T cell receptor alpha-, beta-, and gamma-genes
in T cell and pre-B cell acute lymphoblastic
C A Felix, … , P Goldman, S J Korsmeyer
J Clin Invest.
We examined alpha-, beta-, and gamma-T cell receptor (TCR) gene activation within acute
lymphoblastic leukemias (ALLs) that represent early stages of B and T cell development.
We wished to determine if TCR rearrangement and expression was lineage restricted,
showed any developmental hierarchy, or could identify new subsets of T cells.
Rearrangement of gamma and beta TCR genes occurred early in development but in no set
order, and most ALLs (22/26) were of sufficient maturity to have rearranged both genes.
T-ALLs preferentially rearranged C gamma 2 versus the C gamma 1 complex; no preference
within the beta locus was apparent. Once rearranged, the beta TCR continued to be
expressed (11/13), whereas the gamma TCR was rarely expressed (3/14). The alpha TCR
was expressed only in more mature T-ALLs (8/14) that usually displayed T3. The 3A-1 T
cell associated antigen appeared earliest in development followed by T11 and T3. Within
pre-B cell ALL a higher incidence of lineage spillover was noted for gamma TCR
rearrangements (8/17) than for beta rearrangements (3/17). This also contrasts with the only
occasional rearrangement of immunoglobulin (Ig) heavy chains (3/25) in T-ALL. However, in
pre-B ALL the pattern of gamma TCR usage was distinct from that of T cells, with the C
gamma 1 complex utilized more frequently. Almost all ALLs could be classified as pre-B or
Find the latest version:
Pre-B Cell Acute Lymphoblastic Leukemia
Carolyn A. Felix, John J. Wright, David G. Poplack, Gregory H.Reaman,DianeCole, Paula Goldman, and Stanley J. Korsmeyer
Metabolism and PediatricBranches,National Cancer Institute, Bethesda, Maryland 20892; Department ofHematology-Oncology,
Childrens Hospital National Medical Center, George Washington UniversitySchoolofMedicine, Washington,DC20010; Departments ofMedicine, Microbiology, andImmunology, Howard Hughes Medical Institute,
Washington University School ofMedicine, St. Louis, Missouri 63110
We examineda-,,(-,and'y-Tcell receptor(TCR)geneactivation
within acute lymphoblastic leukemias (ALLs) that represent
earlystagesof Band T celldevelopment.We wishedtodetermine
ifTCR rearrangement and expression was lineage restricted,
showedanydevelopmental hierarchy,orcouldidentifynew sub-setsof T cells.Rearrangementof'yand,( TCRgenesoccurred
earlyindevelopmentbut innosetorder,andmostT-ALLs(22/ 26)wereof sufficient maturity tohaverearranged bothgenes. T-ALLs preferentially rearranged
Cj1complex; nopreference within
the TCR continued to beexpressed (11/13), whereas the y
TCRwasrarely expressed (3/14). TheaTCRwas expressed onlyinmorematureT-ALLs(8/14)thatusuallydisplayedT3. The3A-1 Tcell associated antigen appeared earliest in
devel-opmentfollowedby T111 andT3. Withinpre-Bcell ALLahigher incidence of lineage spillover wasnoted for y TCR
rearrange-ments(8/17) thanfor rearrangements (3/17).Thisalso
con-trastswiththe only occasionalrearrangementof immunoglobulin (Ig)heavy chains (3/25)inT-ALL. However,inpre-BALLthe
pattern ofyTCRusagewasdistinct from that of Tcells,with
C,1complexutilizedmorefrequently.Almost all ALLs could beclassifiedaspre-BorT cell in typeby combining Igand TCR
geneswithmonoclonal antibodies recognizing surfaceantigens, although examples oflineage duality werenoted.Unique
sub-populations of cells were discovered including twogenetically
uncommitted ALLs that failed to rearrangeeither Ig orTCR loci. Moreover, two T lymphoblasts were identified that pos-sessedtheT3moleculebut failedtoexpressaplus,( TCRgenes.
These T-ALLsmayrepresent afortuitous transformation of T cellsubsets with alternative T3-Ti complexes.
uni-versally applicable markers ofT cell clonality. The
character-Address reprintrequests toDr. Korsmeyer, Department of Medicine and Microbiology/Immunology, Washington University School of
Medicine, 660 S. EuclidAve.,St. Louis,MO 63110.
Receivedfor publication 21 August 1986 andinrevisedform13
ization ofthe T cell receptor
improveour understanding of lineage
commitment,clonality, stage of differentiation, and T cell associated chro-mosomal translocations. In this study we examined
rearrange-mentand expression of TCR genes within acute lymphoblastic leukemias of both T cell and pre-B cell type. We wished to de-termine whether the activation of a,
fl,and -yTCR geneswas
lineage restricted and whether this was useful in placing early lymphoid progenitors into T cell versus B cell lineages.
wesought to determine if there was a hierarchy of a, (3, and
TCRgeneutilization, if it correlated with T cell surface antigen expression, and whether this information could pinpoint the
stateofdifferentiation of individual T cell ALLs.
Wepreviously demonstrated that the non-T ALLs that lacked T cell surface antigens were adevelopmental series of B cell precursors (1-3). All of these pre-B ALLs had rearranged im-munoglobulin (Ig) heavy (H) chain genes and- 40% had
pro-gressedtothe later developmental step of rearranging theirKor
X light (L) chain genes. Coordinate with this
hierarchyofIg gene rearrangements wasanorderedexpression of B cell associated surfaceantigens in which B4 and HLA-DRwerepresentonthe mostimmature cells, the commonacutelymphoblastic leukemia antigen (CALLA) followed, and Bl was present on the most maturecells.Approximately 20% of ALLs bore T cell associated surface antigens (4, 5), uniformly retained germline Ig L-chain andusually (90%) germline Ig H chain genes (2), andwere
con-sequentlyfelttorepresent T cell
lineageALLs. A number of T cellassociated surface antigens have been identified, however
none ofthese are
investigatedthe TCR genes in T-ALL to determine ifan
rearrange-ments occursinpre-Tasit does in pre-B development. The classic antigen specific T cell receptor isaTi heterodimer oftheaplus 13 TCR chains that is
complexedintramembranously toform the T3/Ti molecular complex (6). T3-Ti
expressionis noted only in the most mature
peripheralT cells(7, 8). The human I TCR gene is locatedonchromosome 7atband q35 (9) and is
(Cal)segments, 13joining (Jo) segments, two or more
(Do)segments, and -100 variable
(VO)gene segments(10-12). Rearrangements of this locus are
readilydetected and intermediate
(D/J)1.0 kilobase (kb) and complete (V/D/J) 1.3 kb
transcriptscan be discerned (13-18). Thea TCR locus is foundat
14qI1 and is
1.Abbreviations usedinthis paper: CALLA,common acutae lympho-blastic leukemiaantigen;EBV,EpsteinBarrvirus;sRBC, sheep red blood cell;TCR, Tcell receptor,tdt, terminal deoxynucleotidyl transferase;
WBC, white blood cells.
