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MI1-1 I n-uo \ri I ainical oncology @ Springer-Verlag 1981 Leukemia ',I \~itiowad:l',H. Koshiba', K . Sagawa', I . Kubonishi' \ I s.I,ok I , E. Tatsumi T. Hail2, B. 1. S . Srivastava3, arid T. Ohnuma4 I)c.,,~, ,>I`nitnunology I. Medic:il OncologyL,and Experimental Therapeutics3 I ~ , ~ . U C I I Park Memori:il Institote, New York State Ikpartment of llealth. ,,,,o I IIII Street. Dulralo. N Y 14261, USA and 1 )<pi (11 Neoplastic I)iseases4. M o ~ iStinai School of Medicine. New Y o l k , NY 10029. IJSA Suliiinary. Ry means of the multiple marker analysis, a total of 55 human leukernia-ly~ilphiiiilai~iiici~ell lines which included IS T-cell. 30 H-cell, four myelomonocytic-cell, arid six non-T, non-B cell lines was characterized for their Inarker profiles. The multiple markers used included a number of cell surface m;irkers as detected by either rosette or immunofluorescence tests, enzyme ;ISsays. cytogenetic analysis. and certain functional assay. Based on the criteria previously defined it was found that all the cell lines were proved to rcprcscrtt original 1eukemi:i-lyniptiorna of ALL, AM I,, CLL. C MI , in blastic phasc or variety of lyrnphomas. The monciclonality, a "fro7eti" state at ;I specific stape of di fferent iation-mat ti ration, and cytogenetic marker i n each IC u k emia-Iymphoma cell line were remarkable common properties and wcre stable for years ofcultivation. Similar, if not identical, general characteristics were observed in the study on 344 cases of iincultured fresh leukemia-lymphomas by the multiple marker analysis. While n o single marker specific to any type of tumor w;is found, the study offers not only a basis for better understanding of the biology of leukemia-lymphoma but also an insight into normal hematopoietic cell diffcrentiation in man. Key words: I,eiikcmia-lytnphoina -- Cell line ~ Surface marker - Hematopoietic I tiifferentiation Introduction Since a permanent cell line from an African Burkitt lymphoma was established I5 !cars ago for the first time (Pulfertaft 1964), numerous numbers of human hematopoietic cell lines from a variety of sources have been established and Characterized (Moore 1972; Nilsson and Pontern 1975; Minowada 1978). Nevertheless. most of lhcsc cell lines are so-called "normal" B-lymphoblastoid cell lines which are char- -- -_ * Sllpportcd by USPIIS grants (`A-14413. CA-17609. C`A-25865. AI-OX899, and contract 31-Cnl l h 5 from the Nation:il Cancer Institrtte and National Institute of Allcrgy a n d Infectious Iliswses "//prifir rcqirrs~slo: J . Minow:ida. M D (address see above) 017 1-5216/81p l o l p091p2.00 92 J. Minowada et :ti acterized by the presence of immunoglobulin synthesis, Epstein-Barr virus (EBV) infection, and normal cytogenetic constitution. From time to time, true leukemiiilymphoma-derived cell lines were, however, established with great difficulticr (Moore and Kitamura 1968; Minowada et al. 1972). Among many contribution.; resulting from the studies of such proven leukemia-lymphoma cell lines, recogiiition of various immunologic subtypes of leukemias and lymphomas is of spcci;tl significance. It permits not only accurate diagnosis but also leads to a better treatment of these hematopoietic malignancies. In particular, such immunologic idetitifrcation of tumor cells involved in lymphomas would resolve many of the prohlenis encountered in the existing classification