CA2098101A1

CA2098101A1 - A vaccine against cholesterol to prevent hypercholesterolemia and atherosclerosis

Info

Publication number: CA2098101A1
Application number: CA002098101A
Authority: CA
Inventors: Carl R. Alving; Glenn M. Swartz, Jr.
Original assignee: Individual
Current assignee: Casi Pharmaceuticals Inc
Priority date: 1990-12-10
Filing date: 1991-12-10
Publication date: 1992-06-10
Also published as: WO1992010203A1; EP0562020A4; JP3183665B2; AU666473B2; EP0562020A1; AU9155691A; JPH06505714A

Abstract

A VACCINE AGAINST CHOLESTEROL TO PREVENT
HYPERCHOLESTEROLEMIA AND ATHEROSCLEROSIS

ABSTRACT

The present invention relates to immunoreactive compositions and methods for immunizing humans or animals against cholesterol and more particularly to the use of these compositions in methods for reducing the serum cholesterol levels of the immunized individuals and to retard or reduce the severity of atherosclerosis or atherosclerosis plaques caused by ingestion of dietary cholesterol.

Description

~0 92/10203 PCr/~591/09268 2~81~

A Vaccine Aeainss Cholesterol to Prcvent Hv~ercholes~erolemia And Atherosç~rosis 1. GOVERNhlE~'T INTEREST
The invention described herein may be manufactured. Iicensed and used by or for governmcntal purposes without the payment of any royalties to us thereon.

II. RELATED APPLICA ~

~o This applicadon is a condnuadon-in-part of U.S. Patent Applicadon Scrial No. 071444,214 filcd Dccember 1, 1989, which in turn is a condnuadon-in-part of U.S. Patent Applicadon Serial l~.umber 06/875.0~8 filcd June 2, 1988. Addidonally, thc applicadon is a condnuadon-in-part of U.S. Patent Applicadon Serial No. 07/6t)1,090 filcd October 2', 1990.
which in turn is a cantinuation~ part of U.S. Patent Application Serial No.
07n02,S99 filcd Junc 2, 1988 now U.S. Patent No. 4,88S,2S6 issued Decanbe,r S, 1989.

I11. F1ELD OF THE INVE~mON
This invcntion relates to immunoreactive compositions for in~nunizing or hypcrimmunizing humans against cholesterol and more panicularly to the usc of thesc composidons in mcthods for reducing the scrum choksterol levels of the immunized individuals and to retard or rcduce the scvcrity of athcrosclcrosis or atherosclerosis plaques caused b-ingesdon of dietary cholcstcrol or by other factors.

SUEISTITUTE SHEET

, o92~10203 2098101 Pcr/~s9l/o9268 IV. BACKGRO~'~'D OF THE I~ ~10~

It is widely believed that high levels of serum cholestcrol arc a significam causadve effect in the pathogenesis of atherosclerosis and associatcd diseases such as atherosclerotic coronary hcar~ disease atherosclerodc ccrebral vascular diseasc, renal disease, e~c. It is alsc believed that lowcring of blood cholesterol levels is associated with amelioration of atherosclerotic vascular diseases (Goodman~ D.S. er al o Rcport of the national cholesterol education program expert panel on detection, evaluadon, and ~rcatment of high blood cholesterol in adults Arch. Intern. Mcd. 148:36-69, 1988; Kromhout, D. et al.. Serum cholesterol and 25-year incidcnce of and mortality from mvocardial infracdon and canca. Thc Zutphen Study. Arch. Intern. Med. 148:10SI-1055, 1988). In 1984, a Nadonal Institutes of Health consensus dcvelopment conference panel recommended a framcwork of detection and treattnent of hypercholcsterolemia. Based on this the National Cholesterol Education Prograrn has tnade the well-known recommendation to adults:
"Know your cholesterol number" (Luepker, R.V. et al., Recommendations regatding public screening for measuring blood cholesterol. Summary of a Nadonal Heart, Lung, and Blood Institute Workshop, October 1988. Arch.
Intern. Med. 149:26S0-26S4, 1989).

