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production in cloned somatic cell hybrids between two human lymphoid lines. Cell 11: 139 - Rutherford TR, Clegg JB, Weatherall DJ (1979) K562 human leukaemic cells synthesize embryonic haemoglobin in response to haemin. Nature 280:164 - Singer PA, Williamson AR (1979) A novel interaction involving a polypeptide chain (P33) in covalent linkage with IgM on the surface of a Burkitt lymphoma cellline (Daudi). Eur J Immunol 9:224 - Steel CM, Woodward MA, Davidson C, Philipson J, Arthur E (1977) Non-random chromosome gains in human lymphoblastoid cell lines. Nature 270:349 - Steinitz M, Klein G (1975) Comparison between growth characteristics of an Epstein-Barr virus (EBV)-genome negative lymphoma li ne and its EBV -converted sub li ne in vitro. Proc Natl Acad Sci USA 72 :3518 -SteinitzM, Klein G (1976) Epstein Barr virus (EBV)-induced change in saturation density and serum dependence of established, EBVnegative lymphoma lines in vitro. Virology 70: 570 - Steinitz M, Klein G (1977) Further studies on the differences in serum dependence in EBV -negative lymphoma lines and their in vitro converted, virusgenome carrying sublines. Eur J Cancer 13: 1269 - Steinitz M, Klein G, Koskimies S, Mäkelä 0(1977) EB virus-induced B-Iymphocyte celllines producing speeific antibody. Nature 269 :420 - Steinitz M, Koskimies S, Klein G, Mäkelä 0 (1978) Establishment of specific antibody produeing human celliines by antigen preselection and EBV transformation. Curr Top Microbiol Immunol81: 156 -Steinitz M, Seppälä I, Klein G, Koskimies S, Mäkelä 0(1979) Immunobiology 156 :41-Steinitz M, Izak G, Cohen S, Ehrenfeld M, Flechner J (to be published) Continuous production of rheumatoid factor by in vitro EBV-transformed lymphocytes. Nature - Tanaka A, Nonoyama M, Glaser R (1977) Transcription of latent Epstein-Barr virus genomes in human epithelial/Burkitt hybrid cells. Virology 82:63 - Tjian R, Fey G, Graessmann A (1978) Biological activity of purified simian virus 40 T antigen proteins. Proc Natl Acad Sei USA 75: 1279 - Van den Berghe, H, Parloir C, Gosseye S, Englebienne V, Cornu G, Sokal G (1979) Variant translocation in Burkitt lymphoma Cancer Genet Cytogenet 1:9 - Van Furth R, Gorter H, Nadkarni JS, Klein E, Clifford P (1972) Synthesis of immunoglobulins by biopsied tissues and cell lines from Burkitt's lymphoma. Immunology 22: 847 - Yefenof E, Klein G (1976) Difference in antibody induced redistribution of membrane IgM in EBV -genome free and EBV -positive human lymphoid cells. Exp Cell Res 99: 175 - Yefenof E, Klein G, Ben-Bassat H, Lundin L (1977) Differences in the ConA-induced redistribution and agglutination patterns of EBV genome-free and EBV -carrying human lymphoma lines. Exp Cell Res 108: 185 - Zech L, Haglund U, Nilsson K, Klein G (1976) Characteristicchromosomal abnormalities in biopsies and lymphoid celllines from patients with Burkitt and non-Burkitt lymphomas. Int J Cancer 17: 47 - Zeuthen J, Nilsson K (1976) Hybridization of a human myeloma permanent cell line with mouse cells. Cell Differ 4:355 - Zeuthen J, Friedrich U, Rosen A, Klein E (1977) Structural abnormalities in chromosome 15 in cell lines with reduced expression of Beta-2 microglobulin. Immunogenetics 4: 567 - Zeuthen J, Klein G, Ber R, Masucci G, Bisballe S, Povey S, Terasaki P, Ralph P (to be published) Human lymphoma/lymphoma and lymphoma/leukemia hybrids. 1. Isolation, characterization, cell surface and B-cell marker studies. J Natl Cancer Inst - Zur Hausen H, Bornkamm GW, Schmidt R, Hecker E (1979) Tumor initiators and promotors in the induction of Epstein-Barr virus. Proc Natl Acad Sei USA 76:782 190
Haematology and Blood Transfusion Vol. 26 <strong>Modern</strong> Trends in Human Leukemia IV Edited by Neth, Gallo, Graf, Mannweiler, Winkler © Springer-Verlag Berlin Heidelberg 1981 Epstein-Barr Virus: Its Site of Persistence and Its Role in the Development of Carcinomas H. Wolf, E. Wilmes and G. J. Bayliss A. Introduction Epstein-Barr virus (EBV) causes in man infectious mononucleosis as a "primary" disease. Burkitt's Lymphoma and Nasopharyngeal Carcinoma may be considered as "secondary" diseases developing only in persons with long preceeding EBV infection. Following the primary infection with EBV, virus can be isolated from the salivary duct (Morgan et al. 1979) and from throat swabs for months or years (Gerber et al. 1972; Chang and Golden 1971), and B-Iymphocytes are lifelong carriers of EBV genomes (Nilsson et al. 1971) and can be used to establish lymphoblastoid celllines (Pope 1967; Rickinson et al. 1980). For the nasopharyngeal carcinoma, where serology (for review see Henle and Henle 1979) suggested that EBV is regularly associated with the disease, it has been shown by in situ hybridization (Wolf et al. 1973) and other unrelated techniques involving cell separation (Desgranges et al. 1975) or