of lymphoma. The present study describes marker profiles of a total of 55 human leukcmiitlymphoma cell lines which include 29 lines of lymphoma origin. Despite continiious effort at searching an antigen specific to leukemia or lymphoma, all market \ thus far studied turned out to be of normal gene products. Hence, an attempt was made to correlate the observed heterogeneity in the marker profiles of leuketni;ilymphoma cells with a conceptional and hypothetical schcnie of normal hcniniibpoietic cell differentiation (Minowada et al. 1977). Materials and Method5 . A total of 55 proven Icukcmin-lymphotii~ciell lines was used i n the present study. Origin, cslahli.;limctrl and general characteristics of these cell lines were reportcd individually o r in summary in a series of 11 ports (Minowada et til. 1972, 1977: Minowadii 1978: Lok et al. 1979). Many of the cell lines were ~ * I I erously provided to our Iahofiitory by a number of colleagues who cst:ihlished and ch;ir;trtcrii.cd tlietli All ccll lines wcrc niaintuincd in RPMl medium 1640 supplcmcntcd with 5 10% ( v i v ) of heat-itiocli vatcd fctal calf serum. Thccclls in culturc for various analyscs were kept in an cxponctiti;il growth pli;t\, and cell viability was generally nvcr 90% at the time of study. Mycoplasma a n d other iiiirrobi:tl inlu tions in thcccll lines were nionitorcd and all ccll lines were nc&ajivcfor the infection. In :idclition.scvrt~il dozens or so-called normal R-lymphohlastoid cell lines were itlso st ntlied For compariwn. Ovcr 300 cases of various types of leukemias were also studied by the multiple m:irkcr an:ilysi~~ Leukemia specimens were either bonc marrow anpirates or peripheral blood iintl mononuclciir cells liiiai thcm were isolated For examination by standard I-icoll-Hypitquc density gradient centrifupition. '1IN mononuclear cells were washed three times with phosphittc-hufTered sitlinc itnil wcrc used For : t n : i I y ~ A multiple marker analysis used in the present study includcd a number of cell surliicc ni:trkrt \. terminal dcoxynucleotidyl transfcr:ise, and cytogenetic analysis. The surhce m:trkers were dctccted In two major methods. One was the rosette assay, which included rosette formation with shecp erytlrril cytcs (E). with bovine crythrocytcs-lg(i antibody complex ( E A ) and with hovinc erythrocyte-lgM antihody-complement complex (EAC), respectively. Only E rosette was found to he a specific and tlcliniii\t~ marker for T cells. Both EA and EAC were not specific to any single lineage within the hcm:ttopoiclii. cell types, and they were, therefore. considered as non-definitive markers. Another major method *;I\ immunofluorescence using various specific antibody preyarations. Cell surface immunoglohuliil\ (Smlg) or cytoplasmic immunoglobulins (Cylg) was dctcctcd by direct immunofluorcsccncc usiv fluorescein-conjugated goat anti-human immunoglobulin chain-specific nniibody. The presence of iw munoglobulins (Smlg or/and Cylg) was a definitive and specific marker of the B cells. A T-cell-specilic antigen (T-hg) (Tsubota et al. 1977) was detected by indirect membriine immunofluorescence using sprcific rabbit anti-T-cell antibody as a primary reagent followed by fluorescein-conjugated goat anti-rah bit globulins as the secondary reagent. A myelomonocyte-spccific antigen (MAg-I) (Sagiiwa et al. 197')) was delected similarly by indirect membrane immunofluorescence. Both T-Ag and MAg-I were