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1959

Factors modifying the growth of severaltransplantable tumorsLeonard Herbert InkerYale University

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FACTORS MODIFYING THE GROWTH OF SEVERAL TRAHSPLAHTABLE TUMORS

Leonard Inker A.B. Princeton University 1955 n, Submitted to the faculty of the Yale University

School of Medicine in partial fulfillment of the requirements for the degree of Poetor of Medicine

Pepartment of Pathology 1959

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The author wishes to express his appreciation for the aid and encouragement received from Professor Harry S, U. Greene in whose laboratory this work was done. He wishes also to thank Miss Betty Harvey and Mr. Richard Suttkin for their very generous technical assistance.

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TABLE of CONTENTS

Introduction... .1

Materials and Methods........................4

Results.. 6

Discussion...16

Summary and Conclusions...23

Bibliography.. 25

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INTRODUCTION

The transplantation of neoplastic tissue into a new host

offers a very valuable tool for the study of the biologic

characteristics of tumors, as they relate to a selected host,

as well as the reactions of the host to the transplant under

controlled conditions. Of course, it must be realized that these

hosts did not give rise to the tumors they bear and so are

not comparable, in many as yet poorly understood ways, to animals

and humans who have autogenous neoplasms.

The behavior of transplanted tumor3 is subject to many

diverse and complex modifying factors. Among these, two which

are being intensively studied at the present time are the

genetic and immunologic factors in tumor growth. The genetic

concept was originally formulated by Little1 in 1914, and has

been reviewedby Snell1 recently. Its basic assumption is that

a transplanted tumor will grow in a host if the host carries

certain dominant genes which were part of ihe genetic constitution

of the original tumor bearer.

Researchers have learned that the host can be modified

through a process of immunization and the susceptibility of

the host abolished. This immunity is specific for the tumor

for which it was developed and does not become generalized.

Lewis'-1 showed that a tumor which was transplanted to an animal,

and at a later date removed surgically, created immunity in

that host to further implantation with that tumor. Mac Dowell

et. al» by injecting increasing amounts of leukemic cells

into mice succeeded in producing immunity to the turner. Like**

wis$, Marshak and Lrf^, in 1941, and Grcss^ in 1943, succeeded

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in immunizing animals against lymphomata toy the continued

injection of small amounts of tumor tissue intradermally.

Other techniques used to produce immunity have toeen irradiation

of tumor tissue toy Goldfeder7 and the injections of alcoholic

tumor extracts toy Aptekman^ to produce resistance to lymphosarcoma.

Poorly understood immunologic phenomena can also toe used

to make animals more susceptible to transplantation. As long

ago as 1907 Plexner and Jobbing^ noted that they could prepare

an emulsion of the Flexner-Jobling rat sarcoma in such a way

that when injected into rats they could ototain animals more

sensitive to subsequent inoculations. In 1932, in working on the

Brown-Pearce tumor, Casey^Q discovered the "XPZ1' factor,

which when in.iected into the test animals made them more sus¬

ceptible than the controls for months at a time. JLyophilized tissue

from mouse tumors has toeen found to enhance tumor growth under

certain circumstances#In 1942, Law-^ made the very interesting

observation that new born mice of resistant strains could toe

made more susceptible to certain tumors if nursed with sus¬

ceptible foster mothers, establishing a milk factor in certain

tumors.

The host can toe modified in various other ways to become

more responsive to the transplantation of homologous and even

heterologous neoplasms. Toolan--'!, In 1953, reported his ob¬

servation that irradiation of the host reduced resistance of

animals to transplanted tumors# In this way workers have

succesded in overcoming the species barrier, which according

to the genetic theory should toe immutable# The same effects

have toeen shown to occur with steroid treatment as 'v,f :■

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reported "by Bickel arid Holm-Jensen14. Toolan13, with the use

of X ray and cortisone, grew human neoplastic tissue in weanling

rats and hamsters*

The age of the host has been shown to be a factor in the

susceptibility to and growth of tumor transplants* Wood3'0 report*-

ed that except in old animals there was no important age factor

for tiie growth of transplanted tumors* Bristol16, in 1915, re¬

ported that very young and fully grown mice were more refractory

than those of intermediate age. Gussie1®, working in 1912, noted

that during ih e first 8 to 9 days of life, rats were more resist¬

ant to sarcoma, whereas, beyond that age they became more and

more susceptible until the highest rate of successful transplants

was attained in the adult. Their observations run counter to

the reports of more recent observers who have noted an increased

susceptibility in the young. Bunting1® who studied tumor re¬

gression in mice found that among resistant strains he was more

successful in transplanting the tumor to very young(1-2 day old)

mice than to the older animals. Gross1^ observed the newborn

mice of an otherwise resistant strain to be susceptible to

inoculation with leukemic cells.

