AU2004203226B2 - Vaccine - Google Patents

Description

P/00/0 11 28/5/91 Regulation 3.2 AUSTRALIA Patents Act 1990 ORIGINAL COMPLETE SPECIFICATION STANDARD PATENT Name of Applicant: UCL BioMedica Plc Actual Inventors Oscar Adelmo Bottasso, Graham McIntyre & Cynthia Ann Stanford Address for service is: WRAY & ASSOCIATES Level 4, The Quadrant 1 William Street Perth, WA 6000 Attorney code: WR Invention Title: "Vaccine" The following statement is a full description of this invention, including the best method of performing it known to me: 1 FIELD 01FlMENTION Thle present invention relates to an immune m duazr orpoj ad/) s t wh a a ir jg c o p s d n W i h i e c t j i 1 1 8 MI r e e n t a n d / o r P r e v e n t i o n o f np o t h r W e ni g itisystem ic, w ast ig sn d rom e and /cr Porciw der jy j n 10 -BACKGRO~qiND THE MINVENTON POIA-w1eanng mutssei wasting syzidrom (P"vLW) affects piglets post-. weaning from 4 to 16 weeks of age (1:5_50 kg). TypicaHY -PMWS affects piglets ote to two weeks after weanjig and is very different from the was g/poor weaner who fa1s to eat or drink adequately after weai9 'PMWS Piglets are weaners which have 15 stare to Vrow and then collapse quickya dotnh v nete ayp o ep net antibiotics.cl n fe aeA et 5 eyPo epnet POrcirm dermatitis and Deplaropathy syndrome (MIDS) affects pigs from S to 18 weeks of age and the mnost Obvious signs are red-putple blotches on the skin, whi~h 20 become brown and crusted after a few days. Pigs are lethargic and may have swollen legs 'resultizag from their nepbroPathy. This sydrome, also, respon& poorly to The causal agents of both PMWs and ?DNS are at Presey4 unknown. The 25 most likely suspect in both Syndromnes is a pig circovirus .,type ]r t which is antigenically distinct from widely distributed nonnal zon-pathogenic pig Circovi.us "type -r' Circovirus J (l'CV Mi? has been idenifie on UK farmj scrologi ly. PDNS, which is thought to be an ir=MCMun zPle4 mediated disease. m~ay also involve bacteria iz its aemiology, though the paM that they play is not clear. 30 2 PMWS is a problem which iS YecOga ld reported in Canada in 193 ih i eonsdworld-wide- jPMWS was frs (e1o996);j~ Spaain 199, with reported incidents following in France (1995); USA ( 1 9 9 6 ) -; S p a i n ( 1 9 9 7 ) ; A u tId a , D e n ma r j G e r m~ nrl a i t l: je b e ~ ~ n Northern Ireland (1998); Great Britain, Hunay Japan Korea, PorNtgal and Taiwan 5 (1999); Poland (2000); Mexico (2001); and New Zealand (2003). PDNS was .first described in the UK in 1993 as a Spordjc coditon but since that time it has become increasinly associated with heads affected by PMWs, and has been reported all over the world. 10 c With PMWs and PDNS the mortality rate is very high- The clinical 42C).Pis ooth PMWS d c ude an initial high fever (40 42hC) Pigs with PWS develop listlessness; very rapid wasting; development of a haiy coat and rented appearance. In addition, the lymph nodes may be palpable as 15 they are grossly enlarged. With either syndrome eyphgs m ay evplp as cough and ditculty breathing. , pigs may also develop a slight To date, no effective treatmes of or vaccines for pMWS or ?DNS are available. As a general guide the following vcimes have bef used in order to attempt 20 to combat both syndromes: changing of the herd to all-in or all-out protocols; age sectionn; providing eas with a good environment to limit effects of secondary isiectioi; sepeing welter that show signs of the disease to reduce the level of the disease in the pen; good at intoductio protocols to ensure gilts are well acclimatised before 5trY iuto the main unit; and innsing rosS.fotering, 25 These regimes, however, are lirnited in their effectiveness An aim of the present invention is to more effectively protect against anor f eat and/or reduce the level of occurrence of both PMWS and/or PDNS. 30 3 SUM N4pAJy Op THE Z4VENTIQN A semninal finding of the present inventon is that Posqwaji £fl~tisysteri wasting sYndrome (1'MWS) and/or Porcnze derraailis and 1aeplwopay S ndr .~ 5 (PDNs) can be threat arId/or prevented anid/., redcdb d~s~~ ath whdole, CCll of a bacteria from the genra= Rooce Gdo ad4,camdn of awhol ?suikamuwe~ff and Noca'rdloides or an irnmme Modu~ator 1.mp ) Died/or pha zna eutcaiCo Ipo'i jOn co mprsng a w hole cell of a bacteium froln the genera 10Rhodococcus Gw-donia, ocadi4z Dietzia lTskare,,pa andjNocadIoi&, DIETAILED ASPEcTrS OF THE ThVNrIX In one aspeCt, the present invention provides the use of an immune modulator compositioni or a Pharaceulcal omposition comprising a w hoecl fab ce i 15 from r the gen er R hodococcus, r do i 'ho~jle ellf Tsaure iu a Noc-ardiot*S in the rnanuactr of a mdcament fo t' eTsuent revea ndo post-weaning Mufrisysez 1 nj wastizg -s zdrozn (PM WS Mor t e trat entOrPrvetion af DePbrOPathY Syndrorne (PO)Nr).WSan/rPciedmttsad 20 InL another aspect, the Present inventing Provides the use of a whole cell of a bacterjun fromn the genera Rhwdococcw, Gordouja, Nocard1a Dfazia arnd Nocardioides, in the manufacture of a medcmen fo the renio of Post-weaning xfl~tisystmc %vasrg syndrome (MWS) ad/or Porcne dermatit s 25 and nephrOPah SYndrome (PDNS). 'a a further aspect, the presenT invention Provides a Method for treating or p r e v e iti n d P O tne h o ani m s y r o m ic w s i g rde (P M WV S ) a n d /o r P o rc in e d e n a~ i a n fl p l~ o p a ~ y y n d o~~ (P D N S ) in a s u b je c t C o m p r i i g a d m i n st. 30 eompositio igu~ o ha m c uical composition or irnnnme modulator 30 comoito cOnlprising a whole cell of a bacterjuf frorn toe gene a, woc Gorom, ocardia, Diezzi TsukajjPurella and Noapioo t asuj c oct Suitably, the effectv mi ftepaa~~cmoiinado the efecve mouja~rcn~~t j mae aditge as a ingle dose. Mltenatively, the effcti ve m ay bex3 of the Pha rM ce txtjl c o p j a d o mo u a r 5o osthore ea b a d m id n i s t e r d i zn m u l t i p& ( e P e a t ) d o s e s , f o r e a ~ l w r i o e 5 treeor more, four Or more, five or more, six or more, te xPmoe twot or inore r'epeat doses.toormr'weYOroe In another asPect, the present invention provj ehdfr rtcig including irnimising a subject from PatWCPn Yul t ho~ Worptetingzm 1 0 ( P M W S ) a d / o r- P o r c i n e d e n n t i t is a n d n e p ir o p a t h y M dn c r oa e ( P D s Cl~ l s admiz~istling a Pl~rMaceutical Composiio or kw u modN cla o mC~riosing Comprising a whole cell of a bacterium fro the gener moduocot osidon Nocrdi Dej'j~Tsuka,,yella and Nocardioldes to the subject. is Preferably. the Pharmceuti c ofjno rnn mdltrC'pst~ ziadinstere in early life, such a's within the first .3 raonthfb rtPerbl v"tiin the first 3 weeks of birth.s f ikpr.al Th 7e tem "Protected" as ued hawei neazthat te subject is less susceptible to 20 the disease/d sore S Compared Ait% a subject not t~ ed r m in wt th c o m p 0 ~ , ~ 1 ~ o t h p r e e n t n v e n i~ O n d o r t h a t t h e s u b j e c t i s m Do r e a b l e t o counter. or overcome the disease/disorer as c~~ae with a subject not treated. or adstered with the comrposion 5 according to the present inventio. 25 The tenn .. immunel~ modulatoe' as used herein uieam asbrrc hc moduate a Cellular immune system Of a subject ah tes"m ubstdato" whic used herein includes a vaccine.n em"M 73mouar"s The p rase"cellua 'a mune system " as use dI hereji , includes a C f.Zl dae '0 immune resAPons which depends upon the Presence of a ~mh c el (and iae anti en res nri cels) Th t enn "T- lY~ y1 ss" includes cytotoxic T lYmocytes' hl TCell' up'resor I' clls and rtgulaatty T1 cells.

