Acta Radiologica

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Studies on the Effect of Roentgen Rays Upon the Intestinal Epithelium and Upon the ReticuloEndothelial Cells of the Liver and Spleen Sv. A. Chrom To cite this article: Sv. A. Chrom (1935) Studies on the Effect of Roentgen Rays Upon the Intestinal Epithelium and Upon the Reticulo-Endothelial Cells of the Liver and Spleen, Acta Radiologica, 16:6, 641-660, DOI: 10.3109/00016923509174802 To link to this article: http://dx.doi.org/10.3109/00016923509174802

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Date: 24 January 2017, At: 05:10

ACTA RADIOLOGICA E D I T A P E R SOCIETATES KADIOLOGICAS D A N I E , F E N N I B , H E L V E T I I E , H O L L A K D I B , NOKVEGIA;, ET S U E C I E ~

VOL. XVI

FASC. 6

10: XI1 1935

N:o 94

STUDIES ON THE EFFEC'L' O F ROENTGEN RAYS TJPON THE INTES'l'INBL EPII'HELIUM AND UPON THE RETICU1,O-ENDOTHELIAL CELLS O F T H E LIVER AND SPLEEN' by

S v . A. C h r o m I n some experimental studies on the course of Breslau infection in universally irradiated mice, the bacteriological autopsy [after the method given by J. ORSKOV(13)] revealed the fact that the organs and blood of these mice were infected with bacteria belonging to the intestinal flora normally present in mouse, predominatingly colon bacilli. Among 52 mice which had been given universal irradiation with doses from 250 r to 450 r, 23 gave growth of colon bacilli and cultures from organs and of blood, whereas such growth could be demonstrated only in 1 out of 35 control mice. As the striking difference between the two groupsof mice with regard to the frequency of demonstrable infection with Bac. coli cannot be merely accidental, it is made the subject of further investigation as presented in the present paper. Technique ant1 Material The radiation was given with a ))stabilivolt))apparatus from SiemensSchuckert, 160 kilovolts, 4 milliamphres. Here the dose of Roentgen-rays is expressed by r , measured in air. Control measuring of the amounts of radiation employed in these experiments is made every week with Kustner's dosimeter. These control measurings have shown that the apparatus has worked very constantly throughout the experimental period. The radiation distance is recorded in every experiment, and in most instances it has been 42 cm., making the radiation intensity 12.4 r/min. ~~~

Submitted for publication. May 20th, 1935. From the state serum institute, Copenhagen (chief: TH. MADSEN. m. d.). The m. d.). The institute of pathology, radium station (physician in chief: J. JUUL. Bispebjmg hoapital (chief: BJ. VIMTRUP.m. d.). 41- 353527. Actn K n d i o l o g a c a . Val. X V Z . 1.935.

sv.

(:42

-4. ('IILLOAI

All the radiations, universal as well as local, are given through a filter of 0.5 mm. copper 3 mm. aluminum. IJnder the given conditions this means a radiation with dimidiation by 0 . 9 mm. copper. As the same filter is employed throughout, its thickness and composition will not be recorded anew in the various experiments. In experiments with universal radiation, the field has been 20 x 24 em.; in experiments with local radiation, the size of the field is recorded in each instance. The aninial material consists in white mice, weighing each about 20 p. As far as practicable the experimental animals have been kept isolated individually, each in its own cage. for at least 8 days prior to the experiment. Whenever such a mouse has shown any sign of illness, especially symptoms of intestinal disorder, it has been removed from the stable a t once. By insisting upon this preliminary quarantine it has been possible t o cut down the incidence of spontaneous infection in the experimental animals t o a nllnimal number. The mice are kept on a diet of crushed oats (with the bran included) and dry wheat bread (soaked in water). The temperature in the stable is kept constantly a t 18-20"C. The bacteriological technique is described in detail in previous works by J . ORSKOV(14) and H. C. A. LASSEK( 6 ) . The autopsy technique follows clowly the method given by ORSKOV,JENSEN & KORAYASHI (13).

