Adaptation of an Amphibian Renal Carcinoma
in Kindred Races
*
KENYON
S.
TWEEDELLt
(Department of Zoology, University of Illinois, Urbana, Ill.)
@
fowl8ri,allfromMassachusetts,
alsoreceivedin
The renal adenocarcinoma
of the common leop
ard frog, Rana pipiens, generally known as the
Luckécarcinoma, is found naturally in about
per
tra-ocular implants of one of the original tumors.
Tumor transplants were also made to geograph
ical race hybrids produced by crossing R. pipiens
of Vermont and Wisconsin. Some of the implants
were made from a naturally
occurring tumor,
cent of the frogs obtained
from the Lake Cham
plain Valley in northern
Vermont (@9, 35, 46). The
carcinoma
is transmissible,
presumably
by way of
RTX1, into the taiffin prior to metamorphosis. A
a virus-like agent (@9,30, 44). The agent from the
Vermont carcinoma is specific to renal epithelium
(80) and also particularly race-specific (3@). It will
not induce tumors in races of Rana pipiens from
New Jersey (@9) or from Wisconsin (43, 44) nor in
alien species of amphibians, fish, and reptiles (33).
Recently, it has been demonstrated
that the
renal agent is modified by passage through sala
manders in such a way that it will induce growth
in other tissues (43, 44).
The present work is a study of transformations
resulting from growth of the Luckétumor from
second group of young adults received intra-ocular
implants of RT91, a tumor induced in a Vermont
R. pipiens from RT43.
Production of hybrid@.—Eggsand sperm were
Rana pipiens of Vermont in eye chambers of a
kindred
report
@
race of R. pipiens
also includes
a study
from Wisconsin.
This
of host reaction
when
the tumor is implanted in eyes of hosts of varying
degrees of relationship to the Vermont donor.
MATERIALS
@
AND
METHODS
Species, races, and tumors employed.—The tu
mors used as a source of transplant
material
were RT37, a renal adenocarcinoma
which oc
curred
naturally
in a Rana
pipiens
from Vermont,
and RT4S, which had been induced
by an intra
ocular tumor implant in another
R. pipiens from
Vermont.
These two tumors were transferred
to
the eye chambers
of other adult Vermont
R. pipi
ens and to R. pipiens of kindred races from Illinois,
Kentucky, and Wisconsin. Heterologous species,
R. clamitans, R. palustris, R. cates&kina and Bufo
obtained from four Rana pipiens, two from Ver
mont and two from Wisconsin. Ovulation was in
duced following a modification of the method de
veloped by Rugh (45). Four groups of animals were
thus produced : Vermont female X Vermont male,
Wisconsin female X Vermont male, Vermont fe
male X Wisconsin male, and Wisconsin female X
Wisconsin male.
Operoiional
procedure.—Implantations
were
made in the following manner. The frogs were
anesthetized
in an iced solution of tricaine meth
anesulfonate
(MS
and
the
tadpoles
were
anesthetized
in a 1/10,000 concentration
of MS
in Holtfreter's
solution. Immediately
after
wards the tadpoles were transferred to a 1/15,000
concentration
for the maintenance
of the anesthe
sia. Carcinomatous renal tissue, biopsied from the
donor animal, was transferred immediately to an
iced
solution
of amphibian
Ringers
and
divided
into 0.5-c. mm. bits of tissue.
For intra-ocular implantations
a small piece of
tumor tissue was selected, together with a pinch of
sulfadiazine
powder,
with a watchmaker's
into
the anterior
and
the
two
were
forceps beneath
eye chamber.
The
inserted
the cornea
implant
was
then lodged between the iris and the cornea by
a Revision of a portion of a thesis submitted in partial ful
gently stroking the outside of the cornea.
flilment of the requirements
for the Doctor of Philosophy
Prior to tail implantation
an incision-excision
degree in zoology at the University of Illinois.
A summary of this work appeared as part of an abstract was made in the dorsal proximal third of the tail
published in Transplantation Bulletin, 1:27—28,
1953.
mesenchyme with a surgical needle according to the
t Present address:Department of Zoology, University technic of Briggs (6, 7). Small pieces of tumor tis
of Maine, Orono, Maine.
sue were then transferred
with a watchmaker's
forceps to the tail and inserted into the subcuta
Received for publication April 7, 1955.
410
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
TWEEDELL—AdaptatiOn
of an
neous tunnel, along with a small portion of sulfa
diazine.
Controls.—Controls
of the homologous
race
from Vermont were maintained
under the same
environmental conditions but were not submitted
to any operation. From these was learned the mci
dence of tumors arising without treatment
under
laboratory
conditions.
Controls which received
implants of normal renal tissue were also observed.
Similar nonoperated
controls were maintained
for each of the four groups of pre- and postmeta
morphosed
Amphibian
411
Tumor
showed tumors (Table 1).
There was no immediate
the eye chamber when the
were both from Vermont
attached to iris or cornea
grow slowly after
reaction to the graft in
donor and host animals
(Fig. 1). Grafts usually
or both and began to
a few days. Occasionally
the im
plant passed through the pupil and attached
to the
lens (Fig. %). Growth from all these sites varied
greatly, but after the greatest growth the anterior
chamber was filled with tumor tissue in 4-5 weeks
(Fig. 6). It is unusual for the initial growth to be
sustained. More often, the tumor implant, after
animals.
