV Vv-v, *.V'>j

. . f

¿V*.<V,3 <»»V?r««' / ' "Õ J•̂' v- '*v ^t - ' ^'«»V*'

4 •r\ ¿% >L f-.l« < ->^ r ' '

"*• •»J*S%fu9^*••>*••

, " ,v>i1"-V' '* '';'.,-'•u '. ' •- ' ' '*^

t>"**

* * -* '-m^l.'<y.-\

AC. *t 'V m iF —' •''•'V*;"'.Vf

»J>¿^^tWr^-»§;•;.••WT^

.îia-••v-

¿•> *.. •*é

•«jf

9

2594 CANCER RESEARCH VOL. 40

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

Transplantability of Lymphoid Cells in Nude Mice

ll It H II II5

ti II II *i i«n u10 11 12

IE« •I Ai14 15 16 17 18

•I ft* ••**19 2O 21 22 XX

il l>i> K v»123 45

ft Ci » U If U !!6 7 8 9 10 11 12

li II H it e«li13 14 15 16 17 18

II ft •••I \ J

19 20 21 22 XX

JULY 1980 2595

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from

1980;40:2588-2595. Cancer Res   Shaw Watanabe, Yukio Shimosato, Masahito Kuroki, et al.   Leukemia in Splenectomized and/or Irradiated Nude MiceTransplantability of Human Lymphoid Cell Line, Lymphoma, and

  Updated version

  http://cancerres.aacrjournals.org/content/40/7/2588

Access the most recent version of this article at:

   

   

   

  E-mail alerts related to this article or journal.Sign up to receive free email-alerts

  Subscriptions

Reprints and

  .pubs@aacr.orgDepartment at

To order reprints of this article or to subscribe to the journal, contact the AACR Publications

  Permissions

  Rightslink site. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC)

.http://cancerres.aacrjournals.org/content/40/7/2588To request permission to re-use all or part of this article, use this link

Research. on November 19, 2020. © 1980 American Association for Cancercancerres.aacrjournals.org Downloaded from