• Asian-Australasian Journal of Animal Sciences
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• v.28 no.9
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• pp.1354-1361
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• 2015

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 $F_2$ animals of a resource population (DUMI: $DU{\times}BMP$) 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.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.571-577
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• 1990

Alkali extract obtained from mycelia of Lentinus edodes IY105 was shown to have potent anticomplementary activity and alternative complementary activity in vitro. It was also shown to activate reticuloendothelial system of ICR mice in the carbon clearance. Amount of carbohydrates and protein of the extract were 17% and 42% respectively. It was found that carbohydrates were consisted of 5 monosaccharides and protein was consisted of 16 amino acids. Antitumor activity was observed in the alkali extract treated group of sarcoma 180 bearing ICR mice.

• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.490-495
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• 1996

The anti-compliment activity of hemolytic complementary assay(TCH50) of rhamnan sulfate fraction obtained from water extracts of Monostroma nitidum was investigated Rhamnan sulfate Fraction, F-4-3 fraction appeared relatively strong anti-complementary activity which decreased TCH50 over 60% than that comparison with control, and F-4-3 considerably inhibited ACH50. F-4-3 inhibited formation of the classical pathway C3 convertase or C4 cleavage. The results also indicate the mode of complement activation by F-4-3 fraction shows not only the classical pathway but also the alternative pathway.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.290-295
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• 2002

Among 12 lactic acid bacteria examined for their abilities to activate the complement system by hemolytic complement assay $(TCH_{50})$, Lactobacillus plantarum previously isolated from Kimchi showed high anti-complementary activity. The anti-complementary activity of the cell wall fraction of L. plantarum was more potent than that of the cytosol fraction, and both activities showed dose dependency. These high activities of the cytosol and the cell wall fractions were relatively resistant to the digestion with pronase, but sharply decreased after the treatment of $NaIO_4$. These results suggested that the complement activation by the cytosol and the cell wall fractions was mainly due to their polysaccharides. By the cross-immunoelectrophoresis using anti-human C3, the C3 activation products from both fractions were identified in $Ca^{++}$-free condition. Anti-complementary activity $(ITCH_{50})$ of the cell wall fraction was retained under the same condition, whereas that of the cytosol fraction was reduced considerably. From these results, it was inferred that the mode of complement activation by the cell wall fraction was mainly via alternative pathway, and that of the cytosol fraction was via both alternative and classical pathways.

• Applied Biological Chemistry
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• v.39 no.2
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• pp.147-152
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• 1996

To screen inhibitors on complement system from natural resources, micro-screening method was established by using hemolytic complement assay. Complement fixation reaction was carried out in the microplate system. For standard hemolysis (50% hemolysis) of the classical pathway (CP), hemolysin and complement serum were diluted to $1/75{\sim}1/100\;and\;1/80{\sim}1/120$, respectively, when sheep erythrocytes were $5.0{\times}10^8\;cells/ml$. In case of the alternative pathway (AP), complement serum was diluted to 1/5 and EGTA and $Mg^{2+}$ were added 4 mM, $4{\sim}8\;mM$, respectively, when rabbit erythrocytes were $4.0{\times}10^8\;cells/ml$. Dimethyl sulfoxide was used for the assay of non-aquous soluble compounds or extracts and its final concentration was not more than 1%. Three phenylpropanoids showed anticomplementary activities in proportion to the concentration for both pathways and rosmarinic acid exihibited the highest inhibitory activities: $5.4{\pm}3.6%(0.063\;mM){\sim}95.8{\pm}0.2%(0.5\;mM)\;and\;35.1{\pm}0.9%(0.063\;mM){\sim}95.6{\pm}1.1%(1\;mM)$ on the CP and the AP, respectively.

• Korean Journal of Food Science and Technology
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• v.25 no.3
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• pp.197-203
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• 1993

Screenings were performed on edible plants to examine their complement-system activating ability (anti-complementary activity) by hemolytic complement assay $(TCH_{50})$. Among 38 kinds of plant extracts, 5 kinds showed relatively strong anti-complementary activity which decreased $TCH_{50}$ more than 60% comparison with control and the order of activity was Zingiber officinale>Colocasia antiquorum>Capsella bursapastoris>Ginkgo biloba>Alium monanthum in $1000{\mu}g/ml$. The anti-complementary activity of ZR-1 prepared from the root of Zingiber officinale which was showed the most potent activity, did not change by pronase treatment, but decreased greatly by periodate oxidation. These results indicate that not protein moiety but carbohydrate moiety in ZR-1 fraction may also contribute to the anti-complementary activity. Also, the anti-complementary activity of ZR-1 was reduced partially in the absence of the $Ca^{2+}$ ion. When crossed immunoelectrophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the ZR-1 in $Ca^{2+}$ free condition, a cleavage of C3 precipitin line was observed. Furthermore this polysaccharide fraction considerably inhibited $ACH_{50}$. These results also indicate that the mode of complement activation by polysaccharide from Zingiber officinale is via not only the classical pathway but also the alternative pathway.

• Applied Biological Chemistry
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• v.35 no.1
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• pp.42-50
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• 1992

We conducted screening on Chinese medicinal herbs to examine their anti-complementary activity by hemolytic complementary assay $(TCH_{50})$. Among 55 kinds of herbs, several herbs showed relatively potent anti-complementary activity which decreased $TCH_{50}$, more than 70% in comparison with control. Then, hot water extracts of the following herbs, Curcuma aromatica, Areca catechu, Gleditsiae spina, Euonymus alata, Acanthopanax senticous. Lonicera japonica, Aconitum carmichaeli, Curcuma zedoaria and Cinnamoum cassia, which were shown relatively potent anti-complementary activity were partially purified and analyzed their chemical properties. These activities were resistant to digestion with pronase but decreased by treatment with $NaIO_4$. These results may indicate that the complement activating ability in their herbs is due to polysaccharide. Furthermore, the anti-complementary activity of Areca catechu which was showed the most potent activity, was reduced partially in the absence of the $Ca^{++}\;ion$. After incubation of the normal human serum with partially purified polysaccharide of A. catechu in the absence of $Ca^{++}\;ion$, a cleavage of C3 in the serum was found to have occurred through immunoelectrophoresis using rabbit anti-human C3 serum. These results indicate that the mode of complement activation by polysaccharide of A. catechu is via both the alternative and classical pathway.