The major methods recommended for achieving reduced serum chole~ol kvds re throul~h reducdon of dietary intake of cholcsterol and other hts, and treatment of hypercholesterolemic individuals with drugs designed to lower blood cholesterol. The blood cholesterol levels are p nicul rly associated with homeostadc mechanisms related to levels of pl sma lipoproteins that serve as carriers of cholesterol. The dangerous lipoproteins, from the standpoint of atherosclerodc risk are the low density lipoproteins (LDL), and the levels of LDL are regulated by the functional acdvida of LDL receptors on the surfaces of cells, particularly in the liver (Lrown, M.S. and Goldstein, J.L. A receptor-mediated pathway for cholesterol hon~costasis. Science 232:3~47, 1986). Many of the strategies for using drugs to reduce blood cholcsterol involve interference ~ i~h the processing of cholesterol derived frorn LDL (Brown and Goldstcin. 19g6).

SUBSTITUTE SHEET

092/1020~ 2098101 PC~/LS91/09268 The exIenmhat cholesterol can be reduced by diet is linmed by numerous factors, and the reducdon of cholesterol bv drugs could be associated with side effects of the drugs as well as cost. In any case. a vanetv of additional variables can influence cholestcrol levels, such as genetic background.
s stress effccts. and age. Additional methods for reduc~ion of cholesterol might be expcc~ed to havc beneficial health effects. particularly in individuals who might receive such treatrnent before signif~cant progrcssion of atherosclerotic disease has occurred.

o The present invendon describes the use of a vaccine forrnuladon that would be used to immunize humans against cholesterol and thereby lower the concentradon of serum cholesterol, either by itself or in combination with other methods commonly used to lower cholesterol. A variety of immunization procedures might be used to induce antibodies to cholesterol, and the presence of antibodies to choJesterol would be detected either by a solid-phase immunoassay using crystalline cholesterol or a cholesterol conjugate or by a complement-dependent assay such as complemem-dependent immune damage to liposomes containing cholesterol as taught by Swanz et al. lAntibodies to cholesterol. Proc. Nat. Acad. Sci. 85: 190~-1906, 1988] and Alving et al. [U.S. Pa~ent No. 4,885,'S6 issued S
December 19891-To our knowledge, humans have never been acdvely itnmunized against cholesterol and the safety of doing this, as well as the potential consequences relating to serum cholesterol levels or progression of atherosclerosis due to intake of dictary lipids, has not been established. It has been demonstrated that human sera usually do contain varying quanddes, depending on thc individual serum, of "naturally-occurring"
andbodies to cholestol l~lving et al., Naturally occurring autoandbodies to cholesterol in humans. Biochem. Soc. Trans. 17:637-639 (1989)].
However, there has not been any correladon of such naturally-occurring anibodies with sen m chokstaol lewls or with atherosclerosis.

The possibility has been discussed that naturally-occu~ing 3S andbodies to cholesterol might be a normal part of the aging p ocess and might contribute to (ratha than ameliorate) atherosclerosis (Alving. C.R.

