NO170199B - PROCEDURE FOR RELEASABLE FIXING OF A VIEW ELEMENT IN A HOLDER IN A VIBRATION VIEWING APPARATUS, AND VIBRATION VIEWING APPLIANCE WHICH SUCH A VIEWING ELEMENT CAN BE INSTALLED IN - Google Patents
PROCEDURE FOR RELEASABLE FIXING OF A VIEW ELEMENT IN A HOLDER IN A VIBRATION VIEWING APPARATUS, AND VIBRATION VIEWING APPLIANCE WHICH SUCH A VIEWING ELEMENT CAN BE INSTALLED IN Download PDFInfo
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Description
Fremgangsmåte for fremstilling av en vaksine mot rubella. Method for making a vaccine against rubella.
Nærværende oppfinnelse vedrorer en fremgangsmåte for fremstilling av svekket levende vaksine mot rode hunder-virus, (rubella virus) med evne til å indusere aktiv immunitet mot denne sykdom. The present invention relates to a method for producing a weakened live vaccine against rubella virus, (rubella virus) with the ability to induce active immunity against this disease.
Uttrykket "svekket levende virus" anvendes i nærværende beskrivelse med henvisning til en rubella virus-stamme, hvis virulens er blitt svekket ved minst 21 passasjer i serie på primære kaninnyrevevkulturer. The term "attenuated live virus" is used in the present description with reference to a rubella virus strain, the virulence of which has been attenuated by at least 21 serial passages on primary rabbit kidney tissue cultures.
Ifolge fremgangsmåten som beskrives i det folgeride, oppnås en modifisert virus og derved en vaksine som har hby antigenitet og som ikke gir noen vesentlig ubnsket reaksjon når inokulert på barn. According to the method described in the following, a modified virus and thereby a vaccine is obtained which has high antigenicity and which does not give any significant unwanted reaction when inoculated into children.
Nærværet av uonskede reaksjoner, og blant dem mer spesielt den mulige disseminering av virus hos vaksinerte personer, er en av de store problemer ved utviklingen av en rubella virus av levende type. The presence of unwanted reactions, and among them more particularly the possible dissemination of virus in vaccinated persons, is one of the major problems in the development of a rubella virus of live type.
Det er kjent at ved siden, av komplikasjoner, slik som hjerne-betennelse og trombosytopen purpura, som er alvorlig men ganske sjelden, og også artritis som ikke er uvanlig men vanligvis selvbegrensende og av kort varighet, er de mest alvorlige komplikasjoner ved denne sykdom av teratogen natur. Rubella er meget alvorlig tilstand for kvinner i den forste tredjedel av graviditet, på grunn av sin virkning på kogenitale misdannelser hos fosteret, dodfbdsel eller abort. It is known that besides complications such as encephalitis and thrombocytopenic purpura, which are serious but quite rare, and also arthritis which is not uncommon but usually self-limiting and of short duration, the most serious complications of this disease are of teratogenic nature. Rubella is a very serious condition for women in the first third of pregnancy, due to its effect on congenital malformations of the fetus, stillbirth or abortion.
I denne henseende er det vesentlig at et barn som mottar en rubella vaksine av levende type ikke danner en potensiell fare for en gravid kvinne, med hvilken det kan komme i kontakt. In this regard, it is essential that a child receiving a live rubella vaccine does not pose a potential danger to a pregnant woman with whom it may come into contact.
Inntil nu var de eneste mulige midler for å forebygge slike rubella foster-komplikasjoner enten å administrere gamma globulin i store doser til utsatte svangre kvinner eller ut-sette unge piker for rubella for de når kjonnsmoden alder. Until now, the only possible means of preventing such rubella fetal complications were either to administer gamma globulin in large doses to susceptible pregnant women or to expose young girls to rubella before they reach puberty.
Administrasjonen av gamma globulin betraktes langt fra av hver spesialist som en effektiv behandling-, og av selvfølgelige grunner er det å utsettes for sykdommen i ung alder ingen aksepterbar losning av problemet. The administration of gamma globulin is far from being regarded by every specialist as an effective treatment, and for obvious reasons, being exposed to the disease at a young age is not an acceptable solution to the problem.
