DE1492083A1 - Method for selecting a virus or other organism with the ability to induce protective resistance in an animal against a pathogen that is immunologically unrelated to the virus or organism - Google Patents

Description

Method of selecting a virus or other organism with the ability in an animal to resist one with the virus or organism to cause immunologically unrelated pathogens.

The invention is based on a method of just selecting one Virus or other organism old of the ability to Sohuti resistance in an animal against a pathogen that is immunologically unrelated to the virus or organism evoke to which the animal is susceptible and a process to an animal against pathogens caused by an infectious virus or bacterium, for example, to which it is sensitive, can be caused by using a heterotypic, immunologically unrelated pathogen and foreign to the animal resistant close.

The use of the ballpox virus to protect individuals against the Leaf virus Doroh Jenner in 1798 was the first example of its use of a virus sum protection against another virus. These "smallpox" viruses were that Reported leaf virus neutralized as sera from vaccinated individuals and serums of cowpox, leaflets and vaccine pox obtained by alastrim (i.e. of course occurring, weakened leaves) -infected patients were neutralized, immunologically related to each other.

Many other vaccines have been developed since Jenner included but always use homologous viruses or bacteria that are virulent inactivated or were changed.

In all cases, im @ unity was caused by the appearance of antibodies announced, and a vaccine was only considered to be immunologically effective when after sera from vaccinated individuals show significant levels of neutralizing antibodies for the homologous virus or bacterium.

The invention provides a method of selecting a virus or other organism with the ability to provide protective resistance in an animal a pathogen that is not immunologically related to the virus or organism to evoke, to which the animal is susceptible, created its peculiarity consists in sifting out the virus or other organism as long as until a positive complement fixation antibody is obtained and the one so selected Virus then used to conditioning animal cells in such a way that they move faster Generate antibodies and thereby make the animal more resistant to the pathogen do.

In the investigations in this area with the viral diarrhea virus, a Bladerkranibeit, and the swine choleravirus, a swine disease found that the live virus diarrh @ evirus pigs against lethal doses from swine cholera virus s. This was something unexpected because of the two Viruses are not immunologically related to one another in that they are not cross-linked immunization occurs, namely vaccination of pigs with the viral diarrhea virus does not generate neutralizing antibodies against swine cholera virus. The protection, which the heterotypic viral diarrheal virus creates against the swine cholera virus, therefore has no direct immunological basis.

The induced resilience or altered susceptibility is apparently based on the fact that pigs vaccinated with viral diarrhea virus are thereby protected erseugen, dnrs they i @ comparison to control animals that received no viral diarrhea virus, after vaccination with pig cholera virus, pig choler @ antibodies formed very quickly, while the control animals after receiving the fatal pig oholeravirus vaccination no pig choler @ antibodies formed. The benefits of using viral diarrheal virus as something alien to pigs, namely not causing any symptoms of disease in them Virus as a means of making pigs resistant to swine cholera virus to make are obvious.

In this definition, the patent attached herewith is referred to ... es ...... (Akt.Z .: P 30 671 IVa / 30 h referenced in a process for generating a hyperimmune serum @ gq.n pig cholera vaccination first with Virusdiarrho @ virus and then asked for one with swine choleravirus.

In the course of further research it was also found that living aden @ svirus dogs against the immunologically different and unrelated canine hepatitis virus pours. As is the case with the viral diarrheal virus in pigs, vaccination called With the iUr Bunde foreign adenovirus in the dogs no symptoms but also did not produce neutralizing antibodies against infectious canine hepatitis virus.

Here, too, the induced resistance is apparently based the adenovirus-vaccinated Sunde after vaccination with canine hepatitis virus s @ hr quickly generate canine hepatitis antibodies.

The studies have shown that it is important that the heterotypic or foreign pathogens, namely the virus or bacterium, that to make the animal resistant to the infectious, immunologically not related pathogen is used, in the animal complement-binding (however no noticeable neutralizing) antibodies against the latter developed. The complement binding test is based on (Carmich @ el et al. in "The Cornell Veterinaria" 52 (1961), 125 has been described. The complement-binding antigen of infectious canine hepatitis virus was made from infected canine kidney culture cells. Noticeable cytopathic Cultures showing changes were 5 to 7 days after inoculation with 100,000 TCIto harvested and then frozen and thawed six times. @ after 10 minutes Long clarification at 4 ° C. by means of a centrifuge at 7000 g were the supernatant liquids combined and stored at -50 ° C.

