IE881121L

IE881121L - Use of antigenic substances for the prophylaxis or therapy¹of disorders and diseases within the gastrointestinal tract¹of animals and humans

Info

Publication number: IE881121L
Application number: IE881121A
Authority: IE
Inventors: Reinhard K Prof Dr Teichmann; Walter Prof Dr Brendel; Hansgeorg Prof Dr Liebich
Original assignee: Spillers Ltd
Priority date: 1987-04-15
Filing date: 1988-04-14
Publication date: 1988-10-15
Also published as: ES2065894T3; JP2656063B2; AU684824B2; DK185988A; IL85963A; JPH01268644A; AU1370088A; NO881629L; CA1325592C; IE66559B1; IL85963A0; AU7595194A; DE3851971D1; ZA882504B; EP0286847B1; DK185988D0; EP0286847A3; NO881629D0; DE3712890A1; AR243384A1

Abstract

Disorders and diseases in human and animal digestive systems, for example diarrhoea, ulcerative disorders and carcinomas, are a serious, economically important problem worldwide. It was possible to show that immunological mechanisms exhibit a protective effect in the gastrointestinal tract of animals and humans. Antigens contained in the food induce the said protective mechanisms if the body has previously been sensitised to this antigen. The use of antigenic substances in the prophylaxis and therapy of ulcerative disorders in humans is preferred. The antigenic substances according to the invention are furthermore particularly highly suitable in the raising of young cattle, which are subjected to a change of food in the first weeks of life. <IMAGE> [EP0286847A2]

Description

1 6 6 5 5 9 Prophylaxi s or a therapy of disorders and diseases within the gastrointestinal tract of aniaals amd humans using antigenic substances. 5 PESCRIPT20W The invention refers to the prophylaxis and therapy of disorders and diseases within the gastrointestinal tract 10 of animals and humans using an antigenic substance- forbid disorders within the gastrointestinal tract of animals and humans represent a worldwide serious,, ecorsosnical ly significant problem. In human nedi ci ne, among 15 chronic disease,, peptic ulcer disease, for instance, shows a life long prevalence of about 8 - 10% in Europe and North America- Thus, the total costs for peptic ulcer patients in U.S.A., The Netherlands, Italy and Sueden, including working deficit are about 1% of the respective 20 national annual expenses for health care. In Germany, for instance, the aortality because of peptic ulcer disease within the period from 1952 to 1980 has decreased only i nsi gi f i cant ly frotn 7 to 6 deaths per 100,000 inhabitants CSonnenberg R., Fritsch, A.: Changing mortality of peptic 25 ulcer disease in Germany. Gastroenterology 84, 1553 <198355. Within this period, however, peptic ulcer disease, as a deadly disease of aiddle aged and elderly people, shifted to disease of mainly old and high-aged people.. Besides the Mortality because of gastric and 30 duodena 1 ulcers, in women, has increased-, The mortality in ■en because of a duodenal ulcer remained constant; those of gastric ulcers slightly decreased.

Currently*, mainly H2~antagonists are used for the treatment of peptic ulcer disease (8auernfeind et al„,*> UI custherapi e^ JAMA, Heft 2, 135 (1986)). The principle of the therapie with H2-antagoni sts is based on the reduction of the aggressive effect of acid within the gastrointestinal tract- This therapy is effected with a number of unpleasant side-effects- A significant problem of a peptic ulcer disease still is the prophylaxis of a recurrent ulcer because, as already mentioned, peptic ulcer disease is among the chronic diseases with life-long prevalence. For instance., a long term study of three years among 44 Centres within 13 countries with a total of 1,423 patients yielded that only 81% of patients were without recurrence after 12 months*. 73% after 24 months and 65% after 36 months for duodenal ulcers (DM 111. Jig. Nr. 3, 17,, (1986 J). Therefore, there is the urgent necessity for alternative drugs for the prophylaxis or treatment of peptic ulcers without side effects which especially could be taken for a long time and as prophylaxi s- Diarrhoaea and peptic ulcer disease are a serious problem not only in human medicine but also in veterinary medicine,, especially in pig and cattle breeding as well as in poultry farming* It is a yell known phenomenon, for instance*, that piglets after weaning and a change to serai™Iiquid high energy ratio will develop ulcers which may partially be lethal,. Within the Common Market there 3 If* ^ 120 to 160,000^000 pigs (estimated values)*, among which three to four percent will be affected., Based upon these figures*, the loss of pigs will account for about 4 million per year, resulting in a financial loss of up to 1 *. 2 billion German Marks per year- It is obvious that effective measures against these disorders are necessary. Nearly the same is true for cattle breeding and poultry fa run ng.