Det er allerede tidligere kjent (James R. Prier, Basic Medical Virology, U.S.A., side 561, 1966 og Parkman et al. J. Immunol. 93, side 595-607 og 608-617, 1964) å dyrke rubella virus i forskjellige vevkulturer og blant dem primære kaninnyrevevkulturer. It is already previously known (James R. Prier, Basic Medical Virology, U.S.A., page 561, 1966 and Parkman et al. J. Immunol. 93, pages 595-607 and 608-617, 1964) to grow rubella virus in various tissue cultures and among them primary rabbit kidney tissue cultures.
Det er nå overraskende blitt funnet at ved valg av en spesiell vevkultur - som primær er kaninnyrevevkultur - i forbindelse med definerte behandlingsbetingelser, og mer spesielt definert antall av passeringer, er det mulig å fremstille en rubella-vaksine hvis administrasjon overraskende ikke oppviser noen vesentlig ubnsket reaksjon og ikke oppviser de bivirkninger som opptrer ved administrasjon av rubella-vaksiner fremstilt i andre vevkultursystemer. It has now surprisingly been found that by choosing a special tissue culture - which is primarily rabbit kidney tissue culture - in connection with defined treatment conditions, and more specifically defined number of passages, it is possible to produce a rubella vaccine whose administration surprisingly does not show any significant undesirable reaction and do not exhibit the side effects that occur when administering rubella vaccines produced in other tissue culture systems.
Ved uttrykket "ikke noen vesentlig uonskede reaksjoner" som angitt foran skal det fastslås ikke bare at inokulering av vaksine ifolge nærværende oppfinnelse stimulerer immunitet uten å indusere de vanlige patologiske symptomer for vanlig rubella, men også at ved kliniske forsbk som er utfort med en vaksine ifolge oppfinnelsen er ingen serologiske eller virolo-giske tegn på infeksjon blitt fastslått ved utsatt intim kontakt. By the expression "no significant adverse reactions" as stated above, it must be established not only that inoculation of a vaccine according to the present invention stimulates immunity without inducing the usual pathological symptoms for common rubella, but also that in clinical trials carried out with a vaccine according to invention, no serological or virological signs of infection have been established by exposed intimate contact.
Nærværende oppfinnelse vedrorer således en fremgangsmåte The present invention thus relates to a method
til fremstilling av en vaksine mot rubella som inneholder en svekket, levende rubella virus som aktiv bestanddel, hvilken svekkede, levende rubella virus er oppnådd ved dyrkning av en rubella virusstamme på en primær kaninnyrevevkultur og fremgangsmåten karakteriseres ved at rubella virusstammen svekkes ved i serie å passere minst 21 ganger gjennom primære kaninnyrevevkulturer ved en temperatur mellom 28 og 36°C, idet varigheten for hver passering ikke overstiger 5 dager og at den oppsamlede, svekkede, levende rubella virus tilsettes som aktiv komponent i en vaksine. for the production of a vaccine against rubella containing a weakened, live rubella virus as active ingredient, which weakened, live rubella virus has been obtained by growing a rubella virus strain on a primary rabbit kidney tissue culture and the method is characterized in that the rubella virus strain is weakened by serially passing at least 21 times through primary rabbit kidney tissue cultures at a temperature between 28 and 36°C, the duration of each passage not exceeding 5 days and the collected, attenuated, live rubella virus being added as an active component of a vaccine.
En vaksine fremstilles derfor fra en egnet passasje i primære kaninnyre-celler under bruk av enhver teknikk som er kjent på området for slik fremstilling. A vaccine is therefore prepared from a suitable passage in primary rabbit kidney cells using any technique known in the art for such preparation.