Such preparations provided sufficient antigens that were free from anti-complementary or lytic properties was.

The experiments were carried out in such a way that one series of Twice dilutions of warm activated serum or necessary in with veronal produced a pH of 7.3 buffered saline. 0.25 com each of each Dilutions were mixed with the same volume of antigen or serum according to previous "box" titration contained four units. @this antigen serum then two full units of guinea pig complement in 0.5 com liquid added, and the mixture was incubated on the water bath for two hours at 360C.

Prewarmed 2 percent sheep erythrocytes were then collected together with the same volume of hemolysin containing 2 units for 15 minutes Incubated at room temperature.

0.5 com of this mixture was added to each sample tube. The tests were read after an additional 30 minutes of incubation. As endpoints the strongest serum or antigen dilution was recorded, which according to visual Observation produced a rose-tinted complement fixation. All tests were made appropriate controls switched on. This (or the positive gel diffusion) test represents the critical indication that the heterotypic pathogen in spite of its foreign character to the animal, this to the infectious and Makes immunologically unrelated pathogens resistant. It is It is also important that this is heterotypic or foreign pathogen as well as complement-binding as well as neutralizing antibodies against the heterotypic or foreign pathogens developed. it also depends on the heterotypic pathogen that serves to protect the animal against the infectious Making pathogens resistant to ruin, a) no disease symptoms in the animal causes, is therefore foreign to it, and b) none or only negligible Little neutralizing antibodies developed against the infectious pathogen, is therefore not related to him.

These aforementioned factors are interrelated and the investigations have shown that a foreign pathogen, the complement-binding agent in animals Developed antibodies against an immunologically unrelated pathogen, which makes the animal resistant to the latter at the same time.

The investigations have also shown that one of the excellent Advantages of the present invention is that the first with the foreign and then vaccinated with the immunologically unrelated pathogen neither of the two carriers of the disease is transmitted to other @iere. For example, showed Pigs that were exposed to contact with pigs, which first came into contact with the Virus diarrhea and dan @@ h had been vaccinated with the Sohweineeboletsvirus from both viruses show no titer or disease symptoms. This is an essential one difference versus the use of homologous vaccines in which the vaccinated animal has a Is a source of infection for other animals. For example, there is always the Risk of swine cholera from pigs vaccinated with live swine cholera virus is transferred to receptive pigs.

To make the animal more resistant, it used foreign pathogens must be alive fine, since the use of inactivated pathogens does not can achieve the desired immunization. You can use both modified fixed or weakened, living pathogens al also so-called native or virulent Use trunks.

For example, it can be used to make pigs more resistant used against all live virus diarrhea viruses, such as them in the related patent mentioned above and in it listed prior publications are described. Because you're injecting live pathogens into the animal can be carried out according to conventional subcutaneous or intramuscular vaccination procedures.

The following examples serve to illustrate the invention.

Example I Live Herford Calf Mils Virundiarrheal Virus became after that of Gillespie et al. in "The Cornell Veterinarian" 50 (1960) 75-79 described method in tissue-cultured bovine embryonic cells calmly. Using one cubic centimeter of tissue culture fluid from the first pass was made Pigs vaccinated intramuscularly and soled with virulent pig cholera virus excited. All the pigs stayed alive. On the other hand, control pigs went, which nioht had been immunized with the viral diarrheal virus due to vaccination with the virulent porcine cholera virus to porcine cholera.

Example II Example I was repeated with the modification that the live viral diarrheal virus used was one which, according to Gillespie et al., l.c., had seven passes through tissue culture. Pigs intramuscularly with 1 com of undiluted tissue culture each had been sapped, were all at Excitation with virulent porcine choleravirus resistant to porcine cholera.

