bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibody and Complement Dependent Damage to Liposomes Prepared with Bacterial Lipopolysaccharides
Authors: Kinsky S. C. , Kataoka T. , Inoue K. , Lüderitz O.
Published: December 2022

Liposomes were made with either lecithin or sphingomyelin in the presence of various Salmonella minnesota S and R form lipopolysaccharides. These “actively” sensitized liposomes release trapped glucose marker when incubated with both an appropriate rabbit antiserum and guinea pig serum. The liposomes lose most glucose in the presence of homologous antiserum although cross-reactivity, which seems limited to liposomes containing lipopolysaccharides closely related in structure, has been observed. Liposomal response differs from other reported phenomena which have suggested that lipopolysaccharides can interact with complement in the absence of added antibodies. Treatment of S or R form lipopolysaccharide with alkali markedly reduces the amount required for half-maximal sensitization. The alkali-treated lipopolysaccharides are also effective in “passive” sensitization of liposomes; in this procedure, the liposomes are first generated in the absence of any antigen and subsequently incubated with lipopolysaccharide. Both actively and passively sensitized liposomes release a greater percentage of their trapped glucose when prepared with lecithin than with sphingomyelin.
Glycine-Rich γ-Glycoprotein (GBG): Evidence for Relation to the Complement System and for Genetic Polymorphism in Man
Authors: Alper C. A. , Boenisch T. , Watson L.
Published: December 2022

Glycine-rich β-glycoprotein (GBG) is a 6.3S protein which occurs at a concentration of 9 to 47 mg/100 ml of normal serum. On storage of serum, particularly at 37°C, this protein fragments very slowly into glycine-rich γ-glycoprotein (GGG) which is 4.3S and glycine-rich α-glycoprotein (GAG) of similar size. Incubation of serum for 1 hr at 37°C with antigen-antibody precipitates or zymosan but not hydrazine results in virtually complete conversion of GBG but little appearance of GAG in the fluid phase. EDTA blocks GBG conversion by antigen-antibody or zymosan.

GBG is highly polymorphic when examined in various human sera and consists of at least five bands in all samples tested. Population and family studies indicate that this polymorphism is determined by two common genes, GbF and GbS, with frequencies of 0.50 and 0.44, 0.28 and 0.71, and 0.13 and 0.87 in Negroes, Caucasians and Orientals respectively, and two rare genes, GbF1 with a frequency of 0.06 in Negroes, and GbS1 with a frequency of 0.01 in Caucasians.
Limited Heterogeneity of Antibodies against the Para-Azobenzoate (Xp) Hapten
Authors: Hoffman Donald R. , Grossberg Allan L. , Pressman David
Published: December 2022

A series of IgG antibodies raised in 21 rabbits against the p-azobenzoate hapten was examined by isoelectric focusing in acrylamide gels. Specifically purified antibodies from 14 of the rabbits gave isoelectric spectra that consisted of only five or fewer patterns similar to those obtained from myeloma proteins, which showed a single pattern consisting of a series of three or more regularly spaced lines. A restricted number of patterns indicates limited heterogeneity. Five of the preparations were very restricted, giving primarily a single pattern. Four of these five preparations also gave a single band in isoelectric focusing of the light chains and four also appeared homogeneous with respect to binding constant. One antibody preparation that gave two simple patterns on isoelectric focusing and two light chain bands was fractionated to give a fraction with a single isoelectric focusing pattern and a single light chain band. For two rabbits the same antibody isoelectric focusing pattern persisted for over 1 year. On the basis that a simple pattern represents a biosynthetically homogeneous antibody species, a large proportion of the rabbits produced primarily one to five different antibody molecular species directed against this hapten.
Macrophage-Antigen Interaction: Uptake, Metabolism and Immunogenicity of Foreign Albumin
Authors: Schmidtke Jon R. , Unanue Emil R.
Published: December 2022

The uptake of radioiodinated mouse and human serum albumin (HSA) by macrophages from non-immunized CBA mice depended on the degree of aggregation of the albumin, and not on cytophilic antibody on the surface of the cell. Autologous and heterologous albumin of comparable physical states were taken up and metabolized equally by macrophages. The HSA taken up by macrophages was rapidly catabolized, except for a small amount that remained associated with the cell both intracellularly and on the surface for long periods in culture. The membrane-bound albumin represented non-pinocytosed molecules. This membrane-bound antigen was maintained independently of intracellular material and was responsible for most of the immunogenicity of macrophage-bound albumin. A portion of albumin taken up by macrophages was released undegraded into the supernatant during culture. The metabolism of HSA was directly proportional to the degree of aggregation of the albumin as increased aggregation resulted in more extensive catabolism.
Antigen Recognition and the Immune Response: the Capacity of L-Tyrosine-Azobenzenearsonate to Serve as a Carrier for a Macromolecular Hapten
Authors: Alkan Sefik S. , Nitecki Danute E. , Goodman Joel W.
Published: December 2022