+

Universal 1rradi:ition of Normal Mice

A group of normal mice were given universal radiation amounting to 550 r. Bacteriological autopsy was performed in from 3 to 10 days after the radiation. The cultural results are given in Table I. In Table 1 and following tables the outcome of the cultures made from the various organs and blood is presented schematically as follows: 0 ((+))

(+I

meails no bacterial growth whatever; )) growth of 1 colony: ') D )) up to 10 colonies;

+ ++ -+++

)) )) ))

))

))

))

))

50

D

200 0 a dense crowd of colonies or confluent growth. 0

))

0

0

The individual bacterial colonies are identified by means of Gram's method of staining and by the biochemical properties of the respective bacteria. As seen in Table I, when normal mice are exposed to universal radiation with 550 r, and bacteriological autopsy is performed on such mice in 310 days after the radiation, it is practicable in most of these mice t o de-

64.3

STUDIES ON THE EFFECT O F ROENTQEN RATS

Table I C'ulbures frotlr Arairtials r i f t e r Cnirersal Kutliaiion iiith .>50 T (= 1 TIED). t i m e 44' 9 min. Skin target distance: 4 2 cm.

1 . . . 2 . . . . . 3 4 . . . 3

Cervical lymph glS.

Meseiitcric lymph gl.

liver

+

(+)

t

0

J

0 0 0 0 0

4 4

0 0

1;

nays after radi ati o II

Mouse No.

.

.

.

. . . . . . . . . . . . . . . . . . . . . . . . . . . . .

.

6 . . . . . . . . . .

7 . . . . . . . . . . S . . . . . . . . .

3 3 ii t>

9 . . . . . . . . . 10 . . . . . . . . 11 . . . . . . . . . . .

li

0 0

6

0

1 2 , . . . . . . . . . 13. . . . . .

s s

1 4 . . .

9

.

.

.

.

.

.

7

16 . . . . . . !J 1 6 . . . . . . . . . . . 9 17 . . . . . . . . . . . 10

+++

0

+ i-+ + -t- +

U 0 0

0

0

0

u

0 0 0

U

U

+ -1-

+ 4-

++

blood 0

(J

0 0 0 -6

-t I0

0

7-

I I cart's

u

+++ k

Ih~diation

. I ~ 1. 4-

+

+++ r + +

+ +++

+++

t++

u 0

+++ 0 t

-1.

++

inonstrate the presence of an infection, and the bacteria isolated from these animals inay all be classified as bacterial species that niay be said normally to inhabit the intestinal canal in mice. During the first days after the radiation, this systemic infection is largely localized to the lymphatic apparatus of the gut; on the 7th day, however, there appears a marked change in the character of the infection, in the form of a very violent progression, going on to generalization. It is only natural to think of the intestinal canal as a source of origin for the infection brought about by exposure to the Roentgen-rays, as the character of the isolated bacteria and the mechanism of the denionstrated infection are uniformly suggestive of this source of origin. A priori, however, the possibility cannot be excluded that the focal infectious process in the niesenteric lymph gland, spleen or liver may turn into a progressive infection when the animal is exposed t o radiation. Rut, in a total of 207 normal mice, I (1) obtained growth of intestinal bacteria from the organs in only 17 mice, i. e., in about 8 per cent. So it seems only reasonable t o assume that the demonstrated infection with intestinal bacteria after universal radiation is in most of the cases attributable to a direct invasion of the bacteria from the intestinal canal. Local Abdomiual Irradiation of Normil Mice

The above experiment was performed with universal irradiation of the mice, implying radical changes in the functional capacity of the

