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The First Component (C1n) of the Complement System of the Nurse Shark (Ginglymostoma Cirratum)

I. Hemolytic Characteristics of Partially Purified C1n¹

From the Department of Microbiology, University of Miami, School of Medicine, Miami, Florida

Abstract

Nurse shark C1 (C1n) was incompatible with rabbit IgG or IgM but could be hemolytically measured with sheep erythrocytes sensitized with natural nurse shark antibody (EAn). C1n was precipitated from nurse serum by dilution with 4.0 M urea to µ = 0.04 at pH 7.3 and 0°C. The precipitate was washed twice in µ = 0.15 Veronal-buffered saline (VBS⁺⁺) and then dissolved in µ = 0.25 VBS⁺⁺. The resulting C1n preparation had high specific activity but was still contaminated with nurse IgM. When optimally sensitized EAn were reacted for 20 min at 30°C with 90 effective molecules of C1n per cell in isotonic dextrose gelatin Veronal buffer (DGVB⁺⁺) (µ = 0.12, pH 6.9), maximum reactivity of the resulting new intermediate cell (EAnC1n) was obtained. These cells could be specifically lysed with guinea pig serum devoid of C1 or whole guinea pig serum dilutions, since EAn did not react with guinea pig C1. C1n transfer from EAnC1n or the corresponding stroma to EAn was mainly dependent on the ionic strength. Antibody transfer could also be demonstrated, but antibody and C1n did not transfer as a unit. Mixed intermediate complexes: EAnC1n4^gp, EAnC4^gp could be formed.

Footnotes

¹ This report represents in part a Doctoral dissertation by Gordon D. Ross submitted to the Graduate School of the University of Miami as partial fulfillment of the requirements for the Ph.D. degree; it was supported by United States Public Health Service Grant AI-08569 from the National Institute of Allergy and Infectious Diseases. Dr. Ross was a recipient of a National Defense Education Act Title IV Fellowship while in Graduate School.

² Present address: National Jewish Hospital, Department of Allergy and Clinical Immunology, 3800 E. Colfax Avenue, Denver, Colorado 80206.

³ Requests for reprints should be directed to Dr. J. A. Jensen, Department of Microbiology, School of Medicine, University of Miami, P. O. Box 875 Biscavne Annex, Miami, Florida 33152.

This article has been cited by other articles:

				J. Jensen, E Festa, D. Smith, and M Cayer The complement system of the nurse shark: hemolytic and comparative characteristics Science, October 30, 1981; 214(4520): 566 - 569. [Abstract] [PDF]