Top

Published in:

01-05-2005 | Original Article

Comparison of antigen constructs and carrier molecules for augmenting the immunogenicity of the monosaccharide epithelial cancer antigen Tn

Authors: Ella Kagan, Govind Ragupathi, San San Yi, Celso A. Reis, Jeff Gildersleeve, Daniel Kahne, Henrik Clausen, Samuel J. Danishefsky, Philip O. Livingston

Published in: Cancer Immunology, Immunotherapy | Issue 5/2005

Abstract

We have demonstrated previously that the optimal method for inducing an antibody response against defined cancer antigens is covalent conjugation of the antigen to keyhole limpet hemocyanin (KLH) and use of the potent saponin adjuvant QS-21. Single molecules of glycolipids (tetrasaccharides, pentasaccharides, or hexasaccharides) and MUC1 peptides (containing between one and five MUC1 tandem repeats) conjugated to KLH have proven sufficient for antibody recognition and vaccine construction. However, cancer specificity of monoclonal antibodies against the monosaccharide Tn and disaccharide sTn comes largely from recognition of clusters (c) of these molecules on the cell surface. Tn consists of a monosaccharide (GalNAc) O-linked to serine or threonine on epithelial cancer mucins which are uniquely rich in serines and threonines. We test here several Tn constructs: Tn monosaccharide, Tn(c) prepared on a triple threonine backbone, and Tn prepared on a partially or fully glycosylated MUC1 backbone. We determine that Tn(c) is more effective than Tn, and conjugation to KLH is more effective than conjugation to BSA or polystyrene beads for inducing ELISA reactivity against Tn, and FACS reactivity against Tn-positive tumor cells. Surprisingly, MUC1 glycosylated with Tn at three or five sites per 20 amino acid MUC1 tandem repeat and conjugated to KLH, induced the strongest antibody response against Tn and tumor cells expressing Tn, and had the additional advantage of inducing antibodies against MUC1.

Livingston PO, Wong GYC, Adluri S, Tao Y, Padavan M, Parente R, Hanlon C, Helling F, Ritter G, Oettgen HF, Old LJ (1994) Improved survival in AJCC stage III melanoma patients with GM2 antibodies: a randomized trial of adjuvant vaccination with GM2 ganglioside. J Clin Oncol 12:1036–1044PubMed

Chapman PB, Morrissey DM, Panageas KS, Hamilton WB, Zhan C, Destro AN, Williams L, Israel RJ, Livingston PO (2000) Induction of antibodies against GM2 ganglioside by immunizing melanoma patients using GM2-KLH+QS21 vaccine: a dose-response study. Clin Cancer Res 6:874–879PubMed

Helling F, Shang Y, Calves M, Oettgen HF, Livingston PO (1994) Increased immunogenicity of GD3 conjugate vaccines: comparison of various carrier proteins and selection of GD3-KLH for further testing. Cancer Res 54:197–203PubMed

Ragupathi G, Meyers M, Adluri S, Howard L, Musselli C, Livingston PO (2000) Induction of antibodies against GD3 ganglioside in melanoma patients by vaccination with GD3-lactone-KLH conjugate plus immunological adjuvant QS-21. Int J Cancer 85:659–666CrossRefPubMed

Dickler MN, Ragupathi G, Liu NX, Musselli C, Martino DJ, Miller VA, Kris MG, Brezicka FT, Livingston PO, Grant SC (1999) Immunogenicity of the fucosyl-GM1-keyhole limpet hemocyanin (KLH) conjugate vaccine in patients with small cell lung cancer. Cancer Res 5:2773–2779

Ragupathi G, Slovin S, Adluri S, Sames D, Kim IJ, Kim HM, Spassova M, Bornmann WG, Lloyd K, Scher HI, Livingston PO, Danishefsky SJ (1999) A fully synthetic globo H carbohydrate vaccine induces a focused humoral response in prostate cancer patients: a proof of principle. Angewandte Chemie 38:563–566CrossRef

Gilewski T, Ragupathi G, Bhuta S, Williams LJ, Musselli C, Zhang XF, Bencsath KP, Panageas KS, Chin J, Norton L, Houghton AN, Livingston PO et al (2001) Immunization of metastatic breast cancer patients with a fully synthetic globo H conjugate: a phase I trial. Proc Natl Acad Sci U S A 98:3270–3275

Sabbatini P, Kudryashov V, Ragupathi G, Danishefsky S, Livingston PO, Bornmann W, Spassova M, Spriggs D, Aghajanian C, Soignet S, Peyton M, O’Flaherty C et al (2000) Immunization of ovarian cancer patients with a synthetic LewisY–protein conjugate vaccine: clinical and serological results. Int J Cancer 87:79–85CrossRefPubMed

Adluri S, Gilewski T, Zhang S, Ramnath V, Ragupathi G, Livingston PO (1999) Specificity analysis of sera from breast cancer patients vaccinated with Muc1-KLH plus QS-21. Br J Cancer 79:1806–1812CrossRefPubMed

10.

Gilewski T, Adluri S, Ragupathi G, Zhang S, Yao TJ, Panageas K, Moynahan M, Houghton A, Norton L, Livingston PO (2000) Vaccination of high risk breast cancer patients with Mucin-1 keyhole limpet hemocyanin conjugate plus QS-21. Clin Cancer Res 6:1693–1701PubMed

11.

