Biodistribution and elimination kinetics of systemic Stx2 by the Stx2A and Stx2B subunit-specific human monoclonal antibodies in mice

BMC Immunology

Saul Tzipori¹  Pradeep Singh³  Anthony Wiffin¹  Jean Mukherjee¹  Kwang-il Jeong²  Abhineet Sheoran¹ 

[1] Department of Biomedical Sciences, Cummings School of Veterinary Medicine, Tufts University, 200 Westboro Road Building 20, North Grafton, MA, USA;Vaccine Research Center, 401 North Middletown Road Pfizer, Pearl River, NY, 10965, USA;Department of Mathematics, Southeast Missouri State University, 401 North Middletown Road Pfizer, Cape Girardeau, MO, 63701, USA

关键词: Human monoclonal antibody;    Pharmacokinetic;    Toxin concentration;    Toxin elimination;    Antibody;    Radiolabel;    Shiga toxin;   

Others  :  1077913
DOI  :  10.1186/1471-2172-13-27

 received in 2012-03-08, accepted in 2012-06-01,  发布年份 2012

【 摘 要 】

Background

Hemolytic uremic syndrome (HUS) leading to acute kidney failure, is a condition linked to the production of primarily Shiga toxin 2 (Stx2) by some E. coli serotypes. We have previously shown that Stx2 A subunit-specific human monoclonal antibody (HuMAb) 5C12, and B subunit-specific HuMAb 5H8 inhibit cultured cell death, and protect mice and piglets from fatal Stx2-intoxication. We have also shown that 5H8 blocks binding of Stx2 to its cell-surface receptor globotriaosyl ceramide (Gb₃), whereas Stx2 when complexed with 5C12 binds Gb₃ with higher affinity than Stx2. The mechanism by which 5C12 neutralizes Stx2 in vitro involves trapping of Stx2 in the recycling endosomes and releasing it into the extracellular environment. Because of the clinical implications associated with the formation of Stx2/antibody complexes and the potential for trapping and clearance through a severely damaged kidney associated with HUS, we investigated the likely site(s) of Stx2/antibody localization and clearance in intoxicated mice treated with antibody or placebo.

Results

Mice were injected with radiolabeled Stx2 (¹²⁵I-Stx2) 4 hours after administration of 5C12, 5H8, or phosphate buffered saline (PBS) and the sites of localization of labeled Stx2, were investigated 3, 24 and 48 hours later. The liver recorded statistically much higher concentrations of labeled Stx2 for groups receiving 5C12 and 5H8 antibodies after 3, 24 and 48 hours, as compared with the PBS group. In contrast, highest levels of labeled Stx2 were detected in the kidneys of the PBS group at all 3 sampling times. Mice receiving either of the two HuMAbs were fully protected against the lethal effect of Stx2, as compared with the fatal outcome of the control group.

Conclusions

The results suggest that HuMAbs 5C12 and 5H8 promoted hepatic accumulation and presumably clearance of toxin/antibody complexes, significantly diverting Stx2 localization in the kidneys, the target of Stx2 and the cause of HUS. This is in contrast to the fatal outcome of the control group receiving PBS. The results also confirm earlier observations that both HuMAbs are highly and equally protective against Stx2 intoxication in mice.

【 授权许可】

   
2012 Sheoran et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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【 参考文献 】

