Expression and Regulation of the Leukemia Inhibitory ... · ically associated with HTLV-1 infection (1, 2). ATL frequently causes hypercalcemia, which may become an important factor

[CANCER RESEARCH 52, 6961-6965, December 1992]

Advances in Brief

Expression and Regulation of the Leukemia Inhibitory Factor/D Factor Gene in Human T-Cell Leukemia Virus Type 1 Infected T-Cell Lines

Tomoyo Umemiya-Okada, Toshiki Natazuka, ~ Toshimitsu Matsui, Mitsuhiro Ito, Taizo Taniguchi, and Yoshinobu Nakao Third Division, Department of Medicine, Kobe University School of Medicine, Kusunoki-Cho 7-5, Chuo-ku, Kobe 650, Japan

Abstract

The expression of the leukemia inhibitory factor/D factor (LIF) gene in human T-cell leukemia virus type 1 infected T-cell lines was examined. Human T-cell leukemia virus type 1 infected T-cell lines MT-1, MT-2, H89-59, H89-79, and H109 expressed LIF mRNA, but the T-cell lines MOLT-4 and TALL-1 did not. LIF mRNA expression was enhanced by interleukin 2 or 12-O-tetradecanoylphorbol-13-acetate in MT-2 cells. The biological activity of LIF was detected in culture medium enhanced by interleukin 2 in MT-2 cells. The expression of LIF mRNA was suppressed by la,25-dihydroxyvitamin D3 and dexamethasone. These results imply that the expression of the LIF gene is involved in the development of hypercaicemia and abnormalities of the immune system observed in patients with adult T-cell leukemia.

Introduction

ATL 2 is an aggressive, usually fatal T-cell neoplasm etiolog- ically associated with HTLV-1 infection (1, 2). ATL frequently causes hypercalcemia, which may become an impor tant factor in managing the course of the disease (3, 4). Several investiga- tors have reported that HTLV-1 infected leukemic T-cells produce OAF-like activities which can induce bone resorption (5-7). HTLV-1 infected T-cells abnormally express IL-I , parathyroid hormone related protein, and tumor necrosis factor

mRNA, which have OAF-like activities (8-11). HTLV-I infected T-cells secrete these factors in the condit ioned media of cell lines and ATL patient sera (12, 13). The clinical symptoms, i.e., bone lesions and hypercalcemia, thus might represent a direct consequence of the product ion of such factors by HTLV-1 infected T-cells.

LIF is a pleotropic cytokine that affects the growth and differentiation of various cell types, including hematopoietic, he- patic, adipogenic, renal, neuronal, and embryonic cells (14). Recently, Abe et al. (15) have reported that LIF produced by osteoblastic cells (MC3T3-E1) st imulated bone resorption by promot ing osteoclast formation. LIF is mainly secreted by activated T-cells and monocytes in the immune system (16, 17). Furthermore, it has been reported that HTLV-1 infected T-cells have phenotypic similarity to activated T-cells (18). I f LIF is secreted by HTLV-1 infected T-cells as other OAFs, LIF may contribute to the hypercalcemia and/or the immunological ab- normali t ies of ATL.

Received 9/28/92; accepted 10/23/92. The costs of publication of this article were defrayed in part by the payment of

page charges. This article must therefore be hereby marked advertisement in accord- ance with 18 U.S.C. Section 1734 solely to indicate this fact.

l To whom requests for reprints should be addressed. 2 The abbreviations used are: ATL, adult T-cell leukemia; LIF, leukemia inhib-

itory factor; HTLV-1, human T-cell leukemia virus type 1; OAF, osteoclast acti- vating factor; IL, interleukin; TPA, 12-O-tetradecanoylphorbol-I 3-acetate; IFN, interferon; 1,25 (OH)2D3, la,25-dihydroxyvitamin D3; cDNA, complementary DNA; SSC, standard saline-citrate; SDS, sodium dodecyl sulfate.

We examined the expression of LIF mRNA in HTLV-1 infected T-cell lines and analyzed the levels of LIF mRNA regulated by various cytokines and steroid hormones.