Rubella virus som anvendes for å utfore denne oppfinnelsen isoleres fra et typisk klinisk tilfelle under anvendelse av f.eks. halspenslinger eller urin- eller gurgleprover. Provene fryses enten umiddelbart og holdes ved -60°C inntil anvendelse eller inokuleres umiddelbart i et egnet vevkultursystem, f.eks. primær afrikansk gronn apenyre (GMK) eller annet vevkultursystem som er kjent på området for slik isolering. Nærværet av rubella virus kontrolleres ved å pode kulturer f.eks. med en entero-virus slik som Echovirus 11 eller Coxsackievirus A 9 på den 9. eller 10. dagen etter inokulering. Rubella virus used to carry out this invention is isolated from a typical clinical case using e.g. throat swabs or urine or gargle samples. The samples are either immediately frozen and kept at -60°C until use or immediately inoculated into a suitable tissue culture system, e.g. primary African green monkey kidney (GMK) or other tissue culture system known in the art for such isolation. The presence of rubella virus is checked by inoculating cultures e.g. with an enterovirus such as Echovirus 11 or Coxsackievirus A 9 on the 9th or 10th day after inoculation.
Ved å bruke spesielt antiserum fremstilt i kaniner mot en kjent RV-stamme er det mulig å identifisere rubella virus ved spesifikk noytralisasjonsprove i mottagelige celler slik som GMK-celler. Fraværet av tilfeldig forekommende apelignende virus kontrolleres i GMK-celler etter nbytralisasjon ved spesifikt antiserum, fremstilt i et annet cellesystem. By using special antiserum produced in rabbits against a known RV strain, it is possible to identify rubella virus by specific neutralization test in susceptible cells such as GMK cells. The absence of incidental monkey-like virus is checked in GMK cells after neutralization by specific antiserum, prepared in another cell system.
De primære kaninnyrecellekulturer for seriepassasjer fremstilles fortrinnsvis fra nyrer av dyr ikke eldre enn 3 uker. Foretrukket vekstmedium for primære kaninnyre-kulturer er Hanks' balanserte saltmedium, supplert med inaktivert kalveserum, lactalbumin hydrolysat og tryptose fosfat-suppe, men andre media som er kjent for fagmannen på området kan også anvendes. Inkubasjonstemperaturen er ikke over 36°C. Serie-passasjene utfores ved å inokulere kaninnyre-lag med alikvoter av den ovenstående væske fra den tidligere passasje og fortrinnsvis samle inn virusen mellom den 3. og 5. dag etter inokulering. Titrering av den innsamlede virus kan utfores ved anvendelse av interferensmetoden i GMK-kulturrbr under anvendelse av f.eks. Echovirus 11 eller Coxsackievirus A 9 som angitt foran for isoleringstrinnet. The primary rabbit kidney cell cultures for serial passages are preferably prepared from kidneys of animals no older than 3 weeks. The preferred growth medium for primary rabbit kidney cultures is Hanks' balanced salt medium, supplemented with inactivated calf serum, lactalbumin hydrolyzate and tryptose phosphate soup, but other media known to those skilled in the art can also be used. The incubation temperature is not above 36°C. The serial passages are performed by inoculating rabbit kidney layers with aliquots of the supernatant from the previous passage and preferably collecting the virus between the 3rd and 5th day after inoculation. Titration of the collected virus can be carried out using the interference method in GMK culture rbr using e.g. Echovirus 11 or Coxsackievirus A 9 as indicated above for the isolation step.
Ved et visst passasjenivå, ikke for den 21. og fortrinnsvis mellom den 21. og 60. passasje, kastes den ovenstående væske av de inokulerte kulturer på den 5. dag etter inokuleringen og lagene vaskes med en pufret saltopplosning, f.eks. Eagle's opplbsning eller Hanks' opplbsning, og inkuberes videre med et opprettholdelsesmedium, f.eks. Hank<1> medium supplert med casein hydrolysat. Etter en ytterligere inkubasjon på 3 dager, samles den ovenstående væske og klares ved filtrering eller sentrifugering. At a certain passage level, not before the 21st and preferably between the 21st and 60th passage, the supernatant of the inoculated cultures is discarded on the 5th day after inoculation and the layers are washed with a buffered saline solution, e.g. Eagle's solution or Hanks' solution, and further incubated with a maintenance medium, e.g. Hank<1> medium supplemented with casein hydrolyzate. After a further incubation of 3 days, the supernatant is collected and clarified by filtration or centrifugation.