For the pathogeny for peptic ulcer disease, aggressive factors during digestion, (Like the secretion of acid and pepsin, seem to be of special signif i cance CGrossman, K-1.: Regulation of gastric acid secretion. In: Physiology of the Gastrointestinal Tract™ Ed„: L-R- Johnson. Raven Press#. New York 6S9-671 (1981))- Opposi te to these aggressive factors are protective physiologic digestive mechanises. Among these protective mechanisms is the mucus secretion in the stomach, which together with the bicarbonate secretion is regarded as the first layer of a Mucosal barrier.

The various secretion components of the stomach, as well as the cell regeneration,,, are controlled by neurohormonal mechanisms. The hormone, which initiates the above-mentioned aggressive digestive processes is gastrin™ it uas isolated in 1964 (Gregory, R. A- , Tracy, H.4.: The constitution and properties of two gastrins extracted from hog antral mucosa- Gut 5,,. 103-117, (1964)).

Until recently, one aspect concerning studies ©m the stimulation of gastric functions was totally not taken into account: with each raeal a mucosa of the stomach also cones into contact with substances which slight be considered antigenic- Although there were hints for immunological structures in the stomach, however, there was no idea as to their functions™ Recently, it could be shown, in experimental studies, that following preceding immunization with an antigen, oral challenge with sa id antigen induces an immune reaction in the antrum of the stomach, which initiates the complete gastric cascade of digestion. This is mainly the increase of local blood flow 4 in the antrum and duodenum, increase of mucus secretion of th® stomach as well as stimulation of gastrin release® The proof of an immunologi ca I stimulation of gastric functi ons offers a completely _ new view of physiology arid 5 pathophysiology of the digestion- This data came from animal studies in which luminal antigens, in the stomach, after preceding immunization Mere recognized and indcced digestive processes (Teichmann, R-K., Andress, H.J-, Gycfoa, S-, Seifert J - , Qrendel, W«: Immunological 10 mediated gastrin release.. Gastroenterology 84,2 (198533,, Immunofluoresence microscopy revealed binding sites for the antigen applied orally uithin the lamina propria mucosa of immunised animals- Furthermore, there was an 15 increase in the number of cells expressing immune associated antigens on their surface (so-called la-antigens) in the ant rum, following antigen administrati on. Usi ng double immunofluoresence mi croscopy it -could be demonstrated that the binding sites for the 20 antigen seems to be located on la-antigen expressing eel Is.

Furthermore, there was an increase in plasma in mast cells, in the I ami na propria mucosa of immunised animals, 25 after antigen administration and an increase in the mast cell degranulation, as well as an increase in intraepithelial lymphoid cells. CTeichnann, 8.K./ Andress, H.J./ tiebieh, H«, Seifert, J., Brendel, W-: Die Sedeutung iamunkonpetenter Zellen i* Antrum bei der Stimulation von 30 Verdauungsprozessen. Langenbecks Areh„ Chir. Susppl- 1S1, CI9843 J „ % Thus, during immunologically mediated stimulation of <> gastric digestive processes, besides agressive factors like acid also zytoprotective mechanisms like Increase of blood flow and mucus secretion, are i mi ti ated- Based upon these results further experi aents Mere carried out eoncerni ng mediators of the immurtologi cal sti wulati on of gastric functions using the model of isolated antr The supernatant of fluoresic duct lymph,. collected after antigen administration in inmuni zed animals^, sti saulated gastrin release in isolated antrum perfusion- This activity uas »i ssi ng in the lymph of non-i snenuni sed animals when the same antigen was administrated into the stomach, (Tei chnann^ R-K-, Pratschke, E. , G rab, J - ^ Tutert, J . , Enders^. G >,8 rend a I,,. W.: Gast r i nst i ®u I i erende Wirkung von Ductus-thoracicus-Lyraphe nach gastrointestinaler Immunreaktion. Langenbecks Arch. Chir. Suppl. 271 C198S))- Thus,? it has been shown that soluble mediators are responsible for the antigen induced gastrin release.

The mucosa mast cell products Hi stami n and Leukotrien being released during the immunological reactions, significantly inhibit antral gastrin release* Gastrin itself stimulates the release of Histamin so that these results^ for the first ti me, sEiow local antral negative feed-back mechanism between the G-celI and the mast cell CPratschke, 6., Tei chwann, R_ SC. , Grab, J., Tuterty E. , Srendel^. W,: 0er EinfluB von HastzelIprodukten auf die Gastrinfreisetzung. Langenbecks Arclh. Chir. Suppl-, 293^ <1985)).

Simultaneous to the i nM hition of gastrin release by 6 Hi stami a and Leukot ri en C^, there is a release of endogenous prostaglandine E^ ar,c^ ^2" model of Isolated antrum perfusion it could be shown that prostaglandin® inhibits the gastrin release. This night indicate that the inhibition of gastrin,* by saueosa »ast celt products, may be Mediated by endogenously released prostaglandins CPratschke, E-, Tei chmann, R.K., Grab,, J-, Enders, G., Brendel, W, ~ Endogenes Prostaglandin E2 aIs Regulator der Gastrinfreisetzurog. Ins Kongreftbericht der 26, Jahrestagung der osterr* Gesellschaft fur Chi rurgie. Hrsg-: F. Helmer, E. Horcher, Styria-Verlag, Graz, 131., (1985)).