The low molecular weight compound L-tyrosine-azobenzenearsonate (RAT) induced immunity in guinea pigs, as evidenced by delayed cutaneous hypersensitivity and stimulation of DNA synthesis by lymphoid cells in culture. Two different conjugates of RAT and a poly-γ-D-glutamic acid hapten of m.w. 35,000 were prepared. In one, an average of nine molecules of RAT were linked directly to the PGA backbone [PGA-(RAT)9]. In the other, RAT was joined to PGA by means of 6-aminocaproyl (SAC) hydrocarbon chains that extended the RAT groups out from the PGA backbone by a maximum of 10 Å [PGA-(SAC-RAT)12]. Immunization with the two conjugates gave rise to anti-PGA antibody, PGA-(SAC-RAT)12 being somewhat more effective in that regard, and all animals developed delayed hypersensitivity to RAT but not to PGA. Lymphoid cells from animals sensitized to PGA-RAT conjugates demonstrated specificity for the homologous conjugates, assessed by the rate of incorporation of 14C-thymidine into DNA. Prior immunization with RAT potentiated the response to PGA following administration of the conjugates. Thus, it appears that a molecule with an m.w. of about 400 was recognized by antigenreactive cells and could serve as a carrier for a hapten with an m.w. of 35,000.
The Pll Gene and Histocompatibility Genotype in Inbred and Random-Bred Guinea Pigs
Authors: Ellman Leonard , Green Ira , Benacerraf Baruj
Published: December 2022

Previous studies demonstrated a close linkage of the PLL immune response gene and the major histocompatibility locus of the inbred strain 2 guinea pig in both (2 × 13)F1 × 13 back-cross animals and in random-bred Hartley guinea pigs. In the present study, this relationship was confirmed by the demonstration that: 1) anti-strain 2 isoantiserum made in PLL non-responder Hartley animals distinguished between strain 2 and strain 13 lymphocytes, and between Hartley PLL responder and non-responder lymphocytes; 2) Hartley PLL responder anti-strain 2 isoantiserum did not detect these differences; and 3) a strain-specific strain 2 guinea pig leukemia was readily rejected by PLL non-responder (2 × 13)F1 × 13 back-cross animals whereas it was uniformly fatal for PLL responder back-cross animals. Finally, the observation of close linkage of the PLL gene and the major histocompatibility locus of the strain 2 guinea pig was also extended to a second randombred guinea pig strain, the NIH multipurpose strain.
Morphologic Changes in Draining Lymph Nodes and in Lymphocyte Cultures after Sensitization with Complete or Incomplete Freund's Adjuvant. Correlation with Immunologic Events in Vivo and in Culture
Authors: Dvorak Ann M. , Bast Robert C. , Dvorak Harold F.
Published: December 2022

Morphologic changes occurring in draining lymph nodes were studied at two intervals after sensitization of guinea pigs with human serum albumin (HSA) in complete or incomplete Freund's adjuvant (CFA or IFA) and were correlated with the immunologic response of these animals. At 1 week after sensitization with HSA in IFA guinea pigs were primed for the delayed-onset reactions of cutaneous basophil hypersensitivity (CBH) but had little circulating antibody. Their lymph nodes manifested paracortical cell hyperplasia with a striking degree of blast transformation, and cells from such nodes, cultured in the presence of antigen, incorporated 3H-thymidine at an enhanced rate and underwent morphologic blast transformation. At 6 weeks, these animals had lost CBH skin reactivity but had high titers of circulating antibody. Draining lymph nodes contained prominent germinal centers and numerous plasma cells, but paracortical blasts had returned to control levels, and isolated cells were not stimulated to incorporate thymidine by antigen in culture. By contrast, animals sensitized with the same antigen in CFA developed progressively indurated skin reactions characteristic of delayed hypersensitivity (DH), as well as high titers of circulating antibody. In draining lymph nodes, blast cells outside of germinal centers were never as numerous as at 1 week following administration of antigen in IFA; however, a significant number persisted at 6 weeks when excellent antigen-induced thymidine incorporation and blast transformation could be achieved in culture. Taken together, these data provide additional evidence for the existence of two distinguishable forms of cellular immunity, DH and CBH, and define certain properties of each.
Rabbit Secretory IgA
Authors: O'Daly Jose A. , Cebra John J.
Published: December 2022

