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Characterization of an IL-6-like molecule from an echinoderm (Asterias forbesi)
1Department of Pathology, SUNY at Stony Brook 11794-8691, USA.
Our previous studies have demonstrated the presence of two of the three vertebrate inflammatory cytokines in invertebrates, namely, interleukin (IL)-1 and tumour necrosis factor (TNF). We studied the coelomic fluid of the echinoderm, Asterias forbesi, to determine whether it contained the third inflammatory cytokine, IL-6. Coelomic fluids were concentrated and then fractionated by a combination of gel sieve chromatography and preparative isoelectric focusing. The M, observed for the invertebrate IL-6-like molecule was approximately 30,000 and the pI was 5.5. Western blot analysis using a polyclonal rabbit antiserum to human IL-6 detected a band at approximately 30,000 Da. The invertebrate IL-6 was inhibited by an antiserum to human IL-6 when used in the B9 assay. Finally, the coelomocytes were found to be capable or releasing the IL-6-like molecule as early as 12 h following stimulation by LPS. These results, together with our other data, show that echinoderms possess correlates of all three vertebrate inflammatory cytokines.
Our previous studies have demonstrated the presence of two of the three vertebrate inflammatory cytokines in invertebrates, namely, interleukin (IL)-1 and tumour necrosis factor (TNF). We studied the coelomic fluid of the echinoderm, Asterias forbesi, to determine whether it contained the third inflammatory cytokine, IL-6. Coelomic fluids were concentrated and then fractionated by a combination of gel sieve chromatography and preparative isoelectric focusing. The M, observed for the invertebrate IL-6-like molecule was approximately 30,000 and the pI was 5.5. Western blot analysis using a polyclonal rabbit antiserum to human IL-6 detected a band at approximately 30,000 Da. The invertebrate IL-6 was inhibited by an antiserum to human IL-6 when used in the B9 assay. Finally, the coelomocytes were found to be capable or releasing the IL-6-like molecule as early as 12 h following stimulation by LPS. These results, together with our other data, show that echinoderms possess correlates of all three vertebrate inflammatory cytokines.