Materials and Methods

Cell Lines. The established HTLV-I infected T-cell line MT-2 was provided by Dr. I. Miyoshi (Kochi Medical College, Nangoku-City, Japan). H89-59, H89-79, and H109 established from HTLV-1 associated myelopathy patients, were provided by T. Saida (Department of Neurology, Utano National Hospital). MT-I, MOLT-4, and TALL-1 were gifts from the Japanese Cancer Research Resources Bank (Tokyo, Japan). HL-60 was provided by Dr. R. C. Gallo (NIH, Bethesda, MD). These cell lines were maintained in RPMI 1640 (Flow Laboratories, Rockville, MD) supplemented with 10% fetal bovine serum (M. A. Bioproducts, Rockville, MD) and kanamycin (100 #g/ml). Ml T-22 cells were a gift from Dr. M. Tomida (Saitama Cancer Research Center, Saitama, Japan), and were maintained in Dulbecco's minimal essential medium (Flow Laboratories) supplemented with 10% fetal bovine serum and kanamycin (100 ug/ml).

Reagents. Recombinant human IL-1B and IL-6 were purchased from Genzyme (Boston, MA). Recombinant human IL-2 and IFN-a were provided by Takeda Pharmaceutical Co. (Osaka, Japan) and Sum- itomo Pharmaceutical Co. (Osaka, Japan), respectively. Recombinant human IFN--y and TNF-o~ were gifts from Otsuka Pharmaceutical Co. (Osaka, Japan) and Daiichi Pharmaceutical Co. (Tokyo, Japan), respectively. TPA and dexamethasone were purchased from Sigma Chemical Co. (St. Louis, MO). 1,25(OH)2D3 was a gift from Teijin Institute for Biomedical Research (Tokyo, Japan).

cDNA Probe. Human LIF cDNA, 0.6 kilobase, in plasmid pIC2OR (19), was provided by Dr. Nick Gough (The Walter and Eliza Hall Institute of Medical Research, Victoria, Australia). The cDNA insert was removed from the plasmid by digestion with BamHI and HindIII followed by agarose gel electrophoresis and electroelution.

Northern Blots. Cell lines were incubated with various reagents for 24 h. Total cellular RNA was extracted from cells using acid guanidinium thiocyanate-phenol-chloroform (20). Twenty #g/lane of total RNA were electrophoresed in a 1% agarose formaldehyde gel, trans- ferred to nitrocellulose filters (Schleicher and Schuell, Dassel, Ger- many) in 1 m ammonium acetate, and then fixed with 254 nm UV lamp (Stratalinker, Stratagene, CA) to the filters. The filters were hybridized at 42~ in a buffer [40% (w/v) formamide, 5 x SSC (1 x SSC is 0.15 M sodium chloride-0.015 m sodium citrate], pH 7.01 x Denhardt's solu- tion (0.02% bovine serum albumin-0.02% Ficoll-0.02% polyvinylpyr- rolidone)-5 mM NaH2PO4-0.1% SDS-salmon sperm DNA (100 #g/ml)] containing 32p-labeled cDNA probes prepared with the random primer DNA labeling kit (Amersham, Buckinghamshire, United Kingdom; 109 cpm/~g of DNA). After a 16-h hybridization, the filters were washed twice for 20 min in 2 x SSC-0.1% SDS at room temperature and twice for 30 min in 0.1% SDS at 50~ and then autoradiographed.

Determination of LIF Activity (MI Assay). MT-2 cells (1 x 106) were incubated for 3 days in the presence of IL-2 (100 units/ml). The cell supernatants were harvested, centrifuged, and stored under sterile conditions at -20~ until use. The activity of LIF in 2-fold dilutions of MT-2 cell culture medium was monitored by measuring the induction of phagocytic activity in M1-T22 cells essentially as described by To- mida etal. (21). MI cells (5 x 105) were incubated for 2 days in 1 ml of

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LIF GENE EXPRESSION IN HTLV-I INFECTED T-CELLS

Eagle's minimal essential medium containing 10% fetal bovine serum and standard LIF activity. The cells were harvested by centrifugation and suspended in 1 ml of serum free Eagle's minimal essential medium containing a 0.2% (v/v) suspension of polystyrene latex particles (aver- age diameter, 1.02 ~tm; Sekisui Chemicals, Japan). Thereafter, the cells were incubated for 4 h at 37~ The cells that phagocytized more than 10 particles were counted as being phagocytic. Fifty units of LIF were thereby defined as activity giving 50% maximal response in M1 cells under these conditions.