regnrtled each as a definitive marker to respective lineage (T-cell and myelomonocyte, respectively) in the hrmatopoietic ccll difkrentiation. Both antigen specific to an la-like gp 28.30 molecules ( I n ) (Schlossm;in et al. 1976) and antigen associated primarily with common form of acute lymphobl;islic Icukcml;l (CALL)(Greaves et al. 1975) were detected by indirect membrane immunofluoresence with respectibc Marker Profiles of Human Leukemia and Lymphoma Cell Lines 93 ,Illtisera.These two antigens were. nevertheless. of non-definitive marker category, since their exIiression was found heyond a single lineage in the hematopoietic differentiation. Details of production ;rnd characteristics of each rahhit-specific antibody were reported previously (Tsubota et al. 1977: Koyama et at. 1977; Koshiba el al. 1978; Sagawa et al. 1979). Results of terminal deoxynucleotidyl transferase, EBV infection, and cytogenetic analyses were cited from previous reports (Minowada et .it. 1977. 197R. 1980). Results Ilsing the definitive and non-definitive markers as described in Materials and Methods, a total of 55 leukemia-lymphoma cell lines representing original tumors was subgrouped into four groups: ('Table I). (;rorrp I I'-cell leukemia-lymphoma line has a common definitive marker, Le., T-Ag. There were I5 such T-cell lines (nos. 1 -1 5 ) in which E receptor, in spite of being another tlcfinitive marker for T cells, was undetectable in five cell lines. In connection to hypothetical scheme for normal hematopoietic cell differentiation which had been previously proposed (Minowada et al 1978, 1980), the I5 T-cell leukemia-lymplioma lines could be assigned to a respective compartment (Table I , Fig. I). Other tnarkers, such as CALL, la, TdT, and EAC were apparently non-specific to the Tcell lineage, though they were used to divide the entire T-cell differentiation stage itito three compartments. As shown in Fig. I , T-Blast I (nos. 1-6) should have CALLantigen and TdT, but T-Blast It (nos. 7-12) has only TdT. Neither CALL ;intigen nor TdT are expressed in the T-Cell compartment (no.14). Except for MOLT-IO which was an unusual lymphoma-derived cell line, the h e n c e of M i k e antigen was found to be a common feature of all T-cell lines. I:lhle 1. FiTty-five hunian leukentia-lyntpliotna cell lines: A total of 5 5 provcn human leukemiaI\tllphonia cell lines was subcntcgori7ed into b u r categories based on the presence of definitive 1 -lymphoid, B-lymphoid. or myeloid marker. or absence of these three markers. According to the Ivwclive marker profile. each cell line aas further classified for its position of differentiation stage Icl;ltive to a hypothetical scheme of hematopoietic cell differerrtiation (Minowada et a1 1980) CO Cell line Origin Markers detected -.~_ - -- Definitive Non-Definitive Compart merit at differen- tiation stage I [-cell leukemia-lymphoma lines I CCRF-C'EM ALL 7 RPMl8402 ALL 1 HPB-ALL ALL 4 DND-41 ALL 5 HPB-MLT ATL 6 IID-Mar 2 HD'? 7 MOLT 1-4 ALL X JM ALL 'J MOLT-I I ALL II) TALL-I ALL -I I PIZ/lch. ALL T-Ag T-Ag T-Ae T-Ag T-Ag T-Ag T-AB T-Ag T-AB T-Ag T-Ag CALL,TdT CAN. TdT. EAC CALL, TdT. EAC, E CALL. TdT. EAC. E CALL,TdT. EAC.. E CALL, TdT. EAC. E TdT. EAC. E TdT, EAC, E TdT, EAC, E TdT. E EAC. E T-Rl;~stI T-Blitst I T-Blast I T-Blast I T-Blast I T-Blact I T-Bhst II T-Blast II T-Blast 11 T-Blilst I I T-Blast I I c PROLIFERATION 8 DIFFERENTIATION 9s * -.. c- / /\ ;I \ 1 "STEM CELL" .