None of these workers have dealt with the modifying effects

of age on the rate of growth, of successfully transplanted tumors.

Ihe purpose of this paper is to present some observations on

the rate of growth of three hamster tumors in adult and young

hamsters, as well as some observations on the rate of metastases

in the fastest growing of these. Studies were also done on

the effects of multiple tumor implantations and the feeding '

of tumor tissue onthe growth of tumor transplants*

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Materials and Methods

Two sets of animals were used in these experiments* The

"young” group consisted of 3-4 week old hamsters weighing less

than 70 grams* The"adult” group consisted of fully grown ham¬

sters with an average weight of 125- 130 grams. They were all

fed regular laboratory chow unless specified.

The tumors were supplied "by Dr. H* S« 2ST. Greene and were

spontaneously occurring hamster sarcomas which had been carried

for several generations by transplantation. They were a lympho¬

sarcoma, an osteogenic sarcoma, and a melanoearcoma.

The original samples for transplantation were obtained by

sacrificing and adult tumor bearing animal with a large tumor.

A piece of tumor sufficiently small to fit within the lumen of

a #14 or #15 trocar was deposited through a small skin incision

in the right lower quadrant when one implantation was done.

When double implantation was performed a second sample was

deposited through'a left lower quadrant incision. The tumor

was deposited about half the distance to the axilla in ihe

subcutaneous tissue.

In Experiment I the results of which are recorded in Table

I, 24 young and 24 adult hamsters were inoculated with one of

the three tumors, 8 being given each of them. In the cases of

the lymphosarcoma and the osteogenic sarcoma the animals were

sacrificed at 2o days. The samples of osteogenic sarcoma were

rather small and it was decided to allow ih e melanosarcoma

to grow for 30 days before sacrificing the animals. The tumors

were weighed immediately after sacrificing and the volume of

the lymphosarcoma was determined by the displacement of water.

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In Experiment II, recorded in Table II a lymphosarcoma

grow ing very slowly in an adult was transplanted into 5 young

and 5 adult hamsters. The animals were sacrificed after 20

days when the tumors were weighed.

In Experiment III, recorded in Table III and Graph II,

45 young and 45 adult animals were inoculated with lymphosarcoma,

from the second to the tenth day 5 animals were sacrificed in

each group. The tumor dimensions and volume w re determined and

the axillary contents on the side of the transplant were im¬

planted into a normal animal. These animals were permitted to

survive up to six weeks at which time they were autopsied

and observed for grass evidence of tumor growth.

In Experiment IV 18 young animals were inoculated with

tumor bilaterally and 18 with tumor unilaterally. Half of

each group was fed the usual laboratory chow and th e other

half had in addition' 2 to 3 grams of lymphosarcoma daily.

Two of the animals inoculated with two tumors died within

two days of inoculation and are not recorded in the data. The

results of this experiment are recorded in TablelV and on

Graph III.

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RESULTS

Table I and Graph I indicate that in all three of the tumors

tested there was a marked difference between the size of the

tumor in the young and in the old animals after a given period of

'time, 20 days in the case of the lymphosarcoma and osteogenic

sarcoma and 30 days in the case of the melanosarcoma* The greater

growth in each case was in the younger animals.

Table II shows the results of the growth of tumor which

had grown very slowly in an adult animals in both old and young

animals. In this case the animals were sacrificed at the end

of twenty da^s, at which time the growth in the young animals

was about twice as great as in the older ones. Both sets of

tumors were in the same size range as had been observed in

the "well growing'1 tumors in Table I.

Table III and Graph II show the rate of growth over a

ten day period of a lymphosarcoma, in a series of young and

adult animals. The size and volume of these tumors were

recorded at autopsy. In the tally for the day the average for

that day is recorded last."Metastasis" indicates wheti er

a transplant of the axillary nodes of the autopsied animals

showed any growth of tumor tissue when transplanted subcutaneously

into normal animals. The"distance from the axilla" indicates the

proximity of the cephalic end of the tumor to the axillary

tissue taken. This varied from X-g- to 4 centimeters. The tumor

started growing a day sooner in the younger animals. It also

grew more rapidly and appeared to metastasize sooner in these

animals. The lag period of one day appears to account for the

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difference in tlie size of the tumors for the first several

days, for as can he seen on Graph II, the volume of the tumor

in the more mature animals is about the same as that of the

younger animals of one day earlier until the fifth and sixth

days, after which the tumor of the younger animals seem to grow

at a greater rate, there being a lag of about two days after

eight days of growth, and what appears to be a considerably

greater lag at twenty days®

Table IV shows the comparative size of tumors after 14

days, in animals containing one and two inoculums of tumor

tissue. In addition, this table compares the size of tumors

in each of these groups when half the animals are fed on a

diet of lymphosarcoma and regular laboratory chow,while the other

half were fed regular chow only. There appears tc- be no sig¬

nificant difference(in this case less &han one standard deviation)

