Abstract
The complement system helps in the lysis of invading pathogens and modulates the inflammatory as well as the humoral and cellular immune responses. C5 mediates many potent inflammatory and cytolytic events after proteolytic activation by complement convertase enzymes. Hence, to investigate the role of pig C5 (pC5) as a candidate gene for disease resistance in pigs, the complete cDNA of pC5 was sequenced, screened for single nucleotide polymorphisms (SNPs), and an association analysis with various immunological parameters measured in F2 animals of a pig resource population based on a cross of Duroc and Berlin miniature pigs (DUMI) was carried out. In total, 5,422 bp of pC5 cDNA was sequenced, which codes for the 1,677-amino-acid precursor of C5. Four polymorphic sites were detected, one of which was segregating in the DUMI population in three genotypic patterns: AA, AC and CC. Classical (CH50) and alternative (AH50) complement activities, C3c levels, haptoglobin (HP) acute phase protein levels, and antibody titers against Mycoplasma (Mk) and Aujesky (ADV) vaccines were measured in the resource population. Association analysis between C5 and the immunological parameters was carried out using repeated measures mixed and general linear model analysis. The homozygote AA was found to be significantly different from the other two genotypes with respect to AH50 and CH50, whereas genotype CC was found to be significantly different from the other genotypes for C3c and HP levels. No significant difference could be seen between genotypes for antibody titers against vaccinations. Association of C5 with complement activity traits and acute phase proteins promotes pC5 as a candidate gene for innate disease resistance.
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References
Asai T, Mori M, Okada M, Uruno K, Yazawa S, Shibata I (1999) Elevated serum haptoglobin in pigs infected with porcine reproductive and respiratory syndrome virus. Vet Immunol Immunopathol 70:143–148
Bredt W, Wellek B, Brunner H, Loos M (1977) Interactions between Mycoplasma pneumoniae and the first components of complement. Infect Immun 15:7–12
Franchine S, Zarkadis IK, Sfyroera G, Sahu A, Moore WT, Mastellos D, LaPatra SE, Lambris JD (2001) Cloning and purification of the rainbow trout fifth component of complement (C5). Dev Comp Immunol 25:419–30
Greer J (1985) Model structure for the inflammatory protein C5a. Science 228:1055–1060
Haviland DL, Haviland JC, Fleischer DT, Hunt A, Wetsel RA (1991) Complete cDNA sequence of human complement pro-C5: evidence of truncated transcripts derived from a single copy gene. J Immunol 146:362–368
Hughes AL (1994) Phylogeny of the C3/C4/C5 complement component gene family indicates that C5 diverged first. Mol Biol Evol 11:417–425
Karp LC, Grupe A, Schadt E, Ewart SL, Keane-Moore M, Cuoma PJ, Kohl J, Wahl L, Kuperman D, Germer S, Aud D, Peltz G, Karp MW (2000) Identification of complement factor 5 as a susceptibility locus for experimental allergic asthma. Nat Immunol 1:221–226
Kasahara M (1998) What do the paralogous regions in the genome tell us about the origin of the adaptive immune system? Immunol Rev 166:159–175
Kato Y, Nakao M, Mutsuro J (2003) The complement component C5 of common carp (Cyprinus carpio): cDNA cloning of two distinct isotypes that differ in a functional site. Immunogenetics 54:807–815
Komatsu M, Yoshiwara S, Akita T (1998) Immunological characteristics of the Meishan pig: serum complement activity, serum C3 level, immune response and skin structure. Proceedings of the 6th world congress on genetics applied to livestock production. Armidale, N.S.W., Australia, pp 323–326
Law SKA, Reid KBM (1995) Complement. In: Male D (ed) Complement, 2nd edn. IRL Press, Oxford
Lopez-Corrales NL, Beattie CW, Rohrer GA (1999) Cytogenetic assignment of 53 microsatellites from the USDA-MARC porcine genetic map. Cytogenet Cell Genet 84:140–144
Lundwall AB, Wetsel RA, Kristensen T, Whitehead AS, Woods DE, Ogden RC, Colten HR Tack BF (1985) Isolation and sequence of a cDNA clone encoding the fifth complement component J Biol Chem 260:2108–2112