5 The term "whole cell ' as used herein, means a bacteriun which is intact, or whbichi ?c of al he teM "intact" as used herein means a bacterium which s c~npisd Of all of the coponents present in a whole cell, parcully whole, vial cel adoeceprticuarya oe, able cell, and/or a bacterium which has not been specifically treated to remove one or more components from it. By the term "substantially intact" as used herein it is meant that although the isolation and/r pudfcation process used in aining the bacterium may result in' for example, a slight modification to the cell and/or in the removal of onC Or more of the component of the cell, the degee to 10 which such a modification and/or removal occurs is insigunifica , gn Partcular, a substantially intact cell accord.n to a o g to the present invention has not been specifically treated to remove one or more components from it. WO2004/0220 9 3 and UK application number 0404102.6 (both of which 15 references are inorporated herein by reference) diclse an immune modulator composjtio or a phamaceuticaj composition composing a whole cell of a bacterium from the genra Athodococcus C'Ordonzia Vo'cadlal Dielzi, r vukamurella and Ncardoides However, neither of these docwnents teahes Or suggests the use of 20 such an immune modulator composition or pharmaceutical composition for the treatment and/or prevention of the two clinical syndromes PMWS or PDNS. Suitably, the immune modulator composition or phamnaceuical composition used herein may comprise a pharmacetically acceptable carier, diluent or excipient. 25 Suitably, the immune modulator composition and/or pharmaceutical composition may comprise more than one whole cell, and more preferably comprises a plurality of whole cells. 30 In one aspect, the immune modular composition and/or a pharmaceutical Gordorda, omPri ag the whole cell of a bacterium from the genera Rhodococcus, GOsone, orda Dieza, TdjNrc.,diod. ay faat esocompre at least one, or at least One further, antigen or antigenic detrminnt.