between the total tumor mass of one and two inoculums. These

data are presented graphically in Graph III,

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e TABLE I

GROWTH OF THREE HAMSTER TUMORS 'WHEN TRANSPLANTED INTO YOUNG AND MATURE ANIMALS

A. LYMPHOSARCOMA Duration of experiment 20 days YOUNG AN MAES ($$*70Gffl8.) MATURE AN HALS (120-140Gms.)

Animal s Tumor Tumor Ratio Animal s Tumor Tumor Ratio tumor .Wt Wt. in volume T/A tumor .Wt .Wt • in volume T/A in Gms. Gms. In Gms. Gms, 74.5 8.5 8 1:8.7 143.5 9.5 9 1:15 75.5 13.5 13 1:5,6 121.5 7.5 7 1:16 49 10 9 1:4,9 145.5 6.5 6 1:22 53.5 16.5 16 1:3.2 122.5 7.5 7 1:16 72.5 18.5 18 1:3.9 121.5 8.5 8 1:14 74 14 13 1:5,3 1.24 1.0** .8 1:124 62.5 12.5 12 1:5 125.5 4.5 4 1:26 Died c 1; arse tumor, eaten 118 8.5 8 1:14 66 13.5 13,0 1:5 AV.128 6.2 1:19

3.4* 2,5* ** turn or used in "table II

B. OSTEOGENIC SARCOMA YOUNG ANIMALS (50-70Gms.)

Animal s Tumor Ratio tumor .Wt. Wt • in T/A in Gms. Gms. 89.5 3.5 1:26 89 3.5 1:26 92 2 f 0 1:46 97.5 0.5 1:194 92.5 2.5 1:37 104 4.0 1:25 93.5 0.5 1:186 81.5 1.5 1:120 92 2.25/1, 3*1:40

Duration of experiment 20 days MATURE ANIMALS (120-140Gms. )

Animals s Tumor Ratio tumor.Wt. Wt • in T/A in Gms. Gms. 130 0.5 1:260 133 0.3 1:400 125 0.3 1:375 137 1.5 1:91 155 1.5 1:100 130 0.5 1:260 130 ' 1.5 1:88 140 X <* 0 1:140 135 0.9/0, 5*1:150

C. MELANOSARCOMA Duration of experiment SO days YOUNG ANIMALS (50-70Gms.) MATURE ANIMALS (120-140Gms.)

.Animal s Tumor Ratio Animal s Tumor Ratio tumor. Vt. Wt. in T/A tumor. Wt Wt. in T/A in Gms. Gms. in Gms. Gms. 73 7.5 1:9.7 125 0.5 1:250 95 11.5 1:8.3 125 0.5 1:250 82 0.5 1:164 130 0.5 1:260 60 0.5 1:130 135 1.5 1:90 73 4.0 1:18 125 2.0 1:63 95 9.0 1:11 145 3.0 1:48 72 6.0 1:12 142 0.5 1:282 Died and eaten 130 0.5 1:260 80 5.5/3.8* 1:14 AV.132 1.1/0. 9*1:120

* Standard deviation

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GRAPH I

GROWTH of 3 HAMSTER TUMORS in YOUNG (--) and MATURE (—)

animals

LYMPHOSARCOMA OSTEOGENIC SARCOMA

MELANO SARCOMA

10

TABLE Ilia

RATE of GROWTH- and TIME of METASIASES in YOU1TG HAMSTERS using LYMPHOSARCOMA

Lay Animal wt. in Gms.

Distance from Axilla

Size of tumor in mm

Volume in mm'-1

Metast&ses

2 &¥ 2± cm indurated 0 0 45 3 cm indurated 0 0 60 2 cm indurated 0 0 55 2i cm indurated 0 0 40 3 cm indurated 0 0

Average 0 0 3 50 2 cm 5xl.5x.l .75 0

55 3 cm 2.5x2.5x.l .62 0 60 2 cm 4xlx.l .4 0 50 it cm 3x2x»1 • 6 0 45 Ijr cm 2xlx.5 .1 0

Average .49 0 4 65 2 cm 6x2x.1 1.2 0

60 2 cm 6x3x.l 1.8 0 55 3 cm 3xlx.l ,3 0 40 2 cm 5xlx.5 2.5 0 45 1} cm 3x2x.5 3.0 /

Average 1.7 n/5^ 5 35 2 cm 5x5x.5 12.5 0

50 2 cm 7.5x7.5x.5 28 0 ‘40 3 cm 7x3x.5 10.5 0 45 it cm 8x5x.5 20 / 50 2 cm 5x3x.5 7.5 0

Average 15.7 i7eT 6 55 2 cm 7x5x4 140 0

65 3 cm 12x12x4 452 0 50 2 cm 10x7x4 280 / 45 3 cm 8x6x3 144 0 40 ii cm 8x5x4 160 0