During the process of antigen recognition involving antigen presenting cells Iynphok i nes Interleukin 2 and Gamma interferon are released- Both stiraulate gastrin. Thus,, for the first time,* stimulation of a gastrin by mediators of isnmuno competent sells was shown (Prat schke, E., Teichasann,, IR-K., Grab, J., Hammer, C- Srendel, W.: fiastrinfreisetzung durch Kediatoren immunkompetenter Zellen. Langenbecks Arch. Chir., Suppl., 261 (1986)).

Xumunofluoresence ai c roscopi c studies of the ant rum of the perfusion with Interleukin 2 and Gamma interferon, using monoclonal antibodies against class 2-amtigens of the HHC complex, revealed further that, after the perfusion with antigenic structure of DLA Class 2 (Dog-Leukocyte antigens), same are expressed on epithelial cells, macrophages, probably also on intraepithelial T-Lymphocyte. There Mas no expression of those two antigens after perfusion with Interleukin 2, Therefore, Interleuki n 2, but also Prostaglandi ne$ and other substances,, may serve as mediators of this immunological reaction. 9 7 Thus,, the above-mentioned antigen induced react ions are discussed for the development of ulcers arid aay also be of significance for the total gastrointestinal tract.

Whenever antigen induced increase of the antrum blood 5 flow, gastrin and caucus secretion is aissing, or disturbed (Locally,, the resulting acid may not be suff 1 cliently buffered and transported back in case of lack of increase of blood flow,, resulting in penetration of the acid into the mucosa and possibly inducing local ulcers there. 10 Investigations on the immunological stimulation of gastric reactions also showed that after immunological stimulation with a synthetic antigen,, besides the antigens specific in view of the gastrin release, and the strongest stimulator 15 for the gastric acid secretion,, in the sense of an increase of aggressive digestive processes, also an increase of antral secretion of Somatostatin Mas noted which has a protective effect by inhibition of acid and pepsinogen secretion, (Kramli ng,. H.-J., Tei chmann R-K. , 20 Merkle, R., Enders, G-, Brendel, W-: Gastrale Protektion durch immunologi sch indusierte Somatostatinfreisetzung, 153., Tagung Vereinigung Niederrheinisch-WestfIIischer Chi rurgen, Koln, 9.-11., Okt, 1986). 25 This data results in a completely new far reaching, and so far not recognised, possibility to specifically influence digestive processing as a protective mechanism concerning ail disorders and diseases within the gastrointestinal tract- In human medicine, diseases like erosions,~peptic 30 ulcers and, last but not least, cancer are of greatest interests. In veterinary medicine interest is mainly in the critical transition phase of young animals following weaning and subsequent contact with high energy food because of occurrence of erosions and peptic ulcers 8 during this phase- There is a protective Induction of f unctions..of . the. 5 functions and protectsve physiologic digestive processes, which may be used in a broadest .sense; this is the use ot an antigenic substance for the prophylaxis and therapy of disorders and diseases of the gastrointestinal tract of animals and humans inducing protective physiological 10 digestive processes after an oral challenge of the aniaasl or hussars with the antigenic substance following preceding imnunization.

When applying an antigenic substance? as outlined by the 15 invention, the stimulated gastric functions may york as follows" After immunization of the stomach with an antigera which is present in the food the subsequent contact of this 20 antigen, in the food, with the gastric mucosa, the antigen will be recognized by the so-called antigen presenting cells, for instance, macrophages and presented to the T-lymphocytes by the help of class 2 antigens- An indication for a lympocyte mediator immunological 25 stimulation of the above-mentioned gastric functions Mas demonstrated in an experiment in which immunized dogs were irradiated by an 80 Rad of an ultra-hard X-ray resulting in an inhibit ion of the immunologically stimulation of gastrin release based 'Upon destruction of lymphocytes- The 30 capability of the gastrin cell to be stimulated, however, is not effected by the i rradi ation.

Followi ng the contact of the anti gen with the stomach, bei ng immunized as outlined above, there is a type of 9 cascade of release of Interleukin 1 and stimulation of T—Helper-Lymphocytes- The Helper cells themselves Mill release Interleukin 2 and Sasnsaa interferon^ two Lymopbokines uhich were shown to be able to release 5 gastrin., A significant protective physiological aechanisa, namely the increase in mucosa blood flou representing one of severa I steps of the cascade of protective digestive processes/ say be caused by mediators of snast cells,, especially Histamine. Besides the expression of 10 class-2-antigens on antigen presenting cells of the ant rum, Gamma interferon also induced the expression of these antigens on epithel cells. Possibly this additional recruitment of epithel cells represents a potentiation of antigen recognition™ The vagus nerve interferes with this 15 antigen recognition system^ in the sense of immunomodulation. The mentioned inter-relations are shown in Figure 1.