Alternativt utfores innsamling av virus etter frysing, opp-tbing og rystning av kulturen i opprettholdelsesmediumet og Alternatively, virus collection is carried out after freezing, resuspending and shaking the culture in the maintenance medium and
etterfølgende filtrering eller sentrifugering. subsequent filtration or centrifugation.
Det innsamlede RV, supplert eller ikke med et stabiliserende tilsetningsstoff, kan lagres enten i frossen tilstand, f.eks. ved ca. -60°C eller fryses i torr tilstand. Den oppnådde vaksine administreres subkutant eller intramuskulært hvis nodvendig etter gjenfremstilling. The collected RV, supplemented or not with a stabilizing additive, can be stored either in a frozen state, e.g. at approx. -60°C or freeze in a dry state. The obtained vaccine is administered subcutaneously or intramuscularly if necessary after reconstitution.
De fblgende eksempler illustrerer oppfinnelsen The following examples illustrate the invention
EKSEMPEL 1 EXAMPLE 1
En 3 uker gammel kanin fra en oppdrettingskoloni undersbkes på fraværet av patologiske skader etc. Begge nyrer blir aseptisk fjernet og kuttet i små stykker som bringes i kontakt med en pufret saltopplbsning av trypsin (2,5 g/l) og blandingen rores kontinuerlig om i 10 minutter ved en temperatur på 37°C. Væsken helles deretter av og erstattes av samme volum frisk trypsinopplosning. Trypsinbehandlingen fortsettes så under omroring inntil forbruk av vevet, og celler suspendert i væsken fjernes fra tid til annen og sentrifugeres så. A 3-week-old rabbit from a breeding colony is examined for the absence of pathological lesions etc. Both kidneys are aseptically removed and cut into small pieces which are brought into contact with a buffered saline solution of trypsin (2.5 g/l) and the mixture is stirred continuously in 10 minutes at a temperature of 37°C. The liquid is then poured off and replaced by the same volume of fresh trypsin solution. The trypsin treatment is then continued with agitation until the tissue is consumed, and cells suspended in the liquid are removed from time to time and then centrifuged.
Cellesedimentet som er oppnådd, resuspenderes i Hanks' opplbsning og sentrifugeres igjen. Dette siste trinnet gjentas to ganger og det endelige cellesedimentet suspenderes i vekstmedium som består av Hanks' balanserte saltmedium med 10% inaktivert kalveserum, 0,5% laktalbumin hydrolysat og 0,1% tryptose fosfat suppe for å sikre en konsentrasjon på ca. 10<5 >celler pr. ml. The cell sediment obtained is resuspended in Hanks' solution and centrifuged again. This last step is repeated twice and the final cell sediment is suspended in growth medium consisting of Hanks' balanced salt medium with 10% inactivated calf serum, 0.5% lactalbumin hydrolyzate and 0.1% tryptose phosphate broth to ensure a concentration of approx. 10<5 >cells per ml.
Alikvote deler (1 ml) av den oppnådde cellesuspensjonen som inneholder hver ca. 10 5 celler helles i 10 kulturror (12 mm diameter) som inkuberes i en lett skrå stilling ved 37°C i 4 dager. Etter denne inkubasjonsperioden er et fullstendig monolag utviklet. Næringsmediumet erstattes med et lignende volum friskt, like for virus-inokuleringen. Aliquots (1 ml) of the obtained cell suspension containing each approx. 10 5 cells are poured into 10 culture tubes (12 mm diameter) which are incubated in a slightly inclined position at 37°C for 4 days. After this incubation period, a complete monolayer has developed. The nutrient medium is replaced with a similar volume of fresh, equal to the virus inoculation.
Det samme næringsmidiumet anvendes for og etter virus-inokulering og hver primær kaninnyre-cellekultur angitt i dette eksempel fremstilles ifolge den her foran-beskrevne teknikk. The same nutrient medium is used before and after virus inoculation and each primary rabbit kidney cell culture indicated in this example is prepared according to the technique described above.
Alikvote deler {0,1 ml) av en rubella virus-stamme, isolert i primære GMK-kulturer og fort 3 ganger gjennom dette system blir, for videre karakterisering/ inokulert i primære kaninnyre-kulturror som inkuberes ved 34°C i en skrå stilling. Aliquots {0.1 ml) of a rubella virus strain, isolated in primary GMK cultures and rapidly 3 times through this system are, for further characterization/ inoculated into primary rabbit kidney culture tubes incubated at 34°C in a slant position .