Rabbit secretory component (SC) of colostral secretory IgA (sIgA) was isolated following dissociation from the immunoglobulin in 5 M guanidine ·HC1. Two procedures were used, one yielding native SC and the other SC with all half-cystines carboxymethylated. The results of analytic and preparative zone electrophoresis in polyacrylamide gels led to a correlation between the antigenic activities used to define component polypeptide chains of sIgA and the characteristics of their electrophoretic mobility. The native form of a fast-migrating component (F-component) different from alpha, light and SC polypeptides was also isolated by preparative gel electrophoresis. Its cellular localization was compared with that of other components of sIgA.
Recovery of the Antigen Retention Mechanism after Sublethal X-Irradiation in Mice Receiving Cells from Various Sources
Authors: Nettesheim P. , Hammons Anna S.
Published: December 2022

The intravenous injection of spleen or peritoneal exudate cells into mice x-irradiated with 600 R leads to rapid partial restoration of the x-ray damaged antigen retention system. This effect was already clearly discernible when 125I-AHGG was injected 1 day after cell infusion, and became very pronounced when the labeled antigen was given 8 days after cell infusion. In contrast, no significant recovery was observed in mice receiving either thymus cells, bone marrow cells or no cells after x-irradiation. The cell doses were approximately equivalent to one-third spleen, one thymus, one femur and the peritoneal cavity cells recoverable from one mouse. The weight, histologic and autoradiographic studies suggest that spleen and peritoneal exudate cell suspensions contain cells (or their precursors) instrumental in recovery of (and probably also in unimpaired) follicular antigen localization. Further studies are needed to determine the nature of the cells involved and precisely what role they play in the enhancement of recovery of antigen localization after x-irradiation.
Effect of Antimacrophage Serum on Hypersensitivity (Schistosoma Mansoni Egg) and Foreign Body (Divinyl-Benzene Copolymer Bead) Granulomas
Authors: Boros D. L. , Warren K. S.
Published: December 2022

Antimacrophage serum (AMS) was obtained by immunizing rabbits with glassadherent mouse peritoneal exudate cells. The antiserum was repeatedly absorbed until free of antilymphocytic antibodies. Repeated injections of AMS into mice suppressed the level of circulating blood monocytes. AMS was toxic in vitro to peritoneal exudate cells. Repeated injections of AMS suppressed the delayed hypersensitivity granulomas that formed around eggs of Schistosoma mansoni injected into the microvasculature of the mouse lung. Similar injections of the antiserum, however, had no effect on the foreign body granulomas produced in response to similarly injected plastic beads.
Quantitation of Immunoglobulin- and Virus-Producing Cells in Rats Infected with Moloney Leukemia Virus
Authors: Cremer Natalie E. , Taylor Dee O. N. , Lennette Edwin H.
Published: December 2022

Using the fluorescent antibody technique, the numbers of popliteal lymph node cells (PLN) that produce viral antigen and antibody to BSA or ferritin were evaluated in normal rats and those infected with Moloney leukemia virus (MLV). There was a 96% to 99% reduction in cells forming anti-BSA antibody and 64% to 84% reduction in those forming anti-ferritin antibody in infected rats. 47% to 57% of the cells that formed anti-BSA antibody also formed viral antigen and 19% to 43% of those that formed anti-ferritin antibody contained viral antigen. The main class of antibody in the sera of both normal and infected rats was IgGa. In the sera of BSA-immunized rats another class of antibody, either IgGb or γ1, was present. Antibody of the IgM class was seen only in the younger group of infected rats immunized with ferritin. One per cent to 5% of the PLN cells contained large amounts of viral antigen as shown by bright abundant staining with fluorescein-tagged anti-MLV conjugate whereas most cells contained only a fleck or two of specific stain for viral antigen.
Characterization of the Antibody Response to Type III Pneumococcal Polysaccharide at the Cellular Level
Authors: Baker Phillip J. , Prescott Benjamin , Stashak Philip W. , Amsbaugh Diana F.
Published: December 2022

