LIPID A ANALOG/IMMUNOGENIC CARRIER CONJUGATES, THE USE THEREOF AS VACCINES AND POLYVALENT HYPERIMMUNE GAMMAGLOBULINS

Abstract: The invention relates to lipid A analog/immunogenic carrier conjugates and the use thereof as vaccines for the prophylaxis of septic shock in animals. The invention provides for a polyvalent vaccine which contains the lipid A analog/immunogenic carrier conjugate. The invention also relates to a polyvalent hyperimmune gammaglobulin comprising an effective neutralizing amount of an anti-lipid A gammaglobulin and an effective opsonizing amount of an O antigen specific gammaglobulin. Kits containing these materials are also provided.

Description: Title of the Invention LIPID A ANALOG/IMMONOGE IC CARRIER CONJUGATES, THE USE THEREOF AS VACCINES AND POLYVALENT HYPERIMMϋNE GA MAGLOBULINS ' . * 5 The present invention was made with support from the U.S. Government. Accordingly, the U.S. Government has certain rights in the invention. Cross Reference to Related Applications This application is a continuation-in part of 10 U.S. Application Serial No. 07/669,084, filed March 15, 1991, which is a continuation-in part of U.S. Application Serial No. 07/502,555, filed April 2, 1990, abandoned, which is a continuation-in-part of U.S. Application Serial No. 07/385,860 filed July 27, 15 1989, abandoned. This application is also a continuation-in-part of U.S. Application Serial No.07/669,084, filed March 15, 1991, which is a continuation-in-part of U.S. Application Serial No. 07/487,898, filed March 6, 1990, which is a 20 continuation-in-part of U.S. Application Serial No. 07/385,860, filed July 27, 1989. The disclosures of these applications are fully incorporated by reference herein. FIELD OF THE INVENTION: 25 This invention is in the field of medicinal chemistry. In particular, this invention relates to pharmaceutical compositions and the use thereof as * vaccines and polyvalent hyperi mune gammaglobulins. f * BACKGROUND OF THE INVENTION: 30 Each year approximately 194,000 patients in U.S. hospitals develop bacteremia; of these, about 75,000 die. Ma i, D.G., in Nosocomia.1 Infections, R.E. Dixon (ed.), pages 183-196, Yorke Medical Books, New York (1981) . This high frequency of . mortality occurs despite the aggressive use of potent antibiotics. The shortcomings of antibiotic therapy may be attributed to the relative impermeability of the outer membrane of Gram-negative bacteria to the drugs and to the inability of the drugs to counteract the lethal effects of bacterial endotoxin. Ziegler, E.J., et al . , N. Engl . J. Med. 307:1225-1230 (1982); Bogard, .C.Jr et. al . , Infect. Immun. 55:899-908 (1987). One approach toward counteracting the lethal effects of lipopolysaccharide (LPS) endotoxin is the administration of anti-LPS antiseru . Such antiserum may also facilitate the removal of Gram-negative bacteria by the reticuloendothelial system. Several workers have demonstrated successful broad-spectrum protection against challenge by several Gram-negative organisms or endotoxins in different animal models by using rougϊi mutants of E. coli or S. minnesota as .Immunizing agents to generate antisera. Braude, A.I., et al., J " . Jnππunol. 108:505-512 (1972); Braude, A.I., et al . , J. Infect. Dis. 236:S167-S173 (1977); Dunn, D.L., et l . , Surgery .32:212-219 (1982); McCabe, W.R., J * . Immunol . 208:601-610 (1972); McCabe, W.R., et al . , J. Infect. DΪs. 13£T:S161-S166 (1977) ; and Ziegler, E.J., et al . , J. Immunol . 121:433-438 (1973). These mutants lack the 0-specific polysaccharide and express on their surface portions of the LPS core which presumably contains antigenic determinants shared by most Gram-negative organisms. Passive immunization of patients with human antisera raised against the rough mutant E. coll J5 has been shown to * be an efficacious supplement to antibiotics for the control of nosocomi- al infections. Ziegler, E.J., et al . , N. Engl . J. Λfed. 307:1225-1230 (1982) . It is now recognized that most of the biological activity of bacterial endotoxins reside in the lipid A moiety of the LPS molecule. Luderitz, 0., et al., Curr. Top. Membr. Transp. 27:79-151 (1982) . Typical Gram-negative bacterial LPS has three major structural regions: the 0-polysaccharide, the R-core oligosac- 5 charide, and lipid A. The structure of the 0-polysac- charide is highly variable between organisms, even in the same species. Its antigenicity serves as a basis for serotyping the bacteria. The R region is a bridge between the 0-antigen and lipid A; its structure is 10 similar in most Gram-negative bacteria. Antibodies to LPS may promote phagocytosis or the death of the bacteria. The 0-antigen is the most antigenic compo¬ nent of the LPS, yet it has little known toxicity. Lipid A, in contrast, contains the toxic center of the 15 molecule and is remarkably similar in structure across a wide range of bacterial genera. It is believed that the lipid A region of LPS is responsible for a complex array of inflammatory responses l_?p tissue invasion by Gram- egative bacteria 20 by directly stimulating host cells such as macro- phages, neutrophils, and endothelial cells which mediate the inflammatory changes. Responses are both toxic (hypotension, coagulation disturbances, death) and beneficial to the infected host (enhancement of 25 antibody forάvation, mobilization of phagocytes, acute phase protein synthesis) . The structure of lipid A is depicted as Formula (I) below: ' . (I) Lipid A is a glucosamine disaccharide that is phosphorylated at positions 1 and 4' and has six or seven este^ified fatty acids. Four molecules of 3- hydroxytetradecanoate are attached to the glucosamine disaccharide at positions 2, 3, 2', and 3'; the hydroxyl groups of the 3'-OH-14:0 residues at posi¬ tions 2 ' and 3' (and sometimes 2) are substituted with normal fatty acids (dodecanoate, tetradecanoate, hexadecanoate) to form acyloxyacyl groups. In order to gain insight into the structure- activity relationship of lipid A, the biological activity of chemically synthesized lipid A analogs and biosynthetic precursors of lipid A has been examined. For example, Luderitz, O., et al . , Rev. Infect. Dis. 6:428-431 (1984) , disclose the preparation of lipid A analogs and the relationship thereof between the chemical structure and biologic activity. The authors report that phosphate substitution of the glucosamine disaccharide is not essential for the expression of antigenicity, and that the amine-bound 3-hydroxyacyl residues can be replaced by non-hydroxylated fatty acids without reduction of activity. The authors also indicate that the immunodeterminant structure compris¬ es the linkage region of the amide-linked fatty acids and glucosamine. Galanos, C, et al . , Rev. Infect. Dis. 5:546-552 5 (1984) , disclose the preparation of semisynthetic lipid A immunogens comprising glucosamine disaccharide carrying one amide-linked 3-hydroxymyristic acid conjugated to a protein (edestin) for immunization of rabbits. Antibodies were produced in titers compara- 10 ble to those obtained by immunization under similar conditions with acid-treated bacteria coated with lipid A. Behling, U.H., et al . , J. Immunol . 227:847-851 (1976) , disclose the preparation of synthetic glyco- 15 lipid adjuvants comprising N-palmitoyl-D-glucosamine, N-oleyl-D-glucosamine, N-myristoyl-D-glucosamine, N- decanoyl-D-glucosamine, and N-stearoyl-D-glucosamine. The authors report that comparable or superior en¬ hancement elf the iitimune response was obtained with the 20 synthetic glycolipids in comparison to the use of LPS endotoxin in assays measuring anti-sheep red blood cell (SRBC) or gamma-globulin (HGG) hemagglutinin titers. The authors also report that the synthetic glycolipids have no activity in chick embryo lethali- 25 ty, Shwartzman skin assay, and Limulus lysate tests which are characteristic as well as sensitive assays of endotoxicity. Therefore, the authors conclude that the mitogenic and adjuvant effects by otherwise nonendotoxic glycolipids suggest that not all -biologic 30 properties of the endotoxin are related to one struc¬ tural subunit or to one structural feature of this ,.* very complex molecule. V Hodgins, D.S., PCT Application Publication No. WO87/07297, published December 3, 1987, discloses LPS 35 derivatives of reduced toxicity obtained by treatment of lipid A with an acyloxyacyl hydrolase to give a compound of the Formula (II) : (ID This altered bacterial LPS is therapeutically useful as a vaccine to prevent Gram- egative bacterial diseases by^ inducing antibodies to LPS, as an antidote to treat or prevent Gram-negative bacterial sepsis, or as an adjuvant to enhance formation of antibodies to other antigens. The acyloxyacyl hydrolase itself may also be prophylactically or therapeutically useful to detoxify endogenous LPS in patients with Gram-negative bacterial diseases. This enzyme may also be used to remove toxic LPS from therapeutic injectants. A number of researchers have isolated capsular polysaccharides from Gram negative and Gram positive microorganisms for use as vaccines and to raise antibodies in animals which can be used for passive _i_mmunization. Where the capsular polysaccharides are poorly immunogenic, they may be linked to an im unogenic polypeptide to improve the immune response of the immunized animal. Lagergard, T. et al . . Infect. Immun. 58 :687- 694 (1990) , disclose the preparation of a group B Streptococcus type III capsular polysaccharide covalently bound to tetanus toxoid which induces the generation of type III-specific antibodies. Fattom, A. et al . , Infect. Immun. 58:2309-2312 (1990) , disclose the preparation of a Streptococcus pneumonlae type F polysaccharide conjugated to diphtheria toxoid which elicited type-specific antibodies thereto when administered to human volunteers. Guyer, H. et al., ATed. Microbiol . Immunol . 272:135-143 (1982), disclose the preparation of oligosaccharide-protein conjugates from Klebsiella pneumonlae. The conjugates increased the LD 50 of immunized animals by a factor of 10 5 as compared to non-immunized animals. In addition, sera from immunized animals conveyed passive protection in mice challenged by K. pneumonlae. Schneerson, R. et al . , Infect. Immun. 52:519-528 (1986) , disclose the preparation of E. coll, Hemophllua Influenzae and Pneumococcua type 6A (Pn6A) capsular polysaccharides linked to tetanus toxoid which were injected into humans. According to the authors, the conjugate-induced antibodies exerted secondary biological activities that have been correlated with .Immunity. Passive immunization with Pn6A antibodies resulted in about a 100,000-fold increase in the LD 50 of Pn6A organisms in mice. Cryz, S.J. Jr et al . , Infect. Immun.55:1547-1551 (1987) , disclose the preparation of an 0- polysaccharide from Paeudomonaa aeruginoaa covalently linked to toxin A. Rabbit anti-conjugate IgG, when passively transferred to mice, was highly effective at preventing fatal P. aeruginoaa burn wound sepsis. Jennings, H.J. et al . , J. Immunol . 242:3585-3591 (1989) , disclose the preparation of a group B Neiaaeria meningitidis polysaccharide-tetanus toxoid conjugate which elicits antibodies specific for an epitope on Neiaaeria meningitidia and E. coli . Siber, G.R. et al . , Infect. Immun. 45:248-254 (1984) , disclose the preparation of human hyperimmune globulin to Haemophilua influenzae b, Streptococcua pneumonlae and Neiaaeria meningitidia. The respective vaccines comprise polysaccharides which impart an i iminogenic response specific for the respective organism. Porro, M. et al . Mol . Immunol . 23:385-391 (1986), disclose the preparation of an artificial molecule which was synthesized by covalently linking the oligosaccharide haptens derived from Streptococcua pneumonlae type 6A and Neiaaeria meningi tidis group C capsular polysaccharides to the non-toxic mutant protein CRM 197 which is serologically related to diphtheria toxin. The induced serum antibodies recognized the polysaccharides capsule of S. pneumonia type 6A and 6B strains and killed the N. meningitidis group C bacteria by complement-mediated bacterial lysis. Tarkowski, A. et al. , J. Immunol . 244:3770-3778 (1990) , disclose the immunization of humans with polysaccharide Ag {Haemophilus influenzae type b, Neisaeria meningi tidis serogroup A, C, Y, W-135, and Streptococcus pneumonlae capsular polysaccharides) which elicited an immune response in volunteers. According to the authors, the polysaccharide vaccine coupled to a protein carrier induced a higher frequency of anti-polysaccharide antibody-secreting cells. Karakawa, W.W. et al., Infect. Immun. 