TABLE 1
REN@tL TUMOR EYE CHAMBER
KINDRED
IMPLANTh
MADE
INTO HoMoLoGous
RACES AND HETEROL000US
AND
SPEcIEs
PIPI@($RANARazaRazeaBuroVermont
RANA
[email protected]'m.row.To.Total
Wisconsin
Ken
PAL.To
tuckyteltalRenaltumornumber
374343374337No.animalsoperatedupon37
11
43
24
35
37
26
43
20
9
55No.takes
No.survivors
8
22
30
22
14
820
8165120721553Permanentregression
7
21
28
13
14
7
11
18
12
2
4
0
0
0
2
0
0
2
0
0
3@
2
0
211
5
0
2#
2
0
Sustained growths:
Typical
Withhosthypertrophy
Collapsedeye
Primary regrowth:
Typical
1775
6152
227t
721@ 215
585
3#
10
0
0
0
3
3
1
0
10
0
0
5
0
0
0
0
0
0
0
5
4
0
0
0
0
0
0
0
0
0
0
0
4
0
0
02
038
1
8
9
2
3
1
8
Withhosthypertrophy
0
0
0
3
1
0
0
Secondaryregrowths
0
0
0
1
1
0
1
9
4
3
0Percentageofinducedtumors
No.ofinducedrenaltumors
4
57
11
52
15
54
0
0
0
0
09
0
027
011
07
018
8
12
20
0
0
0
0
0
2
10
2
10
0
0
0
10
0
0
0
0
0
0
0
0
00
100
00
00
00
00
takes0000000No.unoperatedcontrols
from
Spontaneous tumors in controls
0S
No.operatedcontrols(normal
0
kid.)0
adulta.)t
(Fifteen
mature
adUlts
; 57 young
metamorphosed.)(Three
(Fourmature
adults;
threere@ntJy
metamorphosed.)*
mature adults; eighteen recently
eyes.if
Theseeye chamberimplantsbecameconvertedintoeitherrainboworcollapsed
chamber.I
Growthfilledeyechamberrapidly,thenhistolysed,followedby a moderatecollapseoftheeye
One eye chamber collapsed.
RESULTS
There was a pronounced
difference resulting
from implantation
of the original Vermont carci
growing for less than a month, was invaded and
covered by pigmented tissue from the iris. It then
nomas
at the most, it remained
in eye chambers
of R. pipiens
of Vermont
and in R. pipiens from other geographical areas.
There was a reaction in the foreign eyes which was
lacking in the Vermont eyes. A further difference
is that
tumors
arose in host kidneys
when the hosts
were from Vermont but not when the hosts were
from other geographical areas.
R.ana pipiens
@
grafts took in
teen of the
kidneys (Figs.
for 15 months,
mental animals
of Vermont
as host.—The
tumor
@8
of 30 hosts, and subsequently flu
developed typical tumors in their
3 and 4). The frogs were observed
and, while over half of the experi
produced tumors in their own kid
neys, only 10 per cent of the unoperated controls
regressed
slowly until
nothing
visible
remained
or,
only as a tiny pigmented
knob on the iris (Fig. 7). Sometimes
the regressed
tumors began to grow again after a period of dor
mancy lasting from 8 to @1weeks and usually
grew at a greater rate than the implants before re
gression (Fig. 8). These rapidly growing tumors
which arise after one period of regression and dor
mancy have a median life of 3 weeks and are called
primary
regrowths.
At the
time
of primary
re
growth there was almost always a palpable and
rapidly growing induced tumor in the host kid
neys. However, tumors were induced in host kid
neys not only in those frogs whose grafts became
active after a period of dormancy,
but also in frogs
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
41@
Cancer Research
in which grafts regressed completely and in others
in which the graft never regressed.
Implantation
into kindred races and species.—
There was a much greater reaction to the tumor
grafts by the host eye tissues when the hosts were
of the same species but not from the same geo
graphical area. The kindred races which received
the Vermont carcinoma were R. pipiens from flli
nois, Kentucky, and Wisconsin. Forty-two Wis
cousin frogs received the Vermont tumor in this
sue does not grow when transplanted
to other
eyes and that it does not induce tumors, it is a rea
sonable presumption that the malignant property
of the original renal carcinoma had not been trans
experiment
ferred
and
38 in a former
experiment
(44),
making a total of 80. A very strong reaction to the
graft was also observed when the grafts were made
to R. palustris, a species closely related to R.
pipiens. Viable hybrids are produced (37—40)
from
@
host reaction
to these implants,
and, although
the
implants took and remained active for 1—@
weeks,
they regressed and remained dormant for the en
tire period of observation, which was 6 months. No
tumors were induced in any of the Wisconsin
hosts. From the facts that this hypertrophied tis
to it.
There is also a striking effect of the Vermont
renal carcinoma on lenses in eyes of animals of
kindred races. The lens may lyse partially or total
ly and a new lens or several new ones regenerate.
Regeneration
of lens after tumor implantation
is
of interest because adult frogs do not naturally
regenerate the lens. This part of the study is coy
ered in more detail elsewhere.
crosses of the above kindred races with each other
and with R. palustris and may account for the
similarity of host reaction.
The immediate reaction by the kindred races
and by R. paluatris was observed within a few
In a control paralleling the transfer of carcino
hours as clouding of the cornea, increased opacity - ma, implants of normal kidney tissue from Ver
mont R. pipiens were made into ten each of the
of the graft, loss of a few cells from the graft, and
inflammation of the periphery of the eye. Although
Vermont and Wisconsin animals. The takes were
actively maintained from
to 4 weeks. While no
this reaction is severe, most of the implants re
covered from it and established a vascular connec
tumors arose from this transfer, there was no re
tion with the host iris. Subsequently,
some grafts
action by the host to the implants of normal tis
continued proliferation,
others regressed perma
sue. The period of active growth should be suffi
nently or remained dormant only to reoccur as cient time for a reaction to take place.
primary regrowths. In contrast to the previous
More dietantly related speeies as hosts.—TheVer
mont R. pipiens tumor was also implanted in eyes
transfer into Vermont animals, tumors were not
induced in any of these closely related hosts.
of more distantly related species, R. cafr,s&iana and
In fourteen of 33 cases where the tumor implant
R. clamil,ans, and in eyes of the toad, Bufofowleri.
grew appreciably
in the kindred race or in R. The more distantly related hosts did not respond
palustris, there was marked hypertrophy
of sur
with either a primary reaction or later hyper
trophy.