passage through nude mice (Klein et al. 1974) that EBV is indeed regularly present in the epithelioid tumor cells. The ability of EBV to induce tumors in nonhuman primates (Shope et al. 1973; Wolfe and Deinhardt 1978) and the proof of EBV DNA in tumors from these animals (Wolf et al. 197 5b) have added further evidence for an etiologic relationship of EBV to the development of neoplasia in man. B. Results I. Site 01 EBV Persistence in the Body Epstein-Barr virus is associated with B-Iymphocytes (Pope 1967). It is, however, not clear whether spontaneous activation of EBV genomes in carrier lymphocytes, whieh are present in the lymphocyte rieh area of the oropharynx, is responsible for lifelong persistence of antibody titers directed against EBV -related antigens and the shedding of virus into the oropharynx or whether EBV resides in addition in specific sites of the body and is produced there, resembling somehow the situation of Marek's disease virus of chicken. That virus is found in T-Iymphocytes and produced and spread from the epithelium of feather follicles (Calnek and Hitchner 1969). In situ hybridizations using sections from the parotic gland as weIl as from tonsils and other tissues of the ear-nose-throat area were performed in an attempt to find cells which harbor EBV genomes. EBV producing cells (P3HR1), EBV carrier Raji cells containing about 50 EBV genomes/cell, and EBV genome negative BJA-B cells were included as controls. Figure 1 shows that only cells in the sections from the parotic gland contained detectable numbers of grains indicating the presence of EBV DNA. The number of grains per cell seems to be in the range of or high er than that seen in Raji cells whieh carry 50 viral genomes per ceIl, though clearly lower than that seen in the virus producing P3HR1 cells. The fact that no producer-type cells were found had to be expected, as the biopsies were taken from healthy adults for reasons not related to EBV infections. The induction of a lytic cycle has to be a rare event, since it leads to cell destruction and would, if it happened on a larger scale, cause necrosis. Because in situ hybridization, in some cases, may give artifactual results, a confirmation of the results with an independent method would 191
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Haematology and Blood Transfusion 2
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Dr. Rolf Neth, Universitäts-Kinder
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Participants of the Meeting Aaronso
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Kurth, R, Friedrich-Miescher-Labora
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Wilsede Scholarship Holders (Grante
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Preface Gut ist eine Lehrart, wo ma
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Personal and scientific discussion
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Acknowledgement We should like to t
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Wilsede, June 21 1978 Memorial Trib
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limiting benign lymphoproliferative
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this most unlikely, however (for re
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longer subject to the same control
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Bregula U, Wiener F, Harris H (1971
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fresh lymph node biopsies from ten
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Fig. 3. Binucleate mitosis in an ob
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y Fig. 4. Schematic diagram of post
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than that among patients with Stage
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Tests of binding affinity, agglutin
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N Engl J Med 297: 963-968 - Green I
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Clinical Aspects
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"ring chromosome". "Mar" is marker,
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191 chromosomally abnormal patients
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tumors of both African and non-Afri
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Haematology and Blood Transfusion V
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Retrospective group 26/93 evolved l
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Treatment
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advantage in terms of duration of f
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leukaemia. An analysis of 168 patie
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11. Treatment Program Chemotherapy
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efractory congestive heart failure
