Hemolytic plaque formation by mouse spleen cells producing antibodies to ovalbumin

The Immune Response of Normal, Irradiated and Thymectomized Mice to Fowl Immunoglobulin G as Detected by a Hemolytic Plaque Technique

“…The PFC responses of mice immunized with alum precipitated F}G and given 2 X 10® Pertussis organisms reached much higher levels than those reported in mice immunized with rabbit immunoglobulin [24] or protein antigens such as bovine serum albumin or human gammaglobulin [7,11]. It is not clear whether the low responses obtained with these antigens resulted from poor immunogenicity of the antigen or suboptimal conjugation onto erythrocytes.…”

“…This method does not require the availability of a particular low efficiency hemolysin and is therefore easier to perform than any technique in which erythrocytes are to be sensitized by mammalian immuno globulins, as those described by Ba r t h and M e r c h a n t [1 ] and S e g r e and S e g r e [24]. Conjugation of fowl anti-SRBC yG to SRBC was readily achieved in 1 ^ h and the degree of conjugation required for a particular batch of fowl antibody could easily be worked out from the hemagglutinin titer.…”

“…Among the antigens that have been coupled are polysaccharides [14], immunoglobulins and various protein antigens [1,7,11,13,24], var ious haptens [8,15,16,21,23,29] and synthetic polypeptides [27]. The cellular responses against protein antigens have generally been small in a primary response [1,7,11,13,24], It is not clear whether the weak response resulted from failure to induce vigorous immuni zation or from inappropriate conjugation of the antigen onto erythro cytes.In this paper, we describe a simple method by which mouse lymphoid cells producing antibody against FyG can be detected. The immuni zation of fowls against SRBC allows one to separate from immune sera anti-SRBC immunoglobulin which will readily coat SRBC.…”

IMMUNOLOGIC RECOGNITION AND IMMUNOLOGIC MEMORY. Final Report.

“…After incubation and addition of complement, hemolytic plaques developed around spleen cells producing antibodies specific for rabbit IgG (16). When other antigens, such as ovalbumin (17), ribonuclease, lysozyme, bovine serum albumin or the hapten dinitrophenol (19), were linked covalently to the IgG hemolysin of rabbit origin, hemolytic plaques were obtained with cells producing antibodies against these antigens or haptens. The use of IgG hemolysin from rabbits of defined allotypes allowed the development of plaques by rabbit cells producing anti-allotype antibodies (18).…”

The Immune Response to Hapten-Azo Bovine Serum Albumin as Detected by Hemolytic Plaque Technique

“…The disadvantage of this original technique, however, was that antigens were limited to erythrocytes, although it has then been extended to the detection of anti-polysaccharide- [14], anti-hapten- [9,15,17] and anti-polyaminoacid [25] antibody, never extended to that of anti protein antibody except for the report of Segre et al [21]. However, it was restricted only to the detection of antibodies against the immuno globulins, not applicable to that of antibodies against other proteins.…”