Average 235 ~T75 7 55 2 cm 20x12x7 1680 0

40 3 cm 10x7x7 490 / 45 2 cm 15x8x5 600 0 60 it cm 12x10x8 840 / 65 2f cm 15x10x2 300 0

Average 782 ~275 8 65 It cm 24x10x5 1200 7~ ' '

50 2 cm 24x14x7 2352 / 50 3 cm 20x12x6 1440 / 40 2 cm 15x10x8 1200 0 45 2 cm 25x10x5 1250 0

Average 1488 3ZL

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11

TABLE 11 Lb

RATE of GROWTH and TUBE of METASTASES in MATURE HAMSTERS using LYMPHOSARCOMA

Bay Animal wt« in Gms.

Bistance from Axilla

Size of tumor in mm

Volume^ in mm°

Metastases

2 135 4 cm indurated 0 0 140 5 cm si. indurated 0 0 125 4 cm indurated 0 0

125 3 cm si. indurated 0 0

120 2 cm si. indurated 0 0 Average 0 0 3 118 3 cm indurated 0 0

120 4 cm indurated 0 0 115 3 cm indurated 0 0 120 2 cm indurated 0 0 125 4 cm Indurated 0 0

Average 0 0 4 150 3 cm 4xlx.1 .4 0

110 4 cm 5xlx.1 ® f) 0 125 2 cm •5x.5x.l . 02 0

140 2 cm 2.5x7,5x.l 1*8 0

130 2 cm lx.5x.l ,5 0 Average 0.64 0 5 115 3 cm 5x5x.l 2.5 0

115 r^rcm 4x4x.l 1.6 0 110 2 cm 3x3x.5 4.5 0 120 3 cm 4x2x.5 4.0 0 135 4 cm 4x2x1 8.0 0

Average 4.1 0 6 140 3 cm 5x5x1 25 0

115 2 cm 3x3x1 9 0 125 3 cm 6x5x.5 15 0 130 4 cm 8x3x.5 12 0 120 2 cm 4x8x1 32 . /...

Average 18.6 1/5 7 160 3 cm 12x6x3 216 IT

140 2 cm 15x5x2 150 0 125 1-j-cm 10x7x4 280 / 130 3 cm 15x3x3 135 0 120 2 cm 6x4x2 48 0

Average 164 "275“ 8 150 3 cm 10x10x4 400 0

145 4 cm 10x10x4 400 / 130 2 cm 15x10x2 300 0 125 1-5'cm 12x6x3 216 0 130 2 cm 10x8x5 400 0

Average 343 0

(continued p 12)

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12

TABLE Ilia (continued)

Day Animal wt. Distance in Gms* from

Axilla

Size of tumor in mm

Volume in ram0

Metastases

9 40 3 cm 23x17x5 1955 0

45 2 cm 45x15x5 3375 0

50 2 cm 26x16x6 2496 0

40 3 cm 20x15x5 2250 / 60 li cm 20x20x5 2000 . / ..

Average 2415 2/ef 10 65 3 cm 30x20x10 6000 0

60 3 cm 25x25x9 5625 / 55 2 cm 30x20x7 4200 0

40 2 cm 20x20x10 4000 / 45 2 cm 35x10x10 3500 0

Average 4625 ' * s75 . 20 Average (see Table I) 13000

TABLE IILb (continued)

Day Animal wt. Distance Size of tumor Volume Metastases in Gms. from

Axilla in mm' In imn^

9 150 3 cm 12x10x7 840 0

140 2 cm 10x9x8 720 / 135 2 cm 15x8x5 600 / 130 3 cm 12x10x9 1080 / 125 3 cm 10x10x6 600 0

Average 768 3/5 10 160 2 cm 20x20x7 2800 0

145 3 cm 15x10x10 1500 / 130 2 cm 18x12x8 1728 0

125 2 cm 20x15x5 1500 / 140 3 cm 18x10x8 1400 0

Average 1785 ~275 w Average {see Ta ble I). 6200

TABLE II GROWTH OB SLOW GROWING LYMPHOSARCOMA from ADULT HAMSTER

YOUNG ANIMALS MATURE ANIMALS Animal s Tumor Wt Ratio Animal s Tumor Wt Ra tic tumor wt in Gms. T/A tumor Wt in Gms. T/A