• [1]Boerlin P, McEwen SA, Boerlin-Petzold F, Wilson JB, Johnson RP, Gyles CL: Associations between virulence factors of Shiga toxin-producing Escherichia coli and disease in humans. J Clin Microbiol 1999, 37:497-503.
• [2]Griffin PM, Tauxe RV: The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol Rev 1991, 13:60-98.
• [3]Milford DV, Taylor CM, Guttridge B, Hall SM, Rowe B, Kleanthous H: Haemolytic uraemic syndromes in the British Isles 1985–8: association with verocytotoxin producing Escherichia coli. Part 1: Clinical and epidemiological aspects. Arch Dis Child 1990, 65:716-721.
• [4]Ostroff SM, Tarr PI, Neill MA, Lewis JH, Hargrett-Bean N, Kobayashi JM: Toxin genotypes and plasmid profiles as determinants of systemic sequelae in Escherichia coli O157:H7 infections. J Infect Dis 1989, 160:994-998.
• [5]Fraser ME, Fujinaga M, Cherney MM, Melton-Celsa AR, Twiddy EM, O'Brien AD, James MN: Structure of shiga toxin type 2 (Stx2) from escherichia coli O157:H7. J Biol Chem 2004, 279:27511-27517.
• [6]Friedrich AW, Bielaszewska M, Zhang WL, Pulz M, Kuczius T, Ammon A, Karch H: Escherichia coli harboring Shiga toxin 2 gene variants: frequency and association with clinical symptoms. J Infect Dis 2002, 185:74-84.
• [7]Russmann H, Schmidt H, Heesemann J, Caprioli A, Karch H: Variants of Shiga-like toxin II constitute a major toxin component in Escherichia coli O157 strains from patients with haemolytic uraemic syndrome. J Med Microbiol 1994, 40:338-343.
• [8]Tzipori S, Sheoran A, Akiyoshi D, Donohue-Rolfe A, Trachtman H: Antibody therapy in the management of shiga toxin-induced hemolytic uremic syndrome. Clin Microbiol Rev 2004, 17:926-941. table of contents
• [9]Ling H, Boodhoo A, Hazes B, Cummings MD, Armstrong GD, Brunton JL, Read RJ: Structure of the shiga-like toxin I B-pentamer complexed with an analogue of its receptor Gb3. Biochemistry 1998, 37:1777-1788.
• [10]Shimizu H, Field RA, Homans SW, Donohue-Rolfe A: Solution structure of the complex between the B-subunit homopentamer of verotoxin VT-1 from Escherichia coli and the trisaccharide moiety of globotriaosylceramide. Biochemistry 1998, 37:11078-11082.
• [11]Lingwood CA, Law H, Richardson S, Petric M, Brunton JL, De Grandis S, Karmali M: Glycolipid binding of purified and recombinant Escherichia coli produced verotoxin in vitro. J Biol Chem 1987, 262:8834-8839.
• [12]Waddell T, Head S, Petric M, Cohen A, Lingwood C: Globotriosyl ceramide is specifically recognized by the Escherichia coli verocytotoxin 2. Biochem Biophys Res Commun 1988, 152:674-679.
• [13]Garred O, Dubinina E, Holm PK, Olsnes S, van Deurs B, Kozlov JV, Sandvig K: Role of processing and intracellular transport for optimal toxicity of Shiga toxin and toxin mutants. Exp Cell Res 1995, 218:39-49.
• [14]Sandvig K, van Deurs B: Endocytosis, intracellular transport, and cytotoxic action of Shiga toxin and ricin. Physiol Rev 1996, 76:949-966.
• [15]Garred O, van Deurs B, Sandvig K: Furin-induced cleavage and activation of Shiga toxin. J Biol Chem 1995, 270:10817-10821.
• [16]Endo Y, Tsurugi K, Yutsudo T, Takeda Y, Ogasawara T, Igarashi K: Site of action of a Vero toxin (VT2) from Escherichia coli O157:H7 and of Shiga toxin on eukaryotic ribosomes. RNA N-glycosidase activity of the toxins. Eur J Biochem 1988, 171:45-50.
• [17]Saxena SK, O'Brien AD, Ackerman EJ: Shiga toxin, Shiga-like toxin II variant, and ricin are all single-site RNA N-glycosidases of 28S RNA when microinjected into Xenopus oocytes. J Biol Chem 1989, 264:596-601.