-.\ I \/ 1 I ig. 1. This Schcme of human hcmcctopoieiic cell dtll'erentiation does not include the megitkaryocyie j+ I I I C J ~ C EBch eompitrtmcnt hils a particular marker conibination beside each definitive marker, such I I - A g for T-cell lineiige. Ig for B-cell Iincagc or MAg-I for rnyelaidmonocytoid litwage 't 1 \lOL,T-lO was an exceptional T-cell line with T-Ag in which a fraction (10-30%) i (d`population at a given time expressed both la-like antigen and cALL antigen, 1 tliough the cell line appeared to a monoclonal population on the basis of cyto- licnctic markers. Ikell leuketnia-lymptio1na line has a common definitive tnarker, Le., immunodobulin, such as Smlg, CyIg, or both. There were 30 such B-cell lines (nos. 16-45) listed in Table 1. An la-like antigen was oh!lgaiory it! all 30 B-cell lines, but it \\:IS not the definitive marker for the t3-cell lineage. Other markers, such as CALL, I dT, EA, and EAC were apparently non-specific to the B-cell lineage. though they \ w e useful to divide the entire 8-cell differentiation stage into five separate comIM-tmentsas shown in Fig. I. The Pre-B Cell(nos. 16-21) without detectable Smlg has both cALL and la-like antigen, but not always positive for TdT. The B-Blast I (nos. 22-35) has Smlg, CALL, and M i k e antigens. Many of the Burkitt lymPhoma-derived cell lines (nos. 23-29, African Burkitt lymphomas) had EBV infec[l(w,but it was not specific to either Burkitt lymphoma or B-BIasr I category. B' h t II (nos. 36-42) is similar to B-Blast I except for the absence of CALL. The Mk+l compartment is in fact a compartment indistinguishable in membrane phe- j I W p e from both B-Blast I t and Plasnw Cell and there is no cell line in the B-Cell c(lmpartment. The Plasnra Cell has no specific positive marker; nonetheless, there ' ;Ire three cell lines in this compartment (nos, 43-45). I t was based on the fact that I 96 J . Minowadn ei :II the Ig produced by the lines were identical to those found in patients myelonla ~ I I teins. Group rrI Myelomonocytic leukernis-lymphoma line has a common definitive marker. I I* MAg-I antigen. There were four such myelomonocyte lines as the listed nos. 46 1') i n Table 1 , All four cell lines expressed EA in varying percents. Both f-f 1,-60 ;ttitl ML 1-3 showed positive myeloperoxidase reaction at 100% (data arc not shorrti). whereas KG-I and U-937 were negative or marginal for the myeloperoxitlase rc<it tion. Accordingly, it was decided tentatively that the myelocytic differenti:tltriit lineage can be divided into several compartments on the basis of both the tnari\cr profiles and cytochemistry. The Prornyelocyfe (HL-60). Mjdohknsf (h.1 I. 1 :I Myeloid Precursor (KG-I) and more Mature Grmiulocyfesare those compartmctil\ in order of progressive differentiation (Fig. I). U-937 line was, however, placed i i i l u the Mortohlust compartment, because it derived from a histiocytic lympholnit Group IV Non-T, non-B leukemia-lymphoma line lacks all three above mentioned clefinili\c markers. Six cell lines (nos. 50-55) were in this group. Nevertheless, cell lines no) 51-54 were considered to be in a single compartment, viz., I,jwp/wk/ P r w w w . Since all four cell lines were of ALL origin with common marker profiles ( I n . CAI I.. and TdT). Based on the latest information, K-562 can be placed in a comparttncrtl for an Ery//woid Precursor (Andersson et al. 1979).The SU-HDL- I (no. 5 5 ) w&c4 is another