S~ TE 5~El' wo 92/10203 2 0981 Dl PC~/~S91/09268 Antibodies ~o liposomes, phospholipids, and cholesterol: Implications for au~oimmunity. atherosclcrosis, and aging. In: Horizons in .Uembrane Biotechnology, ~'icolau, C. and Chapman, D., editors. ~'ilev-Liss. pp. ~1-41, 1990). The possible dangers of injecting liposomes containing cholesterol into animals containing antibodies to cholesterol have been illustrated by anaphylactoid effects observed by Wassef et al.
[Anaphylactoid reacdons mediated by autoantibodies to cholesterol in miniature pigs. J. Imrnunol. 143:2990 2995 (1989)]. Therefore it is not obvious that this invemion could have practical use in humans.
Nonesheless. the potential feasibility of this invendon as a possible safe and effecdve vaccine against choksterol has been demonstrated bv experiments in humans in which repeated injecdons of a candidate liposomal anti-malarial vaccine thu contained cholesterol did result in thc production of antibodies to cholesterol. This is cleuly indicated in a U.S. Patent Application Serial No. 07/601,090 entitled: "Encapsulated High-Concentradon Lipid A Composidons as Immunogenic Agents To Produce Human Andbodies To Prevent Or Treat Gram-negadve Bacterial Infcc~ions"
by Alving and Swartz filed on 22 October 1990. In that disclosure, thc example shown in Figure 9 thaein cleuly dernonstrates that antibodies to cholestaol cun be safely induced in certain individuals. The present invention utilizes an antigen that produces higher and more consistent andbodies than in the previous and-malarial disclosure, and produce such antdbodies for the purpose of preventdng diet-inducet serum cholesterol elevuions and athaosclerosis, ~Ithough we have not found in the prior art any teaching relating to immunization of humans with choleste~l, in the literature there has been one anempl described to try to ameliorate hypercholesterolemia and athosclerosis in r bbits by immunological means. Bailey et al.
Ilmmunizadon with a synthedc cholesterol-ester antigen and induced athe osclerosis in r bbits. Nature 201:407-408 (1964)1 immunized rabbits with an andgen consisting of cholesterol conjugated to bovine serum albumin. Bailey et al. stated that the "mean antibody itre measured by an interfacial precipitaion technique was 1 :70û0", but the was no attempt to prodwe or to me#ure andbodies that had specificity against cholesterol.
The ssay andgen consisted of the original conjugate, not cholesterol either SUBST~ TE S~EE~

wo 92/ 10203 2 0 9 8 1 0 1 PC~/~ 591 /09268 alone or as part of ano~her conju~ate. At no place did Bailey el al. teach that they had induced amibodies to cholesterol. and the- did not teach th31 antibodies to cholesterol could have been produced or that such amibodies might have played a role in the lowering of serum cholesterol levels or amelioration of atherosclerosis.

Bailey et al. did observe a reduced hypercholesterolemia and less aor~ic plaque formation in the immunized animals th~t were fed a cholesterol-rich diet. However. in the absence of funher information the 0 antibody titer could have been entirely directed against the bovine serumalbumin component and the cholesterol-albumin conjugate might simpl!
have lowered cholesterol through nonspecific mechanisms. such as by nonspecific adsorption or serum cholesterol by the albumin. This latter explanation could be supponed by the fact that albumin has a considerable degree of hydrophobicity and can be used as a reagent to prornote solubility of cholesterol in an aqueous medium such as serum. The disclosure by Bailey et al. is too insufficient to draw any immunological conclusion regarding the role. if any, that antibodies to cholesterol may have played in the experimental results. It is probably because of this that Baile,v et al. didnot teaeh any such role.

V. SUMMARY OF THE INVEl~,' llO~

This invendon eonsists of a vaecine which is effective in 2S immunizing hurnans against eholesterol. The purpose of this would be to reduee the sen~m eholesterol levéls of the imrnunized individuals and to red or reduee the severity of atheroselerosis or atherosclerosis plaques eauset by ingesdon of dietary eholesterol or by other faetors. The vaecine would eonsist of a fonnulatdon eontaining eholesterol or eholesterol and phosphatidyl ehoUne; or eholesterol and dimyristoyl phosphatidyl eholine together with a suitable delivery vehiele and may also eontain a suitable adjuvant. The reladve molar rado between the eholesterol and phosphatidyl eholine or dimyristoyl phosphatidyl eholine is within the range of 1:1 to I :~.S. An exarnple of a suitable fonnulation would be liposomes containing phosphatidyleholine, eholesterol. and lipid A in molar ratios of liposomes containing phosphatidylcholine. cholesterol. and lipid A in lar r~tios of SUBSTl-rU~E St ~ET

~0 92/10203 2 0 9 81 V 1 PC r/~ S91 /092h8 2/5/0.0~ (where the molafil~ of lipid A is based on the mol3ril- of phosphate in native lipid A). This ra~io is not necessaril~ cri~ical. bccause other ra~ios might be successful in accomplishing the sarne result Deliver~
vehicles other than liposomes would also be suitable, includins microcapsules, microspheres, lipospheres, polvmers. and slow release devices could serve instead of liposomes. An experiment in rabbits ha:.
demonstrated that the stipulated vaccine does ameliorate diet-induced elevations of serum cholesterol, Vl. BR~EF ~?ES~RI~I-ION' OF THE DRAWI~G