Instead
they tolerated
the Vermont
rounding normal tissues. When Vermont tumors
were implanted into the homologous hosts from
pipiens tumors as well as Vermont pipiens do.
However, these grafts were finally reacted against
Vermont, there was never appreciable hypertro
by the host, as are most tumor grafts even when
phy of normal eye tissues.
Vermont hosts are used, and eventually
disap
Irises of the fourteen eyes of the animals of kin
peared. In the case of the R. clamitans, eventual
dred races became extremely hyperplastic
and
lysis of the implant and concurrent lysis of the lens
within 8—1@
weeks grew up over the tumor and the
and other eye tissues led to collapse of the eye
inner side of the cornea, completely lining the cor
chamber, a condition which was permanent
(Fig.
nea with black and gold iris pigment. The over
5). A similar
effect
was occasionally
seen when
growth of pigment cut off the view of all changes
within the eye chamber. At times, however, lysis kindred species were hosts. Although the implanth
grew well for several months, no renal tumors were
of the cornea and evagination of internal prolifer
ating tissues were apparent (Fig. 9). Such an eye induced in the host kidneys. This was also true, as
noted above, when hosts were of kindred races.
shows in section a mass of wildly growing pigment
Only when hosts of the same species and the same
ed cells of iris and a hyperplastic retina which has
been thrown into folds by its overgrowth (Fig. 10). geographical locality were used did the implants
induce tumors in the hosts. In the next section it
No organized renal tumor remained.
will be shown that a tumor can be changed by
To test whether the hypertrophied
tissue had
growth in the eye of a kindred race so that it will
become malignant, hypertrophied
iris and retina
were taken at the @6thweek from the same eye produce tumor agents which can induce tumors in
shown in Figure 10 and implanted in eye chambers
the kindred race.
Regrowth and subcvlture of dormant tumors.—
of ten R. pipiens from Wisconsin. There was no
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
TWEEDELL—Adaptciion
Usually a tumor implanted in an eye chamber
either takes and grows or regresses, and there is no
further change. However, in some cases (Table 1)
regrowth of the tumor followed regression and
dormancy. This happened in all types of hosts, in
those of the homologous race, in kindred races, and
in foreign species. The median time after implan
tation for primary regrowths to appear in eyes of
Wisconsin frogs was @6
weeks, with a median dura
tion of 1@weeks. One of these regrowths was se
lected for further experimentation.
At 35 weeks
after the original implantation
a biopsy was per
formed. Part of the tumor was sectioned (Fig. 11).
It is a typical renal adenocarcinoma. Bits of the
biopsy material were implanted in eye chambers of
eight Vermont R. pipiens and five Wisconsin R.
of an
Amphibian
418
Tumor
secondary regrowth were subcultured in eye cham
bers of ten Wisconsin and in ten Vermont pipiens.
These transfers and the results are summarized in
Chart 1.
It is noteworthy that no Wisconsin host which
received a Vermont tumor implant developed a
tumor in its own kidney. This holds true even for
those hosts which supported primary and second
ary regrowths in the eye chamber. These Wiscon
sin hosts, although immune to the tumor agent,
were harboring in the regrowths a changed agent
which had acquired the new ability to induce tu
mors in the host race while retaining the ability to
induce tumors in the donor race (Table @).
Subcultures of both primary and secondary re
growths induced renal tumors in Wisconsin and in
[email protected] afterwards the remainder of the
Vermont
tumor in the eye of the donor regressed, but by the
45th week the eye chamber was filled with a see
ond regrowth. Complete regression of a primary
regrowth followed by secondary regrowth was ob
served only 3 times. The secondary regrowth, like
the primary, was histologically typical. Bits of the
teen in which the regrowth implants had taken
succumbed to tumors which developed in their
own kidneys. The number of Vermont hosts sue
cumbing to the same type of tumor was six out of
sixteen. Primary and secondary regrowths were
equally effective in inducing renal tumors. The
frogs.
Five
Wisconsin
hosts
out of thir..
IDN@
WISEDNS
VERMONT
@
\IIIEEDNBIN
TUMORS:
CHART 1.—Transplantation
VERMONT
WIBEONSIN
RENRL@@:@OTHERS
of a renal tumor
from a Vermont
Ilana pipiena into the eye chamber of a Wisconsin Rana pipiena
followed by ocular regrowth,
into new Vermont
subculture,
and Wisconsin
and transplantation
hosts.
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
.414
Cancer Research
change from the original Vermont tumor, which
was incapable of inducing tumors in Wisconsin
frogs, must have come during or prior to the first
regrowth of the dormant implant.
A striking difference was noted in the amount
of proliferation of the eye implants derived from
a subculture of the first or second regrowth. For a
comparable period the total growth was far greater
arose after passage and growth of the transformed
Vermont renal tumor agent through salamander
cartilage and frog skeletal tissue (44). These pox
like lesions have been observed only when the
original agent had been changed by growth in a
foreign host or in the donor animals possessing see
ond ocular regrowths.