in Gms 60 15 1:4

in Gms 125 8 1:16

70 10 1:7 130 4 1:32 53 12 1:4.4 150 5 1:30 72 8 1:9 130 6 1:22

48 14 1:3.4 130 7 1:19 61 11.8/2.5 1:5 AV. 133 6/To 4 1:22

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14

TABLE IVa

GROWTH OR TUMOR OHE and T¥0 TRAHSPLAHT S per AHIMAL Duration 14 days

OHE TRAHSPLAHT TWO TRAITSPLAHTS

Animal wt. Tumor wt Ratio Animal wt. Tumor wt iza Gms in Gms in Gms T/A In Gms larp,e/small: -total Ratio 53 12 1:4.5 41.5 17 4 21 1:1.9 73 15 1:4.9 54 9 9 18 1:3 73 12 1:6 52 8 5 13 1:4 63 12 1:5.2 50 12 12 24 1:2.1 77 8 1:9.6 50 17 1 18 1:2.8 61 T4 1:4.3 57 13 7 20 1:2.8 64 11 1:5.7 38 10 2 12 1:3.2 67 18 1:3.8 45 7 3 10 1:4.5 57 18 1:3.2 65.3 13.3/3. 1 1:4.8 AV.48.5 11.8 5.3 17/4.5 1:2.8

TABLE IVb

EJTECT OR INGESTIOH OR TUMOR TISSUE OH GROWTH 2 -3 Gms Lymphosarcoma 4&de& to < diet Duration 14 days Data in Table IVa

used as control OHE TRAHSPLAHT TWO TRAHSPLAHT S

Animal wto Tumor wt Ratio Animal wt Tumor wt in Gms iSP’Gms in Gms T/A in Gms large/small-total Ratio 52 18 1:2.9 75 8 1 9 1:8.3 57 18 1:3.2 38 10 5 15 1:2.5 59 16 1:3.6 55.5 11.5 8 19,5 1:2.8 49 13 1:3.8 61 12 1 13 1:4.8 64 13 1:4.9 47 10 1 11 1:4.3 64 8 1:8 60 12 10 22 1:2.8 59 13 1:4.5 68.5 9 7.5 16.5 1:4 50 10 1:5 62.5 9 8.5 17.5 1:4 68 17 1:4 58 14/3.3 1:41 AV.11875 10.3 5.2 15.tt

4 1: 3.7

TABLE IVc COMBIHED AVERAGE TUMOR SIZE

OHE TRAHSPLAHT a 13.3/3.1 b 14 /§« 3

is. e/HTTs"

TWO TRAHSPLAHT S a 17/4.5 b 15.5/4

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15

GRAPH III

GROWTH of LYMPHOSARCOMA 14 days after

ONE(f^) and TWO ) inoculums per animal

with ( —- ) tumor added without(—) w " average of those with and without ( —- )

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16

Discussion

From the data presented in Table I there appears to be a

definitely more rapid growth of all three types of tumor tissue

when transplanted into young animals# In the case of 1h e Xympho-

sarcoma or osteogenic sarcoma, both of which are relatively

common in the young, one might expect a host“tumor relationship

in which the tumor would grow more rapidly in the young animal

which is a natural host. The case of the melanosarcoma is

entirely different, occurring with great rarity in the young.

It is truly a tumor of the adult. Even when histologically similar

lesions appear in the young they do not behave as malignant

tissue. However, once the tumor has become malignant it can be

transplanted into a young animal in the same manner as tumors

native to the young. The response appears to be a nonspecific

one of the host to malignant tissue not to a specific tumor.

Presumably, it is the trigger mechanism to create a biologic

malignancy which is lacking in the young animal with histologic

malignancy, not the ability to support the growth of te mature

tumor. It would be interesting to speculate on what might oc¬

cur if the histologically malignant tissue of the young were

successfully transplanted into mature animals.

A wide range in the size of the tumors in various animals

during these experiments raised the question of whether there

is a host tissue resistance to the tumor. The other possibility

was a defective tumor which had been transplanted. It can be

observed,from Table II that there appeared in the instance in

which a slow growing tumor was transplanted into a series of

new host, that it grew as a normal transplant and not as it had

.

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17

in the former host. If the cause of slow grov/th had been in- /

herent in the transplant this should have been transmitted to

the new host. ^

Antibodies to the transplant may present an explanation

for the slower growth of tumor in the older animals. Bunting10

showed that newborn mice among resistant strains were suscept¬

ible to implantation with tumor. He offered the theory that

1h is was on the basis of defective immunologic makeup in the

younger animals. Lewis3 observed the development of immunity

to further tranplantation in animals in which transplanted

tumors had been surgically removed. Moore20 as quoted by Komburger -

suggests that "there are factual and theoretical grounds for ■ . f ■' - % dcrihg * of aor;

believing that during the conversion of normal cells to cancer

cells distinctive antigenic cytoplasmic material can develop

and that the search for neoplastic antigens and evidence of

auto-antibody formation in the case of autochthonous tumors

warrents further exploration." This statement can safely be

extended to transplantable tumors for here, not only can the

tumor act as an antigen, but it is a foreign antigen as well,

having been derived from another host. Gross6 immunized animals

against lymphomas by repeated intradermallinjection of small amounts

of turner. Other workers have done similar studies.4,3 Studies

in the variations of the immunologic mechanism with age might

prove very rewarding in the search fer an explanation of the

increased growth rate among the young. It would be interesting

to repeat the experiments of Gross using animals of various ages.

