. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. Reference: Biol. Bull. 176: 290-300. (June. 1989) Microscopical, Biochemical, and Immunological Studies of the Immune Defense System of the Horseshoe Crab, Limulus polyphemus PETER SUHR-JESSEN, LEIF BAEK1*, AND PER PLOUG JAKOBSEN Department of Anatomy and Cytology, University ofOdense, Campusvej 55, 5230 Odense M, Denmark, and' University Clinic for Infectious Diseases, Endotoxin Laboratory, Rigshospitalet, Copenhagen, Denmark Abstract. The immune defense system of Limulus polyphemus was explored. Two distinct cells, granulo
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. Reference: Biol. Bull. 176: 290-300. (June. 1989) Microscopical, Biochemical, and Immunological Studies of the Immune Defense System of the Horseshoe Crab, Limulus polyphemus PETER SUHR-JESSEN, LEIF BAEK1*, AND PER PLOUG JAKOBSEN Department of Anatomy and Cytology, University ofOdense, Campusvej 55, 5230 Odense M, Denmark, and' University Clinic for Infectious Diseases, Endotoxin Laboratory, Rigshospitalet, Copenhagen, Denmark Abstract. The immune defense system of Limulus polyphemus was explored. Two distinct cells, granulo- cytes and plasmatocytes, were revealed in the hemo- lymph by light microscopy of live cells, by light micros- copy serial sectioning, and by transmission electron mi- croscopy combined with immuno-gold labelling. The granulocytes (amebocytes) comprised about 99% of the hemocytes. They had a heterochromatic nucleus, dis- tended but poorly developed RER, few free ribosomes, few mitochondria, but many large secretory granules. While the majority of these were uniformly stained, structured granules were also present. Monoclonal anti- bodies revealed that coagulogen is present in both types of large secretory granules. It is suggested that a struc- tured granule is an immature stage leading to a uni- formly stained granule. The plasmatocytes had an eu- chromatic nucleus, extended RER consisting of tubular or flattened cisternae, many free ribosomes, many mito- chondria, but few, if any, large secretory granules. Co- agulogen was not detected in plasmatocytes. Following immunizations of five rabbits for more than one year with cell-free hemolymph, 14 polypeptides were immu- nogenic in rabbits as visualized by crossed immunoelec- trophoresis. However, OTarrell gel electrophoresis com- bined with silver staining identified more than 70 major polypeptides in the cell-free hemolymph. At least 60 of these had pi-values between and , and Mr-values from larger than 200 X 103 to sma
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