Tadpoles and young adults of Vermont, Wiscon
in the Vermont eyes with implant from the second sin, and their reciprocal hybrids as hosts.—Because
@
@
regrowth (Fig. 1@)than from the first. The inverse
was true in the Wisconsin eye chambers, for the
second regrowth implants gave a slow, inhibited
type of ocular growth (Fig. 13) even though the
temperature
(34) was constant at
C.
There was a further change in the tumor be
tween the time of first and second regrowths. Im
plants of the secondary regrowth induced a new
TABLE 2
SUBCULTURE OF PRIMARY AND SECONDARY EYE CHAM
BER REGROWTHS FROM WIscoNsiN
Rana pipiens INIO
EYE CHAMBERS OF Rana pipiena FROM VERMONT AND
WISCONSIN
RANAPIPITNI
Vermont
Total
@
1° 2°
Subculture
10
No.operated
animals
8
8
8
10
9
8
18
17
16
6
3
5
3
0
8
2
0
No. survivors
No.takes
Permanent regression:
Induced renal tu
mors
Other growths
No induction
Sustained growths:
Induced renal tu
mors
Totalnumber
of renal
tumors induced
Percentage of takes:
Inducing renal tu
Wisconsin
Total
2°
11
6
5
5
4
5
8
10
9
9
7
1
15
14
18
12
4
2
2
2
5
0
2
3
8
8
5
8
0
5
0
0
0
2
1
2
1
of the difference in behavior of Vermont and Wis
consin frogs toward the original Vermont tumors
and their agents, the behavior of hybrids to the
original tumor was studied. Tumor implants made
into the taiffin of young larvae all regressed before
metamorphosis,
which
began
months
after
the
hosts had received the tumor implants. It was
learned that, contrary to the behavior in mature
adult Wisconsin frogs, the Wisconsin tadpoles, as
well as the Vermont and hybrid crosses, did de
velop the Luckécarcinoma. However, none of the
tadpoles developed renal tumors until the end of
metamorphosis
or thereafter,
from
4—8 months
after implantation
(see Fig. 16).
Ocular implantation
of the tumor into recently
metamorphosed
animals
was accomplished
in all
cases, and all the implants regressed after
months.
These animals were observed
for 8
months, and both the parent races and their hy
brids were found refractory to the Vermont agent
(Table 3).
DISCUSSION
AND CONCLUSIONS
Host resietance.—Generally, the closer the rela
tionship to the Vermont Rana pipiens, the greater
3
3
6
3
2
5
was the immediate response of the host animal to
implantation of the Vermont renal carcinoma; the
Av.
Av.
75 24 49.5 more distant the association, the less the host re
37 37 87
mors
action. Moore's experiments
(37—40) verify the
0
25
12.5
0
88
16.5
Inducing all other
close genetic relationship of the intraspecific ani
growths
mals used in the present experiments. Since they
are quite compatible in hybridization
and exhibit
type of lesion in both Vermont and Wisconsin
resistance
only after tumor implantation,
relative
hosts. These were small growths arising concur
rently with the renal tumors but restricted
to ly great differences in the genomes of the kindred
races are not likely to be the cause for the reaction
those hosts which did not develop renal carcino
The induction of
mas (Table @).They developed in many parts of following tumor implantation.
tumors in the young tadpole hybrids suggests that
the body, in skeletal muscles, liver, kidney, and
tumor resistance might be controlled by a gene(s)
spleen (Fig. 15). These small growths, never more
affecting histocompatibility,
as in mice.
than a few millimeters in diameter, were anaplas
This paradoxical sensitivity response of hosts
tic, grew for only a few days, lysed (Fig. 14), and
has a parallel in normal tissue transplantation.
were replaced by connective
tissue. The time
of embryonic chick tissue to the
course of those which arose in the muscles was Implantation
brain, eye, or muscle of an adult bird (1) is accom
traced. Characteristically,
multiple small, hard
panied by a greater lymphocytic
response than
beans were palpable under the skin for a few days.
when heterologous
tissue is transplanted.
The
These and the skin over them lysed, leaving open
lesions. The frogs often succumbed a day or so same embryonic chick tissues can be transplanted
after the extensive lysis. Histolytic lesions also easily to adult mammalian brain, where they re
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
TWEEDELL—AdaptatI@n
of an Amphibian
415
Tumor
tam their susceptibility to chicken tumor agents.
and regression of the ocular implant in the young
Greene (17) states that embryonic rabbit skin adult frog hybrids (without resultant induction of
transplanted
to the mouse brain does not excite
tumors in either event) may be the result of anti
the foreign-body reaction associated with the in
bodies produced by the host. It is likely that the
troduction
of heterologous
adult tissue. After
renal agent still persists, but because of the newly
maturation,
the implanted
tissue maintains
a acquired host resistance it is prevented from ex
susceptibility to the Shope papilloma agent and is pression. Immunological reactions can occur in the
not influenced by the immunological response of eye chamber (48), and the host is capable of caus
the host. Eichwald et at. (13) transplanted
strain
ing antibodies against alien tumor cells (4@) but
resistant tumors into the anterior eye chambers of not against the agent within the cell.
mice, where they distinguished
first a “natural―
TABLE 3
resistance influenced by species and strain differ
iMPLANTATiON
OF RENAL CARCINOMA INTO THE TAD
ences which prevent a “take―
and an acquired im
POLE TAILFIN AND THE EYE CHAMBERS OF YOUNG
munity, a result of the host's response to the
ADULTSFROMTHE HoMoLoGous AND RECIPROCALHy
tumor.