©TheAmericanSociety for Clinical Investigation,Inc.
particularly prone to interchromosomal recombination in T cell ALLs (19-22). Itconsists ofone
C,region, an exceptionallylong stretch of Ja segments that may contain 50-100 J regions spread over morethan70 kb. No diversity segments are known to exist in the a TCR locus, but there are probably 40 or more
V.regions (23-25). While the markedlylong stretch ofJas makes it currently rather impractical to search for rearrangements in all cells, RNA transcripts of complete recombinations (1.6 kb) and truncated forms (1.3 kb) can be assessed (23). In addition to the a- and (3-chain that
contributeto theTi heterodimer, at least one other TCR gene, yis known to exist (26-29). This rearranging deter-minant is found on chromosome 7 at band p1 5 (30). In humans twoC.s are present, together with three or four Jzs, and perhaps inthe range of 10
V,segments. Despite having a similar
orga-nizationto the other TCR genes the precise function of this locusis uncertain (31, 32). Because of this, we wished to assess its rearrangement and expression in T-ALLs to gain potential insights into its role in T cell development.
Developmental studies inthe mouse suggested a hierarchy of TCR gene
activationin which yTCR expression might pre-cede the(3 TCR. y TCR RNA decreased dramatically during
intrathymic maturation, while(3RNAremained constant, and a RNA
increased(6, 7, 33-36). We sought to determine whether
the T-ALLs representeda
population ofcells early enough
inpre-T development to reveal any ordered usage of TCR genes
examiningT3 andother cell
an-tigens togetherwith a,
(, andywe also
inpre-B and T cell ALLs was
restriction of these developmentalevents and to determine
lymphoid progenitorcells existed.
ALLsexamined.Leukemic cells were obtainedby
lymphocyte fractions from peripheral blood ofpatients with ALLat
cat-egorizedas Tcell ALL and 17patients classifiedaspre-BcellALL.In
addition, threeestablished T-ALLcell lines
(HSB, CEM,and RPMI
(RS, Nalm-6, Nalm-l,and
alsoexamined (2,37, 38).Also studiedwere
lymphocytic leukemia,five Burkittlymphomacelllines
Balm-l, Balm-4) (39-41). Moreover,14clonal
EpsteinBarr Virus transformedBcell lineswereutilized.The Hut 102 cell linewasutilizedas anexampleofa matureTcell(42),while K562 representedan
erythroleukemia(43), HL-60apromyelocyte(44),and SU-DHL-Iahistiocyte (40).
ALLswereinitiallyclassifiedas Tcell type ifthey possessedTcell associatedantigens (3A-1,andattimesT
In contrast, ALLs wereinitiallyclassifiedaspre-Bcell in type ifthey
(B4,HLA-DR, andattimesB.), possessed rearranged IgHchain genes andattimesrearrangedLchain genes(2, 3). Completeexamination of all such markerswereperformedwhenever
possible.Limited numbers of leukemic cellsprevented examination of all markers insomecases
and such cellswere classified aspre-B orTbased onavailable data
TCR geneconfiguration.Highmolecular weight DNA was extracted from leukemic cells or cell lines as well as total white blood cells (WBCs) from normalindividuals. 15ygof genomic DNA was digested to com-pletionwith 5-10U/pgDNAofthe appropriate restriction endonuclease,
sized fractionatedon0.6-1.0% agarose gels, and transferredto nitrocel-lulose paper(2, 45).Such genomic blots were hybridized to 32P-labeled DNAprobes
preparedby a randompriming method (46). These Southern blots were washedatthe appropriate stringencyandautoradiography wasperformed.
Cpprobe was a 700 base pair (bp) Eco RI cDNA fiagment
Cpregionand iscapable ofhybridizingtoboth human
Cpregionsunderroutinestringency(14). In the germline form both Jr
JrC1,2arelocated onasingle 23 kb Bam HI
fiagmentasindicated inFig. I A. Thus, rearrangements of either the
Cp2complex should alter the size of the Bam HI
fragment.Todiscern theidentityof such rearrangements EcoRIandHind IIIdigestswereused.Inthe germ-line form
C#loccupies an 11-kb EcoRI
C,2a4-kb frag-ment.The 11-kb fragmentcontains the
J,0lregion and rearranges when the
rearranges.Occasionallyan8-kb Eco RI band is noted eveningermline sourcesoftissue.This representseitheranEco RI resistant site or perhaps
attimesarestriction fragment length polymorphism, andcannotbe assumedtobeasomaticrearrangement. In contrast, Hind IIIdigestion
is capable of detecting rearrangements of the
an8.0-kb Hind IIIfragmentthat rearranges.
C#lisfound on a 3.5-kb Hind IIIfragmentthatis deleted whenaV, regionrearrangesinto the
C,probewas a300 bpPstI-BamHIfragmentofahuman
containingsubclone(30).Itwasroutinely hybridizedtoBam HIdigested
humangenomicDNAand thosecellsdisplaying germline patternswere
alsoexamined withSac I and PvU II
digestedDNA. Initsgermline configurationtheC7 locusdisplaysa15kb Bam HIfragment containing
C41and a12.5-kbfragmentwith C72 (28). Bam HIdetectsrearrangements ofeitherthe
C71orC72locus.AsinglesetofV7 genes appearstoexist
5'ofC71 asdiagrammedinFig.3 B.Hence, rearrangements into the
CY2locus result in the deletion of the 15-kb
TCR gene expression. In addition RNA was extracted from 14 T cell ALLleukemic samples and cell lines, 4 pre-B cell lines,4mature B celllines,Hut102,HL-60, and K562. Total cellular RNA was prepared
,ggwasdenaturedin form-amide,electrophoresedonagarose-formaldehyde gels, andtransferred
tonitrocellulose paper(47). Northernblotswerehybridized with the C. andC, probes describedaswellaswitha1.1 kb Eco RIfiagment con-tainingCa from ahumanaTCR cDNA clone(23). In addition all Northern blotswerehybridizedwitha-y-actinprobe to insure that equal
amountsofintact, hybridizableRNAwaspresent in each lane(48). Blots werestrippedclean of probeby washing in0.1%sodium dodecyl sulfate
(SDS)at70'Cfor 20 minand autoradiographywasperformedtoinsure acompletewashoff.