56:1090- 1095 (1988) , disclose the preparation of the type 5 and type 8 capsular polysaccharides of Staphylococcus aureus. Antisera to the type 5 and type 8 capsular polysaccharides induced type-specific in vitro phagocytosis of capsulated organisms by human polymorphonuclear leukocytes. Iinuma, H. and Okinaga, K. , J. Infect. Dis. 160:66-75 (1989) , disclose the prevention of pneumococcal bacteremia by immunization with type 6 Streptococcus pneumonlae polysaccharide vaccine in rats. The polysaccharide vaccine elicited significantly higher serum opsonic activity and increased antibody titer to the type 6 polysaccharide compared to controls. There was a close correlation between IgG antibody titer to type 6 polysaccharide and opsonic activity. Baker, C.J. et al . , N. Eng. J. Med. 319.-1180-1185 (1988) , disclose that the immunization of pregnant women with a polysaccharide vaccine of group B Streptococcus gave rise to vaccine-induced immunoglobulin. Serum from infants whose mothers responded to the vaccine uniformly promoted efficient opsonizatic-n, phagocytosis and bacterial killing in vitro of type III strains. Prakobphol, A. et al . , Infect. Immun. 27:150-157 (1980) , disclose the preparation of a polysaccharide from Streptococcus mutans. This polysaccharide was immunogenic in rabbits which were immunized with whole cells or cell walls. Fattom, A. et al . . Infect. Immun. 58:2367-2374 (1990) , disclose amethod of preparing a Staphylococcua aureus CPS/immunogenic polypeptide conjugate and the use thereof as a vaccine. Despite the development of lipid A analogs as immunogens for induction of active immunity to LPS, a need continues to exist for new compounds which can be used in the treatment or prevention of septic shock in animals. In addition, a need continues to exist for the development of multivalent hyperimmune gammaglobulins specific for lipid A, Gram negative and Gram positive microorganisms. SUMMARY OF THE INVENTION The invention relates to a lipid A analog/immuno¬ genic carrier conjugate, wherein said lipid A analog the following Formula (III) : (III) wherein n is 1 or 2; R 1 and R 3 are the same or different and selected from the gtoup consisting of hydrogen, a C 2 -C 18 acyl group, a 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy)-C 3 -C 18 acyl group and a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C.| 8 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. In particular, the invention relates to a lipid A analog/immunogenic carrier conjugate having the following Formula (IV) : (IV) wherein n, R 1 , R 2 , R 3 and R 4 are defined above; A is an immunogenic carrier; m is 0 or 1; p is 1 to 200; L is a linker group which does not interfere substantially with the characteristic ability of the lipid A analog to stimulate an immune response in an animal; R 1 and R 3 are the same or different and selected from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3-(C 2 -C 12 - acyloxy) -Cg-C^ acyl groups; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group or a phosphate group. The invention also relates to a lipid A analog/- immunogenic carrier conjugate having the following Formula (V) : (V) wherein n, m, p, L, R 1 , R 2 , R 3 and R 4 are defined above. The invention also relates to a vaccine for the prophylaxis of septic shock comprising a lipid A analog/immunogenic carrier conjugate of the present invention together with a pharmaceutically acceptable carrier or adjuvant. The invention also relates to a method for treating or preventing septic shock in an animal comprising administering a pharmaceutical composition to an animal comprising a lipid A analog/immunogenic carrier conjugate of the invention; and a pharmaceutically acceptable carrier; wherein said lipid A analog/immunogenic carrier conjugate is present in an amount effective to induce active immunity to LPS in an animal. The invention also relates to intermediates useful for the preparation of the lipid A analog/immunogenic carrier conjugates of the inven¬ tion. This invention also provides for a polyvalent vaccine comprising (a) a lipid A analog/immunogenic carrier conju¬ gate of the invention, (b) at least one Gram negative and/or Gram positive bacteria, or antigenic determinant thereof, optionally conjugated with an immunogenic carrier, (c) a pharmaceutically acceptable carrier, and (d) optionally, an adjuvant. This invention also provides for a polyvalent hyperimmune gammaglobulin comprising an effective neutralizing amount of a gammaglobulin specific to the lipid A/immunogenic carrier conjugate of this invention and an effective opsonizing amount of a Gram negative and/or Gram positive bateria-specific gammaglobulin. This invention also concerns a method of prepar¬ ing a polyvalent hyperimmune gammaglobulin comprising (a) immunizing an animal with the lipid A analog/immunogenic carrier of this invention and at least one Gram negative and/or Gram positive bacteria, or antigenic determinant thereof, optionally conjugated* ith an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said ' bacteria and neutralize LPS. This invention also provides a polyvalent hyper¬ immune gammaglobulin obtained by (a) immunizing an animal with the lipid A analog/immunogenic carrier of this invention and at least one Gram negative and/or Gram positive bacteria, or antigenic determinant thereof, optionally conjugated with an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said bacteria and neutralize LPS. This invention further provides for a method of treating Gram negative and/or Gram positive bacterial sepsis comprising administering an effective neutralizing amount of a gammaglobulin specific to the lipid A/immunogenic carrier conjugate of this invention and an effective opsonizing amount of Gram negative and/or Gram positive-specific polyvalent hyperimmune gammaglobulin to an animal suffering from a Gram negative and/or Gram positive bacterial infection. This invention also provides for a kit for treating Gram negative and/or Gram positive bacterial sepsis comprising a carrier compartmentalized to receive in close confinement therein one or more container means wherein a first container contains a lipid A analog/immunogenic carrier conjugate of the invention. This invention also provides for a kit for treating Gram negative and/or Gram positive bacterial sepsis comprising a carrier compartmentalized to receive in close confinement therein one or more container means wherein a first container contains a hyperimmune gammaglobulin specific for the lipid A analog/.Immunogenic conjugate of this invention. Optionally, the kit may further comprise gammaglobulins specific for at least one Gram negative and/or Gram positive bacteria. Unexpectedly, the inventors of the present application have discovered that acyl derivatives of glucose and gentiobiose, when linked to an immunogenic carrier, induce active immunity to LPS in animals. This discovery allows the ready preparation of inexpensive vaccines which may be used for prophylaxis of septic shock. DESCRIPTION OF THE FIGURES Figure 1 depicts a graph showing the elution profile of FD-4 PS from a G75 gel filtration column. Figure 2 depicts a graph showing the elution profile of FD-7 PS from a G75 gel filtration column. Figure 3 depicts a graph showing the elution profile of INT-3 from a G75 gel filtration column. Figure 4 depicts a graph showing the elution profile of INT-4 PS from a G75 gel filtration column. Figure 5 depicts a graph showing the elution profile of KPO-2 PS from a G75 gel filtration column. Figure 6 depicts a graph showing the elution profile of the FD4-PS-DT conjugate from a CL-6B Sepharose column. Figure 7 depicts a graph showing the elution profile of the FD5-PS-DT conjugate from a CL-6B Sepharose column. Fi ured 8 depicts a graph showing the elution profile of the FD7-PS-DT conjugate from a CL-6B Sepharose column. Figure 9 depicts a graph showing the elution profile of the INT4-PS-DT conjugate from a CL-6B Sepharose column. Figure 10 depicts a graph showing the elution profile of the KP02-PS-DT conjugate from a CL-6B Sepharose column. Figure 11 depicts a graph showing the elution profile of the high molecular weight (HMW) and low molecular weight (LMW) KP02-PS-DT conjugates from a CL-6B Sepharose column. DESCRIPTION OF THE PREFERRED EMBODIMENTS The invention relates to lipid A analog/immuno¬ genic carrier conjugates comprising glucose or a polyglucose molecule such as gentiobiose linked to an immunogenic carrier, wherein at least C 3 position of the glucose or gentiobiose moieties are substituted with an acyl group (R 2 ) . In another embodiment, the C 2 position of the glucose or gentiobiose moieties is not substituted by hydroxy (R 1 H) . The immunogenic carrier may be linked to any free hydroxyl group on the Lipid A analog molecule, so long as the conjugate induces active immunity to LPS when administered to an animal. Preferably, the immunogenic carrier is linked to the C 1 or C 6 position of the glucose or polyglucose moiety. Equivalents of the lipid A analog/immunogenic carrier conjugates of the present invention include all non-amino saccharides and polysaccharides acylated at least at the C 3 position of at least one of the saccharide moieties and linked to an immunogenic carrier, wherein the conjugate induces active immunity to LPS when administered to an animal. A lipid A analog/immunogenic carrier conjugate of the inventfbn is considered to induce active immunity to LPS wherein when the conjugate is administered to an animal, anti-LPS antibodies are produced by the animal. The efficacy of the anti-LPS antibodies can be tested, for example, by administering the sera of an .Immunized animal to a second animal followed by challenge of- ' the second animal with LPS-producing bacteria (see Example 7, below) . Where administration of the anti-LPS sera results in enhanced survival of the second animal, the conjugate is considered to induce .Lmiriunity to LPS. Anti-LPS antibodies can also be tested by inhibi¬ tion of the Shwartzman reaction, a well-known biologi¬ cal response to LPS (Lee, L. et al . , In: Zweifach, B.W. et al . (eds.) THE INFLAMMATORY PROCESS, Academic Press, NY, 1965, p. 791) . Animals, particularly rabbits, actively immunized with LPS or with one of the compositions of the present invention are given an appropriate LPS challenge to induce a localized Shwartzman reaction. Prevention or amelioration of the erythema, hemorrhage or necrotic reaction is evidence for the presence of anti-LPS antibodies. Alternatively, serum from an animal immunized with LPS or with one of the compositions of the present inven¬ tion is transferred to a naive rabbit, which is then tested for a Shwartzman reaction. Inhibition of the reaction is a measure of antibody activity. The vaccines of the present invention are useful for the prophylaxis of septic shock .in an animal which is due to the release of LPS endotoxin by Gram nega¬ tive microorganisms. Such Gram negative microorgan¬ isms include, but are not limited to Salmonella, Eacherichia, Hemophilua and Neiaaeria, Klebaiella, Shigella, Paeudomonaa, Enterobacter, Acinetobacter, and Bacteroidea. See Bergey's Manual of Syatematic Ml crobi ol ogy. The conjugates of the invention are useful as vaccines wϊjich induce active .Immunity toward LPS in animals. Preferably, such animals are humans, however the invention is not intended to be so limiting. Any animal which may experience the beneficial effects of the vaccines of the invention are within the scope of animals which may be treated according to the claimed invention. In particular, the invention relates to lipid A analog/immunogenic carrier conjugates having the Formula (IV) : (IV) wherein A is an immunogenic carrier; L is a linker group which does not interfere substantially with the characteristic ability of the lipid A analog to stimulate an immune response in an animal; R 1 and R 3 are selected from the group consisting of hydrogen', C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy)-C 3 -C 18 acyl groups; m is 0 or 1; n is 1 or 2; p is 1 to 200; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy Cg-C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy)-C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group or a phosphate group. The invention also relates to a lipid A analog/- immunogenic carrier conjugate having the following Formula (V) : (V) wherein n, m, p, L, R 1 , R 2 , R 3 and R 4 are defined above. By the term "immunogenic carrier" is intended any macromolecule which is capable of inducing an .Lmmuno- genic reaction in an animal. Since many small mole¬ cules such as the lipid A analogs of the invention do not induce' active immunity by themselves, it is necessary to conjugate the analog to an immunogenic carrier to induce production of antibodies which are specific for the small molecule. Such immunogenic carriers include, but are not limited to, proteins such as bovine serum albumin, diphtheria toxoid, tetanus toxoid, edestin, keyhole-limpet hemocyanin, Pseudomonal Toxin A, choleragenoid, cholera toxin, pertussis toxin, viral proteins, and eukaryotic proteins such as interferons, interleukins, or tumor necrosis factor. Such proteins may be obtained from natural or recombinant sources according to methods known to