One of the three tumors, the lymphosarcoma was furti er

studied as to the rate of growth. This tumor apparently begins

to grow in the younger animal after a two day lag period in

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18 /

'v. \

which all that can he seen grossly is a hemorrhagic, ,indurated

area about the inoculum. At one day little reaction had occurred. 'A

Perhaps before growth can take place the tumor inoculum acts as

a stimulus to stroma formation with the ingrowth of a vascular

network which then supplies nutrients from the host. This reaction

appears to take place a day later inthe more mature animals.

After twenty days the tumor in the mature animal is only half

the size of that of the younger animal. The lag period might

account for the size variation if the older animals were one

day behind the younger animals in growth. This appears to be

true for the first several days, after which time the lag of

one day no longer seems to account for the difference. Perhaps

the older animals not only delay in producing a stroma but the

one that ih ey produce does not support the tumor growth as well.

The analogy might be made to the general lack of response of

tissue which seems to be a function of aging. Russell^ explained

the resistance of certain mice to transplantable tumors on the

failure of the host to provide a stroma and blood vessels

necessary for its growth. He postulated that the tissues of ti e

host might be altered in such a way that the animal could not

respond.

In an effort to study the effect of stroma formation on

the growth of transplanted tumors, multiple turner inoculations

were done. It was assumed that if this were the key factor in

the growth, a tumor bearing animal with two separate tumors

rather than one should have twice the stroma and hence twrice

the tumor mass after a given period of time. On the contrary,

no statistically significant difference in the amount of total

tumor tissue was present, although there was less volume of

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■ > oainlov sasX saw . x..-.• tx-iva ,Xu?7-tj aaw ©naslf loaruf

19

tumor to "be nourished on each side of the animal. There appears

to he something regulating the amount of total tumor to the

total host. They grew at a slower rate than similar transplants

in an animal possessing only one transplant.

One possible explanation is a nutritional limiting factor.

Ehrlich in his "athrepsia" theory quoted by Woglom2,i, postulated

that tumor tissue was such because it hafi a greater avidity

for foodstuffs than normal tissue and 1h erefore grew at the

expense of the host. That this is the primary difference between

neoplastic and normal tissue is very doubtful,yet as an observ¬

ation of the continued growth of tumor while there is concomi¬

tant loss of weight in the host is well known•2^ Ingle25 observed

that when tumors approach, one third the size of the host the host

develops a negative nitrogen balance while the tumor progresses.

Mider et. al.^5 observed three nutritional stages in rats which

bore transplanted tumors, in this case Walker carcinoma 256.

In the first stage, which lasted about ten days, both the animals

and tumors gained weight. Then the animals began to lose weight

while the turner continued to grow. In the last stage the hosts

lost excessive amounts of nitrogen while the tumors continued

to grow. Ingle25 reported that even forced feeding could not

control the pirating of host nitrogen by ih e tumor. pc

Homburger in reviewing the studies of tumor chemistry

states that ih e general impression may be gained that various

neoplastic tissues resemble each other far mere closely than

they resemble the normal cells of their tissues of origin.

Although with the possible exception of Warburg’s27 observation

of the fermentative nature of energy formation in tumor tissue

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20

no characteristic chemical changes in all cancer tissues has

yet "been found, although ti ere are certain substances which

are found in greater concentration than in normal tissue such

as aldolase and beta-glucuronidase. If the tumor tends to pirate

host nitrogen there is no reason to assume that other essential

nutrients are not also siphoned off into the tumor from the stores

of the host. Perhaps the limiting factor in the growth of the

tumor is a nutritional one being produced at a fixed rate by the

host and absorbed by the tumor in its growth. Thus, it would

not make any difference whether one or two inoculums were intro¬

duced, but only that a given amount of tumor could be produced

from the limiting nutrient. As the tumor grows there is greater

and greater need for whatever might be the limiting factor.

Perhaps the host adapts to the drain on its stores by producing

more of the substance. Horberg and GreenbergSO noted that

tumor bearing animals have a higher turnover rate of labeled

glycine in plasma protein and liver than normals. The different

rate of growth in the tumors of the young might be the result

of an increased abilitjr on the part of the young animals to

produce some substance for growth of the tumor whereas the older

animals might not be able to mobilize or produce if as rapidly.

If there is a nutritional basis for the control of the

grov/th rate in animals, perhaps the feeding of tumor tissue would

effect the growthrate. The results of experiments in which lympho¬

sarcoma was fed to tumor bearing animals in addition to the

regular diet appeared to have no statistically significant

effect on either the size of the host or the tumors after 14

days. It is nossible that those nutrients which, hypothetically,

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are absorbed, by the tumor tissue, may he changed into an un¬

usable form and hence the feeding of the end product would

not stimulate growth. Another possibilty is the destruction

of thissubstance by the process of digestion.

