Production of recombinant C5a from rainbow trout (Oncorhynchus mykiss): role in leucocyte chemotaxis and respiratory burst

Boshra, Hani; Peters, Rodney; Li, Jun; Sunyer, J. Oriol

doi:10.1016/j.fsi.2004.04.002

Cited by 26 publications

(19 citation statements)

References 39 publications

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“…Complement components have also been shown to enhance DC mediated CD8 þ T cell responses especially in antiviral immunity [21]. While in rainbow trout the chemoattracted leucocytes to C5a displayed DC-like shapes [32].…”

Section: Discussionmentioning

confidence: 99%

“…The DNA fragments encoding C5a or G protein were amplified using the plasmids constructed previously [32,33] as templates. To amplify the fragments encoding G-C5a, splice overlap extension (SOE) PCR [34] was applied, viz., using the plasmids described above as templates to amplify the fragments encoding G and C5a respectively, with the primer pairs Forward1/Reverse3 for G and Forward3/Reverse2 for C5a, followed by an amplification using the mixed PCR products for G and C5a as template and primer pair Forward1/Reverse2.…”

Section: Construction Of Expression Plasmids Encoding Trout C5a Ihnvmentioning

confidence: 99%

“…Cell migration was applied using a 96-well chemotaxis chamber (ChemoTX system, Neuroprobe) as previously described [32,35] to assess the chemoattractant activities of recombinant C5a, G and GC5a (10 nM in DPBS, PBS with 100 mg/L CaCl 2 and 100 mg/L MgCl 2 , pH 7.4) to trout periphery blood leukocytes (PBLs). DPBS (pH 7.4) and complement-activated trout serum (diluted 1/1000 in DPBS, pH 7.4) were used as the negative and positive controls respectively.…”

Section: Cell Migration Analysismentioning

confidence: 99%

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Complement C5a acts as molecular adjuvant in fish by enhancing antibody response to soluble antigen

LaPatra

et al. 2014

Fish & Shellfish Immunology

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Section: Discussionmentioning

confidence: 99%

Section: Construction Of Expression Plasmids Encoding Trout C5a Ihnvmentioning

confidence: 99%

Section: Cell Migration Analysismentioning

confidence: 99%

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Complement C5a acts as molecular adjuvant in fish by enhancing antibody response to soluble antigen

LaPatra

et al. 2014

Fish & Shellfish Immunology

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“…Each of the three C3a isoforms stimulated to a significant degree the respiratory burst of trout head kidney leukocytes, suggesting that teleost fish contain a C3aR (39). Recently, recombinant trout C5a has been produced and shown to play a prominent role in inducing leukocyte chemotaxis (40,41) and respiratory burst (40). Thereafter, a C5aR was identified in rainbow trout, representing the first cloned (42,43) and functionally characterized anaphylatoxin receptor in a nonmammalian species (42).…”

mentioning

confidence: 99%

Characterization of a C3a Receptor in Rainbow Trout and Xenopus: The First Identification of C3a Receptors in Nonmammalian Species

Boshra

Wang

Hove‐Madsen

et al. 2005

The Journal of Immunology

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Virtually nothing is known about the structure, function, and evolutionary origins of the C3aR in nonmammalian species. Because C3aR and C5aR are thought to have arisen from the same common ancestor, the recent characterization of a C5aR in teleost fish implied the presence of a C3aR in this animal group. In this study we report the cloning of a trout cDNA encoding a 364-aa molecule (TC3aR) that shows a high degree of sequence homology and a strong phylogenetic relationship with mammalian C3aRs. Northern blotting demonstrated that TC3aR was expressed primarily in blood leukocytes. Flow cytometric analysis and immunofluorescence microscopy showed that Abs raised against TC3aR stained to a high degree all blood B lymphocytes and, to a lesser extent, all granulocytes. More importantly, these Abs inhibited trout C3a-mediated intracellular calcium mobilization in trout leukocytes. A fascinating structural feature of TC3aR is the lack of a significant portion of the second extracellular loop (ECL2). In all C3aR molecules characterized to date, the ECL2 is exceptionally large when compared with the same region of C5aR. However, the exact function of the extra portion of ECL2 is unknown. The lack of this segment in TC3aR suggests that the extra piece of ECL2 was not necessary for the interaction of the ancestral C3aR with its ligand. Our findings represent the first C3aR characterized in nonmammalian species and support the hypothesis that if C3aR and C5aR diverged from a common ancestor, this event occurred before the emergence of teleost fish.

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“…Trout C5a, generated as a recombinant protein, showed potent chemoattractive and respiratory burst-inducing activities, as does mammalian C5a 53 . In carp, two C5a forms that probably represent allotypic variants of C5-I showed different levels of the chemoattractive activity against the head kidney leukocytes 33 .…”

Section: Lytic Pathway Componentsmentioning

confidence: 99%

Diversified Components of the Bony Fish Complement System: More Genes for Robuster Innate Defense?

Nakao

Kato-Unoki

Nakahara

et al.

Advances in Experimental Medicine and Biology

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Production of recombinant C5a from rainbow trout (Oncorhynchus mykiss): role in leucocyte chemotaxis and respiratory burst

Cited by 26 publications

References 39 publications

Complement C5a acts as molecular adjuvant in fish by enhancing antibody response to soluble antigen

Complement C5a acts as molecular adjuvant in fish by enhancing antibody response to soluble antigen

Characterization of a C3a Receptor in Rainbow Trout and Xenopus: The First Identification of C3a Receptors in Nonmammalian Species

Diversified Components of the Bony Fish Complement System: More Genes for Robuster Innate Defense?

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