• [18]Mukherjee J, Chios K, Fishwild D, Hudson D, O'Donnell S, Rich SM, Donohue-Rolfe A, Tzipori S: Human Stx2-specific monoclonal antibodies prevent systemic complications of Escherichia coli O157:H7 infection. Infect Immun 2002, 70:612-619.
• [19]Mukherjee J, Chios K, Fishwild D, Hudson D, O'Donnell S, Rich SM, Donohue-Rolfe A, Tzipori S: Production and characterization of protective human antibodies against Shiga toxin 1. Infect Immun 2002, 70:5896-5899.
• [20]Sheoran AS, Chapman S, Singh P, Donohue-Rolfe A, Tzipori S: Stx2-specific human monoclonal antibodies protect mice against lethal infection with Escherichia coli expressing Stx2 variants. Infect Immun 2003, 71:3125-3130.
• [21]Sheoran AS, Chapman-Bonofiglio S, Harvey BR, Mukherjee J, Georgiou G, Donohue-Rolfe A, Tzipori S: Human antibody against shiga toxin 2 administered to piglets after the onset of diarrhea due to Escherichia coli O157:H7 prevents fatal systemic complications. Infect Immun 2005, 73:4607-4613.
• [22]Psotka MA, Obata F, Kolling GL, Gross LK, Saleem MA, Satchell SC, Mathieson PW, Obrig TG: Shiga toxin 2 targets the murine renal collecting duct epithelium. Infect Immun 2009, 77:959-969.
• [23]Donohue-Rolfe A, Acheson DW, Kane AV, Keusch GT: Purification of Shiga toxin and Shiga-like toxins I and II by receptor analog affinity chromatography with immobilized P1 glycoprotein and production of cross-reactive monoclonal antibodies. Infect Immun 1989, 57:3888-3893.
• [24]Hunter WM, Greenwood FC: Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature 1962, 194:495-496.
• [25]Fuchs G, Mobassaleh M, Donohue-Rolfe A, Montgomery RK, Grand RJ, Keusch GT: Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin. Infect Immun 1986, 53:372-377.
• [26]Davies KA, Robson MG, Peters AM, Norsworthy P, Nash JT, Walport MJ: Defective Fc-dependent processing of immune complexes in patients with systemic lupus erythematosus. Arthritis Rheum 2002, 46:1028-1038.
• [27]Johansson AG, Lovdal T, Magnusson KE, Berg T, Skogh T: Liver cell uptake and degradation of soluble immunoglobulin G immune complexes in vivo and in vitro in rats. Hepatology 1996, 24:169-175.
• [28]Lovdal T, Andersen E, Brech A, Berg T: Fc receptor mediated endocytosis of small soluble immunoglobulin G immune complexes in Kupffer and endothelial cells from rat liver. J Cell Sci 2000, 113(Pt 18):3255-3266.
• [29]Obata F, Tohyama K, Bonev AD, Kolling GL, Keepers TR, Gross LK, Nelson MT, Sato S, Obrig TG: Shiga toxin 2 affects the central nervous system through receptor globotriaosylceramide localized to neurons. J Infect Dis 2008, 198:1398-1406.
• [30]Rutjes NW, Binnington BA, Smith CR, Maloney MD, Lingwood CA: Differential tissue targeting and pathogenesis of verotoxins 1 and 2 in the mouse animal model. Kidney Int 2002, 62:832-845.
• [31]Shen DH, Kloos RT, Mazzaferri EL, Jhian SM: Sodium iodide symporter in health and disease. Thyroid 2001, 11:415-425.
• [32]Cohen A, Madrid-Marina V, Estrov Z, Freedman MH, Lingwood CA, Dosch HM: Expression of glycolipid receptors to Shiga-like toxin on human B lymphocytes: a mechanism for the failure of long-lived antibody response to dysenteric disease. Int Immunol 1990, 2:1-8.
• [33]Menge C, Wieler LH, Schlapp T, Baljer G: Shiga toxin 1 from Escherichia coli blocks activation and proliferation of bovine lymphocyte subpopulations in vitro. Infect Immun 1999, 67:2209-2217.
• [34]Sugatani J, Igarashi T, Shimura M, Yamanaka T, Takeda T, Miwa M: Disorders in the immune responses of T- and B-cells in mice administered intravenous verotoxin 2. Life Sci 2000, 67:1059-1072.