Results

Expression of LIF mRNA in HTLV-1 Infected T-Cell Lines. To investigate the LIF mRNA expression in HTLV-1 infected T-cell lines, Northern blots were analyzed using a 0.6-kilobase fragment of the LIF cDNA probe. As shown in Fig. 1, the LIF mRNA was observed in the HTLV-1 infected T-cell lines MT-1, MT-2, H89-59, H89-79, and H109. Normal human peripheral blood mononuclear cells activated by purified phytohemagglutinin, which expressed high levels of LIF mRNA, were used as a positive control (16). HL-60 cells were used as a

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Fig. 1. LIF mRNA expression in various T-cell lines. A 20-gg sample of total RNA was loaded on each lane and analyzed by Northern blot. LIF transcript (3.6-kilobase) was detectable in HTLV-1 infected T-cell lines. Peripheral blood mononuclear cells were incubated (24 h, 37~ with PHA-P (10 gg/ml) for a positive control (top). Bottom, ethidium bromide stained formaldehyde gel with bands shown as a loading control. PBMC, peripheral blood mononuclear cells; PHA-P, purified phytohemagglutinin.

negative control (16). The LIF transcript was undetectable in the HTLV-1 uninfected T-cell lines, MOLT-4 and TALL-I.

LIF mRNA Enhancement by IL-2. To examine the regulation of LIF mRNA by cytokines, MT-2 cells were cultured in the presence of various cytokines, including IL-I# (250 units/ ml), IL-2 (100 units/ml), IL-6 (200 units/ml), IFN-a (200 units/ ml), and IFN-~, (1000 units/ml), for 24 h and then Northern blotted as described under "Materials and Methods." As shown in Fig. 2,4, IL-2 significantly increased the level of LIF mRNA expression in MT-2 cells. No significant increase in the level of LIF mRNA was detected in the presence ofIL-lfl, IL-6, IFN-a, and IFN--y (data not shown). TPA, 10 nm, significantly increased the expression of LIF mRNA.

Effect of IL-2 on LIF mRNA Kinetics. The time dependent effects of IL-2 on LIF mRNA levels were measured by means of Northern blotting (Fig. 2B). Cells were cultured with IL-2 (100 units/ml) for periods ranging from 3 to 24 h. LIF mRNA started to accumulate at 3 h and peaked at 12 h. LIF mRNA levels decreased at 24 h during continuous exposure to IL-2.

LIF Activity in the Culture Medium of MT-2 Cells. To de- termine whether LIF mRNA expression results in the production of LIF protein, we determined the biological activity of LIF in MT-2 cell supernatants. LIF activity in the culture medium o f 1 • 10 6 MT-2 cells was assayed by means of the differenti- ating activity of a subline of murine myeloid leukemia, M1-T22 cells (Fig. 3). Culture medium treated with IL-2 (100 units/ml) for 3 days induced M1-T22 differentiation. The medium of MT-2 cells cultured with IL-2 contained 40 units/ml of LIF activity.

Down-Regulation of the Expressed LIF mRNA by Steroid Hormones. We then determined whether the expression of LIF mRNA was modulated by 1,25(OH)2D3 and dexamethasone (Fig. 4). Dexamethasone and 1,25(OH)2D3 suppressed the expression of LIF mRNA. This effect of 1,25(OH)2D3 was more potent than that of dexamethasone. TPA in combination with 1,25(OH)2D 3 decreased the accumulation of LIF mRNA, com- pared to TPA alone.