The cslcer protective effect of antigen specific stimulation of digestion has been demonstrated in an animal model- 'Using the model of the ""SHAV'-Ulcer (Shay. H., Komarov, S.A., Pels, S.S.^. Mekanze, Gruenstein^ flm, Siplet, H.; A simple method for the uniform production of gastric ulceration in the rat., Gastroenterology 5, 43,,. 25 C194S))^ the influence of antigen stimulated regulation of the digestion was investigated. Male Histai—Rats were immunized systemically with a 'Hapten. Afterwards, the Hapten was administered into the stomach and as control sodium chloride Mas given.. In further control studies, the 30 Hapten or sodium chloride, respectively was also - administered in non-immunized animals® These rats being treated in such a way were investigated aacroscopicatly and Microscopically to check gastric lesions after their death but not later than 18 hours- There was a significantly less expression or number -of perforations 10 and large wall necroses in the sore ventri culous in the group of animals which had been immunised with the Hapten (Figure 3,5.

Thus, using the model of the "SHAY-Ulcer" the protective effect of an ant1 gen specific stimulation of the digestion could be demonstrated. Although there Is an ulcerogenic effect by release of gastrin and, consecutively of acid, as an aggressive factor, simultaneously there Is also stimulation of protective mechanisms, foe* instance, an Increase in blood flow, mucus secretion, possibly through release of Somatostatin, Prostaglandines and other mediators which finally preverst the occurance of severe* secretions of the gastric mucosa or gastric wall* This ulcer protective effect of an antigen induced regulation of the digestion, as shown here, may teleologicatly well be regarded as a "logical"4 regulation within physiological processes, as it results simultaneously in a stimulation of the digestion and a reduction of the risk to develop an ulcer- Thus, the develoment of pepti € ulcer may well be understood as a Local failure of the stomach to be ready for antigenic reactions beside other known causes. The stimulation of gastric immunological mechanisms by immunization with antigens, therefor®, resuits in a completely new possibility for the prophylaxis and therapy of disorders and diseases within the gastrin intestinal tract of animals and humans offe ri ng broadest 1ndi cations. The essential, other advantage of use of antigenic substances, for this purpose, is based on the fact that the anti gens being used are biological substances.with which the saaasal as well as the human being comes into contact naturally and, thus, there is no aggressive effect on physeological processes but, moreover, an induction of a logical protective physiological process in a natural aanner and, thus, same is without side effects- In respect of the mentioned mechanisms of the immunologi cat stimulation of the digestion, it is, of course, of special advantage to use such substances as antigens which later will also be included within food. Following immunization of the animal or the human with the antigenic substance a level of factors, within protective effect, will be increased in such a way that disorders or diseases of the gastrointestinal tract wilt be prevented or treated,. Thus, plant proteins, preferrabty from ®sai £e or soya bean, but also animal proteins, are especi ally suitable for the immunization because these proteins are often found in food or food products of animals or humans.

Whey protein can, for example,, be used as an ami mat protein., It became apparent that the prophylactic influencing of gastrat mucosal changes (erosions),, e-g- in pigs,, which were additionally administered whey protein via the drinking water during the entire fattening period until slaughtering of the pigs after a fattening of 10 weeks, entailed that in particular serious changes of the mucous membrane in the animals treated, as described above, occurred less frequently by 70Z than in a control group., The whey protein should preferably be present in a 3 to 9% whey protein solution and especially preferably in a 6X whey protein solution,, Human protei ns, which are components of blood or which may be found in serum or other blood derivates, for instance, hemoglobin are suitable to be sased as antigenic substances in the sense of the invention.

Also other antigens, which do not necessarily correspond with substances being present in the i ngested food, nay be suitable for the induction of the immunological situation 12 which is responsible for the abowe-mersti®ned induction of mechanisms. for instance,, in case of immunization with proteins derived from the envelopes of virus, an immunization ^ay be possible, which will work in the sense of the above-described stimulation of the digestion, but also may cause an immunization against virus., Same is true for antigenic substances which are present in mieroorganisms with which the gastrointestinal tract way come into contact In one way or the other. The stomach might induce the above-mentioned protective processes as well- Specially suitable seem to be lipopolysaccharides of Escherichia soli, a natural commensal bacteria of some mammals which is always found ia the food as contamination and, thus,? seems to be suitable.

The use of an antigenic substance being present as a synthetically-produced protein, preferably as a short peptide which contains antigenic determinants for the induction of an immune response, is especially preferred because then a special and directly applicable prophylaxis and therapy will be possible.