A plaque-inhibition test was used to assess the relative avidity of the IgM antibody specific for type III pneumococcal polysaccharide (SSS-III). No changes in avidity were demonstrable at various times after immunization and over a 20,000-fold range of antigen dose. Prior immunization, regardless of the dose of SSS-III employed, resulted in no change in avidity. These results indicated that a restricted population of lymphoid cells participates in the antibody response to SSS-III and that an antigen-mediated cell selection process does not play a significant role.
Immunochemical Studies of Infectious Mononucleosis
Authors: Fletcher Mary Ann , Woolfolk Betty J.
Published: December 2022

A method is described whereby glycoprotein antigens may be extracted from hemoglobin-free stroma of mammalian erythrocytes with hot 75% ethanol. The infectious mononucleosis (IM) receptors of horse erythrocytes have been isolated and partially characterized for the first time. The material was heterogeneous in molecular size but relatively homogeneous by electrophoresis. The carbohydrate content was 40% with a molar ratio of hexose: hexosamine: sialic acid of 1:1:1. The peptide portion was greatly enriched (43% of the total amino acids) in acidic and in hydroxylamino acids. By the same method closely similar substances were isolated from sheep and human erythrocytes. The former was a potent IM antigen and the latter had blood group M and N specificity. An even more potent IM receptor was extracted by aqueous ethanol from the hemoglobin-free stroma of bovine erythrocytes. This extract had only one-third the sialic acid and one-half the total carbohydrate but one and one-half times the nitrogen of the other three preparations. The bovine antigen retained some IM activity after neuraminidase treatment, which destroyed the IM or MN activity of the sheep, horse and human antigens. Gel diffusion and quantitative precipitation studies indicate that the bovine IM antigen may react with a fraction of heterophile antibody which does not react with sheep or horse antigen.
Cell-Mediated Cytotoxicity: An Assay Using 125I-Iododeoxyuridine-Labeled Target Cells
Authors: Cohen Alfred M. , Burdick James F. , Ketcham Alfred S.
Published: December 2022

An assay of cell-mediated cytotoxicity against monolayer cultured cells by pre-labeling target cell DNA with 125I-iododeoxyuridine is reported. Results are presented demonstrating tumorspecific cellular immunity in two animal tumorhost systems. The technique, as currently being applied to the study of peripheral blood lymphocyte cytotoxicity against human tumor cells, is presented.
A Simplification of the Radioactive Antigen-Binding Test by a Double Label Technique
Authors: Gotschlich Emil C.
Published: December 2022

The radioactive antigen-binding test developed by Farr is a relatively simple quantitative technique to measure primary antigen antibody interaction (1). The principle of the test is that radioactive antigen will remain bound to antibody when the latter is precipitated with ammonium sulfate. To quantitate the test one may either determine the radioactivity of the precipitated globulins after washing them with half-saturated ammonium sulfate or, alternatively, one may determine the amount of radioactive antigen remaining in the supernatant. Both of these methods become technically difficult when one is dealing with very small volumes of sera, which may become necessary when working with small animals or with human specimens obtained by fingerstick.

It is the purpose of this communication to present an alternative method that requires neither the removal of an accurate sample of the supernatant nor the washing of the precipitate. The basis of this method is to add radioactive 22Na to the 125I-labeled antigen as a volume marker.
Immunoglobulin D Concentration in Pregnant Women
Authors: Klapper David G. , Mendenhall Hiram W.
Published: December 2022

Statistical analysis revealed a significant elevation in IgD concentration in the sera of pregnant women collected at delivery. Sera collected at other stages in gestation and from women using oral contraceptives showed considerable variation in IgD levels but with less significant elevation when contrasted with nonpregnant controls.
Studies on the γA Protein from MOPC-315
Authors: Eisen Herman
Published: December 2022

Protein 315 (the A-myeloma protein produced by MOPC-315) binds ε-DNP-lysine and menadione with high affinity (1 × 107 M-1 and 5 × 105 M-1, respectively, at 4°C). Over 90% of residues of the light chain of this protein (L315) have been sequenced by E. Schulenburg, E. Simms, R. Bradshaw and H. Eisen. Of the residues in the C-terminal half, 30% differ from the corresponding residues of the MOPC-104 light chain (L104) and about 65% differ from those of mouse κ chains. In its N-terminal half, L315 differs in only about 10% of the residues from L104. A rabbit antiserum specific for L315 has been prepared by R. Lynch and used in a double antibody assay (125I-315 plus rabbit anti-L315 plus goat anti-rabbit Fc) to demonstrate L315-like chains in normal sera from BALB/c and C57BC/6 mice. Both sera contained these chains at the same low level, about 0.02% to 0.1% of the total immunoglobulin.
The Amino Acid Sequence of Four Mouse Lambda Chains
Authors: Appella Ettore
Published: December 2022