6 Suitably, the antigeni or antigewnj determinnt may be an antigen or antigenic determiat frOza one or more of the foflowbWi BCG bC il~ fCltet Gueziu) vaccine, diPhtheria toxoid vaccine Y e(iatesoflmtean/pru(dTPo , rple) vaccine, pertussis vaccine, tetaUs toxoid vaccine, measles vaccine, rlmp~s vaccine rubella vaccine OpV (oral pomyelitis vaccine) , ~vcao patthereof (as aught in 01900256 94 1) and a generic plsradju anig en, for e,18mple a mral aria parasite antigen. 10( Suitably, the immne modulator cmoj~ n/rPa~acuia COMPO~itOn m ay comnprse two Or more s c tiges orainj dR n nzphrmctica Ile pharmaceutical cOmPOsition and/or an inunune mo~duator comnposition used in accordance with the present invetinmyb odjns.dotesbjc 15 with an antigen Or antigeyjic deemnlt._t mybcO2riseedothsujt WhIen the composition is co-ad ministered with an antigen or antigenic determinant in accordance 'With the present invention thle antigen o niei determinant maY suitably be an antigen or antigeii ecmrfo or author of 2 0 t~ e f ll o w n g : o r ci e ci c air y p ) e 11I, o th e r P o rc in e v in us e s , b a c t e i a o f th e g e n e a StrepH foo~nJaphila Salmnonella AfycoPlasma; AdclhObacillI, BordegeL-a ad Partewella that have been associated with either syndrme, BCQY (bacillus of Calrn tte and Guerin) vaccne, diPhth zi toxoid vaccine, diphhwateu/pert 5 .i 25 ( DTP or Tziple) vaccine, part sss vaccine, W =ta toxid vaccine 0eases v ci, 25 mu ps vccine ub~ a V cine. 01W (oral poliOMYcltis vaccine), a~ccine,~ vaccae, or part thereof (as taught int GB300256 94 .1) and a genetic plasmodium antigen, for example a malaria paraite antigel'L Suitbly two or more. or three or more, of such antigmns or anageznic determ.iants may be coadiiten with a pharmaceutical Composition or an inlImIe modulator composition accordng to the presentivjjj 30 n n~ln 7 The 'rrmuzne modulator Conpostjz Used ;in cot whtepesr en ti4O Inay be a vaccine. Tfhe vaccine Mayb accr'ftyJ 0 u accte rsan Therapeutic vaccine. yb rpblc'vcie1 5 S Suitably, the coiliPosition -for use in accordance hepea~jzejo a conprse two or more, or three or more, baczi thez psn ne o meeay Ahodococcz, Gordoi 4 Nocpdi Jeef a frozUi anyeo h Preferably, the bacteria for use 1, accorrdw, ,f the presen invej re n 10 secies frO nX any Of the genr ae Rhod oc~ Iod p Noc r i D eany andNo ardiod~ssul2 s odonia b rdOncia, N. a ~ G. 2, G. terra c, N ocard a astero ld ,~ D ietz j ra r T S12r pel a .o m ta ol 1?Aodococcus rube, Akodococcut rhodnfl, ~popJ A. pau Pwo. eihol 1 5a d ~ o ~ j u s e. ~ad f r o mm ~ r l e a c pfo r e x M P e S u t a b l y , t h e s3 e c i e s 15 use fro m a rtiepax rJ g enu s are ones w h ich ~ can b e g r u wm o n m ~d iu r W h is lo ,P rfe mbly no-n-. anligelnjc nedium,, 13y Way of eaIple ornj ac s i s n a antigetrn edium is Saut'ox's med iuzn.y Sialent 20 More Preferably, the bcte~i for use in accordance ith the Present ivntion 20 are from the genus AOdococu including Ahodococcuswuber (Peouykn a MVO cardia nib.ra), Abodococcus rhodocro, 1 , Rpoocc~ roly ]mzowoc, a coProph ilus A hodoco cu: opacur, Ahd -oc ehopolis ro*, adCC MOre Preferably, the bactera for usein, Bccordnz With the Present McO 25 is Rthodococw COproph; The temi "Subject"' as used herin, rnea' a" animal. Preferably, the subject is a Mma~al, bird, fsh Or Crustacean including for example livestck and hamrnjs. 0 Preferably., the subject refened to herefti is a pig or a pigletPM SadPN spe ifcalydiseases of Pigs' although oth er S pci es M AY be s b e t t eae Syn drones It is it eded that be Pres et irven rji 2 Co ul ubje t t re la e dt relate yndrOMC in subjects other than pigs. gence th e se in ej is efect v 8 for the treatment and/or Prevention and/or reduction of (including piglets). HoWever, should this syd n of a Or PDNS in 'pigb identified in another subject, such as a differydoent or a sitniar syndrome be immune modulator composition and/or plvestock, it is visaged that the 5 would be effective to treat and/or prevencusucal coVmposition taught herein other livestock. The ten "lvsPrevent such a syndrome in Other subjectsucasi Preferabl, livestock Thetivestock as used herein refers to any farmed animal. (including lambs), s o e r ncrd Poultry, Pigs (including piglets), sheep (inluinglarib),cows or bu~ls (including calves). fish and crsens.~ More 10 Preferably, livestock means pigs - including piglets,. cm e cra viues and agents probably associate with PDNS and PMWS are comMensal or latent viruses carried by the mnaiorityofig and ulyoyenptW and produce a diseased state under e Cmority of Pigs and usually only erupt and status.