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A B c -----95 IK ------ 7 I OK Fig. 1. SDS polyacrylamide gel electrophoresis of (A) FOCMA-L moleeules purified by immunoaffinity chromatography of a feline LSA (FeLV nonproducer ) over a column of Sepharose containing bound IgG from cat FOCMA serum, (B) FOCMA-S moleeules purified over the same column from a lysate of GA-FeSV mink cells (64F3C17 cells), and (C) P95 gag - x molecules purified on an anti-FeLV IgG immunoaffinity column from a lysate of 64F3C17 cells. Aliquots of material binding to the columns and eluted with 4 MMg0 2 were radioiodinated and subjected to electrophoresis in a 5 %-20% acrylamide slab gel and visualized by autoradiography. cipitation with specific antisera to individual proteins, (2) glycosylation and phosphorylation (Snyder et al. , to be published a), (3) similarities in peptides genera ted by partial digestion with S. aureus V8 protease or chymotrypsin (Snyder et al., to be published b), (4) associated protein kinase activity (Collett and Erikson 1978; Sen and Todaro 1979; Snyder et al. , to be published b), and (5) associated glucose binding and/or transportstimulating activity (Lee and Lipmann 1977). In terms of this preliminary characterization (Table 1) the purified p70 moleeules from LSA cells and GA-FeSV-transformed mink fibroblasts were indistinguishable. However, the gag-x polyprotein was distinguishable by its labe1ing with 32p-orthophosphate, its association with a protein kin ase activity, and its precipitability with antibodies in a hyperimmune antiserum to FeLV structural proteins but not with antibodies in a well-characterized cat-derived natural FOCMA serum. C. Discussion Previously, two independent investigators showed that absorption of certain cat FOCMA sera with FeLV structural proteins did not diminish immunoflourescent antibody reactivity for FOCMA-S on FeSV-transformed mink cells, while absorption with partially-purified gag-x polyproteins was effective (Stephenson et al. 1977; Sherr et al. 1978a). This was the most direct evidence for a link between FOCMA and the "x" portion of the FeSV polyprotein. Indirect supportive evidence came from an experiment wherein hyperimmune rabbit antibody to FeSV gag-x-containing pseudotype virions, made specific for "x" by absorption with helper virus proteins, stained the surface of FL 74 LSA cells (Sherr et al. 1978b). Khan et al. (1978) showed that many FeLV -absorbed cat FOCMA sera were capa- FOCMA-Lp70 GA-FeSV Table 1. Comparison of proteins induced in cells transformed by FeLV and FeSV gag-x p70 Celliocalization Plasma Plasma Plasma membrane membrane membrane Molecular weight 65-70K 95K 65-70K Associated FeLV Ag None p15p12(p30) None Protease digest maps Similar ? Similar Phosphorylation Glycosylation + Associated protein No Yes No Kinase activity Associated glucose No ? No Binding activity Associated glucose transport stimulating No activity ? No 489
le of reacting in sensitive radioimmunoprecipitation assays with a highly purified gag-x protein probe. However, the fact that not aB such sera displayed this reactivity was the first suggestion that there may not be complete concordance between recognition of FOCMA and recognition of "x". Recent experiments by Barbacid et al. (1980) and Ruscetti et al. (to be published) as weIl as those described in the present report are consistent with this lack of complete concordance. While we have not ruled out the possibility that the p70 molecule we have isolated from GA-FeSV-transformed mink cells is a breakdown product of unstable P9S gag -X, for the present p70 appears to be a good alternate candidate for a FOCMA-bearing protein in these cells. By extrapolation from all of the available evidence one might speculate that FOCMA, as defined in terms of a tumor regression-correlated cat antibody re action with the surface of FL 74 LSA cells, may actually be a family of antigens, not all of which cross react with FeSV polyprotein antigens. Acknowledgments This research was supported by grants CA-16599 and CA-24357 and by NCI core grant CA-08748 from the National Institute of Health. The work was carried out while H.W.S., Jr. was a Special Fellow of the Leukemia Society of America. References Barbacid M, Lauver AV, Devare SG (1980) Biochemical and immunological characterization of polyproteins coded for by the McDonough, Gardner Arnstein, and Snyder-Theilen strains of feline sarcoma virus. J Virol 33: 196-207 - Collett MS, Erikson RL (1978) Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci USA 75 :2021-2024 - Essex M, Sliski A, Cotter SM, Jakowski RM, Hardy WD Jr (1975) Immunosurveillance of naturally occurring feline leukemia. <strong>Science</strong> 190:790-792 - Essex M, Cotter SM, Stephenson JR, Aaronson SA, Hardy WD Jr (1977) Leukemia, lymphoma, and fibrosarcoma of cats as models for similar diseases of man. Cold Spring Harbor Conf Cell Prolif 4: 1197-1214 - Essex M, Sliski AH, Worley M, Grant CK, Snyder HW Jr, Hardy WD Jr, Chen LB (to be published) Significance of the feline oncornavirus-associated cell membrane antigen (FOCMA) in the natural history