Ig geneconfiguration.HumanIggeneprobes were used to assess the
statusof H chainaswellas Kand A Lchain genes.Ahumangenomic
6.0 kb Bam HI-HindIII
fragmentcontainingtheentire JH regionwas
usedto assessrearrangements in both BamHI andHindIIIdigested DNA(49). Kgene rearrangementsweredetected inBam HIdigested
(50). A0.8 kb BglII-Eco RI fragment containing C,, wascapableof detecting anyXrearrangements in EcoRI digestedDNA(50).
Phenotypic characterization.Whenadequate numbers of leukemic cellsexisted,theywereassessed for cellsurfaceantigens.Onemillion viable cells pertest werereacted withapredetermined concentration of
a mousemonoclonalantibody at4VC for 30 min, washedtwice,and thenreacted withafluorescein-labeled goat mouse Ig second anti-body. After three washes the cellswereassessedby flowmicrofluorometry
witha fluorescence-activated cellsorter(51). Cellswerereacted with normalmouse serum and thefluorescein-labeled secondantibodyas a
ofthe percentageofcellswithin the leukemicpopulationthatbore the antigen. Alternatively, a few samples were assessed by direct visualization withconventional immunofluorescence microscopy. If > 50%o of the cells bore the antigen, the monoclonalantibodywas scored aspositive (+), whereas<10% was scored asnegative, and values between 10and50% were listed in Tables I andII.
Monoclonal antibodies used here include the 3A-Imonoclonal that recognizes CD-7 (52),T1I (CD-2) (53), T3 (CD-3) (54),T4(CD-4) (54), T8 (CD-8) (54), T6 (CD-l) (54), B4(CD-19) (3), Bl (CD-20) (3), J5 (CALLA, CD-10)(55), and I2(HLA-DR) (56).Inaddition many of the cellswere analyzed with BA-1 (CD-24) (57),BA-2 (CD-9) (58),T12
(CD-6)(54), and for terminaldeoxynucleotidyl transferase(tdt)(data
notshown). CellslackingIg and TCR gene rearrangementswereexamined for myeloid-associated antigens with M02 (59), LeuMl (60), and LeuM3 (61).
patients cells that would phenotypically beclassified as Tcell
type ALLpossessed rearranged(3TCR genes(23/26) (Table I,
Fig.1). Most cells rearranged both alleles
(19/23) of their
1-chain genes, whileoneexceptionalcasedisplayedan
unantici-pated three rearrangements
of1 TCR restriction
thiscell revealed numerous
translocation, t(7: 11) (q35-36; q14)at or near
TCR, 7q35.Such an
breakpointthat fell within the
Alternatively, this mightrepresent clonal evolution or a dupli-cated TCR gene that
subsequently rearranged.This leukemia
possibilities. By utilizingthree
restrictionendonucleases, Bam HI, Eco RI, and
discriminatebetween rearrangements of
A-D).All three rearrangements in
(Fig.1 B). Incontrastboth rearranged
germ-line (Fig.1 C). Rearrangement
patient2 and several others
C62usage and the
TCRgeneexpression in T
cellALL. MostTcell ALLs with rearranged13 TCR genes also expressedthis locus (11/13) (Table I and
Fig.1E). Nearly all cases that expressed13produced the
transcript (10/11) signifyingthe presence
completed V/D/Jrearrangement. Four cellsexpressed the 1.0-kb RNA
compatible withthe product
ofanintermediate D/ J rearrangement. Three
ofthese also expressed a
lineage restrictedas Raji, a Burkitt lymphoma cell line, produced a truncated 1.0 kb
(notshown). Four of the Tcells that expressed13
failedtoexpress theaTCRsupportingasequential use of these genes.
a TCR gene expression in T
cellALL. The extraordinary
genomic Ja regionsmakesit currently rather
conclusivelydetermine the status ofa TCR rear-rangements in every T cell. For this reason we concentrated uponaTCR locus
expres-sion, slightlyoverone-half (8/14)of the T-ALLsexpressedthe aTCR locus (TableI,Fig. 2). Most oftheseTcells (7/8)possessed
the full length 1.6 kb transcript. Most T-ALLs with a TCR expressionwereofthe most mature type displaying surface T3. Mostof these (7/8) also expressed a 1.3-kb1 mRNAand pre-sumably possessedafunctional T3-Ti complex. One important exception expressed yandaunique 2.0 kbaTCRgenebutnot 13 (patient 7). In addition one case expressed a, 13, and y TCR genes(patient 4). No individuals expressed a alone, compatible with a developmental sequencewherea-chain production isa
later eventin T cell ontogeny. a TCR gene expression is not lineage restricted as RPMI 8432 a B cell lymphoblastoid line (Fig. 2), SU-DHL-6 a mature B cell and RS a pre-B cell (not shown) were all noted to make a truncated 1.3-kb form of the
yTCR gene rearrangement in TcellALL.MostTcellALLs
(23/26)have rearranged their -y TCR locus
Fig.3 A, B). Moreover the vast majority of T-ALLs that utilized y,
rear-ranged both alleles (17/23). Examples
(4/23)occurred in which
either one allele had been deleted or twoseparate rearrangements fortuitously createdthe same sized Bam HIfragment. Inspection of y TCR rearrangementin T-ALLs revealed thatapparently all hadrearranged into the Cz2 complex. In general T-ALLs
C.y1 containing 15.0 kb BamHIgermline fragment and no remaining
C72containing 12.5 kb Bam HI
germline fragment (Fig. 3 A). This findingresults from
C.Y2locusonboth alleles(Fig. 3 B). This
rearrangement pattern in T cells is different than the y TCR rearrangementsseenin pre-B cells
presentedbelow. Unusual patterns included
patient16 with three
patient16. Overall 22 of 26 T-ALLs
rearrangedboththeiry aswellas 1 TCRloci and the
importantexceptions will bedetailed later.
y TCR gene expression in TcellALL.
frequentrearrangement ofy TCR genesinT-ALLs this genewasonly
occasionallytranscribed atdetectable levels
analysisof total RNA.