På den 5. dagen etter inokulering innsamles den ovenstående væske og den innsamlede virus identifiseres og titreres ved å anvende interferensmetoden beskrevet foran. On the 5th day after inoculation, the supernatant is collected and the collected virus is identified and titrated using the interference method described above.
Titeret i GMK-celler etter denne forste passasjen i kaninnyre-2 83 The titer in GMK cells after this first passage in rabbit kidney-2 83
celler er 10 ' InD5oPr* m^* cells are 10 ' InD5oPr* m^*
Alikvote deler av den samlede ovenstående væske fra den forste passasjen inokuleres i andre primære kaninnyre-kulturror som inkuberes ved 34°C. Aliquots of the pooled supernatant from the first passage are inoculated into second primary rabbit kidney culture tubes incubated at 34°C.
På den 5. dagen etter inokuleringen samles den ovenstående væske. Virusen titreres som angitt ved slutten av den forste passasjen. Denne prosessen gjentas opp til den 21. passasjen, og inkubasjonsperioden av hver individuell passasje varierer mellom 3 og 5 dager. On the 5th day after inoculation, the supernatant is collected. The virus is titrated as indicated at the end of the first passage. This process is repeated up to the 21st passage, and the incubation period of each individual passage varies between 3 and 5 days.
Fra den 9. passasje av iakttas en cytopatisk effekt i primære kaninnyre-celler. From the 9th passage a cytopathic effect is observed in primary rabbit kidney cells.
For den 21. passasje i kaninnyre-celler fremstilles primære kaninnyre-cellekulturer i 500 cm 3 Roux flasker, under anvendelse av teknikken beskrevet foran. For the 21st passage in rabbit kidney cells, primary rabbit kidney cell cultures are prepared in 500 cm 3 Roux flasks, using the technique described above.
Etter en inkubasjonsperiode på 3 dager ved 36°C oppnås et fullstendig monolag. Den ovenstående væske kastes derpå og erstattes av samme volum av samme vekstmedium og inokuleres med alikvote deler av virusmateriale fra den 20. passasje. Etter videre inkubasjon i 5 dager Ved 34°c, kastes den ovenstående væske og erstattes av et like volum av opprettholdelsesmedium som består av Hanks' opplbsning supplert med 0,3% After an incubation period of 3 days at 36°C, a complete monolayer is obtained. The supernatant is then discarded and replaced by the same volume of the same growth medium and inoculated with aliquots of virus material from the 20th passage. After further incubation for 5 days at 34°C, the supernatant is discarded and replaced by an equal volume of maintenance medium consisting of Hanks' solution supplemented with 0.3%
casein hydrolysat. Etter videre inkubasjon i 3 dager ved 34°C, samles den ovenstående væske og klares ved sentrifugering. casein hydrolyzate. After further incubation for 3 days at 34°C, the supernatant is collected and clarified by centrifugation.
Prover er blitt tatt for titrering, identifisering og sikker-hetsprbving og virusen lagres ved -60°C. Samples have been taken for titration, identification and safety testing and the virus is stored at -60°C.
Virustiteret er lO^InD^ pr. ml. provet i primære GMK-celler ved interferenssystemet. The virus titer is lO^InD^ per ml. tested in primary GMK cells by the interference system.
Dette virusmaterialet blir så utsatt for sikkerhetsprbving som inkluderer bakteriesterilitet og fravær av fremmedstoffer ved inokulering i kaniner, hamstere, marsvin, mus og aper. This viral material is then subjected to safety testing that includes bacterial sterility and the absence of foreign substances when inoculated into rabbits, hamsters, guinea pigs, mice and monkeys.
Ytterligere sikkerhetsprbving utfores i forskjellige vevkultursystemer. Further safety testing is carried out in different tissue culture systems.
Preliminær potensprbving utfores ved å inokulere kaniner og aper med 10 3 * 5 inD^. Serumsnbytralisasjonsprbver utfores i GMK-celler. Preliminary potency testing is carried out by inoculating rabbits and monkeys with 10 3 * 5 inD^. Serum catalysis tests are performed in GMK cells.