(i14-

S V . A . CLlROhI

irradiated organism. It is easy to imagine that an infection originating from the intestinal canal will find a good soil for spreading in an animal after universal irradiation, by which treatment the reticulo-endothelial apparatus and lymphatic tissues besides the nucleated elements of the blood have been damaged severely, if not destroyed altogether. In order to elucidate whether the general cellular injury causes a decrease in the resistance of the organism to the demonstrated infection with colon bacilli, some experiments were made in which the effect of the lloentgen-rays was limited as far as possible to the intestines alone. Previously, WARREN& WHIPPLE(1922) (9) have shown that Roentgen-radiation of the thoracic half of the body in dogs with amounts of up to 512 milliampBre minutes gives no clinical symptoms of intoxication, whereas a dosage of 320 milliampirre minutes applied t o the abdomin a1 half of the body in dogs produces violent gastro-intestinal disturbances with fatal outcome. CORI (1924) (2) found similar conditions in mice exposed to local radiation of the abdomen.

Author’s Expeyirnents.

A group of 24 mice were given local radiation with 550 r, applied t o the abdomen, to a field of 1 x 1.5 em., immediately above the symphysis; the rest of the body was covered by 2 nim. of lead-foil. Bacteriological autopsy, performed as above, revealed in about half of the animals a moderate infection with colon bacilli, corresponding t o the lymphatic apparatus of the intestine, while cultures from the organs and blood of the remaining animals showed no bacterial growth. Thus there was a distinct difference in the occurrence of enterogenous infection in the animals exposed to local abdominal radiation, contrary t o the findings after universal radiation. When the amount of local abdominal radiation was doubled, making a dose of 1 100 r, the bacteriological autopsy showed that the infection with colon bacilli still kept localized t o the mesenteric lymph gland in about half of the animals. There is also a striking difference in the mortality of the irradiated animals in the two groups: 50-80 per cent of the animals exposed t o universal radiation with 550 r died ))spontaneously)), whereas local abdominal radiation with 1,100 r proved fatal to only 4 out of 24 animals. These findings appear to be in conflict with the findings reported by WARREN& WHIPPLE in experiments with abdominal radiation applied to a variety of animals. But, in their experiments with thoracic and abdominal radiation, these authors divided the body of the animal into two halves by a transversal a t the level of the xiphoid process of the

STUDIES OX T H E EFFECT 01’ 1:OESTGFN R A Y S

(i4

?)-

sternum. On abdominal radiation in this manner, the spleen and liver besides a large amount of the circulating blood are exposed directly t o the Roentgen-rays, effacing largely the picture of the results from a local radiation that is limited chiefly to the intestines. So the two series of experiments are not commensurable. The abdominal radiation em& WHIPPLEgives similar findings on bacteriological ployed by WARREN autopsy of the animals as are obtained after universal irradiation. Effect of Roentgen Rays iipon the Reticula-endothelial App:iritt1ls in the Liver arid Spleen

Extensive abdoniinal irradiation, by which the liver arid spleen are exposed directly to the action of Roentgen-rays, differs from local abdominal irradiation by giving rise to the same features of progressive enterogenous infection as is found after universal irradiation. As early (12) demonstrated that the fixed phagocytes as in 1886, WYSSOKOWITSCH (the reticulo-endothelial system) play an important r d e in t h e organism’s combating of bacterimnia. ORSKOVand collaborators have demonstrated that the fixed phagocytes in the organism, especially those located in the liver anti spleen, are capable of phagocytosis of bacteria injected into the blood stream, YO that the blood gives no bacterial growth in cultures that) are made within a certain length of time after the injection of the bacteria. On injection of 1 million Breslau bacilli into the blood st,r.eam in mice. ORSKOV,JENSEN & KOBAYASHI (13) found the blood t o be perfectly sterile as soon as 3 hours after the injection, while cultures from the liver and spleen showed growth of Rreslau bacilli. This sterile condition of the blood kept for about 20 hours, after which there was again a dcmonstrable condition of bacterkmia besides an increase in the iinnhcr of Breslau colonies in cultures from the liver and spleen. . It seemed reasonable to expect that by eniployrnent of the rxperinieiital technique given by these authors it might be practicable t o settlv whether the functional capacity of the reticulo-eiidothelial apparatus in the liver and spleen is lowered after Roentgen-radiation. It would be natural then for the manifestation of a normal or an altcrtd fnnction to employ the time interval in which the blood is usually sterile in t’t e experimental animals after intravenous injection of Breslau bacilli. 1 f this interval be shortened considerably, or completely absent, in animals with irradiated liver and spleen, it will be reasonable to conclude that the phagocytic power of the reticulo-endothelial system in these aniinals has been impaired. The Breslau strain (Br. 206) employed in these experiments h a s been cultivated numerous ))generations))by K. A. JENSE~Y (5), with incubation