those skilled in the art. When obtained from recombinant sources, the immunogenic carrier may comprise a protein fragment comprising at least the immunogenic portion of the molecule. Other known immunogenic macromolecules which may be used in the practice of the invention include, but are not limited to, polysaccharides, tRNA, nonmetabolizable synthetic polymers such as polyvinylamine, polymethacrylic acid polyvinylpyrrolidone, mixed polycondensates (with relatively high molecular weight) of 4'4'-diamino- diphenyl-methane-3,3'-dicarboxylic acidand4-nitro-2- aminobenzoic acid (See Sela, M., Science 166:1365-1374 (1969)) or glycolipids, lipids or carbohydrates. Preferably, the immunogenic carrier is a protein. By the term "linker group" is intended one or more bifunctional molecules which can be used to link the immunogenic carrier to the lipid A analog and which do not interfere with the production of anti- lipid A antibodies in vivo. The linker group may be attached to any part of the glucose or gentiobiose moiety so long as the point of attachment does not interfere with the production of anti-lipid A antibod¬ ies in vivo and thus interfere with the induction of active immunity. Examples of linker groups which can be used to link the lipid A analog to the immuribgenic carrier may comprise (VI) - { CH 2 ) q -NH- f wherein q is 2-10; O O II II - ( CH 2 ) q -NH-C- (CH 2 ) x C ( ii ) wherein q = 2-5, x = 2-12; and 0 ( CH 2 ) y -C- ( viii ) wherein y = 1-3 Typical acyl groups which can be substituted on the carbohydrate moiety include, but are not limited to, acetate, propionate, butanoate, pentanoate, hexanoate, heptanoate, octanoate, nonanoate, decano- ate, palmitoyl, olelyl, myristoyl, stearoyl, 3- hydroxybutanoate, 3-hydroxypentanoa e, 3- ydroxy- hexanoate, 3-hydroxyheptanoate, 3-hydroxyoctanoate, 3- hydroxynonanoate, 3-hydroxydecanoate, 3-hydroxy- decanoate, 3-hydroxypalmitoyl, 3-hydroxyoleyl, 3- hydroxymyristoyl, and 3-hydroxystearoyl groups. Also included within the scope of R groups include the 3- (C 2 -C 12 acyloxy) -substituted aforementioned C 3 -C 18 acyl groups wherein the C 2 -C 12 acyloxy groups include, but are not limited to, acetate, propanoate, butanoate, pentanoate, hexanoate, heptanoate, octanoate, nona¬ noate, decanoate, and dodecanoate groups. Preferred lipid A analog/immunogenic carrier conjugates are derived from gentiobiose and have the following formula (IX) : (IX) A preferred lipid A analog/immunogenic carrier conjugate is gentiobiose peracetate linked at the C 1 - position to an immunogenic carrier having the follow¬ ing Formula (X) : Another preferred lipid A analog/immunogenic carrier conjugate is an acyl glucose molecule linked at the C 1 -position having the Formula (XI) : Another preferred lipid A analog/immunogenic carrier conjugate is an acyl gentiobiose molecule linked at the C 6' -position to an immunogenic carrier having the Formula (XII) : (XII) Another preferred lipid A analog/immunogenic carrier conjugate is an acyl glucose molecule linked at the C 6' -position to an immunogenic carrier having the Formula (XIII) : (XIII) The lipid A analog/immunogenic carrier conjugates linked at the exposition may be prepared (see Scheme I) by treatment of an appropriately substituted and protected carbohydrate with a reagent capable of linking the carbohydrate to the i_mmunogenic carrier having Formula (XIV) either directly or through a linker group. For example, gentiobiose or glucose peracetate (Formula (XV), n = 1 or 2; R 1 , R 2 , R 3 and R 4 = Ac) may be treated with HBr in glacial acetic acid to give the acetobromosaccharide derivative (XVI) . The acetobromo derivative (XVI) may then be reacted with a linker such as 2-aminoethanol (XVII) , 3-amino- propanol, 4-aminobutanol, or 5-aminopentanol, to give the aminoethyl peracetate saccharide derivative (XVIII) which may be coupled to the immunogenic carrier (XIV) with, for example, 1-ethyl-3- (3- dimethyl-aminopropyl) carbodiimide (EDAC) to give (IV) . The use of EDAC to form conjugates between amino containing substances and proteins is disclosed in U.S. Patent No. 4,526,714 to Feijen et al. Alternatively, the immunogenic peptide may be derivatized with a second bifunctional spacer group, such as succinic acid or any of the other linker groups having two carboxyl groups, to give the derivative (XIX) . The derivative having Formula (XIX) may then be condensed with (XVIII) in the presence of EDAC to give (IV) . Where R 4 is a phosphate group, the hydroxy group may be phosphorylated prior to preparation of the conjugate by reacting an appropriately protected glucose or gentiobiose compound with the product of phosphorous acid and N-methylimidazole in the presence of HgCl 2 . This reaction allows the preparation of the phosphate ester at the free hydroxyl group. The ratio of lipid A analog molecules per immuno- genie carrier molecule may vary considerably according to the molecular weight of the immunogenic carrier, the number of binding sites on the immunogenic carrier capable of being coupled to the lipid A molecule, and the antigenic characteristics of the particular lipid A molecule. In general, the ratio of lipid A analog molecules to immunogenic carrier molecules may be about 1:1 to about 200:1. Preferably, the ratio may range from about 5:1 to about 100:1. More preferably, where the immunogenic carrier is diphtheria toxoid, the ratio of lipid A analog molecules to diphtheria toxoid molecules may range from about 5:1 to about 40:1. Scheme I H0-CH 2 CH 2 -NH 2 (XUII) _l_mmunogenie Carrier (XIV) Where the immunogenic carrier is linked to the lipid A molecule at the exposition, the conjugates may be prepared according to scheme II by the selective tritylation of, for example, the C 6 '-position of gentiobiose (XX) with trityl chloride (XXI) to give the C 6' -trityl gentiobiose derivative (XXII) . Acyla- ion of the remaining hydroxyl groups with, for example, acetic anhydride gives 6'-O-tritylgentiobiose heptaacetate (XXIII) . Removal of the trityl group with, for example, HBr gives the C 6 '-hydroxygentiobiose heptaacetate derivative (XXIV) which may be linked to an immunogenic carrier such as the succinate deriva¬ tive of diphtheria toxoid to give (XXV) . Scheme II The invention also relates to intermediates useful for the preparation of the lipid A analog/- immunogenic carriers of the invention. In particular, the invention relates to a 1-bromo-saccharide having the Formula (XXVI) : (XXVI) wherein n=2. The invention also relates to the intermediate having Formula (XXVII) : (XXVII) The invention also relates to lipid A analog/linker conjugates having the Formula (XXVIII) : ~CH CH ~NH (XXVIII) In particular, the invention relates to a lipid A analog/linker conjugate having the Formula (XXIX) : (XXIX) The invention also relates to a lipid A ana¬ log/linker conjugate having the Formula (XXX) : (XXX) The invention also relates to a lipid A ana¬ log/linker conjugate having the Formula (XXXI) : (XXXI) The invention also relates to a lipid A ana¬ log/linker conjugate having the Formula (XXXII) : (XXIII) The conjugates of the invention may be purified by any method known to those of ordinary skill in the art. For example, the conjugates may be purified by reverse phase chromatography, ion exchange chromatog- raphy, size exclusion chromatography, or by dialyzing the reaction product against water followed by freeze- drying. Alternatively, the conjugates may be purified by passing a solution of the conjugate through a column having anti-lipid A antibodies immobilized on a solid phase support. (See the Examples section of the application for a method of preparing antibodies to lipid A.) _. The invention also provides for a polyvalent vaccine comprising (a) a lipid A analog/immunogenic carrier conju¬ gate of the invention, (b) at least one Gram negative and/or Gram positive bacteria, or antigenic determinant thereof, optionally conjugated with an immunogenic carrier, (c) a pharmaceutically acceptable carrier, and (d) optionally, an adjuvant. Preferred Gram negative bacteria are the Pseudo- monadales and Eubacteriales. As Pseudomonadales, included are the genera Paeudomonas, Xantohmonas, Zoogloea, and Gluconobacter . Preferred are the Pseudomonas which include bacteria of the species Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas pseudomallei , Pseudomonas mallei, and Pseudomonas saccharophilia. Most preferred are the bacteria of the species Pseudomonas aeruginosa. Included in the Eubacteriales are the families Azotobacteraceae, Rhizobiaceae, Achromobacteraceae, Enterobacteriaceae, and Brucellaceae. Preferred are the Enterobacteriaceae which include the Escheri- chieae, Klebsielleae, Proteeae, Yersinieae, and Erwinieae. Among the Escherichieae, the genera Eachericbia, Edwardaiella, Citrobacter, Salmonella, and Shigella are preferred. Included as preferred in the genus Eacherichia are the bacteria of the species Eacherichia coli . Among the Klebsielleae, the genera IQebsiella, EnteroJacter, Hafnia, and Serratia are preferred. Included as preferred among the genus Klebsiella are the bacteria of the species Kebsiella pneumoniae^; Klebsiella ozaenae, Klebaiella oxytoca, and -Klebsiella rhinoacleromatia. Preferred Gram positive bacteria are the Micro- coccaceae and the Streptococcaceae. Included in the Micrococcaceae are the genera Micrococcus, Staphyl- coccus, and Planococcus. Among the Staphylococci , bacteria of ' the species Staphlycoccus aureus and Staphlycoccus epidermidis are preferred. Included in the Streptococcaceae are the genera Streptococcus, Leuconostoc, Pediococcus, Aerococcus, and Gemella. Preferred are bacteria of the species Streptococcus faecalis, Streptococcus faecium, Streptococcus mitis, and Streptococcus pyogenes. This invention also provides for a polyvalent hyperimmune gammaglobulin comprising an effective opsonizing and neutralizing amount of a gammaglobulin specific to the lipid A/immunogenic carrier conjugate of this invention and an effective opsonizing amount of an 0 antigen specific gammaglobulin. The polyvalent hyperimmune gammaglobulin can further comprise at least one antigenic determinant of a Gram positive bacteria. The hyperimmune gammaglobulin can be separated from ^animals which have been hyperimmunized. An animal can be hyperimmunized by administration, preferably by intramuscular or subcutaneous injection, of one or more conjugates of this invention. Typical¬ ly, the conjugates can be effectively administered in a range of between about 10 and 100 μg of polysaccha¬ ride as a component of each conjugate. A range of between about 12.5 and 50 μg of polysaccharide, as a component of each conjugate, is preferred. Approximately 4 to 6 weeks following the initial administration of a conjugate of this invention, the animal can be reinjected with between about 10 and 100 μg of polysaccharide as a component of each conjugate. Preferably, the animal receives an injection of between about 12.5 and 50 μg of polysaccharide, optionally as a component of each conjugate. Generally, a second injection exponentially increases the number of B cells bearing antigen-specific cell- surface antibodies. A third or subsequent injection can also be administered which duplicates the second injection. As is well known in the art, this repeated boosting of antigen results in a hyper_Lmmunized animal. An adjuvant suitable for injection may also be used to enhance the immune response to the conjugates. Plasma separated from an immunized animal contains a relatively large concentration of antibody to the injected antigens and is appropriately called a hyperimmune serum. Methods for separating the hyper_b__rmune serum from a hyperimmunized animal are known in the art as exemplified by Harlow et al. , Antibodies, Cold Spring Harbor Laboratory, 1988. Hyperimmune gammaglobulin preparations can be prepared from the hyperimmune serum using various methods known in the art such as the Cohn fraction method See, Conn, E.J. et al . , J. Amer. Chem. Soc. 68: 4.59 (1946) and U.S. Patent No. 4,965,068. In general, plasma may be separated into fractions which are immunologically active (immunoglobulins) and immunologically inactive (albumins and clotting factors) by fractional precipitation using ammonium sulfate. The immunologically active fraction is then briefly digested with pepsin to remove the complement- activating component of the molecules to render them less immunogenic. The immunologically active component then is recovered by dialysis and fractional precipitation or centrifugation. * See Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, (1990) . The terms "opsonizing" and "neutralizing" concern activity which relate to both humoral and cellular immunity. * For example, with respect to humoral immunity, lipid A-specific antibodies "neutralize" lipid A, but not any other type of toxin. In contrast, cellular immunity is not dependent entirely upon circulating antibody. For example, if