~ ~ ~ ~ ' cti hmoeevsgdertain circumstances, such as an altered imrnune 15 phatns. t is cmfore envisaged that the innUne modulator composition and/or and/or prevent other iestio adording to the present invention may be used to treat totrat and/or prevent other di ahidor dsorders caused by Similar latent vimses and/or I rea' and/or prevent other idiopathic eruptions. Pref-rably, the bacterium according to the Presern invena'on is killed prior to 20 use. Preferably, the bacterium according to the prsen invention is killed by heat O ent thereof' forexam ple hefo -irealentin an autoclave at 121"C for 15 minutes. Other suitable treatments for kil tle bacterium may include ultraviolet or ionising radiation or Treatmient ,ith 1 chemiuc~4 hal s pheol aloo orfnnjr aspenol, alcohol or formalin. 25 Prferably, the bacterium according to the Present invention is purified isolated. and/or Preferably, the bacterium according to the Present invention ia suspended in 0 'water or buffered saline, suitably borate buffered at pr i. Preferably, the pharmaceuical compogjlio or im w modulator composition Sadministered (for the frst tie f more than one administration is to be made) after the livestock -has suckled for the *fizzlie. In particular frsm Pletosi a be important to allow the inf=n to take in, a, for dg st me re ctost itr may adMiniLste:ing the Pharmaceuical comnposli 0 or lm oua~cmoj~ o priorto an~Instring the first dose of the pb ac..eUtca com~positi 0 or 0, rnn d o u at or aorposition (w ere there is m ore tha n on'oe. F rt e a o dn ce o dobt orsoeappliaions the first adnstatf sboud not be, ive on the actual day of birth (-day). For son emlbodzw the frt a m li a on f th bepenabou C- dyposti or iuu odulator composi~on should be give be we1ao0 - days Pot b~ : orebir !1 preferably 1-3 days Po st-birth o e p f ra i 1 1 2 10 ay p stbjr1~ mo e reerably 2-3 days Post-birth. SUbs equ,,t ad2 0 mor -P e e a y. be gven7 das ad/or 14 days and/Or 8- 12 weeks after, the lirt injection VACCNES 15 The preparation of vaccinL~ which conti oroio us~Mnlatv iflgred'ent(s), is knOwvn to one skilled in the art T~pically, such vacir ar reae as injcabes, either as liquid solutions or Suspesions solid form~s Sia o Solution in, or susPenson inl, liquid prior to injection sauaitb eae foe Preparation may also be emulsified, or the 23ctiale bnrd ent 5 epsaed ine 20 I Pos mes he ctiv in redi~ t 5 are O tlen m ixed with eXcipiet w hich are phannaccutical 4 accepale and coznpatible with the active ingredient Suitable recipients are, for ecmple, water, saline, dextrose, oycerot, ethanol, or the lk n comnbinations thereof. Alternatively, the vaccine May be prepa~j lik and~tob 25 orallY ingested and/or capable Of inhataion.ed F xaptob In addition, if desired. the Vaccine mlay contain minor amount of auxiliary substances such as wttng or ezzlls'ing agents And PH buffering agents. 30 TYPically,- a physician will determine the actual dosage o ac~hzin ftA~~ zlaoz c m postion and h a c al com po uion w hch W Ml be niOst suitable 10 for am inivdual subject and it minl vary with the age, wigt and response of the parhiculr Subject. The dosages below are exe~p1wy of the average case. There caT, of course, be individual inistanie where higher or lower dosage ranges are mneied. 5 Preferably, the actual dosage that is 'Used results in mdimal toxjcjy to the subject. T he com p osition f e pr eg n en oay be d dnSt d by direct inj cti n T e C~ fl)O~~ 0 ma befozrnz at 0 fr pa rezteral. nucoa innla uscu 1ax 10 ntravno~ subcutaneous, firtraocular, intraden.,W or. trasdmla adnjistmtio Suitably, the comnPositjon according to the preset invention may be adrmi~stered at a dose of 103 - l01l organisms, preferably i0o'- 1010 Otganisms more preferably 106 - 10 5 101 orgaisms, 81d even more Preferably 1 0" -1 9 organisms. 15 ypialI' the composition accOrding to the presenz invenhtioy may be ad nied a10 dose Of]I O - 109 bacteria for huina and auimal use. diitrda rf thecpoiions o'f 41e pjresaet ivention ame to be acmnsme asin u enhuzcrs then I os - 10" o'raniss per dose, preferably I0 D. _ra~ir per 20 dose, mole preferably 106 -10 - 5XIO 9 orajprdsadee O8WM s pr erfla 16-]07 oranisrs per dose, and even more Preferably, 1 09 bacteria per dose for humaan and animal uISe may be .6.d Mni~Stered at regular. intervals. A6 will be readIly apprecte by a skIlleA pezMO. tedoaeam eewlle 25 dependent upon the Organism to whc theag dosestr~ isl bebe 7he taim "adv*99credp' includes delivery by defivery .mcn~ cldg Mcjctiork lipid mediatedi tmnsfectiM, liposone, irnn1pomsocS cini c 30 for cdl p ivery (nC Fh a nd om bi a 0 the reo Or even~ Vital delivery. Te routes 30 fr"A gastroei te st~ e isn include but am e not limited to mnUCOW j Masal. oral, pare t~g gasroine 5