Discussion

In this study, we demonstrated that HTLV-1 infected T-cell lines produce LIF and express the LIF gene. Various human tumor cells, e.g., melanoma, lung adenocarcinoma, bladder car- cinoma, and the T-cell lymphoma, express the LIF gene and secrete LIF into the culture medium (22-24). Five of five HTLV-1 infected T-cell lines expressed LIF mRNA, but uninfected T-cell lines did not express detectable levels of LIF mRNA (Fig. 1). LIF has been purified as human interleukin DA produced by HTLV-1 infected T-cells, C10-MJ2 (25). There- fore, our results suggest that HTLV-1 infection is closely associated with LIF gene expression in T-cells.

The Tax-1 product, which is encoded by the pX region of HTLV-1, activates various cytokine genes, IL-2, IL-3, and granulocyte-macrophage-colony stimulating factor, and the IL-2 receptor a-chain, mediated to the NF-KB-Iike site (26). However, the 5'-flanking region of LIF does not contain the NF-KB-Iike sequence (27). Further studies are required to investigate the role of Tax-1 on LIF gene activation.

A key role of IL-2 in LIF production has been reported in alloreactive human T-clones (28). In our study, IL-2 increased the level of LIF mRNA on MT-2 cells, which peaked at 12 h, and secreted LIF protein. The IL-2 induced LIF mRNA level was increased upon treatment with cycloheximide, suggesting that LIF mRNA stimulated by IL-2 did not require new protein

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Fig. 2. A, effect of IL-2 and 12-O-tetradecanoylphorbol-I 3-acetate (TPA) on LIF mRNA expression in MT-2 cells. MT-2 cells were incubated (12 h, 37~ with recombinant human IL-2 (100 units/ml) and 12-O-tetradecanoylphorbol-13-acetate (10 nM). Total RNA was isolated and analyzed by Northern blot (top). Bottom, ethidium bromide stained formaldehyde gel with bands shown as a loading control. Data are representative of two independent experiments. B, kinetics of LIF mRNA accumulation. MT-2 cells were incubated with recombinant human IL-2 (100 units/ml). Total RNA was isolated at the indicated times and analyzed by Northern blot. Densitometry readings are shown of the blot in B, normalized for the amount of/~-actin in each lane.


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Fig. 3. LIF/D-factor activity secreted by MT-2 cells. MT-2 cells (1 x 106) were incubated for 3 days in either culture medium alone (cont.) or medium with IL-2 (100 units/ml) as indicated. The culture medium was assayed with M1-T22 cells as described in "Materials and Methods." Data are representative of three independent experiments. Bars, SD.

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synthesis . 3 O n the o the r hand, IL-2 and the IL-2 receptor a -cha in are trans-activated by Tax-1 and are cons ide red to de- velop an au tocr ine loop in the early stage o f A T L (29, 30). The induc t ion o f L IF by IL-2 may be no t i m p o r t a n t only for reac- t ions o f the i m m u n e and h e m a t o p o i e t i c sys tems but also for the pa thophys io logy o f A T L pat ients .

Bo th 1 ,25(OH)2D3 and g lucocor t ico id exer t mul t ip le i m m u - noregu la to ry effects (31-33) . Act iva ted T-cells are inh ib i ted by 1 ,25(OH)2D3 and d e x a m e t h a s o n e (31, 32). Previously, we and o thers have repor ted tha t bo th 1 ,25(OH)2D3 and dexa- m e t h a s o n e inhibi t the express ion o f IL-2 or I F N - 7 in act ivated T-cells (33, 34). Moreover , bo th 1 ,25(OH)2D3 and d e x a m e t h - asone suppress pro l i fe ra t ion in s o m e H T L V - 1 infected T-cell l ines (35). We d e m o n s t r a t e d tha t the express ion o f the L IF gene was suppressed by 1,25(OH)2D3 and d e x a m e t h a s o n e (Fig. 4). The suppressive m e c h a n i s m s o f 1 ,25(OH)2D3 and dexame tha -

3 T. Umemiya-Okada and T. Natazuka, unpublished data.

a b c d e f Fig. 4. Effect of steroid hormones on LIF mRNA expression in MT-2 cells.