A well suitable substance for the induction of the immune situation to induce the protective mechanisms are i iapte ns, preferably nitrophenyl acetic acid (NSP5. This substance has shown its speci al usefulness in the experiment to induce the immunization, which prevents the development of mucosal lesions after intraluminal challenge of an antigen specifi c stimulation of the stomach. Especially with this substance, an icnmunologisally mediated ulcer protection was shown for the first time.

Preferably, the above-mentioned nitrophenyl acetic acid 13 is used by coupling same to a carrier protein, namely ovalbisrai n» As out Lined in the description above, immune modulatory substances are in the equilibrium state between the aggressive and protective digestive snechani ssas. The addition of "immune modulatory substances together with the antigenic substances may, therefore^ cause an increased effect of the induced protective digestive mechanisms.

The use of combinations of one or several of the above-mentioned substances may also result in an improved prophylaxis or therapy™ The pharnaceutical coniectioning of one or several of the above-ia en tioned antigenic substances,, perhaps by adding of immune modulators^ may be performed for the enteral as yell as for the parenteral application- Concerning the enteral confectioning, aerosols,, being applied to the nose or the bronchial system,, are preferably used- The confectioning of the parenteral application of antigenic substances preferably concerns substances to be used for intramuscular and/or sub~cutanous applications.

In the veterinary medicine, a preferred use of the antigenic substances is to immunize piglets with the ■antigenic substance about two weeks after birth. As has aLready been mentioned above, piglets suffer from peptic ulcers after the weaning and the change to semi-liquids energy-food. They may even die from these ulcers.^ Treating these piglets in this critical phase with antigens which later aill be present in the food prevents, or at least, reduces damages- The proteins being used as antigenic substances are biological substances which, after application, will be metabolized completely without 14 residue or being excreted in only small amounts.

An especially preferred treatment of the piglets is to immunize theaa intramuscularly, preferably the first 5 injection being about two weeks after birtti; a second injection about four to five weeks after birth and a third injection shortly before changing of food- In respect of their practicability, the animals could then be immunized when they receive iron-injections duri ng breeding., The 10 .antigenic substances may be packed in a kit which already contains iron- U hen the piglets receive the food which contains said proteins.* with which the animals were iamuni zed, ^ the antigen may also be given to the drinking water in order to further induce and maintain the protection of the gastric mucosa by the described immunological mechanisms. 20 In human medicine, the us® of antigenic substances is preferably suitable to prevent or reduce erosions*, peptic ulcers or cancer in the human gastrointestinal tract. The increased production of mucos may prevent the mucosa of oncogenic substances. Furthermore,, oncogenic substances, 25 which are known for the organism, may become inaffacted by the induction of the digestion- Al so, there is a new preferred and broad spectrum of indications in the prophylaxis concerning diarrhoea in 30 travellers who cofae into contact with food products during their trip which are, so far, unknown by their gastrointestinal tract, this means that the gastrointestinal tract, so far, had not been in contact with such food products* The lack of induction of digestion may cause passage of protein Into deeper parts 15 of the got with subsequent possible overgrowth of b® cteri a- It is also a well known problem in the breed i ng of domestic animals that following transportation of the domestic animals to places at which the breeding will be continued after the first week of life* the changes in the envi ronnent and food way cause significant diarrhoea which* again* may be the cause of other bacterial or viral diseases of the whole gastrointenstinal tract.

Immunisation of these animals with antigenic substances^, beirsg keiown to be present later in the foodmay prevent these disorders and diseases- One possible use of an antigenic substance is to sensitise piglets parenterally as of the third week at intervals of fourteen days with 2 ml each of the 6% whey protein solution without the addition of adjuvant that is to say on the whole with 4 ml. After weaning of the piglets in the second week and gradual change over from pig breeding feed to coarse grain fattening fodder the piglets were brought into another pig sty and kept in groups in loose housing boxes., from this time on approximately as of the tenth ueek* all pigs were exclusively fed with very finely ground fattening feed., In literature the administration of such feed is i ndi cated* besides stress factors* as the decisive cause of the formation of mucosal changes* such as hyperkeratoses* erosions or ulcere in the gastric mucous membrane of the pig..

The pigs were administered 3 to 6% uhey protein with the drinking water during the entire fattening period until the slaughtering of the pigs* after a fattening of ten weeks. 16 In animals which have been treated in this way changes of the uucosa-1ike erosi ons appeared much less frequently- In the same way as the antigenic substances have.been ased for.the treatment of piglets, calces may be immunized parenterally and/or enterally in their critical, phases of life. It is preferred to give intramuscular injjestions of the antigen in calves within about the first four weeks after births a second injection at about the sixth week and a third injection in about the eighth to tenth week when changing to calf food.