Only3 of 14 T ALLs examined
(patients4and7and the HSB cell
cells possessedy TCR
withouty TCR rearrangement. Several cells
possessedrearrangement patterns thatargue
fixedorder to13andyTCR gene rearrangement. Two rearranged
-yTCR genes in the presence of
germline1 TCR genes were
evident in patient7
(Fig.4 A,Table I).
dem-onstrates a rare y TCR pattern fora Tcell in thata
germline12.5 kb Cz2 band
isretained arguing that one
ClyIlocus. It also expresses
(Fig. 3 C). Thiscell also
light chain loci. However, thisT-ALL
I). Thisillustrates that13 rearrangement is not
required fora TCR gene
activation.Moreover, the presence of cell surfaceT3 in the absenceof13 TCR gene rearrangementor
expressionin-dicates the presence ofaT3 receptor
oppositeTCR geneconfiguration was demonstrated by
e I + f NM
I~ I I I
o I I I
l- N C<4 + + +
+ ++ I+
+ + I++N + I 1+
+ I + I+ n +I I+
+ + + + + +
+ + + + + 't +
en + + + C 1 +
00CM++IC 0en 'IT
+ en + + + + + + + + + + + + + + + + + +
+ + + + + + + + + + + N + + + + + + + + + +
0 0 0 0
z z z z z z
~~0000~ 0 0000000
000 0 0 0000000
ucuU-M " 4 C4 N
9 0 9
-" " -4
CM CM CM CM - CM
N N - N
---C -- -4 e
-- C14Me1'u '' 10 N- 00 ON 0 C 4M 'It tn NC 00 0%O 0 - CM Cm
.-" " -"-" -l -4-4
+ + +
z Z w Z
00 00 0
CM C CM NCMC eq CM CMl
- - - U
rc c c c
U 0 0
CM CM4 CM
00 u =
.-" n C4 " - C4 eq C
Hindi11 Hindill EcoRi
EcoRI Hindill EcoRI BamHiHindill
cot D02 J32 CP2
D - COProbe -0
- -11Kb -.
_ -1l1Kb _
- t_4w -23Kb _
-.ag _ -6.5Kb
-!i w ~~~~~~~~~~~-35Kb
1.Dkb-Figure1.(A) Schematic presentationof the human,BTCR locusand
theutilizedBamHI,Eco RI, and HindIll restrictionsites. Variable
(V), diversity(D),joining(J),andconstant(C)regionsareshown. (B)
, TCR configuration in patient4 that shows threereangments
(ar-rows) of the
C,92complex when comparedtothe germline configura-tion(dash marks) revealed by thecontrol. The additional8 kb Eco RI
fragmentisseeninthecontrol andmayrepresentanEco RIresistant
t' 4t 't <
i z-- -13kb
5 w ~~~~~-1.0kb
site.(C) , TCR pattern ofCEMdemonstratestworearrangements of
theC~l complex. (D)
PTCRpatternofpatient 2 illustratesone rear-ranged
C,2allele.(E) Northern blotanalysis of total RNA from T ALLpatientsatdiagnosis(Dx)orrelapse (Rel)
and three T-ALL lines. Fulllength (VDJ)1.3kb and intermediate (DJ)1.0 kbtransciptsarenoted.
~,I..* .. E
e. e0 e
.,*-- 4~", !
eee e e e e ~~~~~A,
4t: 4- 4t 4t' t4 t C' Z !~j
Figure 2. NorthernanalysisofT-ALL atdiagnosis(Dx) plus three
T-ALLcelllines (CEM, 8402, HSB),one mature Tcellline (Hut-102),
and an EBVtransformedBcellline(8432).Fulllength 1.6-kbaTCR
transcriptsarenotedbydashmarks,and the 8432Bcellline revealsa
truncated 1.3 kb RNAyactinhybridizationis shown below.
kb Bam HI
patient 3 suggestingthat
ofthe y locus
either.No rearrangement was seen
withSac I or PvU II
that(3 can rearrange
noting (Table I).T-ALL
expressed both full
lengthaand (3 TCR RNA yet
defectivea or (3 TCR
of the T3 molecule.
andIg genes. Two ALLs
of either IgorTCR genes.
heavy,K, X genes as well as
with threeenzymes Bam
(91%),but lacked themore
definitive antigens ofTcell
T1IorT3. Asmall percentage of the
myeloidassociated markers Mo2
Patient18 also demonstrated
entirely germline configurations
of Igand TCR genes and
(Fig.5 B, Table II). The presence of y and (3 TCR gene rearrangement in several T-ALLs with 3A-1, butno T
developmentalsequence is not fixed as
T.,is present in the absence
ofgene rearrangement. Patients 18 and5represent very
Igand TCR gene level.
andTCR gene rearrangements. We
inwhat had been
pre-Bcell ALL than
chaingenes and lacked
antigens (Table II).A
displayedBI and had
pre-Bcell ALL group had
(Table II, III). Moreover,the presence
2, 4,and 17
However,the overall pattern
TCRrearrangement in the
observed in theTcell ALLs
is retained in the
TCR alleles retainedone or
pre-BALLs rearrange the C 1
fre-quentlyincontrast tothe T ALLs that
Cz2. Moreover, pre-Bcells
coordinate rearrangement ofboth (3
in which, was
leukemic subset ofcellsor
rearrangements within the entireB
phenotypeaswell as 14
transformedmature B cell
allele of their(
TCR rearrangements in
found in thematureB celltumors.This
leukemicsubset and donot
progenitorcells that generate themature
restrictedeither. Truncated 1.3-kb amessageswere
12.5Kb-0~) ,%. ~
k- a | = L
O 0)i ~
Pi PI VA
m- CyProbe -[
t~°qN q< zo
". _" -12.5Kb
-15- _ ra
Figure 3. (A) yTCRgeneconfigurationinT-ALLsatdiagnosis (Dx)
orRelapse(Rel)andintwoT-ALL lines. BamHIdigestsprobed with
aC'y probe placetheC1.l complexona15-kbfragmentand
12.5-kbfragment. Rearrangements(arrow)of bothallelesare
com-mon.Deletionof both germline
uniformusageof the C72complex.(B) Schematic presentation ofthe
human yTCRlocus and the BamHIrestrictionsites. (C)Northern
analysisof T-ALLs andTALL cell linesplusHut 102 for
marks. 7y-actinhybridizationis shown below.(D)yTCR
configura-tion inBamHIdigestedDNAfrompre-Bcell ALLpatients.In
con-trast totheT-ALLs, thepre-Bcells retainagermlinecopyof
F.3 PA Probe
Bam HI EcoRI
11 Kb- sw- 8.0Kb-- VW
TI,and usually T3 expressed the a TCR. The T ALLs that expressed theaTCRalso expressed
,3TCR with one important
I).Most T-ALLs areof sufficientmaturity to
I have rearranged both theiryand(3TCR genes and these events
expression.Most T-ALLs that
rear-ranged(3 in our study and in that by Sangster et al. (62) also
expressed it suggestingthat once rearranged, the(3 TCR locus
only occasional T-ALLs that rearranged the y TCR actually
ispossible that this gene
reflect. This is consistent withaproposed murine se-quence
expressionprecedes a, while( is expressed
throughout all of thymic development (33-36). Curiously, the
CTPT-ALLsusually haverearranged both y TCR alleles and have
Cyalleleswerealso observed. Such deletion events have
previouslybeen noted within IgHchain or K L chain loci and may be
isotypic exclusion(2, 50).
