(From the Laboratories of The Mount Sinai Hospital, and the Bacteriological and Serological Divisions of the Beth Israel Hospital, New York)

Transcription

1 Published Online: 1 Octber, 1935 Supp Inf: Dwnlded frm jem.rupress.rg n Octber 21, 218 DISTRIBUTION OF BLOOD GROUP PROPERTIES AND BLOOD GROUP PROPERTY DESTROYING FACTORS IN THE INTESTINAL TRACT OF MAN BY ERNST WITEBSKY, M.D., AND ERWIN NETER, M.D. (Frm the Lbrtries f The Munt Sini Hspitl, nd the Bcterilgicl nd Serlgicl Divisins f the Beth Isrel Hspitl, New Yrk) (Received fr publictin, July 23, 1935) Investigtins crried ut during recent yers hve shwn tht the bilgicl chrcteristics knwn since Lndsteiner's discverys bld grups cncern the entire rgnism nd nt nly the bld. Grup specific substnces hve been fund in lchlic nd in wtery extrcts f rgns f mn nd nimls (1-3). The quntittive distributin f grup prperties, hwever, is quite different in vrius rgns (4). Especilly rich in grup substnces re, fr instnce, kidneys, lungs, pncres, nd the mucus membrnes f the intestinl trct; n the ther hnd the brin cntins nly trces f the substnces, if ny. Mrever, they re fund nt nly in the tissues f the rgnism, but ls in its secretins. Here ls we encunter gret differences in the cncentrtin f grup prperties. Thus the spinl fluid is lmst free f grup substnces, while sliv nd stmch juice generlly re rich in them. They re cnstntly excreted with the urine. Hence, cntinuus regenertin f grup substnces must tke plce. In view f the high cntent f the stmch juice in grup specific substnces it hs been surprising t find tht the feces re lcking in these prperties. 1 Schiff nd cwrkers (7, 8) shwed tht queus extrcts f feces (s well s sliv) destry the grup specific prperties. It is pssible t btin Berkefeld filtrtes f feces which ls destry the grup specific substnces (9, 1). The ctivity f feces extrcts twrd the grup specific prperties is ttributed by Schiff nd his cwrkers t the presence f specific bld grup prperty destrying enzyme which is prduced by the bdy itself. Mny fcts, hwever, seem t spek ginst 1 This sttement is nt ffected by the fct tht smll munts f grup specific substnces cn be demnstrted in feces by mens f specil methds s described recently by Hdy (5) nd Mhrrem (6). 589

2 59 BLOOD GROUP PROPERTY DESTROYING FACTORS the ssumptin tht the bld grup destrying gent is secretin prduct f the bdy such s pepsin r trypsin. Witebsky nd Sth fund tht n increse f the bld grup prperty destrying gent btined frm sliv tkes plce in prgressive trnsplnttin (chin experiments) in which mixture f serum nd egg white ws used s medium. An increse up t 2, times the riginl munt f the bld grup destrying fctrs ws fund. In crrbrtin f these findings Sievers (11, 12) reprted n increse in effectiveness f the bld grup prperty destrying gent up t 1 millin f the riginl strength. Sievers used brth s the medium. Althugh we cnnt mke ny definite sttement cncerning the rigin f the bld grup destrying gent we hve t cnsider the pssibility f its derivtin frm micrrgnisms r their grwth prducts. Very recently the destructin f bld grup specific substnces by micrrgnisms hs been described. A Myxbcterium islted frm vegetble surces by Mrgn nd Thysen, ws fund effective ginst the grup specific substnce A by Lndsteiner nd Chse (13). Schiff (14) bserved the destructin f bld grup specific substnces by B. welchii. The fllwing experiments del with whether the gent destrying the bld grup prperty ccurs in the entire intestinl trct r nly in prts f it. Insmuch s there ppers t be nly brief reference in the literture t the presence f bld grup specific substnce A in the cntent f the jejunum (7), n ttempt hs ls been mde by us t determine in which prt f the intestinl trct the bld grup prperties themselves re present. Mteril Extrcts f intestinl cntent were prepred in the fllwing wy: One prt f the mteril ws mixed with 1 prts f physilgicl slt slutin. When the specimens were slid the mixture f sline nd feces ws shken thrughly in shking pprtus fr but 2 t 3 minutes nd then centrifuged. Methd Severl methds re vilble t determine the presence r bsence f grup prperties. In the fllwing experiments the s clled hemlysis inhibitin test ws mstly used. This test is bsed n the fct tht grup specific A-ntiserum is rich in sheep cell hemlysins. By mixing dilutins f grup specific A-ntiserum with A cntining mteril, the grup specific ntibdies re fixed nd simultneusly the sheep cell hemlysins re bund. Hemlysis f subsequently dded sheep cells is thus prevented. T determine the presence f bld grup destrying gent, its ctin twrd peptne hs been used. Witte peptne (1 per cent) knwn t cntin A substnce ws prepred, heted in wter bth, filtered, nd kept under sterile cnditins.