A more complete appreeiation of the invention and many attendant advantages theof will be readily obtained by reference to the following s DETAILED DESCRIPllON OF THE INVENTION when considered in conjunetion with the aeeompanying drawing, wherein:

Figure 1 shows IgG respon#s 2 weeks after initial immuniza~ion in the 6 human volunteers that eonstituted Group IV in the experimental immunization eholesterol from the above patent applieation. The individuals were immunized with 43% eholesterol liposomes as taught in the presenl diselosure and in the previous patent applieation. Each of the components was individually tested by ELISA for the appearanee of IgG antibodies agunst the purified individud eomponent. In the ease of lipid A, the 2S indiv du ls were injeeted with liposomes eontaining monophosphoryl lipid A. Tl# d-ta ue shown with preimmunization values, if any, subtraeted from the postimmunization vdues. Eaeh serum was diluted Ill00 for ELISA andysis. Three of the six immunized individuals developed signific ntly increased levels of antibodies to eholesterol. Figure i conesponds to figure 9 from the U.S. Pa~ent Applieation Serial No.
07/601,090, entitled: UEnc-psulaled High-Coneentration Lipid A
Compositions as Inununogenic Agents To Produee Human Antibodies To Prevent Or Treat Grun-negative Bacterial Infeetions" by Alving and Swartz filed on 22 Oetober 1990.

SUBSTITU~ F~s wo 92~1020~ 2 0 9 8 1 () 1 Pcrr/~S91/092~#

VII. DETAILED DESCRIPTlO!~ OF THE I~VE~TIO~ A~D
EXAMPLES

The working example set forth below illustrate. withou~ anv implied limitation a vaccine useful for the immunization of humans agains~
cholesterol. This vaccine is useful for immunizing or hvpenmmunizing ~
human against cholesterol. which vaccine comprises as an active ingredient A. a delivery vehicle and B. either. (i) cholesterol; or (ii) cholesterol and ~nadjuvant: or (iii) cholesterol. phosphatidyl choline on an adjuvann or ( i- ~
0 cholesterol, dimyristoyl phosphatidyl choline and an adjuvann or (v) choles~erol and phosphatidyl choline; or (vij cholesterol and dim~Tisto~l phosphatidyl choline EXAMPLE
An experiment is currently underway to determine the possible feasibility of ameliorating diet-induced hypercholesterolemia and atherosclcrosis in rabbits. Groups of rabbits are being immunized while oth groups are not being immunized against cholesterol; at least one eroup ~o of immunized and one group of nonimmunizcd rabbits will be fed a diet rich in cholesterol. It is our prediction that the immunization process will ameliorate the hypercholesterolemia and athosclerosis that is cxpected lo be produced by the cholesterol-rich diet and that this will reduce to practice the inwndon that is herein disclosed. The experimental results from the rabbit expenment described below provides substantive evidence in support of our prediction by demonstrating that the 1% cholesterol diet causes a dramadcally increased serum choksterol level within I week (6 weeks sfter immunizadon in those rsbbits thst were immunized), and the cholesterol condnues to rise ov the second week (7 weeks sfter initial immunization was started in the irnmunized snimals). Howev. the increased level of diet-induced choles~erol was the 30% less elevated in the snim~ls (Gtwp 11 !
thst were irntnunized against cholesterol (sce the Table herein).