Whilemost T-ALLs had rearranged bothyand(3TCR genes important
4-Bam Hl EcoRI HindIII BamHI
*oat F S~s
eCJ e d
rearrange-ment,butgermline TCRs.(B)Patient3 reveals the unusual TCR
configurationinwhich rearranges,but yisretained in thegermline
CTOCT3 CTP JH
alater step inBcelldevelopment. Thus,thefrequencyofyTCR
rearrangementspilloverintothepre-BcellALLswasalsomuch greater thanthe frequency ofIgrearrangements within T cell ALLs.
ofTCR activation andTcellantigen expression. Thedetermination ofa, (3, yTCRgenestatusplus
Iggeneconfigurationcanbecombined withlineageassociated surfaceantigenstoclassify nearlyall ALLsaspre-BorTin type. The extensive characterization of TCRgenesinthisstudy
re-vealed thattheiractivationisnotsolely restrictedtotheTcell lineage,justasIggenerearrangement isnotrestrictedtoBcells.
However,the T cell ALLs did revealarough hierarchyof TCR activationeventsand cellsurfaceantigen expressionwithin the Tlineage (Fig. 6). Onlythemost matureT-ALLs which had
Ne A -23Kb -# -11Kb
CTY CTP CTTP CT3 JH
wellasIgH-chaingeneswhenexaminedwith theJHprobe. (B)
552 Felix, Wright, Poplack, Reaman, Cole, Goldman, andKorsmeyer
15Kb -* 12.5Kb-- -* is
15Kb- J4#$- A
R.. * -
23Kb-Table IL B-Precursor ALL Patients
TCR Genes Ig Genes Phenotype*
No. CTY No. CT# No. JH No. Ck No. CA No. 3Al T1 1/SRBC-R T3 B4 DR Calla BI
1 G R 1 R 1 R 1 - + + 11
-2*R/D 1/1 G R 2 D 2 R 1 - - - + + +
3 R 1 G R 2 G - - + + + +
4 RID 1/1 G R 1 R 1 - - +
5 G G R 1 G - - + + +
6 G G R 2 G - - 49 + +
7 R 2 G R 1 G - - - + + + 22
8 G G R 2 - - - + + + +
9 R 2 R 2 R 1 G - - - + + + 49
10 R 1 G R 1 G - 32 - + + 35
-11 G G R 2 G G - - - + +
-12 G G R 1 18 22 45 + -
-13* G G R 2 R 1 G - - + + +
14 R/D 1/1 G R 2 + + + 46
15 G G R 1 R 1 - - - + + -
-16 G G R/D 1/1 G G 12 30 - + + +
17"§R 2 R 2 R 2 R/D 1/1 G 32 25 + + +
*R,rearranged, D, deleted, G,germline,No., number of alleles. Data in whicha(-) is<10%of cellsshowing reactivity,a(+) is>50%
reactiv-ity,andspecificvaluesaregivenfor the 10-50% range. IPatient 17's WBCwasonly7,800and but53% blastsatthetime ofthisstudy. The3A-1 reactivity (32%) and T3 reactivity (25%)werecontributed bycontaminatingnormalTcells. *TheBcellassociated monoclonalantibody BA-I
was substitutedfor B4 in these cases.
,3TCR rear- that thereis no strict orderfor the initiation ofy and
#3TCR rangementand also
,3without My TCR rearrangement were noted rearrangement. However, we can not exclude thepossibility that here
(Fig.4 A,B).Similar configurations were present in the B unanticipated regionsof the yTCRregion mightexist that can cells
(TableII). These cells argue not be detected at present. Theearliest T cellassociated antigen
toappear is the3A-1 molecule although this antigen isnot
(Fig. 6) (52).The
sheepred Table III.Lineage spillover blood cell (sRBC) rosette receptor follows and T3 is present
expressionof theaTCR and the
patientsandcell lines of the
Rearranged JH: RearrangedCk:
B-Precursor ALLpatients RearrangedTCR y:
Rearranged 2 alleles TCR y: Rearranged 1 allele TCR y:
Rearranged I allele TCR -y, deleted I allele TCR y:
Rearranged TCR -y, butnotTCR A: Rearranged TCR
f,butnotTCR A: Rearranged TCR -y and TCR
Rearranged TCR y: Rearranged TCR
Rearranged Iallele TCR A: ClonalEBVcell lines
Rearranged TCR y: Rearranged TCR
Rearranged 1 allele TCR
1/1 Figure 6. Proposed coordinate sequence of TCRrearrangement and expression plus cell surface antigen expression in T-ALL. 3A-I
ap-0/14 pears to
0/14earlyin development, but in no fixed order. Most T ALLs are mature 3/14 enough to have rearranged both
cell subsets.Theexceptional T-ALL cases presented in Fig. 4 A, B may representunanticipated subpopulations of T cells
immortalizedby atransformation event. Patient 7 expresses a
uniquea and y TCRtranscript and displays T3 yet neither expresses nor even rearrangesthe
#TCR complex. This suggests
(ay,or ax, or
single chains mightbe complexed with the T3 molecule
within certaincells. This leukemia provides an opportunity to
potentiallyunique receptor complex that has
in nonleukemicTcell subsets (31, 32). Con-versely, patient 3 rearranges
y,suggesting that y rear-rangement
isnot a necessary
spillover ofTCRrearrangement. Only
10%ofT cell ALLsin this and prior studies rearranged their Ig H chain genes. Even
provides occasional evidencefor lineage
patient16 who rearranged his Ig Hchain gene also had the unusual presence
ofHLA-DR (usually seen onpre-B
(Table I). Perhaps
pathways is reflected by this cell's failuretoprogress
eitherB or T
ALLs that we
would havecategorized as
beingpre-B cell in type
20% ofpre-B ALLs rearranged
Fig. 3 D). Several
lines of evidenceargue that these
than being mis-classifiedearly Tcells.