MT-2 cells were incubated (24 h, 37"C) with control (Lane a), 1,25(OH)2Da (VDs), 10 nM (Lane b), 1,25(OH)2D3, 100 nM (Lane c), 12-O-tetradecanoylphorbol-13-acetate (TPA), 10 nM (Lane d), 1,25(OH)2D3, 10 nM+ 12-O-tetradecanoylphorbol-13-acetate 10 nM (Lane e), or dexamethasone ( Dex), 1 l~l (Lane j'). LIF mRNA expression was determined by Northern blot analysis (top). Bottom, ethidium bromide stained formaldehyde gel with bands shown as a loading control. Data are representative of two independent experiments.

sone on L I F gene express ion may be s imilar to tha t on IL-2 or IFN-~ in act ivated T-cells. However , ne i the r 1 ,25(OH)2Da nor d e x a m e t h a s o n e affected M T - 2 cell g rowth (36). The effect on

6963



LIF GENE EXPRESSION IN HTLV-1 INFECTED T-CELLS

the LIF gene suppression of 1 , 2 5 ( O H ) 2 D 3 and dexamethasone to MT-2 cells remains unclear.

LIF gene expression is induced by TPA in several T-cell lines 14. (22, 25). TPA also induced high levels of the LIF gene in MT-2 cells (Fig. 2). TPA and 1,25(OH)2D3 synergistically induced 15. LIF mRNA accumulation in human monocytes but not lymphocytes (16, 37). In this study, no synergistic effect of 1,25- ( O H ) 2 D 3 and TPA was observed in HTLV-1 infected T-cells 16. (Fig. 4).

With regard to hypercalcemia in ATL patients, the production of TNF-~, IL-I~, and parathyroid hormone related protein 17. in HTLV-1 infected T-cells has been reported and implicated in accelerated bone resorption (8-11). Abe et al. (15) have shown that LIF as well as IL-1, TNF-a, TNF-/3, and PTH-rp have 18. OAF activity. The bone resorption activity of LIF is synergistically increased by combination with IL-1 and IL-6 (38). We therefore consider that LIF is an additional factor which in- duces hypercalcemia, in conjunction with other cytokines and abnormalities in the immune systems of ATL patients.

Recently, it has been reported that HTLV-II infected T-cells 19. produce Oncostatin M (39), which is related to LIF in structure, chromosomal localization, biological activity (40), and receptor

20. systems (41). These cytokines may provide another explanation for the pathophysiology of ATL.

Acknowledgments 21.

We are grateful to Dr. N. Gough for providing LIF cDNA; to Dr. I. Miyoshi for the MT-2 cell line; to Dr. M. Tomida for the M I - T 2 2 cell line; to Dr. T. Saida for the H89-59, H89-79, and H I 0 9 cell lines; and to Dr. T. Tsukamoto and Dr. H. Nakata for helpful discussions.

22.

23.

References 1. Uchiyama, T., Yodoi, J., Sagawa, K., Takatsuki, K., and Uchino, H. Adult 24.

T-cell leukemia. Clinical and hematological features of 16 cases. Blood, 50: 481-492, 1977.

2. Robert-Guroff, M., Nakao, Y., Notake, T., Ito, Y., and Gallo, R. C. Natural antibodies to human retrovirus HTLV in a cluster of Japanese patients with 25. adult T cell leukemia. Science (Washington DC), 215: 975-978, 1982.

3. Takatsuki, K. Adult T-cell leukemia-lymphoma. Prog. Immunol., 5:1103- 1108, 1984.

4. Popovic, M., Reitz, M. S., Jr., Sarngadharan, M. G., Robert-Guroff, M., 26. Kalyamaraman, V. S., Nakao, Y., Miyoshi, I., Minowada, M., Yoshida, M., Ito, Y., and Gallo, R. C. The virus of Japanese adult T cell leukemia is a member of the human T cell leukemia virus group. Nature (Lond.), 300: 27. 63-66, 1982.