Mallard BA, Wilkie BN, Kennedy BW (1989) Influence of major histocompatibility genes on serum hemolytic complement activity in miniature swine. Am J Vet Res 50:359–363
Mayes JT, Schreiber RD, Cooper NR (1984) Development and application of an enzyme-linked immunosorbent assay for the quantitation of alternative complement pathway activation in human serum. J Clin Invest 73:160–170
Mollenss TE, Redl H, Hogasen K, Bengtsson A, Garred P, Speilberg L, Lea T, Oppermann M, Gotze O, Schlag G (1993) Complement activation in septic baboons detected by neoepitope-specific assays for C3b/iC3b/C3c, C5a and the terminal C5b-9 complement complex (TCC). Clin Exp Immunol 91:295–300
Nilsson UR, Mandle RJ, McConnel-Mapes JA (1975) Human C3 and C5: subunit structure and modification by trypsin and C42-C423. J Immunol 114:815–822
Nonaka M, Natsuume-Sakai S, Takahashi M (1981) The complement system in rainbow trout (Salmo gairdneri). J Immunol 126:1495–1498
Peter G, Weigert MB, Bissel AR, Gold R, Kreutzer D, McLean RH (1981) Meningococcal meningitis in familial deficiency of the fifth component of complement. Pediatrics 67:882–886
Ponsuksili S, Wimmers K, Yerle M, Schellander K (2001) Mapping of 93 porcine ESTs preferentially expressed in liver. Mamm Genome 12:869–872
Rekitt M, Hiss S, Dickhöfer D, Wimmers K, Schellander K, Petersen B, Sauerwein H (2001) Haptoglobin responses towards vaccination with three different antigens in pigs. Proceedings of the 11th International Conference on Production of Diseases in Farm animals, Frederiskberg, Denmark, p62
Rosenfeld SI, Baum J, Steigbigel RT, Leddy JP (1976) Hereditary deficiency of the fifth component of complement in man. II. Biological properties of C5-deficient human serum J Clin Invest 57:1635–1643
Ross SC, Densen P (1984) Complement deficiency states and infection: epidemiology, pathogenesis and consequences of neisserial and other infection in an immune deficiency. Medicine 63:243–273
Song WC, Sarrias MR, Lambris JD (2000) Complement and innate immunity. Immunopharmacology 49:187–198
Sunyer JO, Zarkadis IK, Lambris JD (1998) Complement diversity: a mechanism for generating immune diversity? Immunol Today 19:519–523
Tyler JW, Cullor JS, Osburn BI, Parker K (1988) Age-related variations in serum concentrations of the third component of complement in swine. Am J Vet Res 49:1104–1106
van den Berg CW, Hofhuis FM, Rademaker PM, van Dijk H (1991) Induction of active immunological hypo/non-responsiveness to C5 in adult C5-deficient DBA/2 mice. Immunology 74:380–385
Wetsel RA, Ogata RT, Tack BF (1987) Primary structure of the fifth component of murine complement. Biochemistry 26:737–743
Westel RA, Lemons RS, LeBeau MM, Barnum SR, Noack D, Tack BF (1988) Molecular analysis of human complement component C5: localization of the structural gene to chromosome 9. Biochemistry 27:1472–1482
Westel RA, Flesicher DT, Haviland DL (1990) Deficiency of the murine fifth complement component (C5). A 2-base pair gene deletion in a 5′-exon. J Biol Chem 265:2435–2440
Wimmers K, Mekchay S, Schellander K, Ponsuksili S (2003) Molecular characterization of the pig C3 gene and its association with complement activity. Immunogenetics 54:714–724
Acknowledgements
We like to acknowledge DAAD for support in form of a scholarship to G.K. Kumar. The experiments described in this study were performed in accordance with all appropriate regulations.
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Nucleotide sequence data reported are available in GenBank database under the accession numbers AY332748 and AY349423
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Kumar, K.G., Ponsuksili, S., Schellander, K. et al. Molecular cloning and sequencing of porcine C5 gene and its association with immunological traits. Immunogenetics 55, 811–817 (2004). https://doi.org/10.1007/s00251-003-0637-y
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DOI: https://doi.org/10.1007/s00251-003-0637-y