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Mouse antisera to DNP conjugates were fractionated on agar block electrophoresis, and each fraction was tested for ability to give anaphylactic reactions in vivo in mice and guinea-pigs, and in vitro on mouse mast cells. Concentrations of IgG1 and IgG2 globulin were measured for each fraction, and the results indicate that mouse IgG1 globulin mediates both the in vivo PCA reaction in the mouse and the in vitro mast cell sensitization. PCA reactions in guinea-pigs with mouse antisera are mediated only by electrophoretic fractions containing IgG2 globulin. PCA reactions in guinea-pigs could be blocked by the addition of IgG2a myeloma containing sera to the sensitizing anti-DNP preparation, but not by addition of IgG2b or IgG1 myeloma sera. From six different IgG1 myeloma sera tested on their capacity to block PCA reactions in mice, five showed strong inhibitory activity whereas the sixth was inactive. However, also IgG2a and IgG2b myelomas were found capable of blocking PCA reactions in mice. The passive in vitro sensitization of mouse mast cells by purified mouse anti-DNP antibodies could also be blocked by the addition of specific anti-IgG2 sera. Possible interpretations of these findings are discussed. [ABSTRACT FROM AUTHOR]

Mice of several strains were immunized with DNP-haemocyanin either with Al(OH)3 or with Freund's complete adjuvant. Antibodies of immunoglobulin type γ2 and γM were detected by passive haemolysis, γ2a antibodies by PCA in guinea-pigs, γ1 antibodies by PCA in mice using 2 hours of SP, and reaginic antibodies by PCA in mice using 72 hours of SP. Three of the six strains tested produced reaginic antibodies, i.e. SWR/J, C57BL/6J and A'HeJ, but only when Al(OH)3 was used as adjuvant. All six strains produced γ1 antibodies. Reaginic antibodies were separated by chromatography on DEAE-cellulose column. [ABSTRACT FROM AUTHOR]

Passive cutaneous anaphylaxis in the guinea pig has been studied with rabbit antibody to a series of antigens of differing molecular weight. The results indicated that at a given antibody level the weight of antigen needed to elicit a reaction increases with its molecular weight. Previous observations have been confirmed that the amount of antigen needed to elicit a reaction at a high level of antibody is less than that required at a lower level. The results suggest that extremely small amounts of small molecular weight antigens might be sufficient to produce anaphylactic symptoms in highly sensitive individuals. [ABSTRACT FROM AUTHOR]

Reverse passive cutaneous anaphylaxis experiments were performed in guinea pigs with rabbit gamma globulin and human gamma globulin as antigens and horse, sheep or hen sera containing antibodies against these antigens. These antibodies were unable to directly sensitize the guinea-pig skin but when the reverse technique was used, namely, the antigen (gamma globulin) was injected before the antibody, characteristic anaphylactic reactions were obtained. It is concluded that, if a suitable antigen is used, which can be fixed to the host by the reverse technique, sera from species which cannot directly sensitize the guinea pig can nevertheless give typical anaphylactic reactions. [ABSTRACT FROM AUTHOR]

The immunogenicity of the three mono-DNP-derivatives of bacitracin A as compared with the immunogenicity of the tri-DNP-derivative of bacitracin and of bacitracin itself was investigated in randomly bred guinea-pigs. PCA and an in vitro binding assay (a modified Fart technique) were both employed for antibody detection. All of the DNP-derivatives were able to stimulate formation of anti-DNP antibodies. Mono-DNP(ornithine)-bacitracin and tri-DNP-bacitracin induced antibody production in almost all the animals; mono-DNP(histidine)-bacitracin and mono-DNP(isoleucine)-bacitracin induced antibodies in only one-half and one-fifth of the animals, respectively. No antibodies against bacitracin could be detected in animals injected with this polypeptide or with its DNP-derivatives. Investigation by PCA of the relative reactivities of the anti-DNP-bacitracin antibodies for each of the mono-DNP-derivatives of bacitracin revealed that different populations of antibodies are elicited by each of these substances. Specificity extending to the amino acid on which the DNP group is attached would appear to be present in the populations of antibodies elicited by both mono-DNP (ornithine)-bacitracin and mono-DNP(isoleucine)-bacitracin. Additional evidence would indicate that the specificity of the anti-DNP-bacitracin antibodies may extend beyond the DNP-amino acid determinant. [ABSTRACT FROM AUTHOR]

The formation of different immunoglobulin classes of mouse antibodies to dinitrophenol 9DNP) has been qualitatively determined by biological assays which discriminate between some immunoglobulin classes. Antibody responses to two other antigens were compared in five mouse strains. Mice of different strains immunized with DNP-haemocyanin (DNP-Hcy) in complete Freund's adjuvant made both γG1 and γG2a antibodies. The amounts of antibody are less in DBA and C57 mice and at 80 days after primary immunization a 2-mercaptoethanol sensitive (2MES) antibody is still present in this strains (but not in A, AKR, and C3H). When DNP-Hcy is given in Al(OH)3 very little γG2a is produced in any strain, although γG1 antibodies are produced as in mice given DNP-Hcy in complete Freund's adjuvant. Possible interpretations of these observations are discussed. [ABSTRACT FROM AUTHOR]

Interaction of mouse immunoglobulins with a mast cell tumor HC was studied by the rosette technique. All mast cells formed rosettes, and this could be inhibited by IgG1 myeloma proteins, less efficiently by lgG1, and not by IgA or IgM. Antigen antibody complexes formed under appropriate conditions also inhibit, with at least 100-fold greater efficiency than free immunoglobulin. Antisera containing reaginic activity also inhibit rosettes in the presence of antigen, but on heating the sera, a concomitant loss of reaginic activity and rosette inhibition occurs. It is proposed that there is a specific receptor on the mast cell surface for IgG1 and reaginic immunoglobulins, and that binding to this site in the form of a complex of correct antigen ratio, will initiate the anaphylactic process. [ABSTRACT FROM AUTHOR]

By study of endothelial granulopexy and of increased permeability in local, passive, cutaneous anaphylaxy of the guinea-pig it was demonstrated that in the animals kept under an antihistamine drug (2786 RP.: neoantergan) for a sufficiently prolonged period at the time of the passive transport sensitization was not obtained. This fact is referred to the lack of fixation to the cells of the antibody injected into the skin. [ABSTRACT FROM AUTHOR]