5. Bunn, P. A., Schechter, G. P., Jaffe, E., Blayney, D., Young, R. C., Mat- thews, J. J., Blattner, W., Broder, S., Robert-Guroff, M., and Gallo, R. C. Clinical course of retrovirus-associated adult T-cell lymphoma in the United 28. States. N. Engl. J. Med., 309: 257-264, 1983.

6. Grossman, B., Schechter, G. P., Horton, J. E., Pierce, L., Jaffe, E., and Wahl, L. Hypercalcemia associated with T-cell lymphoma-leukemia. Am. J. Clin. Pathol., 75: 149-155, 1981.

7. Fujihira, T., Eto, S., Sato, K., Zeki, K., Oda, S., Chiba, S., and Suzuki, H. 29. Evidence of bone resorption-stimulating factor in adult T-cell leukemia. Jpn. J. Clin. Oncol., 15: 385-391, 1985.

8. Wano, Y., Hattori, T., Matsuoka, M., Takatsuki, K., Chua, A. O., Gubler, U., and Greene, W. C. Interleukin 1 gene expression in adult T cell leukemia. 30. J. Clin. Invest., 80:911-916, 1987.

9. Shirakawa, F., Ymashita, U., Tanaka, Y., Watanabe, K., Sato, K., Haratake, J., Fujihira, T., Oda, S., and Eto, S. Production of Bone-resorbing activity corresponding to interleukin-la by adult T-cell leukemia cells in humans. 31. Cancer Res., 48." 4284-4287, 1988.

10. Motokura, T., Fukumoto, S., Takahashi, S., Watanabe, T., Matsumoto, T., Igarashi, T., and Ogata, E. Expression of parathyroid hormone-related pro- 32. tein in a human T cell lymphotropic virus type-I infected T cell line. Bio- chem. Biophys. Res. Commun., 154:1182-1188, 1988.

11. Tschachler, E., Robert-Guroff, M., Gallo, R. C., and Reitz, M. S. Human 33. T-lymphotropic virus-infected T cells constitutively express lymphotoxin in vitro. Blood, 73: 194-201, 1989.

12. Fukumoto, S., Matsumoto, T., Watanabe, T., Takahashi, H., Miyoshi, I., and 34. Ogata, E. Secretion of parathyroid hormone-like activity from human T-cell lymphotropic virus type-l-infected lymphocytes. Cancer Res., 49: 3849- 3852, 1989.

13. Ishibashi, K., Ishitsuka, K., Chuman, Y., Otsuka, M., Kuwazuru, Y., Iwa- 35.

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hashi, M., Utsunomiya, A., Hanada, S., Sakurami, T., and Arima, T. Tumor necrosis factor-~ in the serum of adult T-cell leukemia with hypercalcemia. Blood, 77: 2451-2455, 1991. Gearing, D. P. Leukemia inhibitory factor: does the cap fit? Ann. NY Acad. Sci., 628: 9-18, 1991. Abe, E., Ishimi, Y., Takahashi, N., Akatsu, T., Ozawa, H., Yamana, H., Yoshiki, S., and Suda, T. A differentiation-inducing factor produced by the osteoblastic cell line MC3T3-EI stimulates bone resorption by promoting osteoclast formation. J. Bone Miner. Res., 3: 635-645, 1988. Anegon, I., Moreau, J. F., Godard, A., Jacques, Y., Peyrat, M. A., Hallet, M. M., Gordon, W., and Soulillou, J. P. Production of human interleukin for DA cells (HILDA)/leukemia inhibitory factor (LIF) by activated monocytes. Ceil. Immunol., 130: 50-65, 1990. Abe, T., Murakami, M., Sato, T., Kajiki, M., Ohno, M., and Kodaira, R. Macrophage differentiation inducing factor from human monocytic cells is equivalent to murine leukemia inhibitory factor. J. Biol. Chem., 264: 8941- 8945, 1989. Gallo, R. C., Kalynaraman, V. S., Sarngadharan, M. G., Silski, A., Vonder- held, E. C., Maeda, M., Nakao, Y., Yamada, K., Ito, Y., Gutensohn, N., Murphy, S., Bunn, P. A., Catovsky, D., Greaves, M. F., Blayney, D. W., Blattner, W. A., Jarett, W. F. H., zur Hausen, H., Seligman, M., Brouet, J. C., Haynes, B. F., Jegasothy, B. V., Jaffe, E., Cossman, J., Broder, S., Fisher, R. I., Golde, D. W., and Robert-Guroff, M. Association of human type-C retrovirus with a subset of adult T cell cancers. Cancer Res., 43: 3892-3899, 1983. Gough, N. M., Gearing, D. P., King, J. A., Willson, T. A., Hilton, D. J., Nicola, N. A., and Metcalf, D. Molecular cloning and expression of the human homologue of the murine gene encoding myeloid leukemia inhibitory factor. Proc. Natl. Acad. Sci. USA, 85: 2623-2627, 1988. Chomczynski, P., and Sacchi, N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Bio- chem., 162: 156-159, 1987. Tomida, M., Yamamoto-Yamaguchi, Y., and Hozumi, M. Purification of a