Further experimentation on multiple transplants may prove

of value. Will duplication of this experiment with larger

numbers of animals reproduce these results? Foes the regulation

of the growth of like tumor transplants carry over when the

transplants are dissimilar? Foes the process continue or does

the tumor become independent of whatever is limiting its growth

and proceed to grow as rapidly as if if mere the only tumor

present? Will the same hold true if the tumors are transplanted

at different times? It is unfortunate that these questions

cannot be answered in this paper because of the limitations of

time.

Another possible explanation for this dissimilar growth in

the two sets of animals Is a hormonal one. The great difference

in hormonal secretions in mature and immature individuals,

particularly concerning sex hormones is obvious. That hormones

play a part in tumor growth is well known for certain specific

tumors, eg, testosterone and carcinoma of the prostate where

marked regression of tumor often follows castration. The role

of steroid treatment to reduce host resistance has already been

alluded t©*1- That some sought of steroid like mechanism may

be present in young animals and not in old is certainly a possi-

bility.

It is of interest to note that the tumor tissue of the

younger animals not only grows at a faster rate but also appears

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to metastasize sooner. In examining the data presented it is

impossible to determine from the way in which the experiment

was set up whether the metastases are truly "blood "borne or the

result of lymphatic extenticn. A good number of the positive

metastatic transplants were in close proximity to the axilla.

It would be well to repeat this experiment using the axillary

contents on the tumor free side. In only one animal in all

the series was there evidence of gross metastases at death,

yet there were viable tumor cells in the axillary nodes at

an earljr time. GreeneoC) has shown that the tumor cells do

get into the blood stream in the case of the lymphosarcoma

after about six days. The fact that the nodes were positive

and yet no gross metastases could be demonstrated may indicate

that although the cells are getting into the blood stream

and disseminating, the primary tumor in some way inhibits

them from gaining a stroma and developing. They do however,

remain viable for a certain period of time and if transplanted

into a nontumor bearing animal they may take hold and grow.

The possible inhibitory effect of the primary tumor on metastases

may be a possible explanation for the previously discussed

observation that two tumors transplanted at the same time do

not grow as rapidly as if each were the only tumor present.

Perhaps they are mutually inhibitory®

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23

SUMMARY and CONCLUSIONS

1) Three hamster tumors were grown in young and mature

animals with greater growth in the younger animals in all

three cases.

2) A slow growing tumor removed from an adult animal was

inoculated into a series of young and mature hamsters with

the result that it grew as large in a given period in these

animals as had a tumor which had been considered to be growing well

in the animal from which it had been removed.

3) A series of young and mature hamsters were examined

as to the rate of growth of transplanted lymphosarcoma over a

tend day period. It was found that the tumor began to grow one

day sooner in the young animals and that after the first five

days appeared to be growing at r more rapid rate than in the

mature animals. At twenty days the tumor in the young animals

was twice the size of that of the mature ones. In addition,

metastases appeared to occur two days earlier in the younger

animals but it is immpossible to determine whether these are

blood borne or the resultof lymphatic mxtention.

4) A series of animals were inoculated with two samples

of lymphosarcoma and a second series with only one. The total

tumor weight at a specific time was determined. There appeared

to be no statistically significant difference in the weights

of total tumor obtained from animals in which two tumors had

grown and from those with only one tumor.

%) A series of animals with one and two tumor transplants

of lymphosarcoma were fed, in addition to their regular laboratory

chow, 2-3 grams of lymphosarcoma* When sacrificed after 14 days

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24

they showed no significant difference in total tumor weight

from controls fed only the regular diet.

6) Several possible explanations for) these observations

were discussed.

7) The great difference in the rate of growth of these tumors

in animals of different ages should be kept in mind when an

attempt is made to compare the results of two different experiments

as well as different animals in the same experiment.

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BIBLIOGRAPHY

1 Little, C.C. Apossible Mendelian explanation for a type of inheritance apparently non-mendelian in nature* Science 40: 904-906, 1914

2 Snell, G.D. Tr ansplantable tumors, in "The Physiopathology of Cancer"* P. Homburger and W.H# Fishman editors. P.B. Hoeber, Inc. hew York 1953 pp. 338-391.

3 Lewis, M.R. Immunits^ in relation to 1,2,5,6, dibenzanthracene induced sarcomata. Bull. Johns Konkins Hosp. 67; 325-344, 1940.

4 Mac Dowell, E.C., Potter, J.S. and Taylor,M.J. The influence of transplantation upo n immunological properties of leukemic cells. Prop, hat * Acad. Sci. 25: 416-420.