3 ERNST WITEBSKY AND ERWIN NETER 591 Experimentl Experiment 1. Quntittive Determintin f Grup Substnce A in the Cntent f Vrius Prts f the Intestinl Trct.--The experiment ws set up s fllws: Decresing munts (vlume.2 cc.) f extrcts ~ f the cntents f () jejunum, (b) middle, (c) lwer, (d) scending cln, (e) descending cln were mixed with.2 cc. f suitble dilutin f grup specific A-ntiserum. The mixtures were incubted t rm temperture fr 2 minutes. Then.2 cc. f 5 per cent sheep cell suspensin nd.2 cc. f guine pig serum in dilutin f 1 TABLE I Hemlysis f Sheep Cells by Cmplement nd A-Antiserum fter Tretment f the Ltter with Intestinl Cntent Extrcts Amunt f extrct f intestinl cntent () Jejunum tr. c. (b) Middle m. tr. c. c. (c) Lwer # tr. c. In. c. & c.. c. c. c. (d) Ascending cln c. m. c. (e) Descending cln # tr c. c c.... c.. c. c. c. --- hemlysis fter 2 minutes. --- hemlysis fter 4 minutes. e. ~- cmplete hemlysis. c. = lmst cmplete hemlysis. m. = mderte hemlysis. tr. = trces f hemlysis. = n hemlysis. t 1 were dded. The resulting hemlysis ws nted () fter 2 minutes, (8) fter 2 hurs, incubtin t 37 (Tble I). As is seen frm Tble I, the cntents f the jejunum s well s tht f the middle re rich in grup specific substnce A. The lwer cntins nly mderte munt f A substnce, while Befre the experiment ws set up the extrcts were kept in wter bth t 65-7 fr 1 minutes t cunterct nticmplementry effects.

4 592 BLOOD GROUP PROPERTY DESTROYING FACTORS the cln cntents re lmst cmpletely free. The experiment shws stedy decrese in A substnce in the intestinl cntents in dults, frm bve dwnwrds. Similr results were btined in mny cses (with few exceptins which will be described belw) quite independent f the disese. Thus the described distributin f the grup specific A substnce in the cntents f the intestinl trct my be clled physilgicl ne. Experiment 2. Quntittive Determintin f Bld Grup Prperty Destrying Fctrs in the Cntents f Vrius Prts f the Intestinl Trct.--Fr the quntit- TABLE Hemlysis f Sheep Cells by Cmplement nd Grup Specific A-Antiserum fter Tretment f the Ltter with Mixture f Peptne nd Intestinl Cntent Extrcts II Amunt f peptne-extrct mixture () Jejunum (b) Terminl (c) Cecum (d) Sigmid (e) Peptne cntrl tr. m. c. c. c. tr. t c. Ill. & e. C, C, tr. m. m. c. c. = hemlysis fter 15 minutes. t~ = hemlysis fter 2 hurs. tive determintin f the bld grup destrying gent its ctin upn Witte peptne ws used in the fllwing wy: 1 cc. ech f n extrct f cntents frm the () jejunum, (b) terminl, (c) cecum, (d) sigmid, btined frm persn belnging t Grup O s well s (e) sline cntrl were mixed with 1 cc. f 1 per cent Witte peptne slutin nd incubted t 37 The resulting hemlysis is shwn in Tble II. Tble II shws tht the mixture f peptne nd jejunum cntents rects s strngly s the peptne itself upn the grup specific A- ntiserum. The mixture f peptne nd terminl cntent displys nly mderte inhibitin f hemlysis. The cecum nd sigmld cntent-peptne mixtures prved bslutely negtive. This