unusual cell line lacking all markers was tentatively placed of the co~npartment of Monocy/oid Precursor on the basis of its derivation of a histiocylic lymphoma (Epstein and Kaplan 1974). A total of 344 patients with leukemias were studied by the multiple tnarktv analysis. Table 2 summarizes immunologic subclassification of these leokemi;tx In view of our hypothesis that the observed heterogeneity of marker profilcs among 55 leukemia-lymphoma cell lines was interpreted as reflecting normal hcmatopoietic cell differentiation scheme, the heterogeneity of marker prolilcs among these leukemias was similarly correlated with the scheme shown in Fig. I . Of 19 T-cell ALLs, six had cALL antigen expression which can be placed in [tic T-Blust Icompartment in Fig. 1 . The rest of the T-cell ALL were in the 7-Rkrsl I / compartment. Of six B-cell ALLs, two had cALL antigen expression which can Iw placed in the B-Blnsf Icompartment. Since we did not determine systematically for the pre-B-cell phenotype, all 118 common ALLs were in the Lvniphoid Precrrrsof compartment. Of seven null-cell ALLs, four had Ia antigen expression which might be placed alternatively in the Myekjid Prectrrsor. A total of 39 AMLs are shown in Table 2. An attempt at subclassification hp the multiple marker analysis (Minowada et al. 1980) was made to subclassil'y AMLs into three subtypes. The type I AML is characterized by the presence of 1;) antigen but lacking MAg-I antigen and all other markers. While type I1 AML expresses both la and MAg-I antigens with variable expression for EA and EA('. type I l l A M L expresses only MAg-I antigen which commonly accompanies wi(h expression of EA and EAC. Expression of la antigen in the type 111 AMI, cells is l \ C rI1:llkcr. I t l I 4 f Y i l l f ) S lh 1`) h \ I f l Iff 00 :111l1 c tlr t tt.1 w t i:it 14 111 h i t 11 t h t . rnarl i r d h c ! fhll I :I w ~ ~ l ~ l l l pt :l llI Cr l l l \ ,C'Lf't pI:Ic.C(l 1111ll ~,,i,k2. Immunologic suhclassificatlon of 344 patients with leukemias. According ti, 11,c presence of definitive markers with combinations of other non-definitive I,, Itkers,four major types of leukemias were further siibclawified 1 ) Ipnosis fmmunotogical Incidence Remarks (No positive/nn total) 13% (19/150) CALL', 32% (6/19) CALL', 33% (2/6) 20% (R/39) 49% (19/39) Il"/o (12/39) 99% (100/101) I % (1/101) la', 71% (4/7) la', 69% (27/39) MAg-1'. 79% (31/39) CALL', 46% (25154) tit iotvd tlclirii~r\~~ c + h f dl hnr\ ria\ 1'1llp/rfw/ f f 111 \ f l 1 rofilt,\ f 1;). C A I 1 it1 ,i v o i i i p t tiiiwl I (tin iS)\\I,kh I:KtYl of I l l C (`0111 111 of :t lirstioc!IIL IIO longer detected. I t was thus possible to place type I, 11, and I I I AMLs into I II chid Precvrsor. Myeloblast, and Proniyelocyic compartments, respectively. ( MLs in blastic phase were also subtyped into three types according to their memlliltne phenotypes (Janossy et al. 1976).The lymphoid type of CML in BP is charictcrbed by the same marker profile (la, CALL,and TdT positive) with Lymphoid i f m:ir kci prolilr\ cc!itit* ti(irrii;iI I i i s it* ~ \ i t~i ilniI~ig I ,111 lv f't:tccclfflI1)C c In I k I - N h , I/ -+\icvi v hic*h can hc \ \ :twi:itic:ill~{or 1I)tpllrrl,l I'IIV IW\flf Y ~ . I ~\\ VIiich niigltl r h h \ v f i c ; i ! i o r i I>\ :I II(, t ( 3 \ 1I hcfa<siI\ tlw p1CScrlc-C of 1J IC t\pv II A h l l CYftrr 1 1 and FA('. ' :IC( 4~t~lp:llllcw5rfl~ p b I It 2 h i l re115 I' ui For, Mveioid Precursor, or