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period (Sequence II) was gene rally
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children with a multiple drug proto
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DAYS 1 5 10 15 20 25 30 35 , I I I
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Biologically Drug Outcome fit" sens
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may counterbalance the potential be
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Ht (0--0) 50 40 30 20 10 0 Platelet
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10 7 ,-----------------------------
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11. Haematological and Biochemical
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0\ 0\ INTERFERON ... 10 3 rl E "'
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("pre-B" phenotype) has been descri
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human bone marrow cells exhibit the
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10 9.,0 MBKCC .. .,4 B prot:oool 1'
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demann W, Büehner T, Arlin Z, Gee
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-...l 00 a-tLDRENS CAt«:ER S TU>Y
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prognostic characteristics under in
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significantly higher in HR} (14.5%,
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normal or elevated immunoglobulin l
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References Andreeff M, Darzynkiewic
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R GROUP 1 ( 1970 - 1971 ) 17 PTS. R
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Table 3. Influence of initial featu
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LIJ (f) a.. < -1 LIJ Q: u. 0 >- I-
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me nt indicated that additional the
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42:2123-2134 - Chessells JM, Cornbl
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Baum et al. 1979). This study was b
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1. 0 '--'1--" 0.9 e: o .- VI VI E C
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e: o e: 1. 0 ,.--...-- 1'"'\ ......
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In conclusion, we can state that th
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Haematology and Blood Transfnsion V
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INDUCTION VCR + PRED + ASNASE (wk 3
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SCHED.INT 1 MTX • ! Adria MTX •
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~ looh---------ooooc>----o---c>----
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Consoll- Irra .tlon t) .tlon Indu
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Highrisk Standard risk Table 1. Cli
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Cumulo/ive comp/el9 remission 1.0 .
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10° 8 = 6 ö 4 Non T Cell .~ :ö
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% SURVIVAL 100 ~ ~II 0 80 60 40 - -
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a) Percentage of donor cell mitoses
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Table 5. Results of bone marrow tra
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...... w N Table 7. Clinical result
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agent had to be added and that in t
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Page 151 and 152: C. Chronic Myelogenous Leukemia The
Page 153 and 154: Patients with a suitable donor rece
Page 155 and 156: No. Recurrent Leukaemia Other death
Page 157 and 158: followed by dialysis against isoton
Page 159 and 160: Haematology and Blood Transfusion V
Page 161 and 162: HS, HL- Heme >_ 40 0 HS I ;:. =====
Page 165 and 166: knowing at present. These genes may
Page 167 and 168: esistance to antifolates. In Vitro
Page 169 and 170: a DNA probe specific for the gene t
Page 171 and 172: translocation between chromosomes 9
Page 173 and 174: Table 1. Subdivision of 1827 Myelo-
Page 175 and 176: NORMAL CELL ®
Page 177 and 178: some change of the primary type, in
Page 181 and 182: F. Transformation of Mouse Bone Mar
Page 185 and 186: were carried out with 3H-MTX in the
Page 189 and 190: the lysozyme cDNA prepared from ovi
Page 193 and 194: et al. 1976). The BJAB cellline in
Page 195 and 196: dalton bands corresponding to light
Page 197 and 198: 1979). The EBV-carrying cell lines
Page 199 and 200: producer lines (DAUDI and P3HR-1),
Page 201: G, Giovanella B, Westman A, Stehlin
Page 205 and 206: Dc;bt CT + i J.I9 Raji; 1. i .. 100
Page 207 and 208: c. Discussion During infectious mon
Page 211 and 212: C. Nonepisomal Viral DNA We have us
Page 213 and 214: A 260 3.0 A B 1670 70 N2 2.0 c: E .