of feHne leukemia. Cold Spring Harbor Conf Cell Prolif 7 - Frankel AE, Gilbert JH, Porzig KJ, Scolnick EM, Aaronson SA (1979) Nature and distribution of feline sarcoma virus nucleotide sequences. J ViroI30:821-827 - Grant CK, DeBoer DJ, Essex M, Worley MB, Higgins J (1977) Antibodies from healthy cats exposed to feline leukemia virus lyse feHne lymphoma cells slowly with cat complement. J Immunol119: 401-406 - Hardy WD Jr, Zuckerman EE, MacEwen EG, Hayes AA, Essex M (1977) A feline leukemia and sarcoma virus induced tumour specific antigen. Nature 270:249-251 - Khan AS, Stephenson JR (1977) Feline leukemia virus: biochemical and immunological characterization of gag gene-coded structural proteins. J Virol 23: 599-607 - Khan AS, Deobagkar DN, Stephenson JR (1978) Isolation and characterization of a feline sarcoma virus-coded precursor polyp rote in : competition immunoassay of nonstructural components. J Biol Chem 253: 8894-8901 - Lee SG, Lipmann F (1977) Isolation from normal and Rous sarcoma virus-transformed chicken fibroblasts of a factor that binds glucose and stimulates its transport. Proc Natl Acad Sei USA 74: 163-166 - Ruscetti SK, Donner L, Sherr CJ (to be published) A model for FOCMA expression in cells transformed by feline leukemia and sarcoma viruses. In: Fields B, Jaenisch R, Fox CF (eds) ICN-UCLA symposia on molecular and cellular biology, Vo118. Academic Press, New York - Sen A, Todaro GJ (1979) A murine sarcoma virus-associated protein kinase : interaction with actin and microtubular protein. Cell17: 347-356 - Sherr Cl, Sen A, Todaro GJ, Sliski A, Essex M (1978a) Feline sarcoma virus particles contain a phosphorylated polyprotein (pp85) with p15, p12 and FOCMA antigens. Proc Natl Acad Sci USA 75:1505-1509 - Sherr CJ, Todaro GJ, Sliski A, Essex M (1978b) Characterization of a feline sarcoma virus-coded antigen (FOC MA-S) by radioimmunoassay. Proc Natl Acad Sei USA 75: 4489-4493 - Sherr Cl, Fedele LA, Oskarsson M, Maizel J, Vande Woude G (1980) Molecular cloning of Snyder-Theilen feline leukemia and sarcoma viruses: comparative studies of feIine sarcoma virus with its natural helper virus and with Moloney sarcoma virus. J Virol 34:200-212 - Snyder HW Jr, Hardy WD Jr, Zuckerman EE, Fleissner E (1978) Characterization of a tumour-specific antigen on the surface of feline lymphosarcoma cells. Nature 275:656-658 - Snyder HW Jr, PhiIIips KJ, Hardy WD Jr, Zuckerman EE, Essex M, Sliski AH, Rhim J (to be published a) Isolation and characterization of proteins carrying the feline oncornavirusassociated cell membrane antigen (FOCMA). Cold Spring Harbor Symp Quant Biol44 - Snyder HW Jr, Phillips KJ, Dutta-Choudhury M, Hardy WD Jr, Zuckerman EE (to be published b) Biochemical and serological aspects of proteins induced in cells transformed by FeLV and FeSV. In: Hardy WD Jr, Essex M, McCleIland AJ (eds) FeHne leukemia virus. Elsevier, New York - Stephenson JR, Khan 490
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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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C. Chronic Myelogenous Leukemia The
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Patients with a suitable donor rece
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No. Recurrent Leukaemia Other death
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followed by dialysis against isoton
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HS, HL- Heme >_ 40 0 HS I ;:. =====
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knowing at present. These genes may
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esistance to antifolates. In Vitro
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a DNA probe specific for the gene t
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translocation between chromosomes 9
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Table 1. Subdivision of 1827 Myelo-
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NORMAL CELL ®
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some change of the primary type, in
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F. Transformation of Mouse Bone Mar
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were carried out with 3H-MTX in the
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the lysozyme cDNA prepared from ovi
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et al. 1976). The BJAB cellline in
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dalton bands corresponding to light
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1979). The EBV-carrying cell lines
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producer lines (DAUDI and P3HR-1),
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G, Giovanella B, Westman A, Stehlin
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Dc;bt CT + i J.I9 Raji; 1. i .. 100
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c. Discussion During infectious mon
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C. Nonepisomal Viral DNA We have us
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A 260 3.0 A B 1670 70 N2 2.0 c: E .