SUBSTITUTE SHEET

uo 92/ 1 0203 '~ 0 9 ~ Pc r/- S91 /09268 METHODS
Liposomes Liposomes are being manufactured bv standard methods in which liposomes loaded with cholesterol (containing ~19c cholesterol) and also containing lipid A as an adjuvant are prepared for injection as taught b~
Swartz et al. [Amibodies to cholesterol. Proc. Nau Acad. Sci. 85:190'-1906, 1988] and Alving et al. [U.S. Patent No. 4,885.256 issued 5 December 1989].
The liposomes used for immunization contained dimyristovl phosphaddylcholine (DMPC~/cholesterol (chol)/dimyristoyl phosphadd~l glycerol (DMPG)llipid A (molar ratio 0.9/2.5/0.110.0' for rabbits, or 0.910.75/0.1/0.02 for humans, where the molarity of lipid A refers to lipid A phosphate). The total dose of lipid A injected as part of the 71*
cholcsterol liposomes was 50 ug lipid A. Thc liposomal cholesterol concentradon is described as a percen~age. and this is calculated as mol c~
with reference to (DMPC + DMPG); e.g., a cholesterol/(DMPC + DMPG) ratio of 0.7S/1 is 43 mol%, and 2.5/1 is 71 mol'rc.
Enzy~ne-lip~ orbent Assav (ELISA).

ELISAs were performed by using crystalline cholesterol as an andgen on the bottoms of the wells of microtiter plates. Chrystalline cholcstesol w s coated onto the surface of wells in polystyrene plates ~Immunlon 96, "U" bottom, Dynatech Laboratories, Alexandria, VA) by addition of an etlunolic solution and evaporation of the solvent by air under a fume hood. Pl ces were funhcr dried under high vacuum and stored at -30C when not used the same day. Plalcs werc blocked by addition of phosphate-buffercd saline (PBS: 137 mM NaC112.7mM KCI/9.6mM
phosphate, pH7.2) containing 10% heat-inactivated fctal bovinc serum (FBS) (M.A. Bioproducts, WallccrsvUlc, MD). This was accomplishcd by washing thc wclls thrce titnes for 10 min cach. Fifty microliters of ascites fluid containing monoclonal antibodies, diluted in PBS containing 10~
FBS, was added to thc wclls and incubated I hr at room tcmperaturc. Plates were thcn washcd three timcs for 5 minutes each with PBS. Fifty SU~sTlToTE S~EET

~o 92/10203 PC'r~591tO9268 20981~1 microliters of goat and-mouse Ig~ (micro-chain) alkaline phosphatase conjugate (Kirkegaard and Perry Laboratones, Gaithersburg. ~D) al 1 rnicrogram ml in PBS containing 10- FBS was added to the wells and incubated 1 hour a~ room temperature. Plates werc again washed thrce dmes for 5 minutes each PBS. Fifty mioliters of the substrate, p-nitrophen-l phosphate at 2 mglml in diethanolamine buffer (Kirkegaard and Perry Laboratories) was added to the well and incubated 30 minutes at room temperature. Plates were scanned for optical acdvity at 405 nm using a Titenek Multiscan (Flow Laboratories). Values reponed wcre adjusted by subtracting value in blank wells that lackcd both antigen and monoclonal antibody.

Imrnunizanon Protocol Four groups of rabbits were either immunized with liposomes containing 71 1% chol, or were not immunized. Immunization was performed eith intrarnuscularly or intravenously every two weeks for 6 weeks~ The immunization procedure routinely induced andbodies to cholesterol in rabbits. as determined by ELISA or by complement-induced im nune damage to high-cholesterol liposomes as taught by Swartz et al., Antibodies tocholesterol. Proc. Nat. Acad. Sci. 85:190~-1906, 1988, and Alving et al., U.S. Patent No. 4,885,256 issued 5 December 1989.

The immunizadon of human subjects with 43% cholestcrol 2S liposomes w s conducted as part of the testing of the efficacy of a vaccine for ioducdon of andbodies to rnalaTia andgen and andbodies to lipid A, as taught by the previous disclosure entitled: "Encapsulated High-Conccntruion Lipid A Composidons as Imtnunogenic Agents To Produce Hum n ~dbodies To Prevent Or Tre t Gnun-negadvc Bacterial Infccdons"
by Alving and Sw rtz that is cur ently being prepared as a U.S. patcnt applicatdon, the htter of which is a continuation-in-part of U.S. Patcnt Applicadon Serial Number 07n~.sos filcd lune ~, 1988 (A Vaccine For Inducdon of bnmunity to Mal ria. And-cholcstcrol andbodics induccd wcre deteacd in Group IV and are illustrated in thc accompan,ving Figure.