~~

8 jtopical or subling,,Q routes.

The tenn in terd includes but is 'tot limited to delivery by'a mucosj roue, or xaap1 as a nasal spray or aerosol ±br inhalation or as an ingestable solution; 'a Pare nft route where delivery is by an Jje ctble for , s c s o example, an intravenos -rf sj in la~tadenna O Subaeosrut.o The term Cc-diitrobmeans that the site arid tfrne of aduln~tmtj 0 j of each~ of the aduvants(s), atigenls) and/or antigei deterinantS) o h rsn invention are such that the necessary mnodu~fladfn O th i sste , j.i a'iv Thus, whilst the auittln(s) and adj uv 5 oay be a m tu tr d ?t e rese n t 10 ini ti me and at the sam e site there m y be ad antabe d. d i ? rn h a tg n a~ n~' anr g~j~ d et~ ~ t~~ at a d iffre t til e a rd to a diff ere t e f o t e adjuvart®) The entigen(s) and/or antigenic deemlf 5 ai ~ua toina e b de l i v e r d i th e S a rn e d e i v e r y v e h ic le - a rd th e 5 n i e v) a ri d / o r a yi eg e n i ise i~ 5 and adj-uvant(s) may be coupe Jig n s and/or. Uraftd~ ' S ntigerd IS corap lsii a cdor dingco hed By w yOf exam ple only ,. the imm e m odulator SaCof t m orpostj a c rdir to t ePresent nvent on Jm ay be ad mn i nste fr e t h same ~,j,. 0 t eOrota m fat On f one or mnore antigens or fourth beore , t h Th - e ant igen, antigen c de, h, , pe td o om e or imeshe eo 20 May be administered Peptl oc~ tide orth host...o Sbetn ineti herof or in Multiple dosesSprtl rC3.niiee otehs "'ieta igeds The irnmuf. modulator c~f ij~ado hIIO JS ompositio for 5e in~ accordance with the LUnteion MY be adjm~b paa umber cof~itO dfr 25 routes SUCb as WrectiOn (which ~ inisds ae tm-ed 8 by xa number rdWand intm~uscular ection) intra"4 ruucosal, o= ~ral,4 'taera n admiistmion.urera4 or Ocular Preferably, 'A The Present invention, adnmj~sraio is by izteetion.Mr 30 preferably the -?)Cctior i 12 Preferably, in the present invenion, admib r Composition. tution is by an Ora Y acceptable 5 ou o ccjo he composition can be provided in 0.1 to 0.2 al of aqueous S soUtjO1 prTefarably buffered physiological saline, and ad nstere'd PArennmU, fo example by intradennal incltogia anan dii.p Preferably injected intraderlay The vaccine according to the invention is may befobl i injection ad u Slight Swelling and redness, sometimes also itching mbe o a niection sie. The ode of ad station, the dose and the 10 ma ber. e optimised by those skilled it the art in a kmown ANTIGN As used herein, an "antigen" means an entity which, when inrodd into an 15 inmunocompetent host, modifies the poduc tity o w ic wniodced ntbodi that cncmieittheni poeonof a specific antibody or antibodies cha TI2bine tu/ the entity, and/or modifies the relevant T-helper cell response, such as T2 and/or Th. he antigen may be a pure substance a Mixre of sb or soluble or particulate material stane, cetr bstances thissene, he trm ncldes (including cells or celn fmgmnts or cell sonxicate). In 20 this sense, the te includes any suitable antigenic detaminant, cross reacting antigen, 210 antigen, xenoautigeft tolerogen, allergen, lhaptera, and inmuZ0og06 Or Pauts thereof, as well as any combination thereof, and these tern are used i t chrne throughout the tet. ably The ter antigenicc deters ant or epitope" as used herein refer to a size on an 25 antigen which is recognizes by an antibody or Tce receptor, or is responsible for pavtof in nT-elpe cell response teferbly it is a short peptide derived from or as ad carote apite. owev the ter lso i l intended to include glYcopeptides or carbohydrates whichstimulaTetem lsoincludes modified sequences of amino acids 30 responses which recognse the whole organism. It is advantageous if the antigenic detemia is n antienic deniinantof the infectious agent which causes the inn t inous disease A "'preventajveB Or "Pphylic- vaccine is a vaccjDe WhIch is administere to inaive individual to prevent dev01OPM~t of a coadition, suh .s by siuai Protective immunity. 5 A "therapeutic, vaccine is a aiewIi s nstrjOndids~, a existig condition to 'reduce Or inimi~,se the conditions or1 o abrogate the imunpatiologica consequces of the eondiiob 10 ADANM iS The teri 'adjuvan, as used herein men= an entity capbef grItigo Psrticipating in the irtfluencitng of arn immne~respose. Anablev of aunbtance O Or ixture of substances thet assists, increases, dow~ 1 jmd~so iesfe 15 the imnmune response to an anatigen. neuls 40k odirifs The immune izodulator compoiton and/for phamcual cmoioN according to the present invenion may comprise one or mIn, adjaz Cooiionhc the effectiveness of the immn ,ouao copoiton ano Whcehanmcei 20 co position l~XaPles of additional adjuvants which., May be effcuve include bt: arenotllrkied o JAlUMumhdode aluminiurm Phosphate, aluminium potassium sulphate (alum), berylim Sulpate, silic, kaolin, carbon, wfe-ad emulsions oil in-water~ emnulsion, 'ZlrmYl diPePtide, bacterial endoto~ej,, ipid X~ CoI-ynebacteri pav'm (?'$opihobarenm acess, Borderelja PPe1Sv. kfYcobacteplum, vaccae 25 POlYribonucreotid. sodium alginate, lanolin, 1 'sOlecin, vitMj~A neluc s u c a s i n t r~ e ~ 2 a n d 1 1 2 er~ ~ & w .1 2 s a p o n jn4 lip o s o m e s , le v a m is o l ., D E A E dextrau, blocked copolymrrs or other syntic adjuvatsc~ajy~aeBaZl comp cay, Inc o, R a ri oics for e xmple, Mecrck Adjuvant 65 (M erck and Com pa y~ In ., Rmway, NVJ ) or Freund's Incom pet A djuvm and Com plete 30 Adjuvant (J7ifco Labmtois D~etroit, ficbigin). Only almiW~UM hydroxide is approved for human use. Some Of the Other adjunnl, such s Af vac oreaml have been proved for clinical trias, acefreape 14 'Suitably, the adjuvant may be a whole cell of a bacteium froyn any orne of the genera RhOdococcu,~ Gordoni, Nocardia Dgi hcJwl n ,cgdOd 5 I n t h e a r t , i t i s j m o w t h t D N A v a c c n e s w ei c h a n d e s s ec J j ydN Al e Sequences Wtclhc to gold particles and ,c va in s which are fie not e s en ialy hg n are effiient vaccine delivz system Unlike c 1 tenla skcin by aheseu DnA v