Within the first weeks of breeding, the ealves receive milk and calf food and,, generally^ energy food from the eighth to tenth week- From the time of changing to the high-energy food,, the antigenic substance is added,, preferably to the food and/or drinking water to use and/or maintain the I eveI for the protection of the intestinal tract by the described imaiunologicaI mechani sias.

The immunization is preferably performed also with adult ruminents. Bulls and cows increasingly suffer from gastric ulcers of different stages during the high lactation period. These ulcers are also especially considered as the cause of subsequent infectious diseases of the whole gastrointestinal tract in adult cattle.., Enteral or pa rentera I treatment of animals with the antigeni e substances way prevent these damages.

Another field of using the antigenic substances is the treatment of domestic poultry, for instance,* chicken, turkeys., geese,, clucks and turkey~cock$~ The major sources of food^ for the breeding of domestic poultry,, are grain granules,, fish flour, soya and grinded peanuts- To induce 17 the uni sat ion, in respect of the above-ment i oned food substances, a parenteral injection of the antigenic substance will be performed on the first day after hatching because of praci cal reasons™ Later, 'annunizations for maintenance of a preceding immunisation taay be performed by application of the antigen.via the d ri nk i ng water within the early days or weeks of life.

The current invention will nou be described, in more detail, in the following Figures and Examples., It is shown in Figure 1: The regulation of antigen stimulated gastrin release.

Luminal antigen is recognised by antigen presenting cells and may stimulate ssast cells as well as T-lymphocytes among others. The T-cei.I products Gamma interferon and Interleukin 2 cause release of gastrin swhi ch, in turn, stimulates Hi stami n. The mast cell products Histamin, Leukotrien and Prostaglandins £■? again inhibit the gastrin release (from negative feedback)™ The vagus nerve may also influence the T-cell systems and the antigen presenting cells. £.i.9M£&»i: • Ulcer index using the model of the alcohol ulcer of the IT <8 *£ ® Rats being immunised with NIP show a significantly lower 18 ulcer i ndex following intragastric application of the 3rnt T3 Fioure 5: Percentage of animals with lesions,*. induced by the SHAY~Ulcer model in controls and in animals immunized wi NIP-ov®Ibumin- The intragastric application of NIP-human Gamma-globi n (NIP-HGG) results in a significantly lower expression of peptic ulcers in immunised animals.

Fiojure 4; Quantitative changes of the evaluation scales CO and +++ after a pathological-anatomic evaluation of the gastric mucous membrane of pigs in the area of pars proventricuiaris after a 10 week feeding of very finely ground feed rations (%).

Example 1 The purpose of the described examples was to investigate the effect of the i mmunologi eel processes on the other induction concerning stimulation or protection™.

Hale Mi star-Rats C2S0~3S0gS were immunized systemieally with NIP-OA CNitrophenyl acetic acid-Ovalbumin)- To induce ulcers, 1 oil of absolute alcohol was administered into th® stomach under ether anesthesia, fifteen a»1 nutes prior to the alcohol application, one group (13 of the 19 animals Cn=12) , .received the anti gen NIP C coupled to HGG as carrier-protein), Group II (n=12), the carrier protein HGG alone. Non-i mmuni zed ani ssals, six having received NIP-HGG (III), a further six HGG CIV), proved as further controls. An hour after th® administration of alcohol, the lesions were determined after the animal was kilted and the stomach removed. The lesions were measured bsc roscopi cat ly by esti matati on of the length of the hemorrhagic lesions.

Animals havi ng been i tnmuni sed systeai catty ui th NIP developed significantly less caucosal lesioos after oral NlP-application and subsequent administration of alcohol (p ~ 0,02) than all control groups.

Table 1 Immuni sati on Intragastric Length of Administration of hemorrhagic Lesions I (n=12) NIP KIP 24 p-0,02 II (n=12) NIP HGG 57 III Co=6) non~i««- NIP 72 IV Cn=6) non-i mm- HGG 78 .uninal antigen specif i c stimulation. of the stomach in imsaunised Wistar-Rats significantly reduced the development of mucosal lesions. Therefore, for the first time, an iamunoLogi cally mediated ulcer protection is' shown (Figure 2). 20 Eyagple 2 The i ointuni sat *3 on of aa Le Uistar-Rats (250~350g) was performed using ^SP-OA (NitrophenyI acetic 5 acid*-0vaIbunin)• Pylorus ligated rats (SHAY~Ulcer) served as ulcer aodel- following ligation of the pylorus under chloralhydrate anesthesia, the immunized rats received the antigen KIP (coupled to human gamma globulin) directly into the stomach (test group)™, The control animals 10 received only the carrier protein HGG „ Another control group consisted of sham operated animals (without pylorus ligation)- Eighteen hours after the operation,.- the changes in the stomach wall were investigated macroscopically and microseopically„ The lesions were 15 defined as follows: Perforations^ confluent transmural ulcerus and all necroses of more than 0,5 cm in diameter., Animals which were systeraically immunised with HIP showed significantly less lesions after pylorus ligation and intragastric application of NaP~HGG (p = 0^05) than controls which had received HGG. Sham-operated rats showed no mucosal lesions. 30 21 j3pjtenmz 5 INIIP-specificatly intragastric Preportion of ■3aimuni $ed anisals Application of Annuals showing Lesions 10 Test group (n~24) 5*? .IP-HGG 7/24 Control group Cns23) HGG 14/23 Sham-operated For the present immunization of the piglets whey proteins, namely as a 6Z whey protein solution,? were administered to the piglets without the addition of an adjuvant or during the later course of immunisation as a 3 to 61 addition to the drinking water.