Resultater angis i de fblgende tabeller I og II. Results are shown in the following tables I and II.
Viruspreparatene fordeles i glassampuller av hvilke hver inneholder 1 ml væske. The virus preparations are distributed in glass ampoules, each of which contains 1 ml of liquid.
Ampullene fryses, torres og forsegles. The ampoules are frozen, dried and sealed.
Etter gjenfremstilling ved å tilsette 1 ml destillert vann, inokuleres vaksinen subkutant til mottagelige individer og de individuelle doser er ca. 125 InDc^. After reconstitution by adding 1 ml of distilled water, the vaccine is inoculated subcutaneously into susceptible individuals and the individual doses are approx. 125 InDc^.
oo OE OE
Resultater av serologisk responsprbving utfort på en gruppe av 25 seronegative barn (15 mottar vaksine og 10 kontroller lever i nær kontakt) gjengis i tabell III. Results of serological response testing carried out on a group of 25 seronegative children (15 receiving vaccine and 10 controls living in close contact) are reproduced in table III.
Fra de 15 vaksinerte barn, hadde 13 stigning i motstoff på opp til 1/32 eller mer (2 prover - angitt ND - var ikke til-. gjengelige på prøvetidspunktet). Ingen viste kliniske tegn på infeksjon. Alle 10 kontakter forble serologisk negative. From the 15 vaccinated children, 13 had a rise in antibody of up to 1/32 or more (2 samples - indicated ND - were not available at the time of the test). None showed clinical signs of infection. All 10 contacts remained serologically negative.
EKSEMPEL 2 EXAMPLE 2
Arbeidsmåten er den som er» beskrevet i eksempel 1, men passasjene fortsettes opp til den 30. passasje i primære kaninnyre-cellekulturer. The procedure is as described in Example 1, but the passages are continued up to the 30th passage in primary rabbit kidney cell cultures.
En vaksine fremstilles fra disse og sikkerhetsprovning utfores som beskrevet i eksempel 1. A vaccine is produced from these and safety testing is carried out as described in example 1.
Potensen for prepareringen undersokes på dyr (aper). The potency of the preparation is investigated on animals (monkeys).
Resultater angis i tabell IV Results are given in Table IV
EKSEMPEL 3 EXAMPLE 3
Ved å gå ut fra virusmateriale fra den 21. passasje ved 34°c som oppnådd i eksempel 1, utfores 9 ytterligere seriepassasjer i primære kaninnyre-cellekulturer, men ved 28°C inklusive det siste trinn for vaksinepreparering. Starting from viral material from the 21st passage at 34°C as obtained in Example 1, 9 further serial passages are performed in primary rabbit kidney cell cultures, but at 28°C including the final step of vaccine preparation.
Den oppnådde vaksine sikkerhetsproves og inokuleres til kaniner og aper for potensprovning. The obtained vaccine is tested for safety and inoculated into rabbits and monkeys for potency testing.
Resultater angis i de fSigende tabeller V og VI. Results are given in the following tables V and VI.
EKSEMPEL 4 EXAMPLE 4
Arbeidsmåten er den som er beskrevet i eksempel 1, men passasjene fortsettes opp til den 51. passasje i primære kaninnyre-cellekulturer og det endelige opprettholdelsesmedium består av Hanks<1> opplbsning supplert med 0,3% casein hydrolysat og inneholder 50 meg kloramfenikol pr. ml. Hanks' opplbsning. The procedure is that described in example 1, but the passages are continued up to the 51st passage in primary rabbit kidney cell cultures and the final maintenance medium consists of Hank's<1> solution supplemented with 0.3% casein hydrolyzate and contains 50 meg chloramphenicol per ml. Hanks' revelation.
