Earthworm Coelomic Fluid Research Articles

An enkephalin-like molecule in earthworm coelomic fluid modifies leukocyte behavior

Substances that were immunoreactive in an RIA specific for met-enkephalin were detected following HPLC fractionation of earthworm coelomic fluid. Earthworm coelomocytes and human granulocytes were analyzed for changes in conformation based on measurements of cellular area and perimeter and expressed mathematically by using the Form Factor (FF). For coelomocytes the FF decreased following exposure to DAMA, a synthetic enkephalin analogue ( d-Ala 2, Met 5-enkephaliamide). DAMA stimulated migration whereas untreated cells and those exposed to the specific opiate blocker naloxone did not move. The enkephalin-like molecule when exposed to human granulocytes stimulated an increased number of activated cells. Our results suggest a relationship between the immune and nervous systems of earthworms.

Molecules in earthworm coelomic fluid that bind anti-IgA and anti-IgG, but not anti-IgM.

Coelomic fluid from the earthworm Lumbricus terrestris is known to contain immunoactive cells and molecules involved in immune defense. To assess the ability of such molecules to form precipitating complexes with anti-human immunoglobulins, radial immunodiffusion experiments were conducted. Anti-human IgG, anti-human IgA, and anti-human IgM were the three immunoglobulins examined. Earthworm coelomic fluid was found to contain molecules that bind anti-IgA and anti-IgG. No precipitation reactions were observed with anti-IgM. Earthworm coelomic fluid diluted 1:8 still formed precipitating complexes with anti-IgA and anti-IgG. Elucidation of the earthworm binding site on anti-IgG and anti-IgA could make earthworm coelomic fluid a valuable reagent in immunological, chemical and biological research.

Protein analysis of earthworm coelomic fluid. III. Isolation and characterization of several bacteriostatic molecules from Eisenia fetida andrei

A bacteriostatic activity in Eisenia fetida andrei cell free coelomic fluid is described. This activity is detected by growth inhibition of a bacteria Bacillus megaterium. Gel filtration analysis revealed eleven coelomic fluid protein fractions designated A, B,.. J. Antibacterial activity was mainly found within fractions B and C. Chromatofocusing resolved fractions B-C into five different peaks named αBC, βBC,...εBC. Antibacterial activity appeared mediated by three different proteins characterized by their molecular weigths (20,000, 40,000 and 45,000) and their isoelectric points (4.9, 5.75 and 6.0). These bacteriostatic proteins possess either hemolysis or hemagglutination activities. The polymorphic aspect of this humoral antibacterial defense is discussed.

Isolation of agglutinins from lysisn in earthworth coelomic fluid by gel filtration followed by chromtofocusing

Agglutinin, hemolysin and bacteriostatin activities have been detected in several fractions of earthworm coelomic fluid separated by gel filtration. Chromatofocusing performed on fractions A (pH gradient 7.4–4.5), B (pH gradient 7.5–5.0) and C (pH gradient 6.2–4.0) have yielded several single proteins characterized by sodium dodecyl sulphate—polyacrylamide gel electrophoresis and analytical isoelectric focusing. Four different molecules are capable of agglutination: MW 11,500 (p I 7.5); MW 20,000 (p I 4.8); MW 32,000 (p I 1.48) and MW 40,000 (p I 6.3). Three molecules have an hemolytic activity: MW 40,000 (p I 6.3); MW 40,000 (p I 5.9) and MW 45,000 (p I 6.0). The molecule having MW 40,000 (p I 6.3) appears to be a multivalent lytic protein also capable of agglutination.

Mitogenic Effect of Earthworm (Lumbricus terrestris)Coelomic Fluid on Mouse and Human Lymphocytes

We have cultured mouse and human lymphocytes with earthworm (Lumbricus terrestris)coelomic fluid and measured their mitogenic responses. Normal fluid was collected from untreated worms, while induced fluid was harvested from worms injected 24 h earlier with rabbit erythrocytes. At low protein concentrations in the coelomic fluid, human and mouse lymphocytes were significantly activated, as measured by incorporation of 3HTdr. The activation index for induced fluid was approximately 2.5 times that for uninduced fluid. Separation of mouse lymphocytes into B- and T-cell populations revealed that primarily T cells were activated. Normal and induced coelomic fluid contains 12 and 28 electrophoretic bands and agglutinin titers of 32 and 1024, respectively. Addition of agglutinin inhibitors or absorption of agglutinins from coelomic fluid did not alter levels of mitogenic activity, thus the relation between earthworm agglutinins and mitogens is problematic. Techniques designed to separate and purify the agglutinins are in progress to elucidate this point.