BRID CROSSES OF VERMONT AND WISCONSIN RACES
The comparable qualities of “natural―resist
OF Rana pipien.
ance, the autonomous
response of some trans
Wiscon.
planted normal tissues, and the present race re
sin
Vermont
x
x
sistance may result from affecting a level of or
ganization
that
logical factors
foreign-body
is relatively
minor
to the immuno
which act in the promotion
response.
Therefore,
a greater
of a
imme
diate conflict would occur from the introduction of
a modified cell or cellular particulate that is for
eign to the local environmental level of the kindred
race. The incongruities caused by the tissue im
plant may emerge at the time cell malignancy is
acquired. This interpretation
would conform with
the views of Green (15) and Murphy (41), and the
modified hypothesis advanced by Rose (43) that
the Luckétumor agent is a mutated normal tissue
specific particle.
At least another factor is involved in this re
sponse, since resistance of the host tadpole to a
foreign implant does change between early and
late larval stages. In the late larval stage, prior to
metamorphosis,
all the growths at the implanta
tion site were resorbed. Changes in the host tad
pole resulting from the process of metamorphosis
are not the cause for this regression (6, 7). Con
ceivably, the taiffin implants which do not induce
tumors until after metamorphosis
may behave as
the ocular implants in the mature adult animals
which regress temporarily,
followed by alternate
periods of growth and regression.
Furthermore,
it was not possible
to induce
renal
tumors in the young adult hybrids of either the
homologous or kindred race. Resistance in young
homologous adult hosts has been previously noted
by Lucké(@9)and Rose (44). It is also known (11,
1@) that, associated with early growth of normal
chickens, neutralizing
antibodies develop in the
blood for viruses of avian tumors such as the Rous
sarcoma.
The repeated growths and regression of the tu
mor graft in the eye chamber of the kindred races
.
Wiseon.
Vermont
Operated
sin
Wiscon.
sin
Var.
mont
animals:
tadpoles
20
20
adults
No. takes:
20
25
tailfin
eye chamber
No. controls:
tadpoles
adults
Tadpole length:5
initial
plus 8 weeks
Animals surviving
metamorphosis
Ocular implants:
18
20
25
20
25
18
12
18
20
16
17
16
25
0
9
9
0
10
0
10
27.7
50.2
24.5
45.8
5
12
11
0
13
1
0
16
0
22.7(25.6)t
43.4 (45.5)
10
(11)
@4.5
48.5
regrowths
regressions
0
14
6
12
lens growths
Induced renal tu
mors:
0
2
tadpoles
4
2
adults
0
0
8
0
2
0
0
0
0
Spontaneous tumors
in controls:
tadpoles
0
adults
0
a Mean average in mm.
t Figures in parenthesesare for tadpole controls.
Eye-chamber subcultures.—Thechange in the re
growths in the eye chamber of Wisconsin hosts
suggests that either the reaction of the host is
modified or the tumor tissue is altered to fit the
host environment,
although both events could
happen.
Snell, ICaliss et at. found that the growth of
mouse tumor homoiografts implanted into mire
lated, inbred strains of mice (in which the tumor
would normally regress) could be enhanced by a
series of pre-injections of lyophilized cancerous or
normal tissue (18,
@,
50, 51) from the homologous
mouse tumor strain, by unfiltered or filtered breis
of fresh tumor or normal liver (19, 49) or from
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
416
Cancer Research
antiserum produced against them (p20). Similarly,
if the XYZ factor is injected into rabbit hosts,
there is a significant increase in both the growth
rate and the number of takes when the rabbits are
inoculated with the Brown-Pearce tumor (8), but
it has no effect when tried on other mouse
suitable host will return the virus to its original
form.
Again, working with phage, Bertani and Weigle
(5) found
the
existence
of a nonheritable
modifica
Pearce carcinoma (16) has overcome refractory
strains, while inoculation of a carcinogen-induced
tumor into alien strains (@5) may result in a
change of both the host and the tumor.
From these examples it is evident that the
specificity of a tumor can be circumvented
in a
way that does not involve the usual genetic modi
tion as a direct result of introducing the phage
into a new host, in which the progeny can follow
one of three rapid courses: adaptation, no change,
or dc-adaptation.
The modification of the specific renal carcinoma
in the eye chamber of the Wisconsin hosts is prob
ably an adaptation
imposed on it by the host's
environment;
each new proliferation or regrowth
further disciplines the implant. If this new toler
ance is temporary (an induced renal tumor in Wis
consin hosts shows a loss of specificity upon fur
ther transfer), then it is similar to that found in
phage.
The concept of tissue adaptation has appeared
recently in tissue culture studies. Embryonic tis
sue fragments of rabbits cultured in adult cat tis
sues (@3, @4)
willassimilate alien proteins into their
cells. It is not unwarranted to suppose that the eye
fications,
implant
tumors (9).
Another successful technic (14) is treatment of
the host with estrogenic hormones before tumor
implantation.
Parabiosis (10) with a susceptible
animal has been used to overcome the natural re
sistance of a normal refractory animal. The use of
x-rays prior to inoculation of the tumor raises the
number of successful implants in mice (53). Im
plantation
of embryonic
such as mutations
tissues
with
or selection
the Brown
of mutant
types. However, since the ocular regrowths of the
implanted carcinoma do not induce tumors except
when the tissue is subcultured to new hosts, the
adaptations
described do not account for the
alteration
in the specificity
of the regrowths.
Serial transfer or “aging―
(5@) of the tumor by
numerous transplant generations through the eye
chamber is not the primary factor in modifying
the carcinomatous
agent. Passage of the frog car
cinoma in the anterior chambers of homologous
Vermont frogs for fourteen serial generations (47)
never altered the specificity of the agent.