5 ERNST WITEBSKY AND ERWIN NETER 593 mens tht the bld grup prperty destrying fctrs first pper in the terminl nd tht their effectiveness increses in the cecum? In few cses, hwever, intestinl cntent extrcts frm the lwer prt f the displyed slight ptency nly, r even prved t be ineffective. In these cses the bld grup destrying gent ws fund first in the cntent f the cecum nd f the cln. The cmprisn f the distributin f the A substnce in the cntent f the intestinl trct (see Tble I) with tht f the bld grup prperty destrying fctrs (see Tble II) displys clse prllelism in the disppernce f the bld grup substnce n the ne hnd nd the increse in bld grup destrying gent n the ther. Experiment 3. Simultneus Presence f Bld Grup Specific Substnce A nd Bld Grup Destrying Fctrs within the Intestinl Trct. --The questin rse whether simultneus presence f bld grup substnce nd bld grup destrying fctrs ccurs nywhere in the intestinl trct. T elucidte this questin bld grup substnce A s well s bld grup destrying gent were determined in the vrius prts f the intestinl trct f persn belnging t Grup A. The titrtin f the A substnce ws dne s in Experiment 1, tht f the bld grup destrying gent s in Experiment 2. The results re given in TMes III nd IV. As in the cse f the findings in Experiment 1, Tble III shws tht the cntents f jejunum nd re rich in grup specific A substnce, while the cecum cntins nly mderte munts nd the cntent f the sigmid is cmpletely lcking in A substnce. Tble IV shws tht the extrcts frm the lrge intestine re nt nly free f A substnce, but they even destry the A substnce f peptne. On the ther hnd the extrcts f the jejunum nd the cntent d nt exhibit ny bld grup destrying pwer. The cecum cntent extrct, hwever, cntins bld grup destrying gent s well s bld grup substnce A, lthugh the ltter seems t be present in reltively smll munts nly. This bservtin resembles Witebsky nd Sth's demnstrtin f the simultneus existence f bld grup substnce nd bld grup destrying fctrs in the feces f rbbits. 8 In ll the experiments the grup specific nture f the inhibitin f the sheep cell hemlysis ws cntrlled by simultneus exmintin with sheep cell ntiserum.

6 594 BLOOD GROUP PROPERTY DESTROYING FACTORS Experiment 4. The Influence f Time n the Effectiveness f the Bld Grup Prperty Destrying Fctrs.--It is knwn frm the study f the bld grup prperty destrying gent tht the time TABLE Hemlysis f Sheep Cells by Cmplement nd Grup Specific A-Antiserum fter Tretment f the Ltter with Intestinl Cntent Extrct Amunt f intestlni cntent extrct 2.1.5,25.125,625 (.).Upper jejunum (.b) Lwer ~ejunum = hemlysis fter 2 minutes. # = hemlysis fter 4 minutes. III (c) Ileum. (d) Cecum m. tr. m. II1. m. In. c. m.,c. ii1. C,. (e) Sigmid {2. C, TABLE Hemlysis f Sheep Cells by Cmplement nd Grup Specific A-Antiserum fter Tretment f the Ltter with Mixture f Peptne nd Intestinl Cntent Extrct Amunt f peptne- (.).Upper (b) Lwer (c) Ileum (d) Cecum (e) Sigmld (f) Peptne extrct jejunum jejunum cntrl mixture = hemlysis fter 15 minutes. t~ = hemlysis fter 4 minutes. IV tr. In. m. c. m. c. c. Ill. m. In. ],c.. c. c TI. [ctr plys n imprtnt r61e in its effectiveness. Therefre, the questin rse s t whether r nt it is pssible t demnstrte differences in the effectiveness f the bld grup destrying fctrs by