T

accines donot require a traditional adjuvant conmponaI Vaccdnes wthes DA fit~ a s p e c t o f t h e p r e s e n t i n v e rI t i o ,, e i n u n C n o d u l a t r c o Itn a d e S , Ve eM JO may sutably be used inj.thsc dNAto " ccjPO t a s~n de e reina in the influencing of the suchnn Drespojn o ugen JIRIA CEUT C C SIONS 15 713he Present Inlvention also Provides a pharmaceutical composition comnprisW a therapeutically effective ainount Of a whbole cell of a bacteria fio~tegnr Ahodococu Grds, AO car i jgj ' ~~l~ad Ncroes gendr OPtioay -Dai phiwll Indi ona ly Ph m af ucawy a ccp able c ari., dijueat or e r ii a ( n dn 20 cotubintiens thereof). xilns(cudg The Plhazmaceutjoal composition may com~pise two componets - a .firt component cozpsjng an angn and a -second copoe&Lt compj n uvn thereof The first a second comnPonent MAY be Binganjvvad 25 Simltaneously or together, and even by differ admiwirat decvmd sequentially, Suitably, the antigen may even be engender witbinteo tsesapatf a d i e a s p r c e s . T u sa n t g e a M a y o r ig in t f ro m a b a c te r ia l , h o s t o r P a r a s itic invasOn r may be a substnc Jreleased from the tissues such as a stress Protein, 0 equvsleylt to the heat-shock proteins of bacteria or a tumour antgen. The phz*acutca comPsti 0 n may be for human or animal usage in. human and Veterinary medicine and WRil typically Oiflpse any one or More Of a is phamaceutically acceptable diluent, carier o Py cep blediluentsfor, or excipient. A acceptable carriers or diluets for therapeutic use are well known in the pharmaceutical art, and are (A.oexample, m Remington's Pharmaceutical Sciences, Mack Publishing co. (A. R Gennaro edit. 1985). The choice of pharmaceutijca rie, excipie t Or diluent 5 can be selected with regard to the intended rout of a&i o d pbrmaceutidii practice. The phamaceutical composition may comprise as - or in addition to - the carrier, excipient or dilucnt any suitable binder(s), lubricant(s) suspending agent(s), coating agentss, solubilising agent(s). 10 Preservatives, stabilisers, dyes and even flavouring agents may be provided in the Phanuaceutjcal composition. Examples of preservatives include sodium benzoate, sorbic acid and esters of p-bydroxybenoic acid. Antioxidants and suspending agents may be also used. 15 There may be different composition/fomulation requirements dependent on the different delivery systems. By way of example, the pharmaceutical composition of the present invention may be formulated to be delivered using a mini-pump or by a mucosa route, for example, as a nasal spray or aerosol for inhalation or ingestable solution Or Par0nterallY in which the composition is formulated by an injectable form, 20 for delivery, by, for example, an intravenous, intramuscular, intradernal or subcutaneous route. Alternatively, the fomulation may be designed to be delivered by both routes. Preferably in the present invention the formulation is of injectable form. More 25 preferably the fornulation is intradetnally injected. Preferably in the present invention the formulation is an orally acceptable composton. 30 Where the agent is to be delivered mucosAHy through lhlgat tin mucosa, it should be able to remain stable during am t through the gastointestinal 16 tract; for exaizpi;, it should be resitat 0 Prtelyftic, dogrdatj 0 n, Sable at acd pJ4 and resista, to the detergent effects of bile, Where aPpropriate, the phuzmaceutical compostion, canbean1iedy S whairaT Oith om of a Suppoitor or Pessary, topieJly in the fourz of a otioi. solution, cre mwelt or dusting Powder, by useofaldiptoray ithfo of tablets con .aining excipint such sS~ o f laco, oAin Pchu, or oY either, alone or in admixture with excipients, or orh omofeii~ sovles the~r sus exaion co tai j~ lav uri Orcolouring aggntS, or they can be v X~ ected 10 P~rantaaay, for xmple intravenously, inranscala. itradexally, or subutneosl- or Paetezaj adiitratLon, th~e composiio= may be best used it, the fozr of a sterile aqueous soluion which may contain other substances, for example enough salts or moksaclaides to make The solution sotonc Ilit1 blood& For' buccal Or sublingual adi.strti the composition mnay be ad nitstered in the fomo 15 tbesor lozenges which c=n be forulated in a Onventiorw maner, or ,the comPostj 0 ns maY be administered by incorporatijon int h odadoedo h subject tefo n/rfe fh 20 C Ile agent of the Present i'rvC IiOnmy be adznjnstexred wMth Ofeorm eohr the ~acuheul ate stance y way of example the present inventon covers thesin~t~~or sequentia treatments wit an immune 1modulator compoitio and/Or Pharmaceutical composition according to the present inlventinadoer !5 More steroid$, analgesics, antivjiJ 5 , interleudw such as !on2, ador t othr Pbazrnaceuticlly active s"Ibstance(s). .,o te It wil1 be understood that tese regie includ, the admnini, ation of the substAhces sequentially, simnultaneosly or toge*er.

