STUDIES ON ANTIBODY PRODUCTION: I. A METHOD FOR THE HISTOCHEMICAL DEMONSTRATION OF SPECIFIC ANTIBODY AND ITS APPLICATION TO A STUDY OF THE HYPERIMMUNE RABBIT

doi:10.1084/jem.102.1.49

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The Journal of Experimental Medicine, Vol 102, 49-60, Copyright, 1955, by The Rockefeller Institute for Medical Research New York

ARTICLE

STUDIES ON ANTIBODY PRODUCTION : I. A METHOD FOR THE HISTOCHEMICAL DEMONSTRATION OF SPECIFIC ANTIBODY AND ITS APPLICATION TO A STUDY OF THE HYPERIMMUNE RABBIT

Albert H. Coons M.D.¹, Elizabeth H. Leduc Ph.D.¹, and Jeanne M. Connolly ¹

¹ From the Department of Bacteriology and Immunology and the Department of Anatomy, Harvard Medical School, Boston

A method for the specific histochemical demonstration of antibodyin cells and parts of cells is described. It consists of carryingout a two stage immunological reaction on frozen sections oftissues: (a) allowing reaction between antibody in the tissueand dilute antigen applied in vitro, and (b) the detection ofthose areas where this antigen has been specifically absorbedby means of a precipitin reaction carried out with fluorescein-labelledantibody. Examination under the fluorescence microscope revealsthe yellow-green fluorescence of fluorescein over those areaswhere a precipitate has formed.

A study of the hyperimmune rabbiton the first few days after the last of a series of intravenousantigen injections reveals that antibody against human $gamma$ -globulinor ovalbumin is present in groups of plasma cells in the redpulp of the spleen, the medullary areas of lymph nodes, thesubmucosa of the ileum, and the portal connective tissue ofthe liver. Because of extensive non-specific reactions, thebone marrow could not be examined. Small amounts of antibodywere occasionally visible in cells in the lymphoid folliclesof the spleen and lymph nodes, so that a minor contributionby lymphocytes to antibody synthesis cannot be excluded.

Submitted on March 2, 1955

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