A Simple Chromatographic Method for Preparation of Gamma Globulin.

In vivo allotypic suppression in rabbits has been described in newborn heterozygotes which were offspring of mothers immunized against the father's immunoglobulin aUotype (1) or which were injected at birth with antiserum directed against the father's allotype (2). This suppressed allotype is either entirely unexpressed or expressed in very low amounts (1).In rabbit in vitro systems, anti-allotypic antibodies induce blast transformation (3) and inhibit the synthesis of allotypes (4) whereas Nocardia water-soluble mitogen (NWSM) is a B-cell mitogen for several mammalian species (5, 6) as polyclonal activator induces the differentiation of rabbit small lymphocytes to plasma cells and their polyclonal activation (7).The aim of the present work was to study the synthesis of allotypes by lymphocytes from suppressed rabbits subsequent to in vitro stimulation by NWSM. Materials and MethodsAnimals. 4 day, 3, 6, and 12-wk-old offspring (b4/b5) were obtained from b4/b4 mothers immunized against the paternal allotype bS/b5 according to methods described by Dray (1), 12-too old offspring which had escaped from maternally allotypic suppression and 12-too old normal heterozygous b4/b5 rabbits (Pasteur Institute, Garches, France) were used in these experiments.Reagents. Rabbit antiallotypic sera directed against the a and b series allotypes were obtained according to Oudin (8) NWSM from Nocardia opaca was prepared according to Ciorbaru et al. (9) Concanavalin A (Con A) was obtained from Miles Yeda Ltd. and [aH]thyrnidine with 1 Ci/mM sp act was obtained from Commissariat a l'Energie Atomique, Saclay, France.Blast Transformation Assay. Single cell suspensions were obtained from spleens according to a previously described technique (7). Lymphocytes (1.5 x 10 e were cultured for 3 days in I ml RPML-1640 medium (Grand Island Biological Company, Grand Island, N. Y.) supplemented with 15% heat inactivated b6/b6 rabbit serum. The cultures were performed in plastic tubes which were incubated in an incubated model 1-H 100 Gallenkamp under a continuous flow mixture of 5% CO2 and 95% Air.[SH]thymidine (1 pCi) was added to each culture tube 12 h before harvesting the cells and incorporation of radioactivity was measured by liquid scintillation counting (7).Measure ofln Vitro Synthesis of AUotype. Splenic lymphocytes (25 x 10 e) were cultured for 72 h in 5 ml RPML-1640 medium supplemented with 10% fetal calf serum (Flow Laboratories, Inglewood, Calif.) in the same conditions described above.The amount of immunoglobulin in 72-h supernatant cultures was measured by a quantitative inhibition radioimmunoassay. IgG fractions were prepared from b4/b4 and b5/b5 homozygous rabbits by DEAE cellulose chromatography (10) and were labeled with ~5I, according to Green-* Present address:

Section: Methodsmentioning

confidence: 99%

Release from maternally-induced allotypic suppression in rabbit by Nocardia water-soluble mitogen.

Bona¹,

Cazenave²

1977

“…Immunosuppression Experiments.--An IgG fraction was prepared from the absorbed rabbit anti-D antiserum by two precipitations with sodium sulfate (18 and 14°-/o) followed by chromatography on diethylaminoethyl (DEAE)-cellulose (12). The rabbit IgG was concentrated to 10 mg/ml, dialyzed against 0.1 R NaCl-phosphate buffer, pH 7.2, and sterilized by passage through a Millipore filter (Millipore Corp., Bedford, Mass.).…”

Section: Methodsmentioning

confidence: 99%

Suppression of Idiotypic Specificities in Adult Mice by Administration of Antiidiotypic Antibody

Hart

Wang

Pawlak

et al. 1972

250

It has previously been shown that there are extensive idiotypic cross-reactions among antiphenylarsonate antibodies of A/J mice. The present work indicates that administration, into normal, adult A/J mice, of rabbit antiidiotypic antibody directed to A/J antiphenylarsonate antibody suppresses almost completely the subsequent production of antibody of the corresponding idiotype. No effect was noted on the formation of antibodies to the protein carrier or of antiphenylarsonate antibody of a different idiotype. The data are consistent with central suppression of production of the idiotypic antibody mediated through interaction with immunoglobulin receptors on lymphocytes.

“…column equilibrated with 0.0175 • sodium phosphate buffer pH 6.3 (14). At 2 mg/ml this preparation had an HAI titer of 1:1600 against WSA and showed a single sedimentation peak with an s20.w value of 6.5 in the analytical ultracentrifuge.…”

mentioning

confidence: 99%

Viral Oncolysis: Increased Immunogenicity of Host Cell Antigen Associated With Influenza Virus

Lindenmann

Klein

1967

215

A2G mice could be solidly immunized against the Ehrlich ascites tumor by single intraperitoneal injections of homogenized and lyophilized tumor cells which had been infected with oncolytic strains of influenza A virus. Similar homogenates from noninfected tumor cells were not immunogenic, even when mixed with egg-grown virus. The immunizing principle in viral oncolysates could not be separated from the oncolytic virus by differential centrifugation or adsorption to and elution from red cells. It could be inhibited by antibody raised in rabbits against the egg-grown oncolytic virus. This reaction showed serologic specificity. Thus, the immunogenicity of an oncolysate produced with the WSA strain of neurotropic influenza virus could be inhibited by rabbit anti-WSA, but not by rabbit antibody to the TUR strain of fowl plague virus. Conversely, the immunogenicity of an oncolysate prepared with the TUR strain could be inhibited by rabbit anti-TUR, but not by anti-WSA. When mice were preimmunized (primed) with egg-grown WSA virus, their antitumor response to a later injection of WSA oncolysate was of the anamnestic type. Priming with egg-grown influenza B virus had no such effect. It was concluded that the immunogenicity of certain host cell components was greatly increased by incorporation into the makeup of the oncolytic virus.