The second interpretation
necessitates that the
alteration in tumor specificity is caused by the
host. Barrett and Deringer (@) studied a mam
mary adenocarcinoma
which grows in C3H mice
but not in a resistant strain C. When the tumor is
transplanted
into an F1 hybrid host (C3H X C)F1
and then retransplanted
into resistant backcross
hosts, there is a marked increase of hosts with
tumors. This “adaptation― is permanent
(3),
acquired after the first generation, and also rca
tively specific (4).
A similar and very significant host-induced
modification of infecting bacterial viruses is de
scribed by Luria and Human (36). The phenotype
of a new virus, reproducing in a bacterial cell, is
molded by the genotype of the bacterial host in
such a way that the virus has the ability to repro
duce in some hosts but not in others. This change
can be accomplished in one generation of growth
but is only temporary;
one cycle of growth in a
is exchanging
certain
species-indigenous
proteins of the host during its period of adapta
tion in the eye chamber.
STTh@LMARY
Intra-ocular transplantations
of a race- and tis
sue-spevific amphibian renal carcinoma were made
into kindred races of the homologous species and
into heterologous species of frogs. The tumor was
established in all races and species, but under the
usual means of transplantation
the implants failed
to induce tumors in any except the homologous
race.
An accompanying ocular reaction to the tumor
implant increased in intensity inversely to the
genetic relationship of the host and the donor.
Implantation
of the renal tumor into the tad
poles of race hybrids induced renal tumors in both
the homologous crosses and reciprocal hybrids, but
intra-ocular implantation
into recently metamor
phosed adult race hybrids did not induce tumors
even though a host reaction was lacking.
An adaptation of the tumor specificity occurred
after a primary ocular regrowth in a normally re
sistant
kindred
race. When a primary
or secondary
regrowth was subcultured and transplanted
into
new hosts of either the homologous or kindred
races, renal tumors were induced in both races.
REFERENCES
1. AimuNx, W. S., and GREENE, H. S. N. The Transplants
tion of Tissues between Zoological Classes. Cancer Re
search, 13:64—68, 1953.
2. BiiutErr, M. K., and DniuNoxx, M. K. An Induced
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
TWEEDELL—Adaptatton
of an Amphibian
Adaptation in a Transplantable Tumor of Mice. J. Nat.
24.
Cancerlnat., 11:51—59,
1950.
-@
@-
8.
. Induced Adaptation
@
in a Tumor: Permanence
of
25. Li@wis, M. R. Myeloid
tion in Bacterial Viruses.J. Bact., 65:118-21, 1958.
Frog Kidney Carcinoma Transplanted into the Tails of
Permanent
and Normal Tadpoles. Cancer Research, 3:
618—20,
1943.
8. CAanv, A. E. Experiments with a Material from the
Brown-Pearce Tumor. Cancer Research, 1:184-35, 1941.
9. [email protected], A. E.; DRYSDALE, G.; Ross, G. L.; and ELROD,
B. A. Filtrates of Fresh Tumor Injected Prior to Trans
Biol. & Med., 1:13, 61—76,
1940.
12. Duna@r-Rai@w@s, F.; Buasmsrant,
B. R.; Comtai.,
C. E.
and Bay@, E. Studies on the Origin of the Naturally
Occurring Antibodies against Tumor Viruses Developing
in Aging Chickens. Cancer Research, 13:408-14, 1953.
18. ExcHww,
E. J.; CHANG, H. J.; and
M. The Sig
nificance of the Anterior Chamber in Tumor Transplants
tion. m. Natural Resistance and Acquired Immunity.
Cancer Research, 12:490-94, 1952.
14. GARDNER,W. U. Hormonal Imbalances in Tumorigenesis.
Cancer Research, 8:897-411, 1948.
15. Gnsmsr, R. G. Parasitic Hybridization as a Cause of
Cancer. Biodynamica, 6:1-21, 1946.
16. Gnaanra, H. S. N. Heterologous Transplantation
of the
Brown Pearce Tumor. Cancer Research, 9:728—35, 1949.
17.
. The Induction of the Shope Papilloma in Trans
plants of Embryonic Rabbit Skin. Ibi4., 13:58—68, 1958.
18. Kauss,
N. Regression or Survival of Tumor Homoio
grafts in Mice Pretreated with Injections of Lyophilized
Tissues.
Cancer Research,
12:879—82, 1952.
19. K&uss, N.; JONAS,G.; and Avira@r,N. L. Growth En
hancement of Tumor Homoiotransplants in Mice Follow
lag Injections of Homogenates and Ultrafiltration Sedi
ments of Mouse Tissues. Cancer Research, 10:228, 1950.
20. Ksnss,
N., and Moumitrr, N. The Effect of Prior In
jections of Tissue Antiserums on the Survival of Cancer
Homoiografts in Mice. Cancer Research, 12:110-12,1952.
21. K&z.iss, N., and Nzw@roN, 0. The Effect of Injection
Dosage Invel of Lyophilized Mouse Tissue on the Subse
quent Growth of a Tumor Hoenoiotransplant.
A.nat. Rec.,
105:585,1949.
About
in
The Occurrenceand Significanceof Metastasis. Ibid., 34:
15—30,1988.
. Carcinoma in the Leopard Frog: its Probable
Causation by a Virus. J. Exper. Med., 68:457-68,1938.
80.
. Kidney Carcinoma in the Leopard Frog: A Virus
Tumor. Ann. N.Y. Aced. Sc., 84:1093-1109,
1952.
31. Lucxa@, B.; Brmwicx, L.; and Nowzu@, P. The Effect of
29.
