A semi-automated microassay for complement activity

Liu, Chau‐Ching; Young, John Ding‐E

doi:10.1016/0022-1759(88)90150-0

Cited by 16 publications

(2 citation statements)

References 12 publications

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“…To distinguish between the classical (CH50) and the alternative-pathway (AH50), the standard CH50 and AH50 hemolytic assays were applied according to the methods of Liu and Young’s (1988) and Demey et al (1993) . Briefly, hemolytic complement activity in the classical and alternative pathway were expressed as the amount of serum that caused a 50% hemolysis of antibody sensitised sheep red blood cells (SRBCs) (CH50, U/mL) and 50% hemolysis of rabbit erythrocytes (AH50, U/mL) in the reaction mixture, respectively ( Wimmers et al, 2003 ).…”

Section: Methodsmentioning

confidence: 99%

Genetic Association of the Porcine C9 Complement Component with Hemolytic Complement Activity

Khoa

Wimmers

2015

Asian Australas. J. Anim. Sci

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The complement system is a part of the natural immune regulation mechanism against invading pathogens. Complement activation from three different pathways (classical, lectin, and alternative) leads to the formation of C5-convertase, an enzyme for cleavage of C5 into C5a and C5b, followed by C6, C7, C8, and C9 in membrane attack complex. The C9 is the last complement component of the terminal lytic pathway, which plays an important role in lysis of the target cells depending on its self-polymerization to form transmembrane channels. To address the association of C9 with traits related to disease resistance, the complete porcine C9 cDNA was comparatively sequenced to detect single nucleotide polymorphisms (SNPs) in pigs of the breeds Hampshire (HS), Duroc (DU), Berlin miniature pig (BMP), German Landrace (LR), Pietrain (PIE), and Muong Khuong (Vietnamese potbelly pig). Genotyping was performed in 417 F2 animals of a resource population (DUMI: DU×BMP) that were vaccinated with Mycoplasma hyopneumoniae, Aujeszky diseases virus and porcine respiratory and reproductive syndrome virus at 6, 14 and 16 weeks of age, respectively. Two SNPs were detected within the third exon. One of them has an amino acid substitution. The European porcine breeds (LR and PIE) show higher allele frequency of these SNPs than Vietnamese porcine breed (MK). Association of the substitution SNP with hemolytic complement activity indicated statistically significant differences between genotypes in the classical pathway but not in the alternative pathway. The interactions between eight time points of measurement of complement activity before and after vaccinations and genotypes were significantly different. The difference in hemolytic complement activity in the both pathways depends on genotype, kind of vaccine, age and the interaction to the other complement components. These results promote the porcine C9 (pC9) as a candidate gene to improve general animal health in the future.

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Section: Methodsmentioning

confidence: 99%

Genetic Association of the Porcine C9 Complement Component with Hemolytic Complement Activity

Khoa

Wimmers

2015

Asian Australas. J. Anim. Sci

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“…As previously described [39], 2.5% v/v normal human RBCs were exposed to various concentrations of GRFN-1 at 37°C for 30 min. All experiments were carried out in triplicate using 96-well microplate (Costar 3596) and the OD at 700 nm was monitored every 30 s. employing the SpectraMAX 250 microplate reader (Molecular Devices, Sunnyvale, CA).…”

Section: Methodsmentioning

confidence: 99%

Grifonin-1: A Small HIV-1 Entry Inhibitor Derived from the Algal Lectin, Griffithsin

et al. 2010

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BackgroundGriffithsin, a 121-residue protein isolated from a red algal Griffithsia sp., binds high mannose N-linked glycans of virus surface glycoproteins with extremely high affinity, a property that allows it to prevent the entry of primary isolates and laboratory strains of T- and M-tropic HIV-1. We used the sequence of a portion of griffithsin's sequence as a design template to create smaller peptides with antiviral and carbohydrate-binding properties.Methodology/ResultsThe new peptides derived from a trio of homologous β-sheet repeats that comprise the motifs responsible for its biological activity. Our most active antiviral peptide, grifonin-1 (GRFN-1), had an EC50 of 190.8±11.0 nM in in vitro TZM-bl assays and an EC50 of 546.6±66.1 nM in p24gag antigen release assays. GRFN-1 showed considerable structural plasticity, assuming different conformations in solvents that differed in polarity and hydrophobicity. Higher concentrations of GRFN-1 formed oligomers, based on intermolecular β-sheet interactions. Like its parent protein, GRFN-1 bound viral glycoproteins gp41 and gp120 via the N-linked glycans on their surface.ConclusionIts substantial antiviral activity and low toxicity in vitro suggest that GRFN-1 and/or its derivatives may have therapeutic potential as topical and/or systemic agents directed against HIV-1.

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