Streptococcus pneumoniae, an encapsulated coccus, is injected into a mouse, death generally follows. However, if antibody specific to S. pneumoniae is injected into the mouse prior to challenge, death will not generally occur. This result is due to the antibody binding to and altering the surface of the challenging organism, thereby promoting phagocytosis. This activity is typically referred to as "opsonizing" activity. Methods for analyzing opsonizing and neutralizing activities are known in the art. Included in these methods are animal models such as the Rabbit Dermal Schwartzman Model, the Murine Hog Mucin Model, and the Rat Neutropenic Model. These models are well known to those of ordinary skill in the art. For example, the Murine Hog Mucin Model, as described by Teng, N.N.H. et al . , Proc. Natl . Acad. Sci . USA 82: 1790-1794 (1985) , can be used to measure opsonic activity. Using the Murine Hog Mucin Model, mice are inoculated with graded doses of an 18-hour culture of Pseudomonas aeruginoaa 134 VA (Fisher Devlon type I) , Paeudomonaa aeruginoaa Fisher Devlon type VI, Klebaialla pneumoniae, Eacherichia coli 0111:B 4 , Escherichia coli 017 or Streptococcus pneumoniae. Graded doses of one or more of the 18-hour cultures are mixed with equal volumes of 14% hog gastric mucin, and 0.5 ml of the bacterial suspension is injected into the mouse peritoneum. Prior to this injection, the hyperimmune gammaglubulin of this invention is injected into the mice, with controls not receiving this injection. Protection against lethal bacteremia can then be determined by calculating the protection ratio. Tliύs ratio is defined as the ratio of LD 50 between the hyperimmune gammaglubulin treated group and the non-treated control group. A protection ration of at least 5:1 is considered to indicate positive opsonic activity. A 10:1 ratio is preferred, with a ratio of at least 15:1 being most preferred. In general, the gammaglobulins specific for the Grama negative and Gram positive bacterial conjugates of this invention will show opsonic activity. As a further example, neutralizing activity can be screened by the Dermal Schwartzman Reaction as also described by Teng, N.N.H. et al . , supra. In this model, rabbits are inoculated in the ear vein with the hyperimmune gammaglubulin of this invention. The rabbits are given challenge doses of lipopoly- saccharide or Iscove's medium ,. as a control. Neutralizing activity is evident when there is a reduction of hemorrhagic necrosis at the site of challenge injection. In general, neutralizing activity will be indicated when the incidence of positive reactions is less than about 90%. It is preferred that the incidence of positive reactions will be less than about 15%, with an incidence of less than about 12% being most preferred. Preferred O antigen specific gammaglobulins are gammaglobulins which are specific to the 0 antigens of the Gram negative bacteria of this invention. Espe¬ cially preferred are gammaglobulins specific to the O antigens of Eacherichia coli, Pseudomonas aeruginoaa, Klebaiella pneumoniae, Klebaiella oxytoca, and Klebai - ella ozanae. This invention also concerns a method of prepar¬ ing a polyvalent hyperimmune gammaglobulin comprising (a) .Immunizing an animal with the lipid A analog/immunogenic carrier of this invention and at least one Gram negative and/or Gram positive bacteria, or antigenic determinant thereof, optionally conjugated with an immunogenic carrier, (b) optionally boosting said animal with the -Immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said bacteria and neutralize LPS. This invention further provides for a method of treating Gram negative bacterial sepsis comprising administering an effective neutralizing amount of a gammaglobulin specific to the lipid A/immunogenic carrier conjugate of this invention and an effective opsonizing amount of 0 antigen specific polyvalent hyperimmune gammaglobulin to an animal in need of such treatment. Treatment can be prophylactic, or it can be to animals actually suffering from a bacterial infection. Generally, an effective neutralizing amount is considered to be between 50 and 1000 mg/kg of gammaglobulin to body weight. Preferably, between 100 and 500 mg/kg of gammaglobulin to body weight is to be administered. With respect to opsonic activity, a range of between 50 and 1000 mg/kg body weight is considered to be effective. Preferably, between 100 and 500 mg/kg of gammaglobulin to body weight may be administered. This invention also provides for a kit for treating Gram negative bacterial sepsis comprising a carrier compartmentalized to receive in close confine- ment therein one or more container means wherein a first container contains a lipid A analog/immunogenic carrier of the invention. The kit can also include at least one other container means which contains at least one Gram negative and/or Gram positive bacteria, or antigenic determinant thereof, conjugated with an immunogenic carrier. The Gram negative and Gram positive bacteria may be killed or attenuated bacteria. The preferred Gram negative and positive bacteria Ure those which are preferred in the polyvalent vaccine. Optionally, an additional con¬ tainer means may be present which contains an adju¬ vant. This invention also provides for a kit wherein the first container contains gammaglobulins specific for the lipid A analog/immunogenic carrier of this invention. In this case, a second container can be provided which contains gammaglobulins specific for said Gram negative or Gram positive bacteria, or antigenic determinant thereof. The preferred Gram negative and positive bacteria are those which are preferred in the polyvalent vaccine. Administration of the vaccines or hyperimmune gammaglobulin may be parenteral, intravenous, intra¬ muscular, subcutaneous, intranasal, or any other suitable means. Preferably, administration is by subcutaneous or intramuscular means. The dosage administered may be dependent upon the age, weight, kind of concurrent treatment, if any, and nature of the antigen administered. In general, the conjugates may be administered at a dosage of 0.0001 to 25.0 •rag/kg of conjugate per animal weight. The initial dose may be followed up with a booster dosage after a period of 4 weeks to enhance the immunogenic response. Further booster doses every 4-6 weeks may be administered for as long as the risk of infection and septic shock exists. The lipid A analog/immunogenic carrier conjugates and other antigenic conjugates of this invention useful in the method of the present invention may be employed in such forms as capsules, liquid solutions, suspensions or elixirs for oral administration, or sterile liquid forms such as solutions or suspensions. Any inert carrier is preferably used, such as saline, or phosphate-buffered saline, or any such carrier in which the compounds used in the method of the present invention ___ ve suitable solubility properties for use in the method of the present invention. Preferably, the lipid A analog/immunogenic carrier conjugates are in lyophilized form or in a weakly acidic buffer solution. The Gram negative and Gram positive conjugations are preferably stored in a weakly basic solution. Such solutions may be kept frozen to improve stability. The vaccines may be in the form of single dose preparations or in multi-dose flasks which can be used for mass vaccination proGrams. Reference is made to Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, Osol (ed.) (1980); and JVew -Trends and Developments in Vaccines, Voller et al . (eds.), University Park Press, Baltimore, MD (1978), for methods of preparing and using vaccines. The vaccines of the invention comprising the lipid A analog/immunogenic carrier conjugates of the invention may further comprise adjuvants which enhance production of lipid A analog-specific antibodies. Such adjuvants include, but are not limited to, various oil formulations such as Freund's complete adjuvant, the dipeptide known as MDP, saponin, alumi¬ num hydroxide, Bordetella pertussis, dephosphorylated lipid A, or an interferon. Freund's adjuvant is an emulsion of mineral oil and water which is mixed with the immunogenic sub¬ stance. Although Freund's adjuvant is powerful, it is usually not administered to humans. Instead, the adjuvant alum (aluminum hydroxide) may be used for administration to a human. The conjugate may be absorbed onto the aluminum hydroxide from which it is slowly released after injection. The lipid A analog/immunogenic peptide conjugate may also be encapsulated within liposomes according to Fullerton, U.S. Patent No. 4,235,877. Having now generally described the invention, the same may be further understood by reference to the following "examples, which are not intended to be limiting unless so expressly stated. EXAMPLES Example 1: Preparation of a Gentiobiose peracetate- BSA conjugate Gentiobiose octaaeetate, obtained from Sigma Chemical Co. (St. Louis, MO) , was converted into the acetobromo derivative by dissolution in dichloro¬ methane (1 gm in 5 ml) and treatment with 33% HBr in glacial acetic acid (10 ml) (Fluka) at 5°C for 45 minutes. A volume of dichloromethane equal to the total volume of the reaction mixture was added and the mixture transferred to a separatory funnel. The organic phase was washed with an equal volume of saturated sodium bicarbonate solution (3-4 times) , followed by two portions of water. The organic layer was then dried over MgS0 4 , filtered, and the solvent evaporated to give a syrup. The acetobromogentiobiose was crystallized from dichloromethane and diethyl ether, filtered and dried in a vacuum desiccator. The crystalline compound gave a single spot on T.L.C. and the H-l NMR spectrum was correct. Acetobromogentiobiose (5 gm in 5 ml chloroform) was coupled to 2-aminoethanol (0.5 ml, approx. 0.5 gm, in 1 ml chloroform) in the presence of drierite (1 gm; W.A. Hammond Drierite Co.). The mixture was stirred overnight at room temperature then filtered. The resulting aminoethyl gentiobiose heptaacetate (AGH) was first purified by chromatography on a column of silica gel (1.6X24 cm, 50 ml total volume) using an ethyl acetate-ethanol solvent mixture (19:1) as the eluting solvent. The fractions were screened by TLC using ethyl acetate-ethanol (19:1) to elute. The fractions containing the aminoethyl derivative were combined, the solvent evaporated in vacuo, and the product crystallized from hot ethanol at 55°C. The crystalline compound gave a single spot on T.L.C. and had a melting point of 176-177°C. Elemental analysis showed C, 49.44%; H, 6.05%; N, 2.1% (calculated for C 28 H 40 O 28 N: C, 49.56%; H, 5.94%; N, 2.06%). In order to increase the number of free carboxyl groups and solubility of bovine serum albumin (BSA) , the BSA (10-50 mg in PBS at 4-5 mg/ml) was slowly treated with succinic anhydride (5-10 mg/mg protein; Aldrich Gold Label) at pH 8-9. The * pH was maintained at 8-8.5 by the addition of concentrated NaOH. The reaction mixture was then extensively dialyzed against water and the resulting succinylated BSA (Suc-BSA) freeze-dried. Dioxane was the solvent found capable of dissolv¬ ing both the protein carriers and the AGH ligand. Suc-BSA (10 mg) was coupled to AGH in 20% 1,4-dioxane (2-3 ml; Fisher Scientific) in the presence of 1- ethyl-3- (3-dimethylaminopropyl) -carbodiimide (10 mg; EDAC) . The pH was monitored using pH paper and maintained1at 5-6 by the addition of dilute HCl. Three additional portions of EDAC (10 mg) were added after 30 minute intervals. The resulting conjugate (Suc-BSA-AGH) was dialyzed against water and then frozen. Analysis for reducing sugar by the phenol- sulfuric acid assay showed that the conjugate consist- ed of 27.4% AGH. This is about 35 moles of ligand/mole of protein. Example 2: Preparation of a Gentiobiose Peracetate-DT Conjugate Diphtheria Toxoid (DT) from SSVI (Bern, Switzer- land) was coupled to AGH (obtained according to Example 1) in 25% 1,4-dioxane and 0.05M NaCl in the presence of EDAC. The resulting conjugate (DT-AGH) was dialyzed against water and freeze-dried. Analysis demonstrated that 16% of the conjugate consisted of AGH. Example 3: Preparation of Gentiobiose Heptaacetate Cl Conjugate Vaccine (Succinγl-DT-APGH) Succinylation of Diphtheria Toxoid The contents of two bottles of diphtheria toxoid lot M (150 ml each, 3.0 mg/ml) were combined, and 200 ml of sterile 0.05 M NaCl solution was added to give a final volume of 500 ml. Succinic anhydride (1.0 g, crystalline, Aldrich) was added slowly with stirring while maintaining the pH between 8.0 and 8.5 with sterile 3.0 M NaOH solution during 2 h. The resultant reaction mixture was then neutralized to pH 7.0 with 1.0 M HCl and was dialyzed against three changes of pyrogen free water (3X10 L) in 48 hours. The dialyzed solution was distributed into 50 sterile bottles and lyophilized under sterile conditions. This ι$_aterial was tested for the presence of primary amino groups by the