17 The tenn "irumue enhancer" as used herein means.one or more bacteziaeither isolated or in culture which when administered to a subject benefit the health of ihat 5 subject. Preferably, this beneht is achieved b he M theah o e response of the subject In accordance with the present invention, immune enhazaem may be used for the treatment of PMWS or PDNS. 10 The immune enhaancers may be administered by consumptioa in special designed food or in animal feeds, for example pig animal feeds suppleiented ith the bacteria of the present invention. 15 The immune enhancers may also be adraistered by other routes - such as direct injection. Preferably, the bacteria are killed so as to avoid the diffculties of maintaining 2ive Products and/or to expose immunologically active substances often hidden in live 20 bacteria. IDE A S A AMOSEF 25 In another aspect, the present invention relates to a method for identing one or more whole cells of bacteria from the genera Rhodococc, Gordoni, Nocarda, Die a, Tsukamurela and Nocardioides that can teat and/or prevent aMWS or PDNS comprising the steps of: (a) administering a first group of test animals with an inmunostimulant (b) administering a second group of test animals with an 30 inmunostimulat mixed with a bacterum (a) nwawing t ber ctOccurances of and/or severity of PMWS or PDNS in each of the test animals; and (d) comparig the results in each of the groups of test animals, wherein, a lower ocu cm of and/or 18 severity of PMWS or PDNS fromthe immuotuan mixed with ba r compaison to the immunostimulant alone is indicative of a bacteria sbee in accordance with the present invention. tu stable for use 5 As used herein, the tenn,"tes animnal,,refer su sedresp t th "rers to any animal that elicits a cellular te immunostimlant. Preferably, the test aimal) is a mazMM. Preferably, the bacterium modifies the T helper Cll response. Sitably, the bacterium may modify the T helper cell response by decreas.g Su Thit and Th2 10 response. Suitably, the bc h h n h h bacteium may modify the T helper cell response by increang the ThI response and decreasing the Th2 response. Suitably, the bacterium may modify the T helper cell response by increasing the Thi response without affecting the Thrsponje. 15 Preferably, the immunostimuant will have a knowjt Thi and IV response. For example, with the inuunostirulant BCG the reaction is usually largest at 24h when it is an indicator of the Thi response; the reaction at 4h is usually less and includes a Th2 contribution. It irsknon that BCG predoinant y stimulatLs a T response. nd 20 By use of such innmostimlants it may be possible to determine the Th1 response of a test bacterium arid, thus, it may be possible to identify one or more bacteria which have a desired Thi1Th response to treat and/or prevent a parteular disease and/or disorder. 25 Preferably, the cellul immune response is measured using the tuberculin skin tst. In Iice, the tuberculin~ skIn test is preferably carried out on the foot pad. In a predominant Thi reaction the positive foot Pad inU-e response is jrjxij~aj at 24 hours and diminishes at 48 hours. However as the Th2 reactivity incmases then the 0 4i hour positive foot p md inmune response increases and can even exceed the tp immune response at 24 hour. ot pad 19 Vciai'o with an 1M ~ -in~n such as BCO - inducs a re onse to 5ki~r st~i w thtuberculizi (a soluble preparation Of ?u er l b cl e ts te later. The local reaction is me asurtrvj 5 fo e p e n 24 ested 4 5 ors ard 72 hours after injection Of tubercuin. Briefy, an im uotm~ (e.g. 5 BCG) is usd thiat induces a positive immune response to tbeculin- Ja the test animal, the tuberculin skin test is Preferably caieouonteftpd.Ia Predominant Thi reaction the positivOt o te foot pad.i~mr~oei Inull m a t 4 hours anp mnsiti at 49 outs. However, as the Th2 reac I ty increases then the 48 ou po ive foot pad im m une response increae and can even exceed the foot 10 pad inrnune response at 24 hour. Thus 2 the assay can be used to assess whether or not the intfroductio Of an imney mloduilator compoitio according to the Present inventijon modulates thle Cellular. irnmu response. Preferbly 2 tile iUMgnuu~tiuant is BCG, ~15 The invention will now be fAmhe described by way Of EXamples and Figures, which are meant to serve to assist one of ordfnary skll in the art in carrying out t .he inv'ention 3nd are not intended in any way to limit the scope of thle invention. 20 ON F1FDS~~~ I IG Figure 1 shbow, the timings for the Occurrence both PMWS and ?DNS during the lifespan Of piglets/PigS together -with the results of treating pigktj With an immune modulator comprising R~ 00Cophu cpaewt 1 apce. 5 rvnto Phjs omare %tfalpacbo Groups of 1 to 3 day old pigles wre injected into the Sin bf the Iheck %t O.1VIJ of a susPensio-u Of I10ng/n1 Of the selected ifltmuine mmodtzlatr e.g. ho do coccs copropholisj or witb saRine as a coutrol. This Is repeated 7 and/or 14 days later, and 20 when aninals are 8-12 weeks old, madng sure that thesate PrEParajo is applied to the same pigs on each occasioz Pigs are then followed until tiey ar 20 Weeks of age with tegular checks for the appearance of the signs of either PMWS oc PDNS. 5 P to 10% Of Pigs ( snaly 3-5%) receiving injections of placebo develop either PMWS or.IPDNS uithift this timeframe in comnparison with less tha 1% of in't, modulated pigs developing the syndromesn The results of I Paiticular experiment can be seen in Figure 1. The timings for both 10 PiWS and tDNS are shown in Figure I and the results obtained showed a 15% increase in the surval to Slaughter rate of pigs treated with k Coprophiis as an mnmune modulator compared with pigs treated with a placebo. Treatment ex 15 Aim~als showing the earliest signs of PMWS or PDNS Wl be randoZjSed to receive either a course of irnmune modulator, or of placebo. Inetion iill be given immediately the diagnosis is suspected, and repeated at 2 weekly intevals until the animal recovers or dies. Samples of blood and biopsies of tissues will be sampled from 20 each animal for confarman of the suspected diagnosis. wilOlp of s0animls rith confnned PMWS or PDNS will be allocted to each treatment n soup. Prelininary investigations suggest that pigs treated with the immune modulator Will increase the survival to slaughter rate of Pigs compared wth Pigs datedr with the 25 placebo. All publications mentioned in the above specificao are herein acorporuted by reference. Various modifications and variation of the described methods and system of the Present invention will be apparent to those skilled in the a m without departing from the scope and 0 S ioit of the present invention r Arthough the present invention bas been described in connection With sPecic Prefer embodimens, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments- Indeed, 21 vayiOu M~odifications of the described modes for carrying out the invention hc r obvious to those skilled in biochenmistfY and biotec>hoogy or related Wlelh areddo be within the Scope of the following claimsfedarinnedt