SUBSTITUTE SHEET

wo 92/10203 PC~/~S91/09268 Ex~enmen~LDiets At week 6. the experimental diets were ini~ated. The diets consisted either of ordinary rabbit chow or a 1% cholesterol die~ (obtained from Bioserve). Four groups and two subgroups of animals were emploved Group I, 4 rabbits, not immunized, fed normal die~: Group lla. 1 rabbi~s.
immunized intramuscularly, fed 1% cholestcrol diet; Group llb, ~ rabbits.
irnmunized inrravenously, fed 1% cholesterol diet: Group III, 4 rabbi~s. not immunized, fed normal diet; Group IVa, 4 rabbi~s, immunized 0 intramuscularly, fed normal diet; Group IVb, ~ rabbits, immunized intravenously, fcd normal diet.

In addition to the above teaching with rabbits, i~ is now evident that even liposomes containing 43% cholesterol (using liposomes as taught above and in the prior att describcd above) also can induce antibodies to cholesterol in a limited numbcr of individual humans.
It appcars tha~ the 71% cholesterol liposomes will bc superior to the 43% cholesterol liposomes as the basis of a vaccine to induce antibodies to cholcstc ol. This conclusion is drawn from the fact tha~ only a small number of thc individual humans immunized with liposomes containing 43'~t chokstcrol developed andbodics to cholesterol Isec Fig. 1. which is derived from tttc prcvious U.S. Patcnt Application Serial No. 07/601,090~ entitled:
"Enc psulatcd High-Conccntradon Lipid A Compositions as Immunogenic 2S Agcnts To Ploduce Hum n Anibodies To Prevent Or Treat Grarn-negative B c~ial Wccions" by Alving and Swartz filed on 22 October 19~0, the lattcr of which is a continuation-in-pan of U.S. Patent Application Serial No.07nO2,S09 filed June 2, 1988 (A Vaccine For Inducdon of Immunin to M l~ia)l. Thc contrast, approximately 70% of splccn cells from mice immunizcd with 71% cholestcrol wcrc producing antibodies to cholesterol Swanz ct al., Andbodics to cholcsterol. Proc. Nat. Acad. Sci. 85:1902-1906, 1988] ant Alving ct al., lU.S. Patent No. 1.885.256 issued December 1989~.

Bascd on the prior art it is evident that cholesterol is highl imrnunogenic and the immunogenicity is enhanced both bv adjuvanls (e.

SUBS TIT U T E S H E E T

3 PCT/~S91/09268 20981~
ll lipid A or other adjuvanls) and by thc epitope densitv of cholesterol used for immunization. Il should be possible to achieve the combination of hiVh epitopc densities of cholesterol togethcr with adju~ants b~ a vanet- Ot carrier mechanisms, including microcapsules, microspheres. Iipospheres.
high density conjugation or association of cholesterol with proteins or orher macromolecules, natural sources of high cholesterol (such as organisms such as mycoplasma that have the capacity to accumulate cholesterol). It is presumed that any established me~hod for inducing antibodies to particulale substances or macromolccules sheoretically could be adapted to inducing ~0 antibodies to choleslcrol.

E2~ncntal Results Table 1. Reduction of Diet-lnduced Hypcrcholesterolemia in Rabbits Immunizcd Against Cholesterol.
High Bk~SngSum Inae~e Rod~
Chobs~ munized Time Chdestelol Canpar~d l~lase G~ Dict~- -- (Wed~s~tm~/dl)~ Weck 5 t~

n - ~ 5 6' m - - 5 ~3 IV - ~ 5 83 n + , 6 797 731 m - - 6 64 IV - ~ 6 68 n ~ , 7 952 790 30 m - - 7 74 ~Data shown are mcans of rcsults (Group 1, 4 rabbits; Il, 6 rabbits: 111, 4 rabbits; IV, 6 rabbits).

e lS~ cholcstcrol diet was initiated at the 5 week time poin~ after starting the cxperiment.

~The immunization againQ cholesterol was ini~ialed at O weel;s.