TNBS reagent method (H.A. Kemp and M.R.A. Morgan; J. Immunol . Meth. 94 : 65 -72 (1986) ) . It was found that 95% of the available primary amino groups were substituted. Preparation of Acetobromo Gentiobiose Crystalline gentiobiose octaacetate (2.0 g, Sigma) is dissolved into dichloromethane (99+%, 5 ml) , and to this solution is added 33% HBr in glacial acetic acid (8 ml) in a 100 ml Round Bottom Flask. The flask is placed on an ice/water bath for 45 min. While stirring the reaction mixture, 30 ml of dichloromethane was added, and the mixture was washed successively with cold water (30 ml) , saturated sodium bicarbonate solution (2 X 40 ml) , and cold water (40 ml), using a separating funnel. The dichloromethane phase was dried with anhydrous sodium sulfate (2.0 g) and evaporated to dryness in a rotary evaporator at or below 40°C. The dry product was dissolved into diethyl ether (5 ml) and allowed to crystallize for one h at room temperature and then.left overnight at 5°C. The crystalline compound was filtered, washed with cold ether and dried under vacuum in a dessieator at room temperature. The yield was 1.4 g (70%) . This material gave a single spot on TLC using ethyl acetate: n-hexane (2:1) as solvent, Rf •= 0.43. This was crystalline acetobromo gentiobiose. Coupling Acetobromo Gentiobiose to 3-amino Propanol Crystalline acetobromo gentiobiose (1.0 g) was mixed with dry chloroform (5 ml, 99.9+%) and to this solution was added dry Drierite (1.0 g) . The mixture was stirred, and 3-amino propanol (400 μl, 98%) was added. The reaction vial was wrapped with aluminum foil and stirred for 16 h at room ' temperature. The reaction mixture was filtered on a sintered glass funnel, and the filtrate was washed once with cold water (10 ml) , and then the chloroform phase was dried with anhydrous sodium sulfate and filtered again. The clear filtrate was loaded onto a washed silica gel G-60 column "' (2.0 X 40 cm) and eluted with ethyl acetate:ethanol (3:1). The column fractions were tested by TLC in ethyl acetate:ethanol (19:1) , and the fractions containing the component moving with Rf value of 0.65 were combined and evaporated to dryness in a rotary evaporator. The dry product was dissolved into ethanol (10 ml) that had been warmed to 50°C. The mixture was then allowed to slowly cool to room temperature for 2 h. The aminopropyl gentiobiose heptaacetate (APGH) crystallized under these conditions. The crystalline material was filtered, washed with cold ethanol (5 ml) and dried under vacuum in a desiccator at room temperature. The yield was 350 mg (36%) , m.p. 164- 165°C. The analysis calculated for C 29 H 42 0 18 N was: C, 50.25%; H, 6.06%; N, 2.02%. Found C, 49.50%; H, 6.12%; N, 1.98%. The final compound was aminopropyl gentiobiose heptaacetate (APGH) . Coupling Succinylated DT to APGH All procedures were performed in a sterile laminar flow hood. To each of six bottles of lyophilized Suc.DT (25 mg in each bottle), 3.0 ml of pyrogen free water was added. The Suc.DT solution from the bottles was combined and filtered through a sterile 0.22 μm membrane. Final volume was 18 ml. 1,4-Dioxane (4.5 ml) was filtered through 0.22 μm membrane and added to the above solution to make 20% dioxane (22.5 ml total volume) . Aminopropyl gentiobiose heptaacetate (APGH, 100 mg) was dissolved into 1,4-dioxane (4.0 ml), and to this solution was added pyrogen free water (16 ml) to make 20% dioxane (20 ml) . This solution was sterile filtered through 0.22 μm membrane and added to the solution of Suc.DT. Combined volume was 42.5 ml. The pH of this mixture was adjusted t ' d 5.8 (autoclaved probe used) with sterile 0.1 N HCl. l-Ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, 1.02 g) was added to the reaction mixture and the pH maintained between 5.5 and 5.6 for 1 h by adding sterile 0.1 N HCl. After one hour 1.10 g of EDC was added and the pH maintained between 5.4 and 5.5 for 1 h. The reaction mixture was then transferred into a sterile (autoclaved) dialysis tubing and dialyzed against four changes of sterile pyrogen-free water (4 X 6 L) in 36 h. The dialyzed solution was distributed into multiple dose sterile vials and lyophilized. This was Suc.DT-APGH lot #AKB X-26. The hapten content of this conjugate was determined by the phenol sulfuric acid method (Dubois et al . , Anal . Chem. 28:350 (1956)) and was found to be 4.2%. Sterility, toxicity and pyrogenicity of this product were tested by standard procedures. Example 4: Binding of Gentiobiose-Protein Carrier Conjugates to LPS specific antibodies Although gentiobiose octaacetate is insoluble in water, the protein conjugates dissolve easily. These soluble conjugates were tested for their ability to bind anti-lipid A human monoclonal antibody (obtained by fusion of the heteromyeloma SHMA6(H4) with Epstein Barr virus (EBV) treinsformed lymphocytes from the spleen of an animal immunized with the J5 mutant of E. coli 0111-B 4 which is deficient in the enzyme uridine 5' " '-diphosphategalactose 4-epimerase which prevents attachment of the side chains responsible for the marked antigenic diversity among Gram-negative bacteria) by enzyme-linked solid phase immunoassay (ELISA) . As shown in Table 1, both BSA-gentiobiose octaacetate and diphtheria toxoid-gentiobiose octaacetate conjugates were able to bind the human monoclonal antibody. These experiments demonstrate that the important binding epitope remained intact during the conjugation process. Therefore, it would be expected by one of ordinary skill in the art that anti-lipid A antibodies would be produced in vivo upon administration of the lipid A analog/immunogenic carrier conjugates to an animal. Table 1: Binding of the lipid A Antibody to Gentiobiose Octaacetate/Pro ein Conjugate Suc-BSA- GH conjugate 2,500 Sue-BSA-Control 640 DT-AGH conjugate 48 DT control 0 AGH in ethanol 1,280 _3. coli Lipid A 2,560 Suc-DT-APGH 3,200 Suc-DT (control) 0_ Example 5: Preparation of the Vaccine Formulation of Alum The alum used was a sterile suspension called "Rehsorpta-S 1 * and was obtained from Armour Pharmaceutical Company. This preparation has 2% A1 2 0 3 (w/v) as specified by the manufacturer. Rehsorptar suspension (104 ml) was diluted with sterile 0.9% NaCl (221 ml) to give 325 ml of diluted alum suspension. The diluted alum suspension was distributed into sterile vials (5.0 ml in each) and was autoclaved. This formulation contains 6.4 mg A1 2 0 3 (w/v) or 3.38 mg of aluminum metal per ml of suspension. Composition of the Vaccine The Suc.DT-APGH lipid A analog Cl-conjugate vaccine has the following composition: Hapten 4.2% Protein 84% Moisture , 12% Molar ratio of Hapten : Protein 4.2:1 Each bottle of lyophilized vaccine (1 ml fill) has 1.5 mg vaccine on a dry weight basis. One human dose will be 100 μg of vaccine. Therefore, each vial of lyophilized vaccine lot AKB X-26 (1 ml fill) has 15 human doses. Formulation of Vaccine Each bottle of lyophilized vaccine will come with a bottle of diluted alum suspension (5 ml) that has been prepared with water for injection and a bottle of sterile 0.9% NaCl (5 ml) for reconstitution of the vaccine. The lyophilized vaccine will be reconstituted with 4.0 ml of sterile 0.9% NaCl. This solution will be mixed with 3.5 ml of diluted alum suspension to give a final volume of 7.5 ml of vaccine formulation. This formulation will contain 200 μg of vaccine/ml and 1.57 mg of aluminum/ml. The human dose will be y.5 ml of this vaccine formulation. Therefore, one human dose will contain 100 μg vaccine and 785 μg of aluminum. The aluminum to protein ratio will be 7.8:1. The reconstituted vaccine will be used within 2 h. Any remaining vaccine in the bottle will be discarded. Exclusion Criteria Vaccine lots failing to meet the following criteria, even if they meet the safety and pyrogenicity standards, will not be used in clinical trials: a. The molar ratio of hapten to protein must be 4.0:1 or higher. b. A 50 μg dose of the vaccine administered to rabbits with alum as adjuvant should give at least a 20 fold rise in titer against the hapten, after two immunizations four weeks apart, as determined by ELISA using the BSA conjugate of the hapten as the plate antigen. Stability The following five tables are concerned with the stability of the gentiobiose-toxoid conjugate. The vaccine is stored at minus (-) 70°C. It is used within 2 hours of thawing and is disposed of after that time. It contains no preservative. Table 2 clearly shows that the amount of free hapten which has dissociated from the toxoid is constant at about 5% over 35 months. In Table 3 evidence is presented for the maintenance of strong immunogenic potential. Tables 4, 5, and 6 show that the succinylated but unconjugated- toxoid does not induce an immune response to the hapten (Table 4) , but that the * lipid A conjugated vaccine is very -Immunogenic after 7 months of storage (Table 5) . Table 6 presents data from a dose response assay in rabbits. This experiment was performed after over 1 year storage at minus (-) 70°C, the required temperature for storage. The data show that the 1 meg dose is not "" effective; the 10 meg dose is more immunogenic; the 50 and the 100 meg doses, which include the dose to be used clinically, are very immunogenic. Thus, although the moisture content is high, at the recommended sub-zero temperature for storage -Lmmunogenicity is retained. The gentiobiose octaacetate protein/diphtheria toxoid conjugate was administered to rabbits according to the immunization schedule outlined in Table 7. Antibody titers were then determined by ELISA. Sera from rabbits 1 and 2 was then assayed for antibodies specific for a series of antigens. Table 8 shows that the rabbit sera has reactivity with LPS and diphtheria toxin, but little reactivity with APGH. "Substitution of succinylated diphtheria toxin by APGH expressed as weight percent, Table 8 a All data is expressed as the dilution giving an optical density reading of 0.4 units. Rabbit sera was also assayed .against a tetanus toxoid-APGH conjugate (TT-APGH) , which does not have a spacer group. The results listed in Table 9 show that the antibodies are also specific for this conju¬ gate, confirming that the antibodies are reactive with the APGH part of the molecule. Table 9 Antibody specificity for LPS on boiled J5 was then determined by specific adsorption experiments. In each experiment, the sera of rabbits was diluted 1:500 and adsorbed twice with boiled J5 organisms at 10% volume of packed cells. The first adsorption was for 2 hours followed by a second overnight adsorption (both at 4°C) . The J5 organisms were analyzed by GLC and found to be free of galactose. The sera from the final bleeds of rabbits 1 and 5 were then assayed pre- and pos -adsorption by ELISA using TT-APGH. The results are shown in Table 10. Table 10 As shown in Table 10, there was significant reduction of antibody titer when the sera was adsorbed to J5 LPS. These data confirm the expectation that the sera of animals immunized with the conjugate is reactive with LPS on boiled J5. Example 6 Preparation of fi'-substituted Gentiobiose Heptaacetate Succinyl-Diphtheria Toxoid (SUC-DT) Conjugate A. Preparation of 6'-O-Trityl-Gentiobiose Gentiobiose (4 gm, Sigma) was added slowly with stirring to pyridine (60 ml) and* to the resulting suspension was added with stirring triphenylmethyl- chloride (4 gm, Aldrich) . The mixture was stirred overnight at-room temperature. The solvent was evaporated off in a rotary evaporator and the resident pyridine was removed by co-distillation with toluene (3 x 15 ml) . The solid product was dissolved into dichloromethane (30 ml) and the solution was loaded onto a washed silica gel column (2.0 x 70 cm) . The excess trityl chloride was first eluted with ethyl acetate. The 6'-0-trityl- gentiobiose was then eluted with methanol. The fractions containing the product were combined and evaporated to dryness. B. Preparation of 6 '-O-Trityl-Gentiobiose Heptaacetate 6'-O-Trityl-gentiobiose was dissolved into pyridine (35 ml) and to this solution was added with stirring 17 ml of acetic anhydride (Aldrich Chemical Corporation, Milwaukee, WI) . The mixture was stirred overnight at room temperature and then poured into 400 ml of crushed ice. The resulting precipitate was filtered, washed with cold water and dried under vacuum (yield 5.5 gms) . C. Preparation of 6' -O-Hydroxy-Gentiobiose Hep¬ taacetate (C-6'-HGH) 1.0 gm of 6'-O-trityl-gentiobiose heptaacetate was dissolved into 5.0 ml of glacial acetic acid and to this solution was added 33% HBr in glacial acetic acid (Fluka) to make 2% HBr. The mixture was poured into 50 ml of iced water and the resulting precipitate (trityl bromide) was filtered out. The aqueous solution was extracted with dichloro- methane (50 ml) and the organic phase was washed with saturated NaHC0 3 solution, cold water, dried with anhydrous sodium sulfate and evaporated to dryness. The solid product was crystallized from diethyl ether. Yield 