Immunology

Dr L J Partridge

The research in my laboratory falls into two main areas: investigations on the structure and function of the tetraspanin super-family of membrane proteins and the application of antibody technology to medical research and therapeutics.

The tetraspanins are a heterogeneous family of proteins with four membrane spanning domains. They have a wide tissue distribution in multicellular organisms and 33 distinct tetraspanins have been identified in humans. Tetraspanins have been implicated in basic cell processes such as motility and adhesion and more specifically in sperm:egg fusion (CD9), lymphocyte activation (CD81), cancer metastasis (CD9, CD63, CD82) and virus susceptibility e.g. Hepatits C virus (CD81), HIV (CD63). Although their precise activities are largely undefined, tetraspanins appear to act as 'molecular organizers', regulating the formation and activity of membrane microdomains. We are using site directed mutagenesis, recombinant protein expression and phage display techniques to further investigate the structure and functions of human tetraspanins proteins (1-5).

In collaboration with Professor Mike Blackburn in the Department of Chemistry we previously generated catalytic antibodies capable of converting an inactive prodrug into an active cytotoxic drug (6-9). Such antibodies may be potentially useful in ADEPT (antibody directed enzyme prodrug therapy) programmes for cancer treatment. We are now using antibody phage display techniques to generate catalytic antibodies with improved properties (10,11). We are also using antibody phage display techniques to examine antibody responses in patients suffering from autoimmune disease (12-15), infectious disease and cancer. We have generated antibody gene libraries from patients with ovarian cancer and have recently isolated human antibodies to the ovarian tumour marker human placental alkaline phosphatase (HPLAP) from a semi-synthetic library.

The diagram shows a giant multinucleated cell. These cells are formed by the fusion of monocytes and are feature of chronic inflammation and reactions to foreign bodies (e.g. implants). Monocytes also fuse to form osteoclasts, cells involved in bone resorption. Our research, using recombinant tetraspanin proteins and specific anti-tetraspanin antibodies, suggests a role for tetraspanins in controlling this process.

Selected Publications

Strategies for targeting tetraspanin proteins: potential therapeutic applications in microbial infections. Hassuna N, Monk PN, Moseley GW, Partridge LJ. BioDrugs. 2009;23(6):341-59.

Distinct roles for tetraspanins CD9, CD63 and CD81 in the formation of multinucleated giant cells. Parthasarathy V, Martin F, Higginbottom A, Murray H, Moseley GW, Read RC, Mal G, Hulme R, Monk PN, Partridge LJ. Immunology. 2009 Jun;127(2):237-48.

Characterisation of receptor binding by the chemotaxis inhibitory protein of Staphylococcus aureus and the effects of the host immune response. Wright AJ, Higginbottom A, Philippe D, Upadhyay A, Bagby S, Read RC, Monk PN, Partridge LJ. Mol Immunol. 2007 Apr;44(10):2507-17.

The role of the N-terminal domain of the complement fragment receptor C5L2 in ligand binding. Scola AM, Higginbottom A, Partridge LJ, Reid RC, Woodruff T, Taylor SM, Fairlie DP, Monk PN. J Biol Chem. 2007 Feb 9;282(6):3664-71.

Recombinant extracellular domains of tetraspanin proteins are potent inhibitors of the infection of macrophages by human immunodeficiency virus type 1. Ho SH, Martin F, Higginbottom A, Partridge LJ, Parthasarathy V, Moseley GW, Lopez P, Cheng-Mayer C, Monk PN. J Virol. 2006 Jul;80(13):6487-96.

Hamidpour M, Behrendt M, Griffiths B, Partridge L, Lindsey N.(2006) The isolation and characterisation of antiplatelet antibodies. Eur J Haematol. 76:331-8.

[1] Higginbottom A, McCullough B, Lanza F, Azorsa DO, Wilkinson I, Partridge LJ, Monk PN. (2000) Antibody cross linking of the human tetraspanin protein, CD9 and the high affinity IgE receptor can stimulate secretion from transfected rat basophilic leukemia cells. Immunology 99 546-552

[2] Partridge, L.J., Moseley, G.W., Higginbottom, A., and Monk, P.N.(2002) Selective anti-tetraspanin antibody activation of RBL cells activated with human tetraspanins. p667-670 In Leucocyte Typing VII. Ed D. Mason et al. Oxford University Press.