7 ERNST WITEBSKY AND ERWIN NETER 595 shrtening f the incubtin perid. set up. The fllwing experiment ws Equl munts f extrcts f the cntent f () jejunum, (b) middle, (e) terminl, (d) cln, (e) physilgicl sline s cntrl were mixed with Witte peptne (1 per cent slutin) nd incubted fr () 3 hurs t 37, (b) 15 hurs t 37 The mixtures were then heted fr 1 minutes in wter bth t 65-7 The titrtin f the A cntent f the vrius mixtures ws dne s in Experiment 2. The results re given in Tble V. TABLE Hemlysis f Sheep Cells by Cmplement nd Grup Specific A-Antiserum fter Tretment f the Ltter with Mixture f Peptne nd Intestinl Cntent Extrct () Incubtin time 3 hrs. t 37 V Amunt f peptne-extrct mixture () Jejunum (b) Middle (c) Terminl (d) Cln (e) Peptne cntrl In. e. In. # c. m. c. t~ m. c t m. # C, C, (b) Incubtin time 15 hrs. t 37 cx In. c. e. c. 8.C, e. c, c. c. # C l m. c. # = hemlysis fter 2 minutes. # = hemlysis fter 2 hurs. As is seen frm Tble V there is mrked difference in the effectiveness f the terminl cntent extrct depending upn the incubtin perid. After n incubtin f 3 hurs the A substnce f the peptne is prtly destryed by the bld grup destrying gent, while fter 15 hurs incubtin the A qulity hs cmpletely disppered. The bld grup destrying gent frm the cln, hwever, destrys the entire A cntent f the peptne fter 3 hurs incubtin. On the ther hnd, there is n trce f bld grup prperty destrying ctivity t be fund in the extrcts f the jejunum nd centrl cntent, even fter 15 hurs incubtin time.

8 596 BLOOD GROUP PROPERTY DESTROYING FACTORS It fllws frm the experiments mentined bve tht there is n brupt increse in the cncentrtin f the bld grup destrying gent in ny given site f the intestinl trct. Its cncentrtin increses rther grdully. It cn be fund first in lw cncentrtin in the lwer prt f the nd reches its highest ptency within the lrge intestine. Experiment 5. Distributin f Bld Grup Prperty Destrying Fctrs in Cses with Intestinl Invlvement.--The bve described chrcteristic distributin f the bld grup destrying gent in the cntents f the intestinl trct differed in few cses with intestinl bnrmlities. Tw re f specil interest. The first ws crcinm f the rectum with clnic fistul, the secnd stricture in the lwer prt f the cln due t lymphgrnulm. Intestinl cntents frm tw tumr cses withut invlvement f intestine were used fr cntrl experiments, ne ws cse f chrinepithelim; the ther cse f tumr f the lungs. The experiment itself ws crried ut s described in Experiment 2. Specimens f the cntent f the fllwing prts f the intestinl trct were exmined: () jejunum, (b) upper, (c) terminl, (d) cecum. These extrcts were mixed with equl munts f 1 per cent Witte peptne slutin nd kept vernight in the incubtr t 37 Then the mixtures were heted nd the A cntent determined s described bve. The results re given in Tble VI. Tble VI shws tht the cntent f the jejunum s well s f the in tw cses with intestinl invlvement (clnic fistul nd stricture due t lymphgrnulmtsis, respectively) displyed mrked bld grup destrying cpcity. On the ther hnd the crrespnding specimens f the tumr cses withut intestinl invlvement were cmpletely ineffective. DISCUSSION The experiments reprted in the present pper shw tht the cntents f the upper intestinl trct (jejunum nd upper ) f mn belnging t Grup A re usully rich in grup specific substnces. The stedy decrese in grup specific substnces in the intestinl trct in dults runs prllel t the increse in bld grup prperty destrying pwer. Thus link exists between the disppernce f grup prperties nd the ppernce f the bld grup prperty destrying fctrs in the intestinl trct. There is zne, hwever,