All of these cells boreB
cell surface antigens thatare
rarelyor never seen in T-ALLs
includingHLA-DR, B4 and
uniformly rearranged theirH
chain genes. Moreover,even
cells than it did in theT-ALLs. They
in pre-B cellswere more
frequently utilized C1, whereas the T ALLs
rearranged into the
complex. Thus, bymany
differs from theT
rearrange-ment was not
witnessed inthe matureB
neoplasms and cell
pro-genitor cells that give riseto thesemore mature
srict order of locus
ofyTCR rearrangement argues that the
unanticipatedevent exist. The
eitherB or Tcell
pathways. Theylack the Ig
might preclude further effectiveTcell
lineage commitment. Alternatively,it
is conceivablethat the y
TCRrearrangements are a
transfor-mation.Perhaps these ALLs
lacking either completeIg or TCR rearrangements and because
proposalthe yTCR locus
vulner-able to further recombination. In this context it would not be surprising ifthe specific pattern ofyrearrangement were different in pre-B or uncommitted cells than in a truly committed T cell.
Standards oflineagecommitment. Considerable spillover
ofIg H chain rearrangement
withinbonafide mature T cells and
activation within bona fidemature B cells have been noted inthis and
(2, 14, 63, 64).Some
may be even more pronounced earlier in differentiation. This lack
lineage fidelity ofthese genetic events necessitates some
usingthese markers to determine a lineage
chainrearrangement appears to be a later event
indevelopment and to date hasbeen restricted to B cell devel-opment.
H-chainrearrangement thepresence of the B cell associated
and B1aswell as HLA-DR are
favorsa Bcell assignment, but some pre-Bcells
willrearrange yor (3andeven express a TCR genes.
cell commitment inALLs
andLchain rearrangement and the lack
ofTCRs isalso strong
evidence favoringa T
com-mitment,although the pattern
(3,andyTCR rearrangement and
antigen isseen on
fully lineage restricted,whereasthe
T1Iand T3molecules appear later in development and are more
patients lacked both Ig and TCRrearrangements
of the leukemiccells. It thus
by several criteria. The other leukemia displayed
committed its TCR loci.These
of commitment in
progenitors. The extensive characterization of Ig loci, TCR loci,
define the developmentalstate
rear-rangement events and clonal
of unique and
Wewishtothank Dr. Mark Davis for the
PTCRclone, Dr. Tak Mak for theaTCRclone,and Dr. JonSeidmanfor the
'yTCRclone used inthisstudy.We also wishtorecognizeMrs.MollyBoschfor her expert assistance in the
1. Korsmeyer, S. J., P. A.
Hieter,J. V. Ravetch,D. G. Poplack,
T.A.Waldmann, andP.Leder.1981. Developmentalhierarchyof
im-munoglobulingene rearrangements in human leukemicpre-Bcells. Proc.
Natl.Acad. Sci. USA. 78:7096-7100.
2. Korsmeyer, S.J.,A.Arnold,A.Bakhshi,J.V.Ravetch,U.
LeBien,J.H.Kersey,D.G. Poplack, P.Leder, andT. A.Waldmann. 1983.Immunoglobulingene rearrangement and cell surfaceantigen
leukemias ofTcell andBcell precursor
3. Nadler,L.M., S. J.Korsmeyer, K. C.Anderson,A.W.Boyd,B.
lymphoblastic leukemia.Amodel for discrete stages ofneoplasticand normal pre-B cell differentiation. J. Clin. Invest. 74:332-340.
4.Foon, K. A., R. W.Schroff,and R. P.Gale. 1982.Surfacemarkers onleukemia andlymphoma cells:recentadvances. Blood. 60:1-19.
5.Roper, M., W. M.Crist,R.Metzgar,A.H.Ragab, S.Smith,K. Starling, J. Pullen, B.Leventhal,A.A.Bartolucci, andM. D.Cooper.
1983. Monoclonal antibody characterization of surface antigens in childhood T-cell lymphoid malignancies. Blood. 61:830-837.
6. Acuto, O., M. Fabbi,A.Bensussan,C.Milanese,T. J. Campen, H. D. Royer, and E. L.Reinherz. 1985. The human T-cell receptor. J. Clin. Immunol. 5:141-157.
7.Royer, H. D., 0. Acuto,M.Fabbi,R.Tizard,K.Ramachandran, J. E.Smart, and E. L.Reinherz. 1984. Genesencodingthe Ti /3 subunit of theantigen/MHCreceptorundergorearrangementduring intrathymic
ontogenypriortosurface T3-Ti expression. Cell. 39:261-266. 8.Collins,M.K.L., G.Tanigawa, A.M.Kissonerghis, M. Ritter, K. M.Price, and S. Tonegawa. 1985.Regulationof T-cell receptor gene expression in human T-celldevelopment. Proc. Natl. Acad. Sci. USA 82:4503-4507.
9.Lebeau, M. M., M. 0. Diaz, J. D. Rowley, and T. W. Mak. 1985. Chromosomal localization of the human T-cell receptorU-chaingenes. Cell. 41:335.
10.Yanagi, Y., Y. Yoshikai, K. Leggett, S. P. Clark, I. Aleksander, and T. W. Mak. 1984.Ahuman T-cellspecificcDNAclone encodesa
proteinhaving extensive homologytoimmunoglobulinchains.Nature (Lond.). 308:145-149.
11.Clark, S. P., Y. Yoshikai, S. Taylor, G. Siu, L. Hood, and T.W.
Mak.1984.Identification of adiversitysegmentof human T-cell receptor /3chain,andcomparisonwith theanalogous murine element. Nature
12.Yoshikai, Y., D.Anatoniou,S.P.Clark,Y.Yanagi,R.Sangster,
P. Vanden Elsen, C. Terhorst, and T. W. Mak. 1984. Sequence and expression oftranscriptsof the human T-cell receptor/3-chaingenes. Nature(Lond.). 312:521-524.
13.Minden, M. D., T. Toyonaga, K. Ha, Y. Yanagi,B.Chin, E.