Page 215 and 216: vivo. Nature 260:302-306 - Kirk JTO
Page 217 and 218: Fig. 1. a Polyacrylamide gelelectro
Page 221 and 222: 1979). The lymphocyte subpopulation
Page 223 and 224: lysin 0 antigens are found, and mix
Page 225 and 226: Pope JH (1978) Long-term T cell-med
Page 227 and 228: Haematology and Blood Transfnsion V
Page 229 and 230: Charaeteristie Morphologie maturati
Page 231 and 232: after TPA-induction (Table 3). The
Page 233 and 234: PA (1978) Reactivity of a rabbit an
Page 235 and 236: Blood Monocyte. B lood Monocytes 1-
Page 239 and 240: and nonhistone proteins can serve a
Page 243 and 244: The findings of the evaluation of s
Page 245 and 246: Fig. 3. Tumor grawth of L 428 cells
Page 247 and 248: Cell biological and Immunological A
Page 249 and 250: consideration. Sanel (1973) obtaine
Page 251 and 252: endotoxin serum, were used. A parti
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monocytic leukemia cell line in cul
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B. Isolation, Characterization and
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don al hemopathies generally displa
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nitors. Blood Cells 6:595-607 - Min
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\I) ....I ....I IU Jl 200 r 100 90
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their haemopoietic cells' ability t
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normal normal ALL ALL ALL persons p
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64.5 { V)
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References Biermann E, Neth R, Gros
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ehambers, the growth pattern observ
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References Anderssen LC, 10kinen M,
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REH CELL UNE TO CALLb~Ö ~q~ V.M. c
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the macrophage series in which the
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Table 1. Production of factor depen
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Table 1. Characteristics of the adh
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Fig. 3. A human marrow culture at 6
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300 250 • HYPERPROLIFERATIVE PATT
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Fig. 9. Nonadherent population from
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The cobblestone areas were often no
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L (1976) Induction of colony format
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B. Materials and Methods J. Tissue
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Fig. 2. Ultrastructural appearance
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Page 307 and 308:
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Table 1. Monoclonal antibody reacti
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al. 1979; Janossy et al. 1979) is e
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to which this is an exact replica o
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inger JL (1976) Distribution of la-
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I region antigens were found to be
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have been reported to be present on
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V. FunctionalAssays Assays for PHA
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Functional studies of UCHT1 separat
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40 ,...... (5 L.. C o u 30 .9 ~ .~
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with immunohistochemieal methods (L
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can be dissected by cloning. Utiliz
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Table 1. Origin and marker profile
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complete identity of gp 100 from no
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Table 1. Reaction of single anti se
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media. Following 24-h incubation, c
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nonleukemic eells (Greaves 1979). L
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Fig. 2. Cytocentrifuged smears of b
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Modification of amino-groups of hum
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disturb this balance in the directi
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The majority of human blood lymphoc
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of the EBV infected B cells. Howeve
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and immunoglobulins. J Immunol 120:
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" K 10 • j ~ ALL RNlL AL wtfh les
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tion akuter Leukämiezellen in "spo
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D. Results and Discussion J. Acute
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selectively responsive to different
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l1J ~ 100 ~ Q. ::) l1J Z 0 ~ z >- .