Etter inkubering på 9 dager ved 34°c, samles den ovenstående væske, klares ved sentrifugering og blandes med et like volum stabiliserende opplbsning bestående av 30 g kalium glutamat, 200 g sukrose og 50 mg kloramfenikol pr. liter destillert vann, og blandingen fordeles i glassampuller som inneholder 1 ml. vann. After incubation for 9 days at 34°c, the supernatant liquid is collected, clarified by centrifugation and mixed with an equal volume of stabilizing solution consisting of 30 g of potassium glutamate, 200 g of sucrose and 50 mg of chloramphenicol per liter of distilled water, and the mixture is distributed in glass ampoules containing 1 ml. water.
Ampullene blir så fryse-tbrret og forseglet. The ampoules are then freeze-dried and sealed.
Etter gjenfremstilling ved å tilsette 1 ml destillert vann inokuleres vaksinen subkutant til 65 seronegative individer, After reconstitution by adding 1 ml of distilled water, the vaccine is inoculated subcutaneously into 65 seronegative individuals,
og titreringen av de individuelle doser når ca. 10 2 " 6InD,-n i GMK-celler eller 10 3 " 7 PFU (plateformede enheter) i kaninnyre 13-celler. En gruppe på 30 seronegative barn ble holdt i intim kontakt med de vaksinerte i en periode på 6 uker. and the titration of the individual doses reaches approx. 10 2 " 6 InD,-n in GMK cells or 10 3 " 7 PFU (plate-shaped units) in rabbit kidney 13 cells. A group of 30 seronegative children were kept in intimate contact with the vaccinated for a period of 6 weeks.
En serologisk responsundersbkelse utfort på gruppen på 65 vaksinerte individer viste at alle uten 1 responderte på vaksinen med et gjennomsnittlig antistoff-titer på 1/128 som provet i HAI (Hemagglutinhemning) forsbk. Ingen viste kliniske tegn på infeksjon. Alle 30 kontakter forble serologiskt negative. A serological response study carried out on the group of 65 vaccinated individuals showed that all but 1 responded to the vaccine with an average antibody titer of 1/128 as tested in the HAI (Hemagglutininhibition) trial. None showed clinical signs of infection. All 30 contacts remained serologically negative.
EKSEMPEL 5 EXAMPLE 5
Arbeidsmåten er den som er beskrevet i eksempel 1, men passasjene fortsettes opp til den 61. passasje i primære kaninnyre-cellekulturer, og det endelige opprettholdelsesmedium består av Hanks<1> opplbsning supplert med 0,3% casein 1 hydrolysat og inneholder 50 meg kloramf enikol pr. ml Hanks' opplbsning.. The procedure is that described in Example 1, but the passages are continued up to the 61st passage in primary rabbit kidney cell cultures, and the final maintenance medium consists of Hank's<1> solution supplemented with 0.3% casein 1 hydrolyzate and contains 50 meg of chloramphenicol enikol per ml Hanks' revelation..
Etter inkubering på 9 dager ved 34°C, samles den ovenstående væske, klares ved sentrifugering og blandes med et like volum av stabiliserende opplbsning bestående av 30 g kalsium-gliitamat, 200 g sukrose og 40 mg kloramf enikol pr. liter destillert vann, og blandingen fordeles i glassampuller som inneholder 1 ml. vann. After incubation for 9 days at 34°C, the supernatant is collected, clarified by centrifugation and mixed with an equal volume of stabilizing solution consisting of 30 g calcium glitamate, 200 g sucrose and 40 mg chloramphenicol per liter of distilled water, and the mixture is distributed in glass ampoules containing 1 ml. water.
Ampullene fryse-tbrres så og forsegles. The ampoules are then freeze-dried and sealed.
Etter gjenfremstilling ved å tilsette 1 ml. destillert vann, inokuleres vaksinen subkutant til seronegative individer, og titreringen av de in3 d9ividuelle doser når ca. 10 2 " 83InD^ i GMK-celler eller 10 * PFU i kaninnyre-13 celler. After reconstitution by adding 1 ml. distilled water, the vaccine is inoculated subcutaneously into seronegative individuals, and the titration of the individual doses reaches approx. 10 2 " 83InD^ in GMK cells or 10 * PFU in rabbit kidney-13 cells.
En serologisk responsprbve utfort i en gruppe på 7 seronegative individer viste at alle responderte på vaksinen med et gjennomsnittlig titer på 1/64, som provet i HAI-forsbk. A serological response test carried out in a group of 7 seronegative individuals showed that all responded to the vaccine with an average titer of 1/64, as tested in the HAI trial.