646

S V. A

.

CLIROM

l'ablr TI

Controls .

C'iiltur~es f r o m Norrrial dnimcils a f t e r Intrarermus Injrcfion of 1 Million Rieslau Bacilli ( i n 0 . 2 C.C. physiol . salt sol.)

Mouse

No .

1s

. . . . . . . . .

19 20 21 22 23 24 25 26 37

. . . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . . .

2s . . . . . . . . . 29 3) 31 32 33

. . . . . . . . . . . . . . . . . . . . . . . . . .

Ilrs. after i n t r:ivenuua in.j . of bact . . . :i . . 6 . . 6 . . 7 . . 8 . . 10 . . 10 . . 11 . . 11 . . 1) . . 12 . . 15 . . 15 . . 26 . . hL'

('ervical lymph g l .

Meseiiteric lymph gl .

+

-1-

+

-c

+

L

+ +

T

+

+ + +

+ -I-

I 1

.

. . . . . . . . . . . . . . . . 72

-t

+ + +

-t

+ +

Liver

+ -t +

++

+++

+++ i -+ +

+++ ++ +

!

'

t t I+ -4-1- i-I-

-t

+

Fl.. ecn

i

++ + + 4+ :-

++ i~+ + !-

..

i-

Heart's blood

++ ++

++ + -1 +i

-I- 1-

++ -I- t-1- t -1-1- I

+

21 15 6 11 14 3 10 :3 4 1

++

1 2:1 1 1

I- erthe radiation shows the presence of a n infection of t,he organs and heart’s hlootl with bacteriii which are normally present in t>lieintestinal canal in mice. Local radiation, limited chiefly t)o the intestines: wit,li Roentgen-ray doses lip to 1,100 r does not give rise t>o;L progressirr cnterogenous infection. Radiat,ion of the entire abdomen, exposing the liver and spleen to direct radiation, with suitable closes of Roentgen-rays gives rise to a progressive h c , terizmia. Mention is made of an experimental technique employed here to study the rGle played by the ret>iculo-endothelialapparatw in the liver and spleen of t,hr niouse in combating infection of t>heblood strea.m with Ijreslnu bacilli. Histological-hacteriolog~cal examination is made of sections from the intestines of mice exposed to universal radiation. The writer thinks he has oliserved that the epithelial cells of the intestines in mice, corresponding in particular to the ileocoecal 1)art of the gut, aft,cr radiation may lose the ability to retain the bacteria normally present in the intestinal lumen.