150 mg. D. Preparation of 6'-Hydrox -GentiobioseHepta¬ acetate Succinyl-Diphtheria Toxoid (SUC-DT) Conjugate. Succinyl-DT (7.2 mg) was coupled to 6'-hydroxy¬ gentiobiose heptaacetate (3.5 mg) in 20% 1,4-dioxane (Fisher Scientific Co.) in the presence of l-ethyl-3- (3-dimethylaminopropyl) -carbodiimide (18 mg, EDAC) . The pH was maintained between 5.7 and 6.0 with 0.1N HCl. Two additional portions of EDAC (10 mg) were added at 30 minute intervals. The resulting conjugate (6'-O-SucDT-GH) was dialyzed against water and then freeze dried. Analysis for reducing sugar by the phenol-sulfuric acid assay showed that the conjugate consisted of 20% 6'-HGH. This is about 30 moles of ligand/mole of protein. Example 7 Immunogenicity Studies of 6' -0-SucDT-GH Vaccine The vaccine was prepared for immunization studies by dissolving the lyophilized conjugate in sterile phosphate buffered saline to a final concentration of 200 ug/ml, followed by adsorption to aluminum oxide (Rhesorptar) overnight at 4°C. The vaccine was administered intramuscularly to NZW rabbits at 50 ug per dose, and sera were collected periodically during the immunization schedule. Vaccine was administered at weeks 0, 3, 6, 9, 12, and 16. Immunogenicity was determined by ELISA using a heterologous tetanus conjugate vaccine (1' -0-TT-APGH) as the plate antigen at a concentration of 2 ug/ml. Sera were run in triplicate beginning at a 1/50 dilution, and data reduced to*-voptical density (O.D.) units, the titer at which the O.D. is equal to 1.0. The results for serial bleeds obtained 1 week after each dose shown in Table 11. Table 11 Immunogenicity of 6' -O-Suc-DT-GH Vaccine The results show that the vaccine induced ligand- specific antibodies in each rabbit immunized. To determine the protective efficacy of these antibodies, their ability to protect mice in a hog mucin challenge model was assayed. Pooled sera obtained at week 14 from immunized rabbits, or pooled sera obtained from control rabbits not receiving the vaccine, were sterile filtered and administered mtraperitoneally to NSA mice at a dose of 1.0 ml per mouse. Two hours later the mice were challenged with from 4.8 x 10 3 to 4.8 x 10 5 Pseudomonas aeruginosa strain 134VA suspended in a 14% preparation of hog mucin. Mice were examined hourly for the next 72 hours and the time of death recorded. Table 12 reports the results of these studies. Table 12 Hog Mucin Challenge Model Protection from Challenge with Pseudomonas aeruginosa These data demonstrate that the vaccine was immunogen¬ ic and produced antibodies that protect mice against challenge with a virulent Gram negative bacteria. Example 8 Active and Passive Protection from a Localized Shwartzman Reaction A. Active Immunization: Three rabbits were immunized twice with Suc-DT-APGH (see Example 3) in aluminum hydroxide gel (20:1, w/w) and subsequently received 4 injections of the antigen in saline. To test development of a localized Shwartzman reaction, rabbits each received an intracutaneous injection of 50 μg E. coli 06 LPS followed 23 hours later with 25 μg LPS in 0.1 ml saline, intravenously. Only 1 of 3 immunized rabbits showed a positive erythema, although no hemorrhagic reaction, whereas 3 of 3 non-immune rabbits were positive for erythema and 2 had hemor¬ rhagic reactions. No necrosis was observed. B. Passive Immunization: Rabbits whose sera were to be tested were immunized once with Suc-DT-APGH (Example 3) in complete Freund's adjuvant, followed by 7 injections of the antigen in saline. Serum obtained from these hyperimmunized rabbits ^ had an antibody titer of 1:64*00 measured in an ELISA with TT-APTGH. Test rabbits were injected intracutaneously with 100 μg E. coli 06 LPS in 0.2ml saline. After 21 hours, rabbits received 15 ml of the hyperimmune serum or normal serum intravenously (in the ear vein) and 2 hours later were challenged by intravenous injection of 20 μg LPS in 0.2 ml. In the control rabbits receiving normal serum, 4 of 5 showed hemorrhage and 1 had tissue necrosis. Only 1 of 5 rabbits receiving hyperimmune serum showed a positive hemorrhagic reaction and none had any signs of necrosis. Therefore both active and passive immunization against Suc-DT-APGH afforded significant protection against the LPS-induced Shwartzman reaction, further confirming the capacity of the composition to induce anti-LPS antibodies capable of acting in vivo. Example 9 Preparation of __7. coli-Toxin A Conjugate A. Preparation of E. coli Lipopolysaceharide E. coli 018 is obtained and grown in Trypticase soy broth. The cells are washed by centrifugation. Lipopolysaceharide (LPS) is prepared by phenol extrac¬ tion from the cell paste according to a modification of the phenol-water method of Westphal et al . , Naturforscher 7: 148-155 (1952) , and then sequentially treated with DNase, RNase, and pronase, followed by a second phenol extraction and ethanol precification dialysis and by lyophilization. B. Preparation of Toxin A Toxin ;A is purified from the supernatant of a culture of Pseudomonaa aeruginoaa 103 as described by Shiloach, J. et al., in Yu, P.L., ed., Fermentation Technologiea : Induatrial Appliationa. C. Preparation of E. coli-Toxin A Conjugate The conjugate is prepared according to the method disclosed in Chu, C.Y. et al . , Infec. Immun. 40: 245- 256 (1983) , incorporated by reference herein. The purified lipopolysaceharide is detoxified and the polysaccharide is purified by gel filtration and sized. The polysaccharide is activated and adipic acid dihydrazide, or other appropriate spacer, is added. The derivatized polysaccharide is conjugated to the carrier protein via activation of the protein. The reaction is continued for 24-48 hours, and the conjugate is separated from the non-conjugated reactants by chromatography. Example 10 Immunogenicity studies of E. coli-Toxin A Conjugate The E. coli-Toxin A conjugate of Example 9 is prepared for immunization studies as in Example 7. The results will show that the vaccine induced ligand- specific antibodies in each rabbit that is immunized. To determine the protective efficacy of these antibodies, their ability to protect mice in a hog mucin challenge model is assayed as in Example 7. The results will indicate substantial opsonic activity. Example 11 Immunogenicity Studies of Klebsiella- Diphtheria Toxoid Conjugate Unencapsulated Klebaiella pneumoniae 01 is grown and prepared as in Example 9 and the LPS is recovered. Diphtheria Toxoid (DT) from SSVI (Bern, Switzerland) is coupled, to the LPS as in Example 9 to form a conjugate. " ' The conjugate is then injected into 10-12 week old mice and antibodies specific to the conjugate are recovered as in Example 7. The conjugate is also separated by electrophore¬ sis using nitrocellulose paper and serum extracted from each mouse is tested for antibody binding accord¬ ing to the Western Blot method. Approximately 70% of the mice tested are found to have antibody specific for the conjugate. The antibody specific for the Klebsiella-DT conjugate is further tested according to the Hog Mucin Model of Example 7. The results will indicate sub¬ stantial opsonic activity. Example 12 Immunogenicity Studies of Pseudomonas aeruginosa-Toxin A Conjugate * Pseudomonas aeruginosa, Fisher Devlon type l, is grown according to Blumentals, I.I. et al., Appl . Environ. Microbiol . 53 : 2013-2020 (1987). LPS arid Toxin are separated and then conjugated as in Example 7. The conjugate is injected into mice and the antibody specific to the conjugate is recovered and immunologically tested as in Example 11. The results will indicate substantial opsonic activity. Example 13 Immunogenicity Studies of Staphlyococcus aureus-Toxin A Conjugate Staphylococcua aureus type 5 and 8 are grown as described by Fattom, Ali et al . , Infection and Immuni ¬ ty 58: 2367-2374 (1990) , the disclosure of which is incorporated by reference herein. Capsular polysaccharide is separated and conjugated to Toxin A as in Example 7 to form Staphylococcus aureus-Toxin A Conjugate. The conjugate is injected into mice and the antibody specific to the conjugate is recovered and .Lmmunoiogically tested as in Example 11. The results will indicate substantial opsonic activity. Example 14 Recovery of Hyperimmune Gammaglobulin From Plasma Donors The conjugates of Examples 6, 9 and 11-13, comprising 50 μg of each polysaccharide and lipid A analog, is injected into human volunteers. Approxi¬ mately 4 to 6 weeks, later the volunteers are reinjected with 50 μg of polysaccharide (in each conjugate) . Plasma is recovered from each volunteer and antibodies are screened by means of the passive hemagglutination test or by other appropriate means such as by ELISA. A volunteer generating a minimum antibody titer of 5 μg/ml is considered a responding donor. Plasma from responding donors is pooled and stored. This plasma will contain a relatively large concentration of antibody to the injected antigen and is appropriately called a hyperimmune serum. Preferably, the plasma is also screened for effective LPS neutralizing and bacteria opsonizing activity in the Swartzman and Hog mucin models, respectively. Hyperimmune gammaglobulin preparations are subsequently made using the Conn fraction method. See Conn, E.J. et al . , J. Amer. Chem. Soc. 68:459 (1946) . If necessary, donors are reinjected with the conjugates to achieve the minimum antibody level. Example 15 Treatment of Patients Suffering From Bacteremia Blood is collected from patients exhibiting bacterimic symptoms. The blood is tested for presence of a high concentration of bacteria using the direct Gram stain procedure. Gram negative and Gram positive bacteria are counted. The patients identified as suffering from bacteremia are* then injected intramuscularly with 250 mg/kg of the hyperimmune gammaglobu-'in per body weight of Example 14. At 6 hour intervals, blood is collected from the injected patients and Gram negative and Gram positive bacteria are recounted. It will be found that after each interval of collection, the presence of both Gram negative and positive bacteria will be significantly decreased. In addition, the symptoms of septic shock will be reduced or eliminated. Example 16 Purification of JCleJsiella and E. coli Polysaccharides and Conjugate Preparation According to Scheme III, the polysaccharides of the following Gram-negative microorganisms were purified: Pseudomonas Aeroginosa tSerotype PA FD-4, PA ED-5, PA FD-7, PA INT-3, PA INT-4) , Klebsiella (KPO-2, KPO-3), and E. coli (0-18). Scheme III Bacterial cell ψ Hot phenol extraction * Crude LPS 4- Hydrolysis Ultracentrifugation Filtration Freeze-drying Crude PS Gel filtration on Sephadox G-75 Freeze-dry Pure PS Figures 1-5 depict graphs showing the elution profile by gel filtration of the polysaccarides obtained from P. aeroginosa (FD-4, FD-7, INT-3 and INT-4) , and Klebaiella (KPO-2) , respectively. Derivatization of polysaccharide Purified pseudomonas polysaccharides were derivatized by EDAC Reaction and purified klebsiella polysaccharides were derivatized by CNBR Reaction using the method described by Chu et al . . Inf. & Immun. 59 : 4450-4458 (Dec. 1991) . According to this method, the purified pseudonomas polysaccharides (5.0 mg/ml of H^O) were brought to pH 11.0 with 1 N NaOH in a pH stat, and an equal weight of CNBr (1 g/ml of acetonitrile) was added. The pH was maintained at 11.0 with 1.0 N NaOH for 6 min at 4°C. An equal volume of 0.5 M ADH in 0.5 M NaHC0 3 was added, and the pH was adjusted to 8.5 with 1.0 M HCl. The reaction mixture was tumbled overnight at 3 to 8°C and dialyzed against 0.2 M NaCl at 3 to 8°C. The outer fluid was changed to HgO, and the dialysis was continued for 2 days with four changes of H^O. The contents of the dialysis bag were freeze-dried, dissolved in H 2 0 (20 mg/ml) , and passed through a G-25 Sephadex column (5 by 50 cm) in H-.0, and the void volume was freeze-dried. Conjugation Derivatized polysaccharides were coupled to protein carriers DT or BSA using the method described by Chu et al., Inf. & Immun. 55:4450-4458 (Dec. 1991). According to this procedure, the derivatized polysaccharide (PS) was dissolved in ELjO to 20 mg/ml, added to an equal weight of TT-20-mg/ml saline, and placed in an ice bath, and the pH was adjusted to 5.6 with 0.1 M HCl. EDAC was added to 0.05 M, and the pH was maintained at 5.6 in a pH stat for 4 h. The reaction mixture was dialyzed against 0.2 M NaCl for 2 days at 3 to 8°C with two changes of the outer fluid daily. This material was centrifuged at 14,700 x g and 10°C fδr 1 h, and the supernatant was applied to a CL-6B Sepharose column, in 0.2 M NaCl-0.01% Thimerosal. The void volume was stored at 3 to 8°C. The following conjugates were obtained according to this procedure: FD4-PS-DT, FD5-PS-DT, FD7-PS-DT, INT3-PS-BSA, INT4-PS-DT, KP02-PS-DT, KP03-PS-DT. The elution profiles of the FD4-PS-DT, FD5-PS-DT, FD7-PS-DT, INT4-PS-DT and KP02-PS-DT conjugates are depicted in Figures 6-10, respectively. The degree of substitution for the particular coupling agents is set forth in Table 13. Table 13 The polysaccharide and protein contents of each of the respective conjugates is shown in Table 14. Table 14 CHARA<TERIZATION OF P.AEROGINOSA CONJUGATES Klebsiella-PS Conjugates Mice were immunized with the following materials: 1. KpOl polysaccharide 2. KpOl-DT HMW Conjugate 3. KpOl-DT LMW Conjugate 4. KpOl-DT Mixed Conjugate 5. KpOl detoxified LPS (dLPS) 6. KpOl dLPS conjugated to DT These materials were prepared according to Chu, Infect. Immun . 