Ingen viste kliniske tegn på infeksjon. None showed clinical signs of infection.
Claims (2)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB858514982A GB8514982D0 (en) | 1985-06-13 | 1985-06-13 | Screen clamping |
| GB858514983A GB8514983D0 (en) | 1985-06-13 | 1985-06-13 | Screen clamping |
Publications (4)
| Publication Number | Publication Date |
|---|---|
| NO862214D0 NO862214D0 (en) | 1986-06-04 |
| NO862214L NO862214L (en) | 1986-12-15 |
| NO170199B true NO170199B (en) | 1992-06-15 |
| NO170199C NO170199C (en) | 1992-09-23 |
Family
ID=26289367
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| NO862214A NO170199C (en) | 1985-06-13 | 1986-06-04 | PROCEDURE FOR RELEASABLE FIXING OF A VIEW ELEMENT IN A HOLDER IN A VIBRATION VIEWING APPARATUS, AND VIBRATION VIEWING APPLIANCE WHICH SUCH A VIEWING ELEMENT CAN BE INSTALLED IN |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0218315A3 (en) |
| AU (1) | AU583127B2 (en) |
| CA (1) | CA1304718C (en) |
| ES (1) | ES8706485A1 (en) |
| NO (1) | NO170199C (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2631255B1 (en) * | 1988-05-16 | 1990-08-24 | Szilvasi Peter | REMOVABLE FRAMES OF CASSETTE OR CARTRIDGE-TYPE CRIBLES |
| US5690826A (en) * | 1996-05-10 | 1997-11-25 | Cravello; William Myron | Shaker screen assembly |
| GB0301509D0 (en) | 2002-10-17 | 2003-02-19 | Varco Int | Vibratory seperator and screen assembly |
| GB2408006B (en) * | 2003-11-13 | 2007-04-25 | Russel Finex | Improvements in screen separators |
| US8245850B2 (en) | 2003-11-13 | 2012-08-21 | Russell Finex Limited | Screen separators |
| PL2209566T3 (en) * | 2007-10-05 | 2013-06-28 | Mi Llc | Screen clamp |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2279042A (en) * | 1940-08-03 | 1942-04-07 | Inland Lime & Stone Company | Screening apparatus |
| FR1282627A (en) * | 1960-12-13 | 1962-01-27 | Socam Sa | Device for sieve clamping |
| FR1326260A (en) * | 1962-06-26 | 1963-05-03 | Device for stretching sieving and filtering fabrics | |
| AT329474B (en) * | 1974-02-25 | 1976-05-10 | Oesterr Amerikan Magnesit | CLAMPING DEVICE FOR SCREEN BOTTOM |
| US4082657A (en) * | 1976-01-19 | 1978-04-04 | Gage Ernest L | Separator apparatus |
| GB2085744B (en) * | 1980-10-20 | 1984-06-13 | Thule United Ltd | Vibratory screening apparatus |
| EP0130744A3 (en) * | 1983-07-01 | 1986-03-05 | Sweco, Inc. | Separator screen and method of making same |
-
1986
- 1986-06-04 NO NO862214A patent/NO170199C/en unknown
- 1986-06-10 ES ES555914A patent/ES8706485A1/en not_active Expired
- 1986-06-10 EP EP86304423A patent/EP0218315A3/en not_active Ceased
- 1986-06-11 AU AU58567/86A patent/AU583127B2/en not_active Expired
- 1986-06-12 CA CA000511418A patent/CA1304718C/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| NO170199C (en) | 1992-09-23 |
| EP0218315A3 (en) | 1988-05-11 |
| AU5856786A (en) | 1986-12-18 |
| EP0218315A2 (en) | 1987-04-15 |
| NO862214D0 (en) | 1986-06-04 |
| AU583127B2 (en) | 1989-04-20 |
| ES555914A0 (en) | 1987-07-01 |
| NO862214L (en) | 1986-12-15 |
| CA1304718C (en) | 1992-07-07 |
| ES8706485A1 (en) | 1987-07-01 |
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