Thirty pigs were taken as a test group and thirty pigs as control animals for an examination of the prophylactic effect of the whey proteins administered as antigens regarding gastral mucosal changes- During the test five pigs of the test group and two of the control animals died as piglets or during change-over.

The test animals were pret reated according to the following scheme. Piglets were sensitized parenterally as of the third week at intervals of 14 days with 2 ml each of a 62 whey protein solution without the addition of adjuvant that is to say on the whole with 4 nil ™ After weaning of the piglets in the second wee^ and gradual change over from pig breeding feed to coarse grain fattening fodder both the test and th® control animals were brought into another pig stay and kept in groups in loose housing boxes™ From this tisae on,, approximately as of th® 10th week,, all pigs were exclusively ted with very finely ground fattening feed. leu literature the ad ministration of such a feed is indicated besides stress factors as the decisive cause of the formation of mucosa I changes, such as hyperkeratoses, erosions or ulcere in the gastric mucous membrane of th® pig. 23 Both groups of animals were adaini stered 3 to 6% uhey protein with the drinking water during the entire fattening period till the slaughtering of the pigs after a fattening period of 10 weeks.

The changes of the gastric mucous membrane of pigs in the area of pars proventricularis was evaluated according to pathological-anatomic criteria..

Table 3 shows the frequency of the evaluation scales CO to **4-) after a 10 weeks feeding of \re ry finely ground feed rations and their percentages £%>« Table 3 0 4- 4"f 6 6 8 21,4% 21,4* 28,6% 10 7 6 40% 28% 24% 4.4.4. 8 control group 28,6% n = 28 2 8% test group n ~ 25« Irs figure 4 it is graphically represented that in the animals treated with whey protein twice as much animals had an intact mucous membrane than in the controls (the two left columns in Figure 4 with the designation "0" which is used for an intact mucous membrane). The two right columns or Figure 4 reveal ora the other hand that serious changes of the mucous membrane, e.g. erosions, which are designated with "occurred by ?0% less frequently than in the control. zu The exact figures of the macroscopic evaluation of the gastric mucous membranes of the experimental animals as they are revealed by table 3 show that low grade, flat hyperkeratoses (designated with ""•f8) or moderate 5 hyperkeratoses with pointed serrations (designated with occurred with the same frequency in both groups. However, the changes with i.e. the erosions, are decisive,? whose prevention in this test with pigs was possible still more clearly than in the experiments 10 described in examples 1 and 2- Example 4 15 Performance of Immunisation of Cattle As antigen.* a protein is used which is not present in the calf food, especially not in the milk replacer with which the calves are fed after birth™ However, the antigen must 20 be present in the high-energy food which is given to the calves in about the eighth to tenth week after births The ulcerative lesions preferably develop during the change to high-energy food, usually at an age of four to ten weeks., These disorders are in connection with the mentioned lack 25 of immunization of animals regarding the proteins being present in th© high-energy food. The main consti tuents of the high-energy food for ruminents are mainly corn, wheat, soya, barley, grassi lage and hay™ These consti tuents may change according to the degree of intensive breeding. 30 The first intramuscular injection of the used antigenic substance will be given in the first four weeks after the birth of the calf. A second injection will follow in the sixth week and a third injection at the tisae when high-energy food will be given in the eighth to tenth 25 week- In the first week of breeding the calves receive milk and starter diet. They will receive high-energy food about the eighth to tenth week., Then., at the time of changing to the high-energy food - the antigenic substance 5 Mill be added to the food and/or drinking water to use or establish the imtnunologi cal protection of the gastrointestinal tract by the above-described iumunologi cal mechanism. 10 The dose of the antigenic substance should be about 0^1g test substance per kg bodyweight by intramuscular application and will be repeated two to three times- To induce the protective mechani sras„, the antigenic 15 substance will then be added to the drinking water in a dose of up to 1g per 100 ml water.

