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STUDIES ON STREPTOCOCCAL HYALURONIDASE AND ANTIHYALURONIDASE. II. THE LOCALIZATION OF SITES OF ABSORPTION OF STREPTOCOCCAL HYALURONIDASE (GROUP C) WITH FLUORESCENT ANTIBODY

E. W. EMMART 1, R. M. COLE 1, E. L. MAY 1, and J. B. LONGLEY 1

1 From the National Institutes of Health, Bethesda 14, Maryland

In an earlier report it was shown that streptococcal hyaluronidase isolated from the culture medium of Group C, strain 7 is antigenic in rabbits (10, lOa). The present report demonstrates the following:

1. Antihyaluronidase globulin, isolated from the sera of rabbits immunized with streptococcal hyaluronidase, is capable of completely inhibiting in vitro the hydrolytic action of the enzyme upon hyaluronic acid. Conjugation of the antihyaluronidase globulin fraction to fluorescein isocyanate does not affect the inhibiting action of the antibody upon the hydrolytic activity of the enzyme.

2. Using the Coons fluorescent antibody technique, streptococcal hyaluronidase has been identified in tissues of mice following the injection of the enzyme in the caudal vein.

3. The treatment of control normal tissues with fluorescent antihyaluronidase globulin solution gave negative results as did the treatment of experimental tissues with fluorescent normal globulin. As an additional and highly specific control the staining action of the fluorescent antibody upon hyaluronidase could be blocked by preliminary absorption of the tissue with unconjugated antibody.

4. Streptococcal hyaluronidase when injected intravenously, is widely distributed in tissues of the body but is absorbed and retained in selected areas and cells. The localization of fluorescent antigen-antibody complex may be nuclear or cytoplasmic or both. The selection of specific cells within any given tissue suggests that the basis for absorption of the enzyme may be the functional or the physiological state of the cell as well as the histogenesis of the cell.

Submitted on September 13, 1957


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