Example III Example II was made using live viral diarrheal virus repeats that in the by Gillespie et al., l. c, described way elfml in two days Spacing through which tissue culture was passed. Pigs that have 1 com of the virus were vaccinated from the eleventh passage, were found to be virulent when excited Pig choleravirus as immune.

Example IV Young pigs about 8 weeks old from a disease-free Herd were divided into three groups. Group 1 became 10 percent with 1 com @Filzemulsion vaccinated from one with the virulent New York l strain of Viruddiarrhoevirus infected calf was obtained. Group 2 was inoculated with 1 com tissue culture, the obtained after nine serial passages of Oregon C 24V strain of viral diarrheal virus and group e 3 was left untreated as a control.

Boi none of the pigs with live viral diarrhea virus, indifferent whether from spleen or tissue culture, disease symptoms were observed. Controls with friends who were exposed to contact also showed no signs of illness. After four weeks the pigs were in all groups each vaccinated with 1 @@ m of a mil @ -emulsion, which from a with the virulent strain A pig choleravirus infected pig had been obtained. All the pigs showed an increase in temperature 2 to 4 days after vaccination with the porcine holeravirus. The control pigs not vaccinated with the viral diarrhea virus retained their elevated levels Temperature and died within 8 to 15 days of the test. In contrast this was shown by the pigs to which the viral diarrhea virus was initially incorporated was, a temperature increase of lesser extent and lasting only 1 to 2 days Duration after which the temperature went back to normal, and the pigs iortiuhren with eating and showed no other symptoms of disease.

Lie pigs infected with viral diarrhea developed 28 days after vaccination against this virus, an average of a neutralizing antibody titer of 27, but not the trace of such a titer compared to the pig cholera virus. After vaccination with the pig cholera virus, the pigs that were previously shown Virus diarrhea virus had given 7 days later the presence of pig cholera antibodies, and 21 days later the titer averaged 515. After vaccination with porcine cholera virus the titer against Vrusdi @ rrhoevirus also increased from 27 to 729 and ran at the accelerated rate Parallel development of the titre against swine holeravirus. In contrast to forming dF in the control pigs until their death neither did the pig cholera nor any antibodies against the viral diarrhea virus.

Example V About 4 to 8 months old litters of purebred SpUrbund puppies, the experiences were sensitive to infectious bundle hepatitis divided into two groups each containing littermates. Group 1 was subcutaneously using Each 1 com live, adenovirus type 4 vaccinated, while group 2 for control without Vaccination was left. After this vaccination, none of the dogs showed inclusion the vaccinated as well as the non-vaccinated, but brought into contact with the vaccinated Dogs have any symptoms of sickness. After 5 weeks, all groups received one intravenous injection of 1 @@ m of a suspension of the Cornell 1 tribe of infectious hepatitis. After this Ispiang, all dogs showed elevated temperature, which lasted longer in control dogs not previously vaccinated with adenovirus. In addition, all of the groups showed viraemia one day after the hepatitis vaccination, namely, viruses in the blood, which were only used with infectious canine hepatitis Control dogs persisted for four to six days and it developed in the Most dogs have corneal opacity while the dogs face adenovirus and then infectious Canine hepatitis virus only showed viraemia on the first day after vaccination without the corneal opacity was observed. All bundles, which first with adenovirus and then vaccinated with canine infectious hepatitis virus, neutralizing ones appeared Antibodies against infectious canine hepatitis virus on the far day after the last Vaccination. The control dogs, on the other hand, which received infectious dog @hepatitis, neutralizing antibodies did not develop until 4 to 6 days after vaccination.

In the same manner as described above, the following animals thus protected against infectious pathogens to which they are susceptible that you are in them by special heterotypic pathogens that are foreign to them Resilience arises.

In all cases, the heterotypic pathogen develops complement-binding ones Antibodies against the immun @ logically unrelated, infectious pathogen.

Example VI A horse can by isspiung with living, foreign, heterotypical bovine phlebotracheitis protected against infectious equine @ hinopneumonitis been.

Example VII Duroh vaccination with the living, strange, heterotypical Mink enteritis virus can be used to protect a cat against infectious feline fever.