Perforating this procedure,,, the ulcerative lesions in the gastrointestinal tract of the calves will be prevented or 20 reduced., According to the above-mentioned handling of the cattle^ they will be much better protected against changes of the climate^ the food and transports ("Crowding Disease").,. 25. especially within the first weeks after birth- The frequently-oceurring disorders or diseases of the gastrointestinal tract, during these environmental changes,*. especially the diarrhoea*, may well be prevented by the new aechani sn- Di arrhoea can be the cause of other 30 bacterial or viral diseases in the gastrointestinal tract.

Concerning the use of antigenic substances in adult ruminants, the required antigenic substances will be applied in bulls and cows during the period of intensive feeding and high lactation. Thus,* also in adult animals 26 the more common gastric ulcers., of various stages,*. siay be prevented or reduced. £xa_t»p5. <& __5 Performance of immunization of poultry Similar to the {maunization of pigs end cattle,, domestic poultry,, for instance^ chickens^ turkeys^ geese^ ducks or turkey-cocks^, will be immunised with antigenic substances in the above-men tiioned way- The antigenic substances will consist of grain,? fish flour, soya or peanuts according to the major sources of food during the later breeding.

The parenteral injection of the antigenic substance is performed on the first day after hatching due to practical reasons. This first immunization will be intensified and established by later application of the antigenic substance by putting the antigenic substance into the drinking water in an sntraI form about one week after the hatching. » 27 -

Claims

1. CLAIMS 1. Use of at least one antigenic substance for the preparation of a medicament for the prophylaxis and treatment of disorders and diseases in the digestive tract of animals and humans, in which method after prior immunization or sensitization by the antigenic substance(s), a subsequent gastrointestinal administration of the antigenic substance(s) in a suitable formulation is carried out, with the following exceptions*;(a) a method for preventing diarrhoea in poultry, as a consequence of a coccidial infection by rearing them with a feed which contains added, live, sporulated oocysts of at least one coccidial species to which the poultry is susceptible, the oocysts being present in a concentration which is sufficient only to induce a subclinical infection in the poultry,* (b) a method for the prophylaxis of intestinal diseases in pigs by oral immunization and additional oral administration of heat-inactivated antigen formulations of E. coli strains, which lead to intestinal diseases; and (c) a method for the prophylaxis and treatment of intestinal diseases in pre-ruminant calves by oral immunization and additional oral administration of heat-inactivated antigen formulations of E. coli and Salmonella strains, which lead to intestinal diseases.

2. Use of at least one antigenic substance according to Claim 1, in which the antigenic substance is vegetable protein, preferably from maize or soya.

3. Use of at least one antigenic substance according to Claim 1, in which the antigenic substance is animal protein.

4. Use of at least one antigenic substance according to Claim 3, in which the animal protein is whey protein.

5. Use of at least one antigenic substance according to Claim 4, in which the whey protein is used as a 3-9% strength, preferably 6% strength, whey protein solution. - 28 -

6. Use of at least one antigenic substance according to Claim 1, in which the antigenic substance is human protein, preferably from blood or blood constituents, namely serum, plasma or haemoglobin.

7. Use of at least one antigenic substance according to Claim 1, in which the antigenic substance is viral protein.

8. Use of at least one antigenic substance according to Claim 1, in which the antigenic substance is a microorganism or a constituent of a microorganism.

9. Use of at least one antigenic substance according to Claim 8, in which the antigenic substance is a bacterium or a component of the cell wall of a bacterium.

10. Use of at least one antigenic substance according to Claim 8, in which the microorganism is a representative of the protozoa.

11. Use of at least one antigenic substance according to Claim 1, in which the antigenic substance is a synthetic protein, preferably a short peptide, which contains antigenic determinants for triggering the immune response.

12. Use of at least one antigenic substance according to Claim 1, in which the antigenic substance is a hapten, preferably nitrophenvlacetic acid (NIP).

13. Use of at least one antigenic substance according to Claim 12, in which the hapten is coupled to carrier proteins.

14. Use of at least one antigenic substance according to one of Claims 1 to 13, in which immune modulators are used in addition to the antigenic substance.

15. Use of at least one antigenic substance according to one of the preceding claims, in which the antigenic substance is produced for administration by the intestinal route, preferably as an aerosol, if appropriate with the addition of pharmaceutical^ tolerated auxiliaries. - 29 -

16. Use of at least one antigenic substance according to one of Claims 1 to 14, in which the antigenic substance is produced for parenteral administration, preferably intramuscular and/or subcutaneous administration, if appropriate with the addition of pharmaceutically tolerated auxiliaries.

17. Use of at least one antigenic substance according to Claim 15 or 16, for the prophylaxis or treatment of erosions, ulcers or carcinomas in the human digestive tract.

18. Use of at least one antigenic substance according to Claim 15 or 16, for the prophylaxis of diarrhoea caused by foodstuffs or feedstuffs with which the human or animal digestive tract has not yet come into contact. 20. Use of an antigenic substance as claimed in Claim 1 substantially as described herein with reference to the Examples. TOMKINS ft CO.