Characterization of serum complement activity of saltwater ( Crocodylus porosus) and freshwater ( Crocodylus johnstoni) crocodiles
We employed a spectroscopic assay, based on the hemolysis of sheep red blood cells (SRBCs), to assess the innate immune function of saltwater and freshwater crocodiles in vitro. Incubation of serum from freshwater and saltwater crocodiles with SRBCs resulted in concentration-dependent increases in S...
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| Published in | Comparative biochemistry and physiology. Part A, Molecular & integrative physiology Vol. 143; no. 4; pp. 488 - 493 |
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| Main Authors | , |
| Format | Journal Article |
| Language | English |
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United States
Elsevier Inc
01.04.2006
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| Abstract | We employed a spectroscopic assay, based on the hemolysis of sheep red blood cells (SRBCs), to assess the innate immune function of saltwater and freshwater crocodiles in vitro. Incubation of serum from freshwater and saltwater crocodiles with SRBCs resulted in concentration-dependent increases in SRBC hemolysis. The hemolytic activity occurred rapidly, with detectable activity within 2 min and maximum activity at 20 min. These activities, in both crocodilian species, were heat sensitive, unaffected by 20 mM methylamine, and completely inhibited by low concentrations of EDTA, suggesting that the alternative serum complement cascade is responsible for the observed effects. The hemolytic activities of the sera were inhibited by other chelators of divalent metal ions, such as phosphate and citrate. The inhibition of SRBC hemolysis by EDTA could be completely restored by the addition of 10 mM Ca
2+ or Mg
2+, but not Ba
2+, Cu
2+ or Fe
2+, indicating specificity for these metal ions. The serum complement activities of both crocodilians were temperature-dependent, with peak activities occurring at 25–30 °C and reduced activities below 25 °C and above 35 °C. |
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| AbstractList | We employed a spectroscopic assay, based on the hemolysis of sheep red blood cells (SRBCs), to assess the innate immune function of saltwater and freshwater crocodiles in vitro. Incubation of serum from freshwater and saltwater crocodiles with SRBCs resulted in concentration-dependent increases in SRBC hemolysis. The hemolytic activity occurred rapidly, with detectable activity within 2 min and maximum activity at 20 min. These activities, in both crocodilian species, were heat sensitive, unaffected by 20 mM methylamine, and completely inhibited by low concentrations of EDTA, suggesting that the alternative serum complement cascade is responsible for the observed effects. The hemolytic activities of the sera were inhibited by other chelators of divalent metal ions, such as phosphate and citrate. The inhibition of SRBC hemolysis by EDTA could be completely restored by the addition of 10 mM Ca
2+ or Mg
2+, but not Ba
2+, Cu
2+ or Fe
2+, indicating specificity for these metal ions. The serum complement activities of both crocodilians were temperature-dependent, with peak activities occurring at 25–30 °C and reduced activities below 25 °C and above 35 °C. We employed a spectroscopic assay, based on the hemolysis of sheep red blood cells (SRBCs), to assess the innate immune function of saltwater and freshwater crocodiles in vitro. Incubation of serum from freshwater and saltwater crocodiles with SRBCs resulted in concentration-dependent increases in SRBC hemolysis. The hemolytic activity occurred rapidly, with detectable activity within 2 min and maximum activity at 20 min. These activities, in both crocodilian species, were heat sensitive, unaffected by 20 mM methylamine, and completely inhibited by low concentrations of EDTA, suggesting that the alternative serum complement cascade is responsible for the observed effects. The hemolytic activities of the sera were inhibited by other chelators of divalent metal ions, such as phosphate and citrate. The inhibition of SRBC hemolysis by EDTA could be completely restored by the addition of 10 mM Ca2+ or Mg2+, but not Ba2+, Cu2+ or Fe2+, indicating specificity for these metal ions. The serum complement activities of both crocodilians were temperature-dependent, with peak activities occurring at 25-30 degrees C and reduced activities below 25 degrees C and above 35 degrees C. We employed a spectroscopic assay, based on the hemolysis of sheep red blood cells (SRBCs), to assess the innate immune function of saltwater and freshwater crocodiles in vitro. Incubation of serum from freshwater and saltwater crocodiles with SRBCs resulted in concentration-dependent increases in SRBC hemolysis. The hemolytic activity occurred rapidly, with detectable activity within 2 min and maximum activity at 20 min. These activities, in both crocodilian species, were heat sensitive, unaffected by 20 mM methylamine, and completely inhibited by low concentrations of EDTA, suggesting that the alternative serum complement cascade is responsible for the observed effects. The hemolytic activities of the sera were inhibited by other chelators of divalent metal ions, such as phosphate and citrate. The inhibition of SRBC hemolysis by EDTA could be completely restored by the addition of 10 mM Ca super(2+) or Mg super(2+), but not Ba super(2+), Cu super(2+) or Fe super(2+), indicating specificity for these metal ions. The serum complement activities of both crocodilians were temperature-dependent, with peak activities occurring at 25-30 degree C and reduced activities below 25 degree C and above 35 degree C. |
| Author | Merchant, Mark Britton, Adam |
| Author_xml | – sequence: 1 givenname: Mark surname: Merchant fullname: Merchant, Mark email: mmerchan@mcneese.edu organization: Department of Chemistry, McNeese State University, Box 90455, Lake Charles, LA, 70609, USA – sequence: 2 givenname: Adam surname: Britton fullname: Britton, Adam organization: Wildlife Management Incorporated, Darwin, Northern Territory, Australia |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/16487736$$D View this record in MEDLINE/PubMed |
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| SubjectTerms | Alligators and Crocodiles - blood Animals Barium - pharmacology Calcium - pharmacology Complement System Proteins - metabolism Copper - pharmacology Crocodilian Crocodylus johnstoni Crocodylus porosus Edetic Acid - pharmacology Erythrocytes - metabolism Fresh Water Hemolysis - drug effects Humoral immunity Immunology Innate immunity Iron - pharmacology Kinetics Magnesium - pharmacology Temperature |
| Title | Characterization of serum complement activity of saltwater ( Crocodylus porosus) and freshwater ( Crocodylus johnstoni) crocodiles |
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