[3] Higginbottom, A., Takahashi, Y., Bolling, L., Coonrod, S. A., White, J. M., Partridge, L. J., and Monk, P. N. (2003) Structural requirements for the inhibitory action of the CD9 large extracellular domain in sperm/oocyte binding and fusion. Biochem. Biophys. Res. Commun. 311 208-214.

[4] Moseley, G. W., Elliot, J., Wright, M.D., Partridge, L.J. and P. N. Monk (2003) CD63 Expression on Human Cancer Cell Lines. Int. J. Cancer 105 613-616

[5] Martin, F, Roth, D, Jans, DA, Pouton, JC, Partridge, LJ, Monk PN, Moseley GW. (2005). Tetrasapnins in viral infections: a fundamental role in viral biology? J. Virology in press.

[6] Wentworth, P., Datta, A., Blakey, D., Boyle, T., Partridge, L.J. and Blackburn, G.M. (1996) Toward antibody-directed abzyme prodrug therapy, ADAPT; carbamate prodrug activation by a catalytic antibody and its in vitro application to human tumor cell killing. Proc. Natl. Acad Sci USA 93:799-803.

[7] Blackburn, G.M., Datta, A. and Partridge, L.J. (1996) The medical potential of catalytic antibodies. Pure & Appl. Chem. 68 2009-2016

[8] Ruzheinnikov, S.N., Muranova, T.A., Sedelnikova, S.E., Partridge, L.J., Blackburn, G.M., Murray, I.A., Kakinuma, H., Takahashi-Ando, N., Shimazaki, K., Sun, J., Nishi, Y. and Rice, D.W. (2003) J. Mol. Biol. 332 "High-resolution crystal structure of the Fab-fragments of a family of mouse catalytic antibodies with esterase activity." J. Mol. Biol. 332 423-435

[9] Muranova, T.A., Ruzheinikov, S.N.,Sedelnikova,S.E., Moir, A.. Partridge, L.J., Kakinuma,H., Takahashi, N., Sun, J., Nishi,Y. & Rice, D.W. (2001) The preparation and crystallization of Fab fragments of a family of mouse esterolytic catalytic antibodies and their complexes with a transition state analogue. Acta Crystallographica 57 1192-1199

[10] Betley, J.R., Cesaro-Tadic, S., Mekhalfia, A., Rickard, J.H., Denham, H., Partridge, L.J., Plückthun, A. and Blackburn, G.M. (2002) Direct screening for phosphatase activity by turnover-based capture of protein catalysts. Angew. Chem. 41 775-777

[11] Cesaro-Tadic, S., Lagos, D., Honegger, A., Partridge, L.J., Blackburn, G.M., Plückthun, A. (2003) Turnover-based in vitro selection and evolution of biocatalysts from a fully synthetic antibody library. Nature Biotech. 21 679-685

[12] Yazici ZA, Beherendt M, Goodfield M. Partridge LJ and Lindsey NJ. (1998) Does the CDR 3 of the heavy chain determine specificity of autoantibodies in systemic lupus erythematosus? J. Autoimmunity 11:477-83.

[13] Yazici ZA, Behrendt, M., Cooper, D., Goodfield, M., Partridge, L.J. and Lindsey, N.J (2000). The identification of endothelial cell autoantigens J. Autoimmunity 15 41-49

[14] Behrendt M., Partridge L.J., Griffiths B, Goodfield M, Snaith M and Lindsey N.J (2003) The role of somatic mutation in determining the affinity of anti-DNA antibodies. Clin.Exp. Immunol. 131 182-189

[15] Clayton, R., Cooke, I.D., Partridge, L.J. and Moore, H.D.M. (1998) A combinatorial phage display library for the generation of specific Fab fragments recognizing human spermatozoa and inhibiting fertilization capacity in vitro. Biol. Reprod. 59 1180-1186