55:4450-4458 (1991) except for KpOl dLPS, where detoxified LPS was employed. HMW and LMW represent high and low molecular weight polysaccharide conjugates obtained by gel filtration (see Figure 11. The detoxified LPS-DT conjugates were obtained according to the following procedure. LPS to dLPS 40 g LPS dissolved in 8 ml 0.1M NaOH Heat at 65°C for 2 hours Cool Neutralize with 6M HCl to pH=7.0 Dialyze for 2 days Lyophilize Coupling dLPS to DT Dissolve 10 mg of dLPS into 1 ml water Check pH and adjust to 5.5 Dissolve 10 mg of DT in 1 ml water Check pH and adjust to 5.5 Add 40mg ..fee to dLPS, and after 1 min add the DT solution Check pH and adjust to 5.5 with 0.1N HCl Bring pH to 5.0 in 30 min Maintain pH 5.0 for one hour Add another 40 mg EDC Bring pH to 5.0 in 30 min Maintain pH 5.0 for one hour Dialyze for 2 days and lyophilize * Purify the conjugate on Sepharose 6B in phosphate buffer Exp. 1 The data in Table 15 represents the ELISA units obtained for animals immunized with the conjugates. The higher the number, the higher the immunogenicity. Immunogen Saline KpOl Ps alone KpOl HMW conjugate KpOl LMW conjugate KpOl Mix DT primed Exp. 2 KpOl dLPS KpOl dLPS-DT The results (Table 16) are Geometric Means of IgG and IgM titers as evaluated in avidin-biotin Elisa. See Sutton et al., J " . Immunol . Meth. 82:215-224 (1985) . A group of mice (20) immunized with HMW conjugate 5ug Ps/injection three time two weeks apart. The sera were screened and the high responders (20) were pooled and assigned 100 Elisa units and used on each plate as a reference. Results show: (1) Ps-DT conjugates are potent, regardless of their molecular size and heterogenicity. (2) Polysaccharides alone are not immunogenic while the conjugates are. (3) We conferred a T cell dependent properties to the immune response as evidence by the carrier priming. (4) The response to dLPS alone or after the procedure of conjugation is the same. Example 18 Animal Immunizations with E. coli-PS Conjugates The following O serotypes of E. coli are the most prevalent isolates from extra-testinal locations, mainly blood: 01, 02, 04, 06, 07, 08, 09, 011, 018, 022, 025, 075. The above data were extracted from the following studies (Table 17) . Table 17 PS from E. coli were isolated according to Westphal, 0. and Jann, K. , Meth. CarJb. Chem 5:83-91 (1965) by extraction with phenol-water and further purification. The resulting PS from E. coli isolates were conjugated to immunogenic carriers as follows: CONJUGATION Step 1 : R-OH + CNBR 6 min- - -* + HgN-NH- CO- (CH^- CO-NH-NI , >RC- HNH 2 CO (CH 2 ) 4 CONH H 2 (PS-AH) Step 2 + Protein-COOH + PS-AH →Carbodiimide (EDAC) Protein-CONHNHCO(CH 2 ) 4 CONHNH-CPS The relative quantity of ADH and protein for each of the Protein-PS conjugates is set forth in Table 18. Table 18 Immunogenicity of Conjugates in Mice The following animal protocol was designed to show the following: 1) Immunogenicity of PS vs Conjugates; 2) Booster response upon 2nd injection; and 3) Carrier priming response. Ten animals per group were immunized with 2.5 ug PS alone or conjugated to DT. The animals were immunized once, twice or three times two weeks apart, and bled one week following the last immunization. Reference sera were obtained by immunizing 10-20 animals with 5 ug/dose X3 two weeks apart. The sera from these mice were pooled and assigned 100 ELISA units. Sera from mice were analyzed separately in Avidin-Biotin based ELISA. Each plate contained the reference and the tested sera in duplicates. Data (Table 19) is expressed as Geometric Means ELISA units. Conclusions A) Polysaccharides alone are not immunogenic . B) Conjugates are immunogenic and demonstrate a T- cell dependent immune response (Booster response and carrier priming) . The data in Table 20 shows the results of a study to determine the extent of cross reactivity of E. coli O-specific PS (antibodies specific for each serotype were tested for crossreactivity with the other serotype) . *This type has an exceptionally high background. Conclusion Conjugates elicit serotype-specific antibodies. If any cross reactivity, it is less than the background. Next, a tetravalent vaccine comprising 4 E. coli PSA-Conjugates, compared to PS alone, were injected into mice. The immunogenic response for each serotype was then determined (Table 21) . Conclusions 1. The PS alone are not immunogenic. 2. Conjugates are: (a) immunogenic; (b) demonstrate booster response; (c) demonstrate carrier priming. 3. Cross reactivity is minimal if any. 4. Multivalent PS vaccine still not immuno¬ genic. 5. In a multivalent vaccine, conjugates demonstrated the same potency as in the case of monovalent vaccines. Having now fully described this invention, it will be understood by those of skill in the art that the same can be performed with any wide equivalent range of compositions, concentrations, formulation and other parameters without affecting the scope of the invention or any embodiment. All patents and publications cited herein are fully incorporated by reference in their entirety.

Claims: WHAT IS CLAIMED IS: 1. A polyvalent vaccine comprising (a) a lipid A analog/immunogenic carrier conjugate; (b) at least one Gram negative bacteria, or antigenic determinant thereof, optionally conjugated with an immunogenic carrier, (c) a pharmaceutically acceptable carrier, and (d) optionally, an adjuvant; wherein said lipid A analog/immunogenic carrier conju¬ gate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 2. The polyvalent vaccine of claim 1, wherein said lipid A analog/immunogenic carrier conjugate has the following formula: wherein A is an immunogenic carrier; m is 0 or 1; n is 1 or 2; p is 1 to 200; L is -va linker group which does not interfere substantially with the characteristic ability of the lipid A analog to stimulate an immune response in an animal; R 1 and R 3 are the same or different and selected from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group or a phosphate group; wherein said conjugate induces active immunity to LPS when administered to an animal. 3. The polyvalent vaccine of claim 2, wherein said linker group comprises -(CH 2 ) q -NH-, wherein q is 2-3. 4. The polyvalent vaccine of claim 2, wherein L is O O 8 8 ( CH 2 ) -NH-C- (CH 2 ) X C, wherein q = 2-5, and x - 2-12. 5. The polyvalent vaccine of claim 2, wherein L is O 8 - (CH 2 ) y -C- , wherein y = 1-3 6. The polyvalent vaccine of claim 1, wherein said immunogenic carrier is a protein selected from the group consisting of bovine serum albumin, diphtheria toxoid, edestin, Toxin A and choleragenoid. 7. The " polyvalent vaccine of claim 1, wherein said lipid A analog/zurπrunogenic carrier conjugate has the formula: wherein A is an immunogenic carrier; is 0 or 1; p is 1 to 200; L is a linker group which does not interfere substantially with the characteristic ability of the lipid A analog to stimulate an immune response in an animal; ■ v R 1 and R 3 are selected from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy) -C 3 -C 18 acyl groups; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group or a phosphate group; wherein said conjugate induces active immunity to LPS when administered to an animal. 8. The polyvalent vaccine of claim 7, wherein the immunogenic carrier is a protein selected from the group consisting of bovine serum albumin, diphtheria toxoid, edestin, Toxin A and choleragenoid. 9. The polyvalent vaccine of claim 7, wherein said linker group comprises -(CH^ q - H-, wherein q is 2-5. 10. The polyvalent vaccine of claim 7, wherein said linker group comprises O 0 - (CIL,) -NH-C II- (CIL,),. CII-, wherein q - 2-5, and x = 2-12. 11. The polyvalent vaccine of claim 7, wherein L is -(CH^ y -C-, wherein y - 1-3. 12. The polyvalent vaccine of claim 1, wherein said lipid A analog/immunogenic carrier conjugate has the formula: wherein p is 1-200. 13 . The polyvalent vaccine of claim 1 , wherein said lipid A analog/immunogenic carrier conjugate has the formula: lnnunogenic Prot in wherein p is 1 to 200. 14. The polyvalent vaccine of claim 1, wherein said lipid A analog/immunogenic carrier conjugate has the following formula: wherein A is an immunogenic carrier; m is 0 or 1; n is 1 or 2; p is 1 to 200; L is a linker group which does not interfere substantially with the characteristic ability of the lipid A analog to stimulate an immune response in an animal; R 1 and R 3 are the same or different and selected from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group or a phosphate group; wherein said conjugate induces active immunity to LPS when administered to an animal. 15. The polyvalent vaccine of claim 14, wherein L is 0 0 -C II-(CH 2 ) X CII-, wherein x = 2-12. 16. The polyvalent vaccine of claim 14, wherein • v said immunogenic carrier is a protein selected from the group consisting of bovine serum albumin, diphtheria toxoid, edestin, Toxin A and choleragenoid. 17. The polyvalent vaccine of claim 1, wherein said lipid A analog/immunogenic carrier conjugate has the formula: wherein A is an immunogenic carrier; m is 0 or 1; p is 1 to 200 ; L is a linker group which does not interfere substantially with the characteristic ability of the lipid A analog to stimulate an immune response in an animal; R 1 and R 3 are selected from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy) -C 3 -C 18 acyl groups; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3-(C 2 -C 12 - acyloxy)-C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group or a phosphate group; wherein said conjugate induces active immunity to LPS when administered to an animal. 18. The polyvalent vaccine of claim 17, wherein the immunogenic carrier is a protein selected from the group consisting of bovine serum albumin, diphtheria toxoid, edestin, Toxin A and choleragenoid. 19. The polyvalent vaccine of* claim 17, wherein said linker group comprises 0 0 II II -C- (CH-,) X C- , wherein x = 2 -12 . 20. The polyvalent vaccine of claim 1, -wherein said lipid A analog/immunogenic carrier conjugate has the formula: wherein p is 1-200. 21. The polyvalent vaccine of claim 1, wherein said lipid A analog/immunogenic carrier conjugate has the formula: Protein wherein p is 1 to 200. 22. The polyvalent vaccine of claim 1, wherein the Gram negative bacteria is Escherichia coli , Pseudomonas aeruginosa, Klebsiella pneumoniae, Klebsi ¬ ella oxytoca, or Klebsiella ozanae, or an antigenic determinant thereof. 23. The polyvalent vaccine of claim 1, wherein said component (b) is a capsular polysaccharide. 24. The polyvalent vaccine of claim 1, further comprising: (e) at least one Gram positive bacteria, or antigenic determinant thereof, optionally conjugated with an immunogenic carrier. 25. The polyvalent vaccine of claim 24, wherein the Gram positive bacteria is a Micrococcus, Streptococcus or Staphylococcus. 26. The polyvalent vaccine of claim 24, wherein said component (e) is a capsular polysaccharide. 27. A polyvalent hyperimmune gammaglobulin comprising a gammaglobulin specific for a lipid A analog/iirimunogenic carrier conjugate, and a gammaglobulin specific for at least one 0 antigen wherein said polyvalent hyperimmune gammaglobulin is effective for neutralizing LPS and opsonizing a Gram negative bacteria; and wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups-. 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an :ϋmmune response in an animal. 28. The polyvalent hyperdiimune gammaglobulin of claim 27, wherein the 0 antigen specific gammaglobulin is specific for Eshereichia coli, Pseudomonas aerugi ¬ nosa, Klebsiella pneumoniae, Klebsiella oxytoca, or Klebsiella ozanae. 29. The polyvalent hyperimmune gammaglobulin of claim 27, further comprising a gammaglobulin specific for at least one Gram positive bateria wherein said polyvalent hyperimmune gammaglobulin is effective for neutralizing LPS and opsonizing at least one Gram negative and at least one Gram positive bacteria. 30. The polyvalent hyperimmune gammaglobulin of claim 29, wherein the gammaglobulin is specific for at least one Gram positive microorganism selected from the group consisting of Micrococcus , Streptococcus and Staphylococcus . 