The amino acid sequence of the mouse λ chain MOPC 104E has been determined. By the isolation of 19 tryptic peptides and 35 thermolysin peptides complete overlappings were obtained. The mouse λ chain contained five half-cysteines of which four were internally linked in two pairs of disulfide bridges as has also been found with the human λ chains. When this sequence was compared with those obtained from three other mouse λ chains (RPC 20, HOPC 1 and MOPC 511), no differences appeared. The identity of four λ chains strongly suggests that there is a peculiar restriction in the pattern of variability of these chains, and provides support for the somatic theories of antibody diversity.
Common Structural Features Among the Light Chains of γG and γM Anti-γ Globulins
Authors: Capra J. Donald , Kehoe J. Michael
Published: December 2022

Benign hypergammaglobulinemic purpura is characterized by a marked elevation of the γ globulin level which on serum protein electrophoresis is broadbased and only rarely has the appearance of an M component. Anti-γ globulins of the γG type were isolated from these sera using specific absorption on BDB cross-linked γ globulin followed by acid elution. Extensive studies on 4 eluates and more limited studies on 12 other sera provided evidence that these anti-γ globulins had restricted heterogeneity in the great majority of cases. The light chains (VκI) of two γG anti-γ globulins were sequenced to residue 40 and were identical. Two VκIII light chains derived from γM anti-γ globulins were also sequenced to residue 40, all four proteins had a serine residue at position 31 the most variable of all positions in κ light chains except for position 96. Only one other κ chain has been described with a serine in this position.
A Waldenström Macroglobulin with Anti-γG Activity
Authors: Stone Marvin J.
Published: December 2022

A mixed γM-γG cryoglobulin was identified in the serum of a patient with classical Waldenström's macroglobulinemia. The isolated macroglobulin, γMSie, contains only κ-type light chains and an iodinated preparation is quantitatively precipitable by human γG at pH 7.4 in the cold. γMSie fulfills the following criteria of an antibody to γG: 1) it has a clearly defined specificity reacting with the Fc portion of human γG as well as with rabbit γG but not with other human immunoglobulins nor with bovine, porcine, sheep, chicken or goat γG; 2) the combining activity resides in the Fabµ fragments. Direct binding studies strongly suggest that all 10 Fab/mol γM are active (KA = 6.0 × 104 M-1) despite the fact that only five are available in the intact pentamer.
Characterization of a BALB/c Myeloma Library
Authors: Hirst Judith Wegman , Jones Gary G. , Cohn Melvin
Published: December 2022

Immunoglobulin-producing myelomas have been classified as shown in Table I. Those producing κ light chain alone have not been included. Of 767 tumors classified, 212 have been saved. Thirty-nine possess antibody activity: anti-5AU, DNP, TNP, purine, AMP, NIP, P-choline, N-acetylglucosamine (NAG), levan, dextran, or Salmonella lipopolysaccharide. With the exception of one IgG3 anti-levan, all proteins were of the IgA class. Other possible antibody activities were looked for in an attempt to test the Jerne hypothesis. Seventy myeloma proteins screened against H-2a, H-2b, H-2k, and H-2d (self) did not react, nor did purified myeloma proteins with antibody activities which have been found more than once. These proteins with repeat antibody activities (anti-levan, anti-dextran, anti-DNP, anti-P-choline, and anti-NAG) were also screened against fetal antigens extracted by the Reisfeld method and showed no direct precipitation or inhibition of precipitation with these antigens.
The Three-Dimensional Structure at 6 a Resolution of a Human γG1 Immunoglobulin Molecule
Authors: Sarma V. R. , Silverton E. W. , Davies D. R. , Terry W. D.
Published: December 2022

A three-dimensional electron density map has been calculated at 6 Å resolution for a human γG1 immunoglobulin molecule. X-ray diffraction data have been collected for the native protein crystals and for three heavy atom derivatives prepared by soaking the crystals in PCMBS, Hg(CN)2 and PtCl6-. Three distinct globular regions have been identified in the electron density map. One of these surrounds the molecular dyad axis and has been interpreted to be the Fc fragment. The other two regions are identical to one another and have been interpreted to be the Fab fragments. There are four ways of joining these globular regions of electron density to make a complete molecule and these lead to four possible models of which a T-shaped model is favored. The dimensions of this model compare extremely well with those reported by other physical and chemical techniques for immunoglobulin molecules.
Rabbit Antihapten Antibodies of Limited Heterogeneity
Authors: Pressman D.
Published: December 2022