5 Marshak, A. and Erf, L.A. Effect of injection of nuclei on"take" of implant© of a lymphoma in mice. Proc. Soc. Exper. Biol, and Med, 46; 428-430, 19417

6 Gross, L. The importance cf dosage in the intradermal immunization against transplantable neoplasms. Cancer Research 3: 770-778, 1943

7 Goldfeder, K. Induced resistance in inbred homozygous rats to a lymphosarcoma autogenous to the strain* Proc. Soc. Exper. Biol, and Med* 59: 104-109, 1945.

8 Aptekman, P.M. , Lewis, M.R. and King, H.D. A method of producing in inbred albino rats a high percentage of immunity from tumors native to their strain. J. Immunol. 52; 77-85, 1946®

9 Flexner, S. and Jobling, J.W. On the promoting influence of heated tumor emulsions on tumor growth. Proc. Soc. Exper. Biol, and Med. 4; 156-157, 1907.

10 Casey, A.F. Experimental enhancement of malignancy in the Brown-Pearce Rabbit tumor. Proc. Soc. Exper. Biol. and Med. 29: 816-818, 1932.

11 Snell, G.D., Cloudman, A.M., Woodworth, E. Tumor immunity in mice induced with lypholized tissue as influenced by tumor strain, hos$ strain, source of tumor, and dosage. Cancer Research 8: 429-437, 1948.

12 Law, L.W. Foster nursing and the growth of .transplantable leukemia in mice. Cancer Research. 2:108-115, 1942.

Toolan, H.W. Successful growth of human tumors in the subcutaneous tissue of laboratory animals treated with cortisone. Proc. Am. A. Cancer Research 1:56 , 1953

13

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‘ ♦ 8'?V-Q?'V ; " :i)"r. 3 '

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i -2 m*- n£ ' ■. 4 .. i

; i o • v ft flrtPi eM 1 , , . 4 - ■ V ' , I, ; r —

ar-. i a if X In t jix 1.1 ojtrc t •: T n i

no ®ftoi;-c.lms$ *zotLv$ to * . r o -. • „

• «

, .. . ' X • - . ..

vlln?* s :• 9lie8X ■’ i:£ori~xI s- r- «£ .'xorruX to 90*x I

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26

Bickel, J and Holm- Jensen, I. Parabiosis and rsistance to transplantation* I The influence of parabiosis on the growth of mouse leukosis in irradiated rats. ACTA, path, et microbiol. Scandinav. 24: 531-538, 1947*

Wood, F.C. Limits of variability of tumor strains. Jour. Cancer Research 2: 50o, Id,»

Bristol, L.B. Cell enzymes and energesis in relation to cancer. Jour. Med. Res* 27: 475, 1915.

Gussio, S. Transmissions neoplastica e ricettivita d’eta di rsz7,a e di species.Tumor! 2; 173, 1912.

Bunting, H. Studies in regression in a transplantable' tumor in mice. Yale Journal Biol, and lied. 13: 513- 522, 1941.

Gross, L. Susceptibility of newborn mice to ah otherwise apparently resistant strain to innculation with leukemia. Proc. See. Exper. Biol, and Med, 73: 246-248, 1950*

Moore, R.H. The relation of Immunology and Heoplasia. "CanadianCCancer Confercnce", Vcl. I., R.W. Begg ed. Academic Press, Inc., Hew York, 1955.

Honburger, P. "The Biologic Basis of Cancer Management". P.B. Hoeber Inc. Hew York, 1957 p218.

Russell, B.R.G, The processes at the site of inoculation in normal mice and in mice resistant to carcinoma. Journ. Pathol, and Bacterid• 12: 436, 1908.

Woglom, ¥.H. Immunity/’ to transplantable tumours. The Cancer Review 4: 129-214, 1929. pX64

White, P.R. Source of tumor proteins; nitrogen balance studies of tumor bearing mice fed low nitrogen diet. J, Hat. Cancer Inst,5; 261-263, 1945

Ingle, D. J. Simple means of producing obesitA^ in ra«ts. Proc, See. Exper. Biol. and Med. 72: 604-605, 1949,

Mider, G.B., Penninger, L,D», Haven, P.L. and Morten,J.J, The energy expenditure of rats bearing Walker carcinoma 256, Cancer Research 11: 731-736, 1951.

Warburg, 0. On the origin of cancer cells. Science 123:309- 314, 1956. .

Homburger, P. "The Biologic Basis of Cancer Management". P.B. Hoeber, Inc., Hew York 1S57 p2C6.

Horberg, E* and Greeberg, D.M. Incorporation of labeled glycine in the proteins of tissues of normal and tumor bearing mice. Cancer 4; 383-386, 1951.

Greene, H.S.H. Personal communication 1959

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YALE MEDICAL LIBRARY

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