factor inducing differentiation of mouse myeloid leukemic M1 cells from conditioned medium of mouse fibroblast L929 cells. J. Biol. Chem., 259: 10978-10982, 1984. Gascan, H., Godard, A., Ferenz, C., Naulet, J., Praloran, V., Peyrat, M. A., Hewick, R., Jacques, Y., Moreau, J. F., and Soulillou, J. P. Characterization and NH2-terminal amino acid sequence of natural human interleukin for DA cells: leukemia inhibitory factor. J. Biol. Chem., 264: 21509-21515, 1989. Gascan, H., Anegon, I., Praloran, V., Naulet, J., Godard, A., Soulillou, J. P., and Jacques, Y. Constitutive production of human interleukin for DA cells/ leukemia inhibitory factor by human tumor cell lines derived from various tissues. J. Immunol., 144: 2592-2598, 1990. Mori, M., Yamaguchi, K., Honda, S., Nagasaki, K., Ueda, M., Abe, O., and Abe, K. Cancer cachexia syndrome developed in nude mice bearing melanoma cells producing leukemia-inhibitory factor. Cancer Res., 51: 6656- 6659, 1991. Moreau, J. F., Donaldson, D. D., Bennett, F., Witek-Giannotti, J., Clark, S. C., and Wong, G. G. Leukemia inhibitory factor is identical to the myeloid growth factor human interleukin DA cells. Nature (Lond.), 336." 690-692, 1988. Smith, M. R., and Greene, W. C. Molecular biology of the type 1 T-cell leukemia virus (HTLV-1) and adult T-cell leukemia. J. Clin. Invest., 87: 761-766, 1991. Stahl, J., Gearing, D. P., Willson, T. A., Brown, M. A., King, J. A., and Gough, N. M. Structural organization of the genes for murine and human leukemia inhibitory factor. J. Biol. Chem., 265: 8833-8841, 1990. Moreau, J. F., Borreville, M., Peyrat, M. A., Jacques, Y., and Soulillou, J. P. Capacity of alloreactive human T clones to produce factor(s) inducing proliferation of the IL-3-dependent DA-1 murine cell line. I. Evidence that this production is under IL-2 control. Ann. Inst. Pasteur Immunol., 137: 25-37, 1986. Maruyama, M., Shibuya, H., Harada, H., Hatakeyama, M., Seiki, M., Fujita, T., Inoue, J., Yoshida, M., and Taniguchi, T. Evidence for aberrant activation of the interleukin-2 autocrine loop by HTLV-l-encoded p40 x and T3/Ti complex triggering. Cell, 48: 343-350, 1989. Cross, S. L., Feinberg, M. B., Wolf, J. B., Holbrook, N. J., Wong-Staal, F., and Leonard, W. J. Regulation of the human interleukin-2 receptor a chain promotor: activation of a nonfunctional promotor by the transactivator gene of HTLV-1. Cell, 49: 47-56, 1987. Provvedini, D. M., Tsoukas, C. D., Deffos, L. J., and Marolagas, S. Dehy- droxyvitamin D3 receptors in human leukocytes. Science (Washington DC), 221: 1181-1183, 1983. Matsui, T., Nakao, Y., Koizumi, T., Nakagawa, T., and Fujita, T. Regulation of activated T lymphocyte subsets by 1,25-Dihydroxyvitamin D3. Life Sci., 37: 95-101, 1985. Arya, S. K., Wang-Staal, F., and Gallo, R. C. Dexamethasone-mediated inhibition of human T cell growth factor and v-interferon mRNA. J. Immu- nol., 132: 273-276, 1984. Matsui, T., Takahashi, R., Nakao, Y., Koizumi, T., Katakami, Y., Mihara, K., Sugiyama, T., and Fujita, T. 1,25-Dihydroxyvitamin D3-regulated expression of genes involved in human T-lymphocyte proliferation and differentiation. Cancer Res., 46: 5827-5831, 1986. Koizumi, T., Nakao, Y., Kawanishi, M., Maeda, S., Sugiyama, T., and Fujita,