Example VIII A cat can be vaccinated with the living, her foreign, heterotypic virus Miyagawanella bovis against infectious Miyagawanella felis are protected.

Example IX By vaccination with live leptespira pomona one can protect a dog against infectious Leptospira canicola.

The @ is an example of the use of an alien heterotypic Bacterium with which the host is immunologically different against an infectious one Bacterium can be protected.

Example X Cows were injected with the commercial preparation of a live porcine cholera virus, that weakened, i.e. modified, by more than 150 passages through rabbits was vaccinated and proved danaoh when excited with virulent Viruadiarrhceviru @ as resistant to it.

Example XI Litters from purebred sniffer dogs were divided into two groups divided. Group 1 was cultured, live intravenously with 1 com suf tissue each Edmonston strain of measles virus, see Anders et al., Proc. Sec. Ezp. Biol. And Med. 86 (1954), 277, while group 2 was left as kentrolls. All with Those who were infected with the measles virus remained well and did not develop a rash.

24 to 28 days later, all dogs were intravenously @ each with 1 com vaccinated to a 100% Mil @@ uspension that contained the Snyder Hill strain of canine distemper virus contained.

The dogs originally vaccinated with measles virus did not show any Symptoms of illness. In contrast, the control dogs showed the typical Features of congestion infection including biphasic temperature and catarrhal Appearances on.

The investigations explained in the preceding examples show that the hetero-typical pathogen, which is foreign to the animal, acts like an antigen behaves by having resistance to or @econdary @@ responding to the promotes infectious, immunologically unrelated pathogens or, differently in other words, produces an effect similar to that produced by a single vaccination is triggered with homologous, inactivated pathogens. As stated above became, an animal prodes into a state of resilience antibodies neutralizing against a pathogen at an increased rate, as soon as it is exposed to these @ pathogens. So this becomes the time span between the secondary and the primary claim esp. which shortens I immunity. Regardless of the exact procedure, @I results in how the explanatory example @ prove, as a @success, that the disease is not at all the less severe.

Claims (11)

P a t e n t a n a p rü c h e 1. Procedure ru Auelection of a virus or other organism with the ability to provide protective resistance in an animal a pathogen that is immunologically unrelated to the virus or organism to which the animal is susceptible, marked by the nn the virus or other organism looks out until a positive complement binding antibody is obtained, and the virus so selected then ru such conditioning of Animal cells used that they produce antibodies faster and thereby the animal make it more resistant to protection against pathogens. 2. The method of claim 1 for making an animal resilient against an infectious virus to which it is susceptible, characterized in that: that you can meet the animal with a living, strange, heterotypical and with the infectious virus unrelated virus input, which is its ability distinguishes in the animal complement-binding antibodies against the infectious virus to create. 3. The method of claim 2 for making pigs resistant against pig cholera, characterized in that the pig with live Viral diarrheal virus vaccinates. 4. The method according to claim 3, characterized in that the virus diarrhea virus is a native, undeatenuated, live viral diarrheal virus. 5. Ancestors according to claim 3, daduroh gekennseichnet that the virus diarrhea @@ viru @ is weakened. 6. The method naoh claim 2 for making a federal government resistant against infectious canine hepatitis, characterized by having the band with living Adenovirus vaccinates. 7. The method of claim 2 for making a horse resistant against infectious equine rhinepneumonitis, characterized by being the horse vaccinated with live bovine rhinotracheitis. 8. The method according to claim 2 for making a cat resilient against infectious cat fever, marked by the fact that you bring the cat with you living Merz enteritis virus vaccinated. 9. The method of claim 2 for rendering a cat resilient against infektiö @ e Miyagawanell @ felis, characterized in that one is the Kaze vaccinated with live Miyagawanella bovis. 19. The method of claim 2 for making a dog resilient against infectious Leptespira cani @ ola, characterized in that the dog with living @ Leptespira pomona vaccinated. 11. The method of claim 2 for making a cow tough against infectious virus diarrhea @ evirus, characterized @ indicating that the cow is it live @ pig cholera virus vaccinates. 12. The method of claim 2 for making a covenant resilient against canine distemper virus, characterized in that one is hungry with live measles virus vaccinates.