Abstract
Mannan-binding lectin (MBL) is an innate immune collectin present in the serum of humans and many farm animals. This oligomeric pattern-recognition protein effectively binds to the glycoconjugate arrays present on the surfaces of microorganisms and activates the complement system to enhance pathogen killing and clearance. MBL deficiency is often associated with immunodeficiency in humans. Although two MBLs (MBL-A and MBL-C) have been characterized in various species, the identity of porcine MBL (pMBL) was not clearly defined. In this study, we purified an MBL from porcine serum by mannose affinity, ion exchange, and size exclusion chromatography and determined many of its characteristics. Based on the N-terminal sequence, multiple sequence alignment, and relative affinities to various carbohydrate ligands, we propose that the MBL purified in this study is pMBL-A. We have generated antibodies to this protein and established an immunoassay to quantify pMBL-A in serum. Using this assay, we found breed differences in pMBL-A concentration distributions and heritability estimates. In the Duroc breed (n=588), pMBL-A concentrations show a unimodal distribution with a mean of 9,125 ng/ml. In contrast, the pMBL-A concentration distributions in the Landrace breed (n=533) show three distinct mean values: 301, 2,385, and 11,507 ng/ml. Furthermore, heritability calculations based on an additive genetic variance model with no fixed effects indicate that serum pMBL-A concentration is highly heritable in the Landrace (h 2=0.8) but not in the Duroc breed (h 2=0.15). These genetic differences may be useful in selecting breeding pigs for improved disease resistance.
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Abbreviations
- DVS:
-
Divinyl sulfone
- MASP:
-
MBL associated serine protease
- MBL:
-
Mannan-binding lectin
- pMBL:
-
Porcine MBL
- PEG:
-
Polyethylenglycol
- SPR:
-
Surface plasmon resonance
- PCR:
-
Polymerase chain reaction
References
Agah A, Montalto MC, Young K, Stahl GL (2001) Isolation, cloning and functional characterization of porcine mannose-binding lectin. Immunology 102:338–343
Dahl MR, Thiel S, Matsushita M, Fujita T, Willis AC, Christensen T, Vorup-Jensen T, Jensenius JC (2001) MASP-3 and its association with distinct complexes of the mannan-binding lectin complement activation pathway. Immunity 15:127–135
Drickamer K (1992) Engineering galactose-binding activity into a C-type mannose-binding protein. Nature 360:183–186
Fornstedt N, Porath J (1975) Characterization studies on a new lectin found in seeds of Vicia ervilia. FEBS Lett 57:187–191
Garred P, Larsen F, Madsen HO, Koch C (2003) Mannose-binding lectin deficiency—revisited. Mol Immunol 40:73–84
Hansen S, Holmskov U (1998) Structural aspects of collectins and receptors for collectins. Immunobiology 199:165–189
Hansen S, Thiel S, Willis A, Holmskov U, Jensenius JC (2000) Purification and characterization of two mannan-binding lectins from mouse serum. J Immunol 164:2610–2618
Hoppe HJ, Barlow PN, Reid KB (1994) A parallel three stranded alpha-helical bundle at the nucleation site of collagen triple-helix formation. FEBS Lett 344:191–195
Jack DL, Turner MW (2003) Anti-microbial activities of mannose-binding lectin. Biochem Soc Trans 31:753–757
Jack DL, Klein NJ, Turner MW (2001) Mannose-binding lectin: targeting the microbial world for complement attack and opsonophagocytosis. Immunol Rev 180:86–99
Janeway CA Jr, Medzhitov R (2002) Innate immune recognition. Annu Rev Immunol 20:197–216
Jensen J, Mäntysaari EA, Madsen P, Thompson R (1997) Residual maximum likelihood estimation of (co)variance components in multivariate mixed linear models using average information. J Indian Soc Agric Stat 49:215–236
Ji YH, Fujita T, Hatsuse H, Takahashi A, Matsushita M, Kawakami M (1993) Activation of the C4 and C2 components of complement by a proteinase in serum bactericidal factor, Ra reactive factor. J Immunol 150:571–578