Temperature on the Growth of Virus Induced Frog Card
noma. fl. The Temperature Coefficientof Growth in Vifro.
J. Exper. Med., 97(4) :505-9, 1958.
82. Lucx@, B., and SCHLUMBERGER,
H.The Mannerof Growth
of Frog Carcinoma,Studied by Direct MicroscopicExami
nation
Cancer Research, 3:47—52,
1948.
11. Dviw@r-RE1@ua,
F. Neutralization
of Tumor Viruses
by the Blood of Normal Fowls of Different Ages. Yale J.
Brought
Mice of Alien Strains. Am. J. Cancer, 29:510-16, 1937.
Lucx@, B. A Neoplastic Disease of theKidney of the Frog,
Rosa pipiens. Am. J. Cancer, 20:852-79, 1984.
27.
. A Neoplastic Disease of the Kidney of the Frog,
Rana pipiens. II. On the Occurrence of Metastasis. Ibid.,
22:326—84,1984.
28.
. Carcinoma of the Kidney in the Leopard Frog:
plantation of the HomologousTumor. Proc. Soc. Exper.
Biol. & Med., 77:221—24, 1951.
10. Cwurns@,
A. M. Reactions of Hybrid and Parabiotic
Pseudo-Hybrid
Mice to Inoculation
of Tumor C198.
Hyperplasia
Mice b@the Growth of Dibenzanthracene Tumors and
Its Relation to the Transplantability of the Tumors into
K.; Dsnn@Gnn, M. 1@; and ILtxsn@, W. H. In
duced Adaptation in a Tumor: Specificity of the Change.
Proc.Am. Assoc.
CancerResearch,
1:8,1953.
5. Bsi@mi@x,G., and Wsio@, J. L float Controlled Varia
6. Biuaas, R. Transplantation
of Kidney Carcinoma from
Adult Frogs to Tadpoles. Cancer Research, 2:809-22,
1942.
7. Baroos, R., and Gnar@r, R. Growth and Regression of
@
. Tissue Culture and Serological Experiments. Ibid.,
pp. 219-27, 1958.
theChange.Ibid.,
12:1011—17,
1952.
4. Rtmtst'r,
417
Tumor
of Living Intraocular
Transplants.
J. Exper. Med.,
70:257—68,1989.
83.
. Heterotransplantation
of Frog Carcinoma; Char
acter of Growth in the Eyes of Alien Species. Ibid., 72:
811—20,1940.
34.
. The Effect of Temperature on the Growth of Frog
Carcinoma. I. Direct Microscopic Observations on Living
Intraocular
Transplants.
Ibid., pp. 821—80.
85.
. Neoplasia in Cold-BloodedVertebrates. Physiol.
Rev., 29:91—126,1949.
86. Lmsi&, S. E., and Huswr,
M. L. A Non-hereditary,
Host
induced Variation of [email protected]. Beet., 64:557—
69, 1952.
87. MoozE, J. A. Developmental Rate of Hybrid Prop. J.
Exper. Zool., 86:405-22,1941.
88.
. Incipient
Intraspecific
Isolation
Mechanisms
in
Rana pipiens. Genetics, 31:804—26,1946.
89.
. Hybridization between Rana paiusfris and Differ
ent Geographical Forms of Rana pipiena. Proc. Nat. Acad.
Sd.,32:209-12,1946.
40.
. Geographic
Variation
of Adaptive
Characters
in
Rana pipiens Schreber. Evolution, 3: 1—24,
1949.
41. Muitpnv,
J. B. Experimental
Approach
to the Cancer
Problem. Bull. Johns Hopkins Hoep., 56:1—81,
1985.
42. Oaaus@nio, C. The Riddle of Cancer,
pp. 158. 2d ed. New
Haven: Yale Univ. Press, 1958.
48.'Rosz,
S. M. Interaction
of Tumor Agents and Normal
Cellular Components in Amphibia. Ann. N.Y. Acad. Sc.,
54:1110—19, 1952.
44. Rosa, S. M., and Roan, F. C. Tumor Agent Transforma
tions in Amphibia. Cancer Research, 12: 1—12,1952.
45. RUGH, R. Induced Ovulation and Artificial Fertilization in
the Frog. BioL Bull., 66:22—29,1984.
40. Scnz.mnnonn,
H. G., and Lucx@, B. Tumors of Fishes,
Mouse Tissues on the
Amphibians, and Reptiles. Cancer Research, 8:957-754,
1948.
47. —.--. Serial Intraocular Transplantation of Frog Card
Growth of Tumor Homoiotransplants in Mice. Cancer Re
search,
11:122—26,
1951.
28. Luioawi, J. Tissue Culture of Embryonic Tissue Frag..
48. Swm@,A. C., and PAvATA,B. V. The Development of Re
sistance to Re-Inoculation and of Circulating Cytotoxins
22. K&Liss, N., and Sr@u@,G. D. The Effects of Injections of
Lyophilized
Normal and Neoplastic
noma for Fourteen
Generations.
Ibid., 9:52-00,
1949.
Media.K. Akad. Wetenschap. Amsterdam.Proc. Sect. SeA.,
in Response to Reterologous Ocular Tumor Transpianta
tion in the Guinea Pig. Cancer Research, 11:885-40,
56(2):214—18, 1953.
1951.
ments in Syngenesiologous,
Homologous and Heterologous
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
418
Cancer Research
49. SNzLt@,G. D. Enhancement
.
ot
.