Gelfand,and T. W. Mak. 1985. T-cellantigenreceptor gene in human T-cellmalignancies. Proc. Natl. Acad. Sci. USA. 82:1224-1227.
14. Waldmann, T. A., M.M. Davis, K. F. Bongiovanni, and S.J.
Korsmeyer. 1985. Rearrangements of genes for the antigen receptor on
Tcells as markersoflineageandclonalityin humanlymphoidneoplasms. N. Engl. J. Med. 313:776-783.
15. Flug, E., P. G.Pelicci,F.Bonetti,D. M.Knowles, and R. Dalla-Favera.1985. T-cell receptor gene rearrangementsasmarkersoflineage
andclonalityin T-cell neoplasms. Proc. Natl. Acad. Sci. USA. 82:3460-3464.
16.Aisenberg, A. C., T. G.Krontiris,T. W. Mak, and B. M. Wilkes. 1985. Rearrangement of the gene for the /3 gene chain of the T-cell receptor in T-cellchronic lymphocytic leukemia and related disorders.
17.Weiss, L. M., E. Hu, G. S. Wood, C. Moulds, M. L. Cleary, R. Warnke, and J. Sklar. 1985. Clonal rearrangements of T-cell receptor genes inmycosis fungoidesanddermatopathic
lymphodenopathy.N. Engl. J. Med. 313:539-544.
18. Bertness, V.,I.Kirsch, G. Hollis, B. Johnson, and P. A. Bunn. 1985. T-cell receptor gene rearrangements as clinical markers of human
Tcelllymphomas.N.Engl. J.Med. 313:534-538.
19.Caccia,N.,G. A. P. Bruns,I.R.Kirsch, G. F. Hollis, V. Bertness, and T. W. Mak. 1985. T-cell receptorachains are located on chromosome
14 at14qI1-14q12inhumans.J.Exp. Med. 161:1255-1260. 20. Barker,P.E., H. D. Royer, F. H. Ruddle, and E. L. Reinherz. 1985. Regional location of T-cell receptor gene
chro-mosome 14.J. Exp.Med. 162:387-392.
21. Croce, C. M.,M.Isobe,A.Palumbo, J. Puck, J.Ming,D. Tweardy,
and J.Erikson. 1985. Gene fora-chainofhuman T cellreceptor.Location
onchromosome14region involvedinT-cell neoplasms. Science (Wash. DC). 227:1044-1047.
Nowell,and C. M.
Croce. 1985. Locusoftheachain ofthe T-cellreceptor is
chro-mosometranslocation in T-cell leukemias. Science(Wash. DC). 229: 784-786.
Minden,and T. W. Mak. 1985.
Analysisof cDNA clonesspecificfor human T-cells and theaand /3 chainsofthe T-cell receptor heterodimer fromahuman T-cellline.Proc.
Natl. Acad.Sci. USA 82:3430-3434.
24. Yoshikai, Y., S. P. Clark, S. Taylor, U. Sohn, B. I. Wilson, M.D.Minden,and T. W. Mak. 1985.Organizationand sequences of the variable, joining,andconstantregiongenes of the human T-cell receptor a-chain. Nature(Lond.). 316:837-840.
25.Yoshikai, Y.,N.Kimura,B.Toyonaga,and T. W. Mak. 1986.
Sequencesandrepertoire ofhuman T cell receptora chain variable
T-lymphocytes.J.Exp. Med. 164:90-103. 26.
and S.Tonegawa.1984.Athirdrearrangedandexpressedgene inaclone ofcytotoxicTlymphocytes.Nature(Lond.). 312:36-40.
27.Chien, Y.,D. M.Becker,T.Lindsten,M.Okamara,D. I.Cohen,
and M. M. Davis. 1984. A third type of murine T-cell receptor gene. Nature
28.Lefranc,M.P., and T. H. Rabbitts. 1985. Two tandemly
human genes encoding the T-cell y constant-region sequences show
multiplerearrangements in different T-cell types. Nature(Lond). 316: 464-466.
29. Quertermous, T., C. Murre, D. Dialynas, A. D. Duby, J. L.
Strominger,T.A.Waldmann,andJ. G. Seidman. 1986. Human T-cell -y chain genes.Organization, diversityand rearrangement. Science(Wash. DC).231:252-255.
30.Murre,C.,R.A.Waldmann,C.C. Morton,K. F.Bongiovanni,
T. A.Waldmann, T. B. Shows, and J. G. Seidman. 1985. Human y-chain genesarerearrangedinleukemic T-cells and maptotheshortarm
of chromosome 7. Nature(Lond).316:549-552.
31. Brenner, M.B.,J. McLean, D. P. Dialynas, J.L.Strominger,
J. A.Smith, F. L. Owen,J.G. Seidman, S. Ip, F. Rosen, and M. S.
Krangel. 1986. Identification ofaputative second T-cell receptor. Nature
32. Bank, I.,R. A. DePinho, M. B. Brenner, J. Cassimeris, F. W. Alt, and L. Chess. 1986. A functional T3 molecule associated witha
novelheterodimeronthesurface of immature human thymocytes. Nature
33.Raulet,D.H.,R. D.Garman,H.Saito,andS. Tonegawa. 1985. Developmental regulation of T-cell receptor gene expression. Nature
34.Snodgrass,H.R.,P.Kisielow,M.Kiefer,M. Steinmetz, and H. von Boehmer. 1985. Ontogeny of the T-cell antigen receptor within the
35. Snodgrass, H. R., Z. Dembic, M. Steinmetz, and H. von Boehmer.
1985.Expression of T-cell antigen receptor genes during fetal develop-ment in the thymus. Nature (Lond.). 315:232-233.
36. Haars, R., M. Kronenberg, W. M. Gallatin, I. L. Weissman, F. L. Owen, and L. Hood. 1986. Rearrangement and expression of T-cell antigen receptor and ygenesduring thymic development. J. Exp. Med. 164:1-24.
37. Minowada, J. 1982. Immunology of leukemic cells. In Leukemia. F.Gunz and E. Henderson, editors. Grune & Stratton, Inc., New York.
38. Stong, R. C., S. J. Korsmeyer, J. L. Parkin, D. C. Arthur, and J. H. Kersey. 1985. Human acute leukemia cell line with the t(4;1 1) chromosomal rearrangement exhibits B-lineage and monocytic charac-teristics. Blood. 65:21-31.