9 ERNST WITEBSKY AND ERWIN NETER 597 in which the effectiveness f the bld grup destrying gent hs nt yet reched its cmplete pwer, s tht certin munt f grup specific substnce my be still present. This usully hppens in the lwer prt f the r in the cecum. It is knwn tht nt ll persns belnging t Grup A excrete the grup prperties in their sliv. It is necessry therefre t find whether r nt similr differences re encuntered cncerning the presence f grup qulities in the cntent f the intestinl trct. TABLE Hemlysis f Sheep Cells by Cmplement nd Grup Specific A-Antiserum fter Tretment f the Ltter with Mixture f Peptne nd Intestinl Cntent Extrct () Lymphgrnulmtsis with stricture in rectum VI Amunt f peptne-extrct mixture () Jejunum (b) Upper (s) Terminl ileur (d) Cecum (e) Peptne cntrl.67 O I'll, C EtC, C, m, c. c c. c. c, m c. C m c. &C tr. m. (b) Rectum crcinm-cln fistul # O. 67 O. 23 O. 78 m. In &c. ~c m. c.,c. c. m. c. ~tc. c c c. c, C, tr. m. = hemlysis fter 15 minutes. # = hemlysis fter 1 hur. A mrked chnge in the bve described distributin f bld grup substnce nd bld grup destrying gent in the cntent f the intestinl trct ws fund in tw cses with intestinl invlvement. It is very likely, hwever, tht the presence f the bld grup destrying gent in the jejunum f these cses is nt due t the ctul disese but t mechnicl bstructin Further studies re necessry t elucidte the questin s t whether there is reltin between n bnrml bcteril invsin f the upper intestinl trct nd the ppernce f bld grup prperty destrying fctrs. Other cses

10 598 BLOOD GROUP PROPERTY DESTROYING FACTORS with intestinl diseses hve nt been vilble thus fr. Specil ttentin shuld be given t pernicius nemi nd relted frms f nemi. SUMMARY 1. The bld grup specific substnce A is present in the cntents f the jejunum nd upper f persns belnging t Grup A. 2. A mrked decrese in the munt f A substnce ccurs in the lwer nd cecum, while cln nd rectum cntents f dults re mre r less free f grup substnce A. 3. The bld grup destrying gent first ppers in the lwer r cecum nd its effectiveness increses in the lwer prts f the lrge intestine. 4. In tw cses with intestinl bstructin the bld grup destrying gent culd be demnstrted in the cntents f the jejunum nd. BIBLIOGRAPHY 1. Witebsky, E., Z. Immunitiilsfrsch., 1926, 49, Witebsky, E., nd Okbe, K., Z. Immunit~tsfrsch., 1927, 52, Kritschewsky, T. L., nd Schwrzmnn, L. A., Klin. Wch., 1927, 9., 281; 1928, 1, I-Iirszfeld, L., Hlber, W., nd Lskwsky, I., Z. Immunittsfrsch., 1929, 61, 61, Hdy, H., Deutsch. Z. gerichtl. Mel., 1933, 2'2, Mhrrem, I., Z. Immunlt~sfrsch., 1934, 83, Schilt, F., nd Akune, M., Miinctt. reed. Wch., 1931, 1, Sc'hiff, F., nd Weiler, G., Bickem. Z., 1931, ~li, 454; 9.39, Witebsky, E., nd Sth, T., Kiln. Wck., 1933~ 1, Sth, T., Klin. Wctt., 1934, 1, Sievers, O., Klin. Wck., 1934, 2, Sievers,., Z. Immunit~tsfrsch., 1935, 85, Lndsteiner, K., nd Chse, M. W., Prc. Sc. Exp. Bil. nd Med., 1935, 32, Schiff, F., Klin. Wch., 1935, 1, 75.