Hosts
Tumor
Homoiotransplants
with
Yarious
by
Preparations
Pretreatnient
of
Normal
of
and
. transplants
@
by
Prior
and Stimulation
Injections
of
Tumor
of Tumor Homoio
Lyophilized
Tissue, as Influenced by Tumor Strain, Host Strain,
Source of Tissue, and Dosage. Cancer Research, 8:42987,1948.
the
Tissue. J. Nat. Cancer Inst., 12:719-29, 1952@
50. SNzu@, G. D.; Cwimswr, A. M@;Fanon, E.; and Douo
LASS, P. Inhibition
Tumor Immunity in Mice, Induced with Lyophuized
and Inhibition of the Growth
Tumor
52. SmoNo, L. C. A. On the Occurrence of Mutations within
Transplantable
Neoplasms. Genetics, 11:295-808, 1926.
53. W@uurais, S. G., and REINHARD, M. C. The Effect of X
Tiasue@J. Nat. Cancer Inst., 6:308-16, 1946.
51. SNXLI4 G. D.; Cwvimwa,
A. M,; and WooDwonTu,
Rays on a Tumor of Known Genetic Constitution. Proc.
E.
Soc. Exper. Biol. & Med.,42:678-76
1989.
Fia. 1.—Earlygrowth ofan implant froma renal carcinoma
transplanted into the eye chamber of a Vermont Rana pipien.
Taken
1 month
FIG. 2.—The
after implantation.
identical
implant
X10.
after
dissection
of the eye.
.. Pro.
8.—An
induced
renal
tumor
appearing
inaVermont
The lens is shown with the adhering iris and tumor growth.
Rana pipiena after intra-ocular implantation of a Vermont
renal tumor.
Fio. 4.—Asection of a typical renal adenocarcinoma in
duced in a VermontRana
pipisn..
X100.
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
@
@
.
!@ ,@
.
@,.
@-
- ,@•1
,
‘I
‘
@
@
@
__@#@
:*
... .
:¼;
i@
:@1
7@.
. ‘
‘J
..;
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
11
FIG. 5.—I)orsal view of the ‘@col1apsed
ey&' condition
Rana ciarni1art@.
FIG. 6—A well vascularized
most of the eye chamber.
The
allil
iris
periphery
pigment
elements
of the implant
can
initial
heavy
he
iii a
sustained
growth filling
lines are blood vessels,
seen
invading
from
(44 months after implantation).
the
X6.
FIG. 7.—Replacement
of the original renal implant
by coIl
nective
tissue prior to regression.
From an eye chamber
of a
Rana palustris, 5 weeks after implantation.
The implant is
lying on the iris at the pupillaiy edge. At one end, part of the
carcinomatous
remiiants
are
vesicle can still he seen, and numerous
(Cli
scattered
throughout
the connective
tissue.
x100.
FIG.
8.—A primary
merit cells in a Vermont
regrowth
which
Rana pipiens
is being
invaded
eye chamber.
by pig
This un
plant started to regrow at the 21st week.
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
@
.;@.@..
•1,@
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
FIG. 9.—An overgrowth
of the iris producing
the “rainbow
eye―in a Rana pipiens from Wisconsin after implantation
of
the renal carcinoma (dorsal-lateral
view).
Fic. 10.—Histology of the iris and retinal elements sub
cultured
from the eye shown
other Wisconsin
FIG.
11.—Histology
from Wisconsin
into
eye chambers
FIG.
after it
growth
months
in Figure
9 and tI'allspiantecl
irlto
R,ana pipien@s. X50.
of a secon(lary
regrowth
frog, which was suhcultured
of Wisconsin
and
in an eye
and transplante(l
\‘ermont frogs.
X 100.
12.—Growth in the eye chaml)er of a Vermont frog
was implanted with a subculture of a secondary re
from an eye chamber of a Wisconsill frog (taken at @24
after implantation).
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
@
-@ @:
@!
1*
j,
.-.,-.
r
•1@
oH
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
Fio.
13.—Growth
in the eye chamber
of a Wisconsin
frog
after being implanted with a subculture froln a secondary re
growth in the eye chamber of a WISCOnSin frog (5@ months
after implantation).
X8.
FIG. 14.—Histoiysing muscle resulting from disintegration
of nodes which arose concurrent with secondary eye chamber
regrowths.
FIG. 15.—Anaplastic
pipiens.
This
area
ocular impialitation
FIG. 16.—Induced
area in the liver of a %%isconsin Rena
is typical
of those
of secondary
found
eye chamber
renal tumor in a recently
only
after
intra
regrowths.
metamorphosed
hybrid from a Wisconsin female and a Wisconsin male. Im
plantation of the carcinoma was made into the tailfin of the
btdpole
prior to metamorphosis.
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
@
: @‘
@d
;@ ‘
@
.‘.
@
@
I'
@
@
@
@
f
.‘
.,@
I
‘@@:‘@‘
i―
.@‘;@‘@•
1@
‘V
.
@
@A
‘@
1@'
‘‘.
.‘@
â€ẫ€¢@‘.‘..
1:
@
@....
‘#@
SS5@S@5
‘4@
@‘.
‘.
.
...
@:•
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.
Adaptation of an Amphibian Renal Carcinoma in Kindred Races
Kenyon S. Tweedell
Cancer Res 1955;15:410-418.
Updated version
E-mail alerts
Reprints and
Subscriptions
Permissions
Access the most recent version of this article at:
http://cancerres.aacrjournals.org/content/15/6/410
Sign up to receive free email-alerts related to this article or journal.
To order reprints of this article or to subscribe to the journal, contact the AACR Publications
Department at [email protected].
To request permission to re-use all or part of this article, contact the AACR Publications
Department at [email protected].
Downloaded from cancerres.aacrjournals.org on June 14, 2017. © 1955 American Association for Cancer Research.