M~'elOhlasPi/romyelocyie compartment. a total number of leukemia-lymphoma cell lines in&eased, it became apparent I h t a considerable heterogeneity exists in the marker profiles among them, even Iliose cell lines within the same lineage of hematopoietic differentiation (Minowada ~ ' 1al. 1977). Similar heterogeneity in the marker profiles was repeatedly docu- lwnted in a large series of study with fresh leukemia-lymphomas (Greaves et al. 1977, 1978; Greaves and Janossy 1978; Minowada et al. 1977, 1980; Thierfelder et 4 1979). Unlike earlier reports for the presence of leukemia antigens, all cell surI x e markers so far studied were considered to be of normal gene products which m be found in the cells during various stages of hematopoietic cell differentiation ( j i m m y et at. 1977; Greaves 1978; Metzgar and Mohanakumor 1978). Moreover, t i m y of the antigens which were restricted in their expression to one type of mature ljcripheral hematopoietic cells were found to be expressed on the leukemia-lymP h n a cells as well as normal immature hematopoietic cells. These antigens inh l e la-like gp 28.30 antigens (Billing et al. 1976; Schlossman et al. 1976; Janossy ct a!. 1977) and CALL antigens (Greaves 1978; Greaves and Janossy 1978; J. Minowada 1'1 I l.;lkerrrcifites of 11, Minowada et al. 1978). The cell types in which these antigens were expressed \\uV !$stein h l Karl.- often beyond a single differentiation lineage and it was concluded that these :ilili. gens are of differentiation-linked natures. I t was in the same situation for 'T'd'l':ICtivity that the enzyme activity is found not only in immature T cells but also in rllc immature lymphoid precursor cells (Bollum 1979). continiiow ccll CI' ?4:1R(I 1872 I , (iraeves MI'( 1978) tl1;ll hacmcpiictic I ticin: its iipplic:itioi I'rcss. ? 5 3 26.1 (;rc:ivc< hf 1'. Nt (11. cells. ('lin lmniiinr Minowada 1978; Thierfelder el al. 1979; Minowada et al. 1980; RcinherT J I I ~ '. (ircavcs hl, l a n o \ % kiicniias. Rioclirnl Schlossman 1980; Nilsson et ai. 1980). 4) (ircaves M I : , I a i i o Our model for a hematopoietic cell differentiation as presented in this SI iitl! cclls and Ieukcmii (Fig. I ) was based on the multiple marker analysis of 55 leukemia-lymphoma (dl lines and 344 cases of leukemias. The monoclonality of human leiikernia-lyphniii;i\ hliirks PA. Till J I . Ilarhor I ~ h o r a t o i :(I. Circiivcs Mf:. J:ini so far studied is a remarkable feature and it seems to represent a particular inarlcr (1977) hlroihranc I profile of normal counterpart at a specific point in the hematopoietic differciiri- k:icmi:ts. II;icmii!~ ation. Assuming a single pluripotential stem-cell compartment, each lymphid. I 1 Jitncwy C;. (;oltl.l myeloid, monocytoid, erythroid. arid megakaryocytoid differentiation can emctg from it. With this concept in mind, each marker can now be distributed with SOIIIC certainty on the scheme. I t is thus possible that each leukemia-lymphoma linc o r 1)iffcrrnti:ition t i t i IInem:itol 37:JY I I ' Jiinossy G . (.irc:i\c MFJ (1976) Rlitst fresh leukemia-lymphoma with the combination of the definitive marker and nuiii- "lyniphoid" anti I' ber of non-definitive markers can be placed at particular stage of hernatopoic,lii, cell differentiation. This attempt with the current scheme may not be totally u)I. rect, but at least it makes reasonable sense in the immunologic membranc phclio- 11 Kohler G , Milstri specificity. Natur! 