31. A method of preparing a polyvalent hyper¬ immune gammaglobulm comprising (a) immunizing an animal with a lipid A analog/immunogenic carrier conjugate and at least one Gram negative bacteria, or antigenic determinant thereof, said bacteria or determinant thereof optionally conjugated with an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said bacteria and neutralize LPS; and wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 32. A method of preparing a polyvalent hyper¬ immune gammaglobulin comprising (a) immunizing an animal with a lipid A analog/zunmunogenic carrier conjugate, at least one Gram negative bacteria, or antigenic determinant thereof, aόd at least one Gram positive bacteria, or antigenic determinant thereof, said bacteria or determinant thereof optionally conjugated with an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said Gram negative and Gram positive bacteria and neutralize LPS; and wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 τC 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 33. A polyvalent hyperimmune gammaglobulin obtained by (a) immunizing an animal with a lipid A analog/immunogenic carrier conjugate and at least one Gram negative bacteria, or antigenic determinant thereof, said bacteria or determinant thereof conjugated with an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said bacteria and neutralize LPS; and wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-|ιydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an -Immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an iirarrune response in an animal. 34. A polyvalent hyperimmune gammaglobulin obtained by (a) immunizing an animal with a lipid A analog/immunogenic carrier conjugate, at least one Gram negative bacteria, or antigenic determinant thereof, and at least one Gram positive bacteria, or antigenic determinant thereof, said bacteria or determinant thereof optionally conjugated with an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said Gram negative and Gram positive bacteria and neutralize LPS; and wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R r and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 35. A method of treating Gram negative bacterial sepsis comprising administering a polyvalent hyperimmune gammaglobulin comprising an effective neutralizing amount of a hyperimmune gammaglobulin specific to a lipid A analog/immunogenic carrier conjugate, and an effective opsonizing amount of 0 antigen specific gammaglobulin to an animal in need of such treatment; wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C*| 8 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 36. A method of treating Gram negative or Gram positive bacterial sepsis comprising administering a polyvalent hyperimmune gammaglobulin comprising an effective neutralizing amount of a hyperimmune gammaglobulin specific to a lipid A analog/immunogenic carrier conjugate, an effective opsonizing amount of Gram negative bacteria-specific gammaglobulin, and an effective opsonizing amount of Gram positive bacteria- specific gammaglobulin to an animal in need of such treatment; wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 37. A kit for treating Gram negative or Gram positive bacterial sepsis comprising a carrier means compartmentalized to receive in close confinement therein one or more container means wherein a first container contains a lipid A analog/immunogenic carrier conjugate and at least one other container means which contains at least one Gram negative or Gram positive bacteria, or antigenic determinant thereof, optionally conjugated to an immunogenic carrier; wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 38. A kit for treating Gram negative or Gram positive bacterial sepsis comprising a carrier means compartmentalized to receive in close confinement therein one or more container means wherein a first container means contains a hyperimmune gammaglobulin specific for a lipid A analog/immunogenic conjugate and at least one other container means which contains a hyperimmune gammaglobulin specific for at least one Gram negative or Gram positive bacteria, or antigenic determinant thereof; wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - acyloxy) -Cg-C-g acyl group or a linkage to an immuno¬ genic carrier; R 2 is selected from the group consisting of C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 39. The kit of claim 38, wherein said first container means further contains a hyperimmune gammaglobulin specific for at least one Gram negative or Gram positive bacteria, or antigenic determinant thereof. AMENDED CLAIMS [received by the International Bureau on 17 August 1992 (17.08.92), original claims 1,27,31-38 amended; other claims unchanged(10 pages)] 1. A polyvalent vaccine comprising (a) a lipid A analog/immunogenic carrier conjugate; (b) at least one Gram negative bacteria, or antigenic determinant thereof, optionally conjugated with an immunogenic carrier, (c) a pharmaceutically acceptable carrier, and (d) optionally, an adjuvant; wherein said lipid A analog/iinmunogenic carrier conju ¬ gate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - acyloxy) -C^-C^g acyl group or a linkage to an immuno¬ genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C -C 18 acyl group, a phosphate group or a linkage to an insmunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. wherein R 1 and R 3 are the same or different and select- ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - ' acyloxy) -C 3 -C 18 acyl group or a linkage to an _Lmmuno- genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - c i 8 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to tin immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an -Immune response in an animal. 28. The polyvalent hyperimmune gammaglobulin of claim 27, wherein the O antigen specific gammaglobulin is specific for Eshereichia coli, Pseudomonas aerugi¬ noaa, Klebaiella pneumoniae, Klebaiella oxytoca, or Klebaiella ozanae. 29. The polyvalent hyperimmune gammaglobulin of claim 27, further comprising a gammaglobulin specific for at least one Gram positive bateria wherein said polyvalent hyperimmune gammaglobulin is effective for neutralizing LPS and opsonizing at least βne Gram negative and at least one Gram positive bacteria. 30. The polyvalent hyperimmune gammaglobulin of claim 29, wherein the gammaglobulin is specific for at least one Gram positive microorganism selected from the group consisting of Micrococcus, Streptococcua and Staphylococcua. 31. A method of preparing a polyvalent hyper¬ immune gammaglobulin comprising (a) immunizing an animal with a lipid A analog/iiπmunogenic carrier conjugate and at least one Gram negative bacteria, or antigenic determinant. thereof, said bacteria or determinant thereof optionally conjugated with an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said bacteria and neutralize LPS; and wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; n is 1 or 2; R 2 is selected from the group, consisting of C 2 - C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy) -C 3 -C 18 acyl groups; and acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an iiranunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an _Lmmune response in an animal. 33. A polyvalent hyperimmune gammaglobulin obtained by (a) immunizing . an animal with a lipid A analog/-bmmunogenic carrier conjugate and at least one Gram negative bacteria, or antigenic determinant thereof, said bacteria or determinant thereof conjugated with an immunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said bacteria and neutralize LPS; and wherein said lipid A analog/-Immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno- genie carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - c 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage - to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an .Immune response in an animal. 34. A polyvalent hyperimmune gammaglobulin obtained by (a) immunizing an animal with a lipid A analog/immunogenic carrier conjugate, at least one Gram negative bacteria, or antigenic determinant thereof, and at least one Gram positive bacteria, or antigenic determinant thereof, said bacteria or determinant thereof optionally conjugated with an .iiπmunogenic carrier, (b) optionally boosting said animal with the immunizing step of step (a) , (c) pooling plasma from said animal; and (d) recovering the gammaglobulin fraction from said animal, wherein said fraction is effective to opsonize said Gram negative and Gram positive bacteria and neutralize LPS; and wherein said lipid A analog/-unmunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 auid R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 -' acyloxy)-C 3 -C 18 acyl group or a linkage to an .Immuno¬ genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 - 0^ 2 ~acyloxy)-C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 35. Amethod of treating Gram negative bacterial sepsis comprising administering a polyvalent hyperimmune gammaglobulin comprising an effective neutralizing amount of a hyperimmune gammaglobulin specific to a lipid A analog/immunogenic carrier conjugate, and an effective opsonizing amount of 0 antigen specific gammaglobulin to an animal in need of such treatment; wherein said lipid A analog/immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl" groups, 3-hydroxy C 3 - C 8 acyl groups, a 3- (C 2 -C 12 -' acyloxy)-C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - χ 8 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy)-C^-C^g acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an .Immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 36. A method of treating Gram negative or Gram positive bacterial sepsis comprising administering a polyvalent hyperimmune gammaglobulin comprising an effective neutralizing amount of a hyperiiπmune gammaglobulin specific to a lipid A analog/iπraiunogenic carrier conjugate, an effective opsonizing amount of Gram negative bacteria-specific gammaglobulin, and an effective opsonizing amount of Gram positive bacteria- specific gammaglobulin to an animal in need of such treatment; wherein said lipid A analog/.Immunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C,-C 12' acyloxy) -C 3 -C 18 acyl group or a linkage to an rijπmuno- genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy) -C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an immune response in an animal. 37. A kit for treating Gram negative or Gram positive bacterial sepsis comprising a carrier means compartmentalized to receive in close confinement therein one or more container means wherein a first container contains a lipid A analog/iiπmunogenic carrier conjugate and at least one other container means which contains at least one Gram negative or Gram positive bacteria, or antigenic determinant thereof, optionally conjugated to an .Immunogenic carrier; wherein said lipid A analog/iiraπunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl" groups, 3-hydroxy C 3 -C 18 acyl groups, a 3-(C 2 -C 12 - ' acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C - c ι acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C 2 -C 12 -acyloxy)-C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an imntune response in an animal. 38. A kit for treating Gram negative or Gram positive bacterial sepsis comprising a carrier means compartmentalized to receive in close confinement therein one or more container means wherein a first container means contains a hyperiπimune gammaglobulin specific for a lipid A analog/immunogenic conjugate and at least one other container means which contains a hyperimmune gammaglobulin specific for at least one Gram negative or Gram positive bacteria, or antigenic determinant thereof; wherein said lipid A analog/_lmmunogenic carrier conjugate comprises a lipid A analog of the formula wherein R 1 and R 3 are the same or different and select¬ ed from the group consisting of hydrogen, C 2 -C 18 acyl groups, 3-hydroxy C 3 -C 18 acyl groups, a 3- (C 2 -C 12 - acyloxy) -C 3 -C 18 acyl group or a linkage to an immuno¬ genic carrier; n is 1 or 2; R 2 is selected from the group consisting of C 2 - c ι acyl groups, 3-hydroxy C 3 -C 18 acyl groups, and 3- (C -C 12 -acyloxy)-C 3 -C 18 acyl groups; and R 4 is hydrogen, a C 2 -C 18 acyl group, a phosphate group or a linkage to an immunogenic carrier; with the proviso that one of R 1 , R 3 or R 4 is a linkage to an immunogenic carrier, wherein said linkage does not interfere substantially with the ability of the lipid A analog to stimulate an .Immune response in an animal. 39. The kit of claim 38, wherein said first container means further contains a hyperiπrαiune gammaglobulin specific for at least one Gram negative or Gram positive bacteria, or antigenic determinant thereof.

Inventors: BHATTACHARJEE APURBA, SADOFF JERALD, TENG NELSON, BHATTACHARJEE, APURBA, SADOFF, JERALD, TENG, NELSON
Applicants: BHATTACHARJEE APURBA, SADOFF JERALD, TENG NELSON, UNIVAX BIOLOG INC, BHATTACHARJEE, APURBA, SADOFF, JERALD, TENG, NELSON, UNIVAX BIOLOGICS INCORPORATED
Application year: 1992
Main classification: A61K39
IPC codes: A61K39, A61P31, C07H13, C07H15, C07K1, C07K14, C12P19