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Table 2. Growth inhibition of S49 e
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Reverse Infectious ASC/2X1Q6 transc
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munized animals to 1316 tumor cells
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Haematology and Blood Transfnsion V
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Fig. 3. Expression of retroviral gp
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pg70. J Exp Med 143: 151-166 - McGr
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Table 1. Expression of tumor antige
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Virological and Molecularbiological
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Table 1. Oncogenic properties of re
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also Fig. 1). It followed that the
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whether this sequence is related to
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zed in infected cells argue that th
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a specific viral transforming prote
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detect viral RNA as the only charac
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Eisen H (1980) Myeloproliferate vir
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p,60- -34K - 1 2 3 4 Fig. 1. In vit
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.. p - • a'T ... I .t'l Fig. 4. D
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Fig. 2. Photomicrographs of NY68 in
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DEAE (o(lJmn 'liI GI -2 2 o e s 1
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vity was deterrnined. For endogenou
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pr 180 - - - P 12/15 - A B c D E F
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85,- 66- -27 - -66 40- 27- -1'9 19-
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= 0.5 o E Q. o ILI 0.4 (J) « ILI .
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Table 1. Oncogenic potential of avi
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100 III GI c: o "8 80 > w
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Expt. No. Infecting virus Proportio
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Haematology and Blood 'fransfusion
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sequences, provided its gag portion
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77:3009-3013 - Hu SSF, Moscoviei C,
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RI 2.2 md and Hind III 2.6 md leuke
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Page 443 and 444:
RELATIONSHIP OF ENDOGENOUS AND EXOG
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E ........ E Cl. U E :::l .... Q) (
Page 447 and 448:
References Astrin S (1978) Endogeno
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the same animal were examined (e.g.
Page 451 and 452:
I) 'IQ a \0 - CL -- Fig. 2. Restrie
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~ ..................... ~ Cell 3' 5
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a :Sa,C! + . ~ 111 i' ( " '~ .J. b
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Table 1 Sampie Tissue TS fragment"
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a b Sac I / rep ._-_.-_ K J nl rep
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Haematology and Blood 'fiansfusion
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Osteopetrosis and osteosarcomas occ
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Lymphoma cellline Table 1. T and B
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Table 2. In vivo growth and oneogen
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Table 1. Transforming aetivity of c
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Table 3. Transformation aetivity of
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- Shoemaker C, Goff S, Gilboa E, Pa
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indieate a shifting of target eell
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Table 2. Expression of Jl and Sourc
Page 481 and 482:
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~ i ,..1---------,,':::::::::::::::
Page 485 and 486:
fragment of pBR322 DNA, a 3.0-kbp S
Page 487 and 488:
Mak TW (1979) Proc Natl Acad Sci US
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Table 1. Transformation of 3T3 cell
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inserts in the genome of rapidly tr
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pulations, it seemed likely that th
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contains antigens which react with
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le of reacting in sensitive radioim
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gic approach. In: Hiatt HH, Watson
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12 Eco R [ Fig. 1. Hybridization pa
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10 - '1~ )( -~ S:! E A U I o 1 2
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Haematology and Blood 'fransfusion
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cultures. These cells did not conta
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indication of malignant T-cells in
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Fig. 3. Thin-section electron micro
Page 519 and 520:
Table 3. Lack of detectable related
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J. HTLV Was Present in the Primary
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specific signals into nonspecific s
Page 525 and 526:
Table 2. Distribution of HTLV-relat
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u 10 ~ 20 ~ :c 30 a 40 o Fig. 1. Ki
Page 529 and 530:
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SSV TrS-gp labeled effectively with
Page 533 and 534:
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60 HFF/SSV cDNA CELL RNAs: 0 HF/SSV
Page 537 and 538:
Antisera a Table 1. Immunofluoresce
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Haematology and Blood 'ftansfusion
Page 541 and 542:
y competition RIA yielded virtually
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SiS'V' gp70: ANTIGEN, ANTI BODY, IM
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type-C virus p30 antigen in cells f
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c. Results The fractionated whole-b
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induced antibodies as well as exper
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- Fig. la-f. Retravirus particles p
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eplication of infecting murine leuk
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immunodeficiency, multic10nal lymph
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Bovine leukemia --, proteins of 499
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Human T- cell - -, characterization
Page 563 and 564:
RNA -, of tumor virus 439 Rous sarc
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Haematology andBlood Transfusion Su
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