T. Suppression of c-myc mRNA expression by steroid hormones in HTLV- 1-infected T-cell line, KH-2. Int. J. Cancer, 44: 701-706, 1989.

36. Nakao, Y., Koizumi, T., Matsui, T., Matsuda, S., Nakagawa, T., Katakami, Y., Fujita, T., Maeda, S., Morikawa, S., and Ito, Y. Effect of 1,25-dihydroxyvitamin D3 on proliferation of activated T cells and established human lymphotropic virus type 1-positive cell lines. J. Natl. Cancer Inst., 70:1079- 1086, 1987.

37. Anegon, I., Grolleau, D., and Soulillou, J. P. Regulation ofHILDA/LIF gene expression in activated human monocytic cells. J. Immunol., 147: 3973- 3980, 1991.

38. Ishimi, Y., Abe, E., Jin, C. K., Miyaura, C., Hong, M. H., Oshida, M., Kurosawa, H., Yamaguchi, Y., Yomida, M., Hozumi, M., and Suda, T. Leukemia inhibitory factor/differentiation-stimulating factor (LIF/D-factor): regulation of its production of and possible roles in bone metabolism. J. Cell.

Physiol., 152: 71-78, 1992. 39. Nair, B. C., DeVico, A. L., Nakamura, S., Copeland, T. D., Chen, Y., Patel,

A., O'Neil, T., Oroszlan, S., Gallo, R. C., and Sarngadharan, M. G. Identi- fication of major growth factor for AIDS-Kaposi's sarcoma cells as Oncos= tatin M. Science (Washington DC), 255: 1430-1432, 1992.

40. Rose, T. M., and Bruce, A. G. Oncostatin M is a member ofa cytokine family that includes leukemia-inhibitory factor, granulocyte colony-stimulating factor, and interleukin 6. Proc. Natl. Acad. Sci. USA, 88: 8641-8645, 1991.

41. Gearing D. P., Comeau, M. R., Friend, D. J., Gimpel, S. D., Thut, C. J., McGourty, J., Brasher, K. K., King, J. A., Gillis, S., Mosley, B., Ziegler, S. F., and Cosman, D. The IL-6 signal transducer, gpl30: an Oncostatin M receptor and affinity converter for the LIF receptor. Science (Washington DC), 255: 1434-1437, 1992.

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1992;52:6961-6965. Cancer Res Tomoyo Umemiya-Okada, Toshiki Natazuka, Toshimitsu Matsui, et al. Infected T-Cell LinesFactor/D Factor Gene in Human T-Cell Leukemia Virus Type 1 Expression and Regulation of the Leukemia Inhibitory

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Expression and Regulation of the Leukemia Inhibitory ... · ically associated with HTLV-1 infection (1, 2). ATL frequently causes hypercalcemia, which may become an important factor

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