Claims (15)

1. A Method far treating Or preventing Post-wean~ng multisystjc wasting Syndrome (pMWS) and/or Porcine deamatis and flephropatl 1 ynrm (?DN) ina subject comprising admnistering ant effective aznount of a 5Phannaceutical composition Or immune ]Uodulator coMPosi"', comPrisiug a whole cell of a bacteajum from one or mnore of the gene= 'Mo Gordorna, Yocardi,, Die za, Tsukarrnurella and Xoadjic to sid ubjc,

2. A method for protecting, including immunIsfrg, a subjcfom otwenn mXliSySte~iC Wasting SYndrome (PMWS) aidfor pci drmaPOSts andn 10 nePhropathiy Mydrome (PDNS) eorns4 Ndmi n dqt a Phmafeti ad bc it i o r i m n oor r o rposjtion comprising a whole cell 6f a baieiu nro One Or More of the genera Rhodococus Gordont 0 , ocardla, J ,eft Tsikamukell and AocardiOids. to said subject. 15 3. A method according to claims 1 or clam. 2 wherein the bacterium is from one or 15 ~~More of the species: Gordoa bronchialfr G. amarae, G. Spue~G ere Nocardia asle-roide Dielzia mars, Tskmtll puo eral, P~wdcoe ruep-Rhodococc, rhodfzzL &. cprop;hol&, . opaczu; . erYthopO~r, Nocardoides albiss and Tsalomuella inchompensi,

4. A metod according to claim I or claima 2. wh6erein the subject is a livestock 20 anmal

5.A method, according to claim 4 wherein the livestock anima] is a pig.

6. A method accordin~gto claim 5 wherein the livemtck animnal is ak piglet.

7. A method according to any one of the preceding claims wherein the bacteria is killed. 25

8. A method according any one of the precding claims wherein the Phaaeutca composition or immune modulator composition is adiftinstee for the fist time after the subject has taken in andlor digested the colostrumn.

9. Use of an immune modUlator composition or a pharmaceutical composition comprising a whole cell of a bacterium :from the genera Rhodococcu, 0 Gordonia, Nocardia, Dielzia Tsukamrnirella and Nocardioid, in the manufacture of a medicament for the tmatment or prevention of Pos t-weaning 23 mufsystoic Wasting syndrome (PMWS) and/or POrcine dezrnatjtis .nd nepbzOPathy syndrome (PDNS).

10. Use of a whole cell of a bacterium from the gen'er kRzodococw odoi m e zdi e t za s k~ zr~ n ~ T cyj~ in the Manufature of a 5 mei~net Or the teatrnent or prevention of post-weanng mulisyem wasting9 Syndrome (PMWS) and/., Por.ijC deanajitjs -~i fl roa Syndrorae (1'DNS). 4 t oah 1 C h use acardig to claim 9 Or claims 10 wherein 1 the bacteimrai rn oneo more of the species: Gordon~bocili .aaaG .'St G. e r 10 Nocardia asterorde, Diaizt mar, Q. Tskmtl a~o erao Rhodocus~ rubg'r' Rhodococus rho dniz R. coprop holly: A& oacw, A. a'IYthoPolIzA Nocardibides albrur and Tszdcamw-ella inchonemi,.,

12. The use according to any One of claims 9-1 1,Vwheeiji the subject is a li'vego~c 15

13.. The use according to clafin12 w1~eren the 1iveso~Jk animal is a pis.

14. The use according to claim 13 wherein the livestock animal is a piglet.

15-. The Use according to any one of claims 9-14 Wherein the bacteriumn is kiled. 16. The Use according any One of claims 9-15 Wherein the pharmamctical Q~mos''ozor immune modulator composition is admninistered for the first 20 lime after the subject has taken in and/or digested the coloftrun

17. A method as substantially hereinbefore described, IS. A use as substantially liereinbefore describe&L Dated this Nineteenth day of July

2004. UCL BioMedica Plc Applicant Wray & Associates Perth, Western Australia Patent Attorneys for the Applicant