ZUSAMMENFASSUNG Nach allgemeinrr Bestrahlung normaler Mguse mit Ront>gcnstrahlen in Dosen zwischen 250 r und 550 r erwies die hakteriologische Autopsie der Tiere 3-10 Tage nach der Bestrnhlung das Vorhandensein einer Infekt,ion der Organe und des Herzblutcs mit Bakterien, die normalerweise im Dnrmka,na,l von Mgiisrn vorkommen. Lokale, hauptsgchlich auf die Eingeweide heschriinkte Bcstrahlung mit Rontgenstrahlen in Dosen bis 1 100 r bewirkt keine progressive enterogene Infektion. Bcstrahlung des ganzcn Abdomens, wohei Leber und Milz direkter Bestrahlung mit Rontgenstrahlen in gecigneteri Dosen ausgesebzt werden, ruft eine progressive Rakteriiimie hervor. Verf. h s p r i c h t eine experimentelle Technik, die hier zur Anwendung kiim, um die Rolle zii shdieren, die der rctikuloendotheliale Apparat der L e h r und Milz dc>r Maus bei Beliiimpfung dcr Infektion tles Blutstroms mit Breslau-Hazillen spielt. Verf. unterzog Schnitte nus Eingeweidcn universal I)cstrahlter Miiuse histologischhakteriologischer Unt,ersuchiing und g1aul)t beol)acht)et zu hahen, class die Epithclzcllcn der Eingeweide dcr 1LIiinse hauptsiichlich im ileozokalen Teil des Darmkanals nach Bestrahlung die Fdiigkeit verlicren, dio normalerweise im Darmlunien vorhandenrn Bakterien zuriickzuhalten.

660

SV. A. CHROM

RESUME Apr&sune irradiation nniverselle de souris normales, avec des doses de rayons X de 250 r B 550 r, l’autopsie bactkriologique montre, 3 B 10 jours apr& l’irradiation, la prksence d’une infection des organes et du sang du cmur avec des bactkries normalement prksentes dans l’intestin des souris. L’irradiation locale, limit6e principalement b l’intestin, avec des doses de rayons X jusqu’b 1100 r ne donne pas naissance A une infection entkroghe. L’irradiation de l’abdomen en entier, exposant le foie et la rate b une irradiation directe, avec des doses convenahles de rayons X donne naissance b une bact6riirmic progressive. I1 est fait mention d’nne technique expkrimentale emp1oyi:e ici pour ktudicr le rGle jouk par l’appareil r6ticulo-endothklial du foie et de la rate de la souris dans la lutte contre l’infection du sang par le bacille de Breslau. I1 est fait un examen histologique et bact6riologique de coupes des intcstins dcs souris universellement irradikes. L’auteur pense avoir observk que les cellules kpithkliales de l’intestin des souris, en particulier celles qui correspondent la partie i1i:o-caecale,peuvent perdre lcur faculti: de retenir la bactkrie normalement pr6sente dans la lumikre intestinale.

REFERENCES 1. CHROM, S. A.: Experinientelle Undcrsegclser over Rentgenbestraalings Indvirkning paa Infektionsmekanismen hos hvide Mus. (Dissert.) copenhagen. 1934. 2. CORI, G. T.: Journal of Cancer Research, 8: 522, 1924. 3. FAURICIUS-M~LLER, J.: Experimentelle Studicr over hccmorrhagisk Diathese frenikaldt ved Rentgenstraalcr. (Dissert.) Copenhagen. 1922. 4. HEINEKE,H.: Miinchener med. Wochenschrift, 50: 2090, 1903. 5. JENSEN,K. A.: Zeitschr. f . Immunitatsforsch., 63: 298, 1929. 6. LASSEN,H. C. A.: Experimental studies on the course of paratyphoid infections in avitaminotic rats with special reference to vitamin A deficiency. (Dissert.) Copenhagen, 1931. 7. MEYER & RITTER:Strahlentherapie. I : 183, 1912. 8. MOTTRAM, I. C. & KINGSBURY, A. IT.: Brit. Journ. of Exp. Path., 5: 220, 1924. S.L. & WHIPPLE,G. H.: Journ. of Exp. &led.,35: 187, 1922. 9. WARREN, 10. -, ibidem, 35: 202, 1922. 11. -, ibidem, 3s: 713, 1923. 12. WYSSOKOWITSCH: Zeitschr. f. Hyg., I : 3, 1886. 13. ORSKOV, J., JENSEN, I(.A. & KOI~AYASHI, K.: Zeithchr. f . Inimunitiitsf. 55: 34, 1928. J. & MOLTKE,0.: I b i d e m , 59: 357, 1928. 14. ORSKOV,