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vivo. Nature 260:302-306 - Kirk JTO
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Fig. 1. a Polyacrylamide gelelectro
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1979). The lymphocyte subpopulation
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lysin 0 antigens are found, and mix
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Pope JH (1978) Long-term T cell-med
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Charaeteristie Morphologie maturati
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after TPA-induction (Table 3). The
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PA (1978) Reactivity of a rabbit an
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Blood Monocyte. B lood Monocytes 1-
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and nonhistone proteins can serve a
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The findings of the evaluation of s
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Fig. 3. Tumor grawth of L 428 cells
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Cell biological and Immunological A
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consideration. Sanel (1973) obtaine
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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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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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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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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
Page 449 and 450: the same animal were examined (e.g.
Page 451 and 452: I) 'IQ a \0 - CL -- Fig. 2. Restrie
Page 453 and 454: ~ ..................... ~ Cell 3' 5
Page 455 and 456: a :Sa,C! + . ~ 111 i' ( " '~ .J. b
Page 457 and 458: Table 1 Sampie Tissue TS fragment"
Page 459 and 460: a b Sac I / rep ._-_.-_ K J nl rep
Page 461 and 462: Haematology and Blood 'fiansfusion
Page 463 and 464: Osteopetrosis and osteosarcomas occ
Page 465 and 466: Lymphoma cellline Table 1. T and B
Page 467 and 468: Table 2. In vivo growth and oneogen
Page 469 and 470: Haematology and Blood Transfusion V
Page 471 and 472: Table 1. Transforming aetivity of c
Page 473 and 474: Table 3. Transformation aetivity of
Page 475 and 476: - Shoemaker C, Goff S, Gilboa E, Pa
Page 477 and 478: indieate a shifting of target eell
Page 479 and 480: Table 2. Expression of Jl and Sourc
Page 483 and 484: ~ 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
Page 489 and 490: Table 1. Transformation of 3T3 cell
Page 491 and 492: inserts in the genome of rapidly tr
Page 493 and 494: pulations, it seemed likely that th
Page 495 and 496: contains antigens which react with
Page 497: Haematology and Blood Transfusion V
Page 503 and 504: gic approach. In: Hiatt HH, Watson
Page 505 and 506: 12 Eco R [ Fig. 1. Hybridization pa
Page 509 and 510: 10 - '1~ )( -~ S:! E A U I o 1 2
Page 511 and 512: Haematology and Blood 'fransfusion
Page 513 and 514: cultures. These cells did not conta
Page 515 and 516: indication of malignant T-cells in
Page 517 and 518: Fig. 3. Thin-section electron micro
Page 519 and 520: Table 3. Lack of detectable related
Page 521 and 522: J. HTLV Was Present in the Primary
Page 523 and 524: specific signals into nonspecific s
Page 525 and 526: Table 2. Distribution of HTLV-relat
Page 527 and 528: u 10 ~ 20 ~ :c 30 a 40 o Fig. 1. Ki
Page 531 and 532: SSV TrS-gp labeled effectively with
Page 535 and 536: 60 HFF/SSV cDNA CELL RNAs: 0 HF/SSV
Page 537 and 538: Antisera a Table 1. Immunofluoresce
Page 539 and 540: Haematology and Blood 'ftansfusion
Page 541 and 542: y competition RIA yielded virtually
Page 543 and 544: SiS'V' gp70: ANTIGEN, ANTI BODY, IM
Page 545 and 546: type-C virus p30 antigen in cells f
Page 547 and 548: 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
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RNA -, of tumor virus 439 Rous sarc
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Haematology andBlood Transfusion Su
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