Zhang S, Walberg LA, Ogata S, Itzkowitz SH, Koganty RR, Reddish M, Gandhi SS, Longenecker BM, Lloyd KO, Livingston PO (1995) Immune sera and monoclonal antibodies define two configurations for the sialyl Tn tumor antigen. Cancer Res 55:3364–3368PubMed

12.

Dicker M, Gilewski T, Ragupathi G, Adluri R, Koganty RR, Longenecker M, Houghton AN, Norton L, Livingston PO (1997) Vaccination of breast cancer patients (pts) with no evidence of disease (NED) with sialyl Tn cluster (sTa(c))-keyhole limpet hemocyanin (KLH) conjugate plus adjuvant QS-21: preliminary results. Proc ASCO 16:1572

13.

Nakada H, Inoue M, Numata Y, Tanaka N, Funakoshi J, Fukui S, Mellors A, Yamashina I (1993) Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). Proc Natl Acad Sci U S A 90:2495–2499

14.

Ragupathi G, Howard L, Cappello S, Koganty RR, Qiu D, Longenecker BM, Reddish MA, Lloyd KO, Livingston PO (1999) Vaccines prepared with sialyl-Tn and sialyl-Tn trimers using the 4-(4-maleimidomethyl) cyclohexane-1-carboxyl hydrazide linker group result in optimal antibody titers against ovine submaxillary mucin and sialyl-Tn-positive tumor cells. Cancer Immunol Immunother 48:1–8

15.

Liang R, Yan L, Loebach J, Ge M, Uozumi Y, Sekanina K, Horan N, Gildersleeve J, Thompson C, Smith A, Biswas K, Still WC et al (1996) Parallel synthesis and screening of a solid phase carbohydrate library. Science 274:1520–1522CrossRefPubMed

16.

Liang R, Loebach J, Horan N, Ge M, Thompson C, Yan L, Kahne D (1997) Polyvalent binding to carbohydrates immobilized on an insoluble resin. Proc Natl Acad Sci U S A 94:10554–10559

17.

Ragupathi G, Koganty RR, Qui DS, Lloyd KO, Livingston PO (1998) A novel and efficient method for synthetic carbohydrate conjugate vaccine preparation: synthesis of sialyl Tn KLH conjugate using a 4-(4-N-maleimidomethyl)cyclohexane-1-carboxyl hydrazide (MMCCH) linker arm. Glycoconjugate J 15:217–221CrossRef

18.

White T, Bennett EP, Takio K, Sorensen T, Bonding N, Clausen H (1995) Purification and cDNA cloning of a human UDP-N-acetyl-α-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase. J Biol Chem 270:24156–24165CrossRefPubMed

19.

Bennett EP, Hassan H, Mandel U, Mirgorodskaya EP, Burchell J, Taylor-Papadimitriou J, Hollingsworth MA, Merkx G, Vankessel AG, Eiberg H, Steffensen R, Clausen H (1998) Cloning of a human UDP-N-acetyl-alpha-D-alactosaminepolypeptide N-acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat. J Biol Chem 273:30472–30481CrossRefPubMed

20.

Zhang S, Graeber LA, Helling F, Ragupathi G, Adluri S, Lloyd KO, Livingston PO (1996) Augmenting the immunogenicity of synthetic MUC1 peptide vaccines in mice. Cancer Res 56:3315–3319PubMed

21.

Rohrer JS, Miller HI (2003) Detecting O-linked oligosaccharides on glycoproteins. Anal Biochem 316:131–134CrossRefPubMed

22.

Ogata S, Chen A, Itzkowitz SH (1994) Use of model cell lines to study the biosynthesis and biological role of cancer-associated sialosyl-Tn antigen. Cancer Res 54:4036–4044PubMed

23.

Huntsberger DV, Leaverton PE (ed) (1970) In: Statistical inference in the biomedical sciences. Allyn Bacon, Boston, pp 138–140, 337–338

24.

Kurosaka A, Kitagawa H, Fukui S, Numata Y, Nakada H, Fuakoshi I, Kawasaki T, Ogawa T, Iijima H, Yamashina I (1988) A monoclonal antibody that recognizes a cluster of a disaccharide, NeuAcα2→6GalNAc, in mucin-type glycoproteins. J Biol Chem 263:8724–8726PubMed

Title: Comparison of antigen constructs and carrier molecules for augmenting the immunogenicity of the monosaccharide epithelial cancer antigen Tn
Authors: Ella Kagan
Govind Ragupathi
San San Yi
Celso A. Reis
Jeff Gildersleeve
Daniel Kahne
Henrik Clausen
Samuel J. Danishefsky
Philip O. Livingston
Publication date: 01-05-2005
Publisher: Springer-Verlag
Published in: Cancer Immunology, Immunotherapy / Issue 5/2005
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-004-0584-y

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 5/2005

Effective induction of antitumor immunity by immunization with plasmid DNA encoding TRP-2 plus neutralization of TGF-β

A phase I vaccination study with tyrosinase in patients with stage II melanoma using recombinant modified vaccinia virus Ankara (MVA-hTyr)

Tumor sensitivity to IFN-γ is required for successful antigen-specific immunotherapy of a transplantable mouse tumor model for HPV-transformed tumors

Paclitaxel probably enhances cytotoxicity of natural killer cells against breast carcinoma cells by increasing perforin production

In vitro dendritic cell generation and lymphocyte subsets in myeloma patients: influence of thalidomide and high-dose chemotherapy treatment

Harnessing host immune responses to preneoplasia: promise and challenges

Keynote webinar | Spotlight on antibody–drug conjugates in cancer