Liu H, Jensen L, Hansen S, Petersen SV, Takahashi K, Ezekowitz AB, Hansen FD, Jensenius JC, Thiel S (2001) Characterization and quantification of mouse mannan-binding lectins (MBL-A and MBL-C) and study of acute phase responses. Scand J Immunol 53:489–497
Lynch M, Walsh B (1998) Genetics and analysis of quantitative traits. Sinauer, p 980 (ISBN 0-87893-481-2)
Madsen PJ (2002) A user’s guide to DMU. DAIS, AGB, Copenhagen, Denmark
Madsen HO, Garred P, Thiel S, Kurtzhals JA, Lamm LU, Ryder LP, Svejgaard A (1995) Interplay between promoter and structural gene variants control basal serum level of mannan-binding protein. J Immunol 155:3013–3020
Matsushita M, Fujita T (1992) Activation of the classical complement pathway by mannose-binding protein in association with a novel C1s-like serine protease. J Exp Med 176:1497–1502
Palaniyar N, Nadesalingam J, Clark H, Shih MJ, Dodds AW, Reid KB (2004) Nucleic acid is a novel ligand for innate, immune pattern recognition collectins surfactant proteins A and D and mannose-binding lectin. J Biol Chem 279:32728–32736
Storgaard P, Nielsen EH, Andersen O, Skriver E, Mortensen H, Hojrup P, Leslie G et al (1996) Isolation and characterization of porcine mannan-binding proteins of different size and ultrastructure. Scand J Immunol 43:289–296
Tan SM, Chung MC, Kon OL, Thiel S, Lee SH, Lu J (1996) Improvements on the purification of mannan-binding lectin and demonstration of its Ca(2+)-independent association with a C1s-like serine protease. Biochem J 319(Pt 2):329–332
Thiel S (1992) Mannan-binding protein, a complement activating animal lectin. Immunopharmacology 24:91–99
Thiel S, Vorup-Jensen T, Stover CM, Schwaeble W, Laursen SB, Poulsen K, Willis AC, Eggleton P, Hansen S, Holmskov U, Reid KB, Jensenius JC (1997) A second serine protease associated with mannan-binding lectin that activates complement. Nature 386:506–510
Thiel S, Petersen SV, Vorup-Jensen T, Matsushita M, Fujita T, Stover CM, Schwaeble WJ, Jensenius JC (2000) Interaction of C1q and mannan-binding lectin (MBL) with C1r, C1s, MBL-associated serine proteases 1 and 2, and the MBL-associated protein MAp19. J Immunol 165:878–887
Turner MW (1996) Mannose-binding lectin: the pluripotent molecule of the innate immune system. Immunol Today 17:532–539
Turner MW (2003) The role of mannose-binding lectin in health and disease. Mol Immunol 40:423–429
Weis WI, Drickamer K, Hendrickson WA (1992) Structure of a C-type mannose-binding protein complexed with an oligosaccharide. Nature 360:127–134
Acknowledgements
The authors wish to thank K. V. Østergaard for critical reading of the manuscript, L. R. Dal, I. M. Jepsen, and K. B. Poulsen for technical assistance, and the Danish Bacon and Meat Council for giving us access to their pigs. This work was supported by grants from the Research Center for the Management of Animal Production and Health (CEPROS), the Danish Ministry of Food, Agriculture, and Fisheries, the Danish Medical Research Council, and The Novo Nordic Foundation. Nades Palaniyar is a recipient of the Welcome Trust, UK and the Canadian Institutes for Health Research postdoctoral fellowships. The experimental procedures were conducted under the protocols approved by the Danish Animal Experiments Inspectorate and complied with the Danish Ministry of Justice Law no. 382 (10 June 1987) and Acts 739 (6 December 1988) and 333 (19 May 1990) concerning animal experimentation and care of experimental animals.
All authors contributed equally to this work.
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Juul-Madsen, H.R., Krogh-Meibom, T., Henryon, M. et al. Identification and characterization of porcine mannan-binding lectin A (pMBL-A), and determination of serum concentration heritability. Immunogenetics 58, 129–137 (2006). https://doi.org/10.1007/s00251-006-0092-7
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DOI: https://doi.org/10.1007/s00251-006-0092-7