SIIBST~TUTE SltEET

o 92/10203 2 0 9 8 ~ Pc~ S91/09268 The above resul~s demonstrate ~ha~ ~he hi~h cholcsterol diet invariably caused elevated serum cholesterol values. However, two eeks aft inidating the diet (week 7) the elevation of cholesterol in the imrnunized group (Group II) was 30% lcss than thc clevation of cholestcrol in the nonirnmunized group (Group I).

eU~STlTUTE S~EET

Claims

We Claim:

1. A vaccine for immunizing or hyperimmunizing a human against cholesterol, which vaccine comprises as an active ingredient A a delivery vehicle and B. either, (i) cholesterol: or (ii) cholesterol and an adjuvant: or (iii) cholesterol, phosphatidyl choline and an adjuvant; or (iv) cholesterol, dimyristoyl phosphatidyl choline and adjuvant; or (v) cholesterol and phosphatidyl choline; or (vi) cholesterol and dimyristoyl phosphatidyl choline.

2. A vaccine according to Claim 1 wherein the adjuvant is lipid A.

3. A vaccine according to Claim 1 wherein the delivery vehicle is selected from the group consisting of biocompatible-biodegradable, or biocompatible-nonbiodegradable liposomes, or polymers; slow release devices; and combinations thereof.

4. A vaccine according to Claim 3 wherein the delivery vehicle is a liposome or polymer.

5. A vaccine according to Claim 4 wherein the delivery vehicle is a polymer.

6. A vaccine according to Claim 4 wherein the delivery vehicle is a liposome.

7. A vaccine according to Claim 4 wherein the delivery material is in the form of microcapsules.

8. A vaccine according to Claim 4 wherein the delivery material is in the form of microspheres.

9. A vaccine according to Claim 2 wherein the deliver material is a slow-release device.

10. A vaccine according to Claim 1 consisting essentially of phosphatidyl choline, cholesterol and a delivery vehicle.

11. A vaccine according to Claim 1 consisting essentially of phosphatidyl choline, cholesterol, an adjuvant and delivery vehicle.

12. A vaccine according to Claim 1 consisting essentially of phosphatidyl choline, cholesterol and a delivery vehicle which contains lipid A.

13. A vaccine according to Claim 1 consisting of dimyristoyl phosphatidylcholine, cholesterol and a delivery vehicle.

14. A vaccine according to Claim 1 consisting essentially of dimyristoyl phosphadtidyl choline, cholesterol, an adjuvant and a delivery vehicle.

15. A vaccine according to Claim 1 consisting essentially of cholesterol and a delivery vehicle.

16. A vaccine according to Claim 1 consisting essentially of cholesterol, an adjuvant and a delivery vehicle.

17. A vaccine according to Claim 3 consisting essentially of dimyristoyl phosphatidylcholine, cholesterol and a delivery vehicle which contains lipid A.

18. A vaccine according to Claim 1 wherein the relative molar ratio between the cholesterol and phosphatidyl choline or dimyristoyl phosphatidyl choline is within the range of 1:1 to 1:2.5.

19. A vaccine according to Claim 18 wherein the relative molar ratio between the phosphatidyl choline and cholesterol is 1:2.5

20. A vaccine according to Claim 18 wherein the relative molar ratio between the phosphatidylcholine, cholesterol and lipid A is 1:2.5:0.02.

21. A vaccine according to Claim 18 wherein the relative molar ratio between the dimyristoyl phosphatidylcholine and cholesterol is 1:2.5.

22. A vaccine according to Claim 18 wherein the relative molar ratio between the dimyristoyl phosphatidylcholine cholesterol and lipid A is 1:2.5:0.02.

23. A therapeutic method for vaccinating a human against cholesterol to prevent hypercholesterolemia or atherosclerosis, said method comprising treating said human prior to the human having hypercholesterolemia or atherosclerosis caused by serum cholesterol, with an amount of the vaccine of Claim 1 to result in passive prophylaxis.

24. A therapeutic method for vaccinating a human having hypercholesterolemia or atherosclerosis caused by serum cholesterol, said method comprising treating said human with an amount of the vaccine of Claim 1 effective to result in the suppression of serum cholesterol or amelioration of atherosclerosis.