1.1. Koshiha 11, Mint1 cell line which cat types of leukemia-lymphomas. lymphohlastic lcrr Recent advances in methodologies, such as the in vitro culture method, intitic- I < . Koyain:i K . N;ik. tion for differentiation, and murine hybridoma technology, would certainly facilt- lines; "hrtrnitn 1i1 late further progress in the study of human hematopoietic cell system atid ICIIkemia-lymphoma (Collins et al. 1978; Mier et al. 1980; Kohler et al. 1975). as dctcrrnined h! 10 1 cik MS. Koshilv :tclcrization of hi! Proper usage and experimentation with these permanent leukemia-lymphoiil;l iition in thc R . w I cell lines would continue to be'responsible for further advances in the leukeriii;l 17 Met7p:ir RS. hl(> membrane phenotyping and i t would provide a realistic basis to resolve somr of the difficulties in the existing classificatio; of human lymphomas. minars iii Ilcni;ic~ 1s Micr J W ,(;:tllo I rroni phstd1ctii;rr n r . k r i o i s k . c ~ c . n ~ ~Tnh/e. iiuthor gralerully acknowledges the SFR I 1 I at Christian Alhrechts Univci\it! 775134 hllR of Kiel, FRG. for its support i n making this presentation possible. 1') Minow:ttl;i J ( 1 9 7 dit'rercnti:rticin 1I to canccr. 1NSi.1 ''1 Mincrw:tcl;i 1. J;ti: References l%prcsgion d :in phoma cell lint:\ I . Andcrsson LC, Jokinen M , tiiihmberg CG (1979) Induction of erythroid difkrentiation in the liw man leukaemia cell line K 562. Naturc 278:364--365 2. Billing R, Rafinideh B, Drcw 1. flartmiin G, Gale R. Terasaki P(IY7h) flitmiin 9-lymphocyte ittillgcns expresscd hy lymphocytic and myclocytic lcukemia cells. I. Dctcction hy riihhit antisera. J I:v ' I . Minow;icl;i J. Oti lishment rind cvic Minowiiciii J , SiiI N ( 1 980)A mixti Mcd 144:167~178 3. Bollurn FJ (1979) Terminal deoxynucleotidyl transferase as a hematopoietic cell marker. Dlo(al 54:1203-121 5 4. Collins SJ, Ruscetti PW, Gallagher RE, Gallo RC (1978) Terminal differcntiation of' hrttti;ln promyelocytic leukemia cells induced by dimethyl sulfoxide and other polar compounds. Proc Niill -Acad Sci 75:24S8--2462 Minowad:) J. T V Sinks LF. S:inctb lines, and nut1 cc man. Ilacrniitol I ,kcr Prnfiles of tluman Leukemia and Lymphoma ('ell Lines 99 f'p\tcin AId, Keplan 11.7 (1974) Biology of the human malignant lymphomas. I . Establishment in c.t,ntinrlotts cell culture and heterotransplantation of diffuse histiocytic lymphomas. Canccr ~ ~ 1 ~ 1 8158172 I . (;r:\cves Ml: (1978) Memhrane antigens and enzymatic phenotypes in leukaemia: Correlates of nor- ,tl;tl Ir:~cn~opoictidciflcrentiation'? I n : Serrou B, Rosenfeld (' (eds) Human lymphocyte differentiati,,n: its :tpplir:ttion ((1 c;tnccr. INSERM Syniposirrni NO, 8. Elsevicr/North llolland Biochemical pres<. 253 264 (ircitvcs MF, Brnwn (i. R;ipson NT. Listcr TA (1975) Antisera to acute lymphoblastic leukcmia c.clln. Clin Immunol Immunopathol 4:67-84 (;rc:ivcs M. Janossy C i (IY78) Patterns of gene cxprcssion and the cellular origins of human leukiiemias. Hiochem Riophys Acta 516:193- 230 ( ircnves MF, Janossy G. Francis Ci. Minowada J (1978) Membrane phenotypes of human leukemic cclls and leukemic cell lines: Clinical correlates and biologicdl implications. In: Clarkson D, hl;trks PA, Till J E (eds) Differentiation of normal and neoplastic hematopoietic cells. Cold Spring I I it rhor 1,a horatory. 823.-841 t i Circ;tvcs Mf;, Janossy Ci, Roberts M. Rapson NT, Ellis RB. 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