39.Lenoir, G. M., J. L. Preud'homme, A. Bernheim, and R. Berger. 1982.Correlations between immunoglobulin light chain expression and varianttranslocation in Burkitt's lymphoma. Nature(Lond.).
40. Epstein, A. L., R. Levy, H. Kim, W. Henle, G. Henle, and H. S. Kaplan. 1978. Biology of the human malignant lymphomas IV. Func-tional characterization of ten diffuse histiocytic lymphoma cell lines.
A. A.Sandberg. 1979. Establishment and characterization of human B lymphocytic lymphoma cell lines Baltn-3,4,5: intraclonal variation in the 13cell differentiation stage. Int. J. Cancer. 24:572-578.
42. Poiesz, B. J., F. W. Ruscetti, A. F. Gazdar, P. A. Bunn, J. D. Minna, and R. C. Gallo. 1980. Detection and isolation oftype C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-celllymphoma. Proc. Natl. Acad.Sci. USA. 77:7415-7419.
43. Anderson, L. C., K. Nilsson, and C. G. Gahmberg. 1979.
K562-ahumanerythroleukemic cell line. Int. J. Cancer. 23:143-147. 44.Gallagher, R., S. Collins, J. Trujillo, K. McCredie, M. Ahearn, S.Tsai, R. Metzgar, G. Aulakh, R.Ting,F.Ruscetti, and R. Gallo. 1979. Characterization of the continuous, differentiating myeloid cell line (HL-60)from a patient with acute promyelocytic leukemia. Blood. 54:713-733.
45.Maniatis,T., E. F. Fritsch, and J. Sambrook. 1982. Molecular
cloning: Alaboratorymanual. ColdSpringHarbor, New York. Cold
46.Feinberg, A.P., and B.Vogelstein. 1983.Atechniquefor radio-labeling DNA restriction endonucleasefiagmentstohigh specific activity.
Anal. Biochem. 132:6-13.
47.Chirgwin, J.W.,A. E.Przybyla, R.J.MacDonald, andW.J. Rutter. 1979.Isolation ofbiologicallyactiveribonucleicacidfrom sources
48.Gunning,P.,P.Ponte, H.Okayama,J.Engel, H.Blau, andL. Kedes. 1983. Isolation and characterization of full length cDNA clones for human a, ,B,andyactin messenger RNAs. Mol. Cell.Biol. 3:787-795.
49. Ravetch, J. V., U.Siebenlist,S. J. Korsmeyer,T. A.Waldmann, and P.Leder.
1981.Thestructureof the humanimmunoglobulin mu
locus: Characterization ofembryonicandrearranged J and Dgenes.
Heiter,P.A., S. J. Korsmeyer, T.A.Waldmann,andP.Leder.
1981. Humanimmunoglobulinkappalightchain genesaredeletedor
rearranged in lambdaproducingBcells. Nature(Lond.). 290:368-372. 51. Zata, M.
M.,B.J.Mathieson,C. Kanelopoulos-Langevin, and S.0.Sharrow.1981.Separationandcharacterizationoftwocomponent
tumorliles within the AKRlymphoma. AKTB-1, by fluorescence-ac-tivated cell sortingand flowmicrofluorometry
52. Haynes, B. F., D. L. Mann, M. E.Hamler, J.A.Schroer,J. H.
Shelhamer,G. S. Eisenbarth, J.L.
Mostowski, and A. S. Fauci. 1980.Characterizationofamonoclonal
antibodythatdefinesanimmunoregulatoryT-cell subset for
immuno-globulin synthesisin humans. Proc. Natl.Acad. Sci. USA. 77:2914-2918.
53. Reinherz, E., K. A.Fitzgerald,R. E.Hussey, J. Ritz, and S. F. Schlossman. 1981. Determination of T celllineageof acutelymphoblastic leukemia tumor populations: Utility of anti-sheep erythrocyte mono-clonalantibody.Blood.58(Suppl. 1): 150a. (Abstr.)
54. Reinherz, E. L., P. C. Kung, G. Goldstein, R. H. Levey, and S. F. Schlossman. 1980. Discrete stages of human intrathymic differ-entiation. Analysis of
normalthymocytes and leukemic
lymphoblastsof T-cell lineage. Proc. Natl. Acad. Sci. USA. 77:1588-1592.
55. Ritz,J., J. M. Pesando,
Notis-McConarty,J.Lazarus, and S. F. Schlossman. i980.Amonoclonalantibody to human acute lymphoblastic leukemia antigen. Nature (Lond.).283:583-585.
56.Brodsky, F. M., P. Parham, C. J., Barnstable, M. J. Crumpton, and W. F. Bodmer. 1979. Monoclonal antibodies for analysis of the HLA system. Immunol. Rev.47:3-61.
57. Abramson, C. S., J. H. Kersey,andT. W.LeBien. i981. A mono-clonalantibody(BA- 1) reactivewithcellsof human Blymphocyte lineage.
58. Kersey, J. H.,T.W.LeBien, C. S. Abramson,R.Newman, R. Sutherland, and M. Greaves. 1981.P-24:Ahumanleukemia-associated andlymphohemopoietic progenitorcellsurface structure
identifiedwith monoclonalantibody.J.Exp. Med. 153:726-731.
59. Todd,R.F., L. M. Nadler, and S.F.Schlossman. 1981. Antigens
onhuman monocytes identifiedbymonoclonal antibodies. J. Immunol.
60. Hanjan,S. N. S., J. F. Kearney, and M. D. Cooper. 1982. A
myelomonocyticcells.Clin. Immunol.Immunopathol. 23:172-188.
61.Dimtriu-Bona, A., G. R. Burmester, S. J. Waters, and R. J. Winchester. 1983. Human mononuclear phagocyte differentiation
an-tigens. I. Patterns ofantigenic expression on thesurface of human monocytes and macrophages defined by monoclonal antibodies. J.
62.Sangster, R. N.,J.Minowada, N.Suciu-Foca,M.Minden,and T. W. Mak. 1986. Rearrangement andexpression of the a,
chain.Tcell receptor genes in humanthymicleukemia cells and func-tionalTcells. J.Exp.Med. 163:1491-1508.
A.,N.Hozumi,M.Minden,T.W. Mak,and E. W.Gelfand. 1985.Rearrangement of the T-cell receptor
Engl.J. Med. 313:
64.Pelicci,P.G.,D.M.Knowles,and R.Dalla-Favera. 1985. Lym-phoidtumorsdisplayingrearrangements of both