Antibodies of very limited heterogeneity appear relatively frequently in certain anti-hapten sera. For example, in the case of anti-Xp antibodies, four of twelve rabbits gave patterns of limited heterogeneity in electrofocusing. Antibodies of limited heterogeneity exhibit binding curves which have been attributed to antibodies of rather extreme heterogeneity. Even an antibody preparation of two homogeneous populations with binding constants differing by a factor of 10 can show binding curves which would be interpreted as being markedly heterogeneous.
Studies on Idiotypic Determinants
Authors: Nisonoff A. , Wilson S. K. , Brient Bruce W.
Published: December 2022

One of these investigations indicated that the combining site of a rabbit antihapten antibody is either part of or close to a major idiotypic determinant. Rabbit antiidiotypic antibodies were prepared against purified antibenzoate antibodies. Reactions of antiidiotypic antibodies with 125I-F(ab′)2 fragments of the antibenzoate were markedly inhibited by benzoate derivatives. There was a direct correlation between the degree of inhibition and the binding affinity of the hapten. This indicates either that the combining site of an antibody molecule is part of a major idiotypic determinant, with the hapten acting through steric interference, or that hapten causes a conformational change. If a conformational change occurs it must be limited since there is little effect of hapten on the reaction of antibenzoate antibody with goat antirabbit Fab or with antiallotype antisera. Two assays were employed to investigate cross-reactions of individually specific determinants in a myeloma protein with nonspecific IgG.
Antibody Formation by Spleen Cells of Immunologically Tolerant Mice Stimulated with Antigen-Antibody Complexes
Authors: Intini Claudio , Segre Diego , Segre Mariangela , Myers Walter L.
Published: December 2022

Mice were made immunologically tolerant to chicken egg lysozyme, rabbit IgG, and pneumococcal capsular polysaccharide type 3. The mice were splenectomized and their spleen cells were exposed in vitro to the tolerated antigen mixed with specific antiserum of rabbit or sheep origin. Spleen cells from control tolerant mice were exposed to antigen alone or mixed with normal serum. The spleen cells so treated were then injected intraperitoneally into the same mice from which they had been derived. Mice whose spleen cells were exposed to antigen-antibody complexes in vitro became immune, whereas control mice remained tolerant. It was concluded that the spleen of tolerant mice contains cells capable of reacting with the tolerated antigen and of giving rise to antibody-producing cells. This conclusion supported the concept that humoral antibody plays a role in regulating the immune response and tolerance.
Further Studies on Migration Inhibitory Factor (MIF): Evidence for its Glycoprotein Nature
Authors: Remold Heinz G. , David John R.
Published: December 2022

Migration inhibitory factor (MIF) obtained from sensitized guinea pig lymph node lymphocytes that had been stimulated by specific antigen (o-chlorobenzoyl-bovine γ globulin) was characterized by enzymatic treatment and isopycnic centrifugation in a CsCl density gradient. MIF incubated with water-insoluble chymotrypsin lost its biologic activity. Further, neuraminidase, which hydrolyzes terminal sialic acid, abolished MIF activity. When MIF was centrifuged in CsCl density gradients, it had a buoyant density (ρ25 = 1.430) slightly greater than that of the protein reference substance (ρ25 = 1.352), which is consistent with its being a glycoprotein. The results indicated that MIF is a glycoprotein and that sialic acid is necessary for its biologic activity.
Regulation of Homocytotropic Antibody Formation in the Rat
Authors: Tada Tomio , Okumura Ko
Published: December 2022

Anti-hapten homocytotropic antibody (HTA) formation was induced in rats by two succeeding injections of dinitrophenylated Ascaris extracts (DNP-As) with a 5-day interval by the use of Bordetella pertussis vaccine as an adjuvant. The suppressive activity of anti-hapten and anti-carrier antibodies on the HTA formation was tested by immunizing rats with various antigen-antibody precipitates in which hapten, carrier or both were covered by an excess of rabbit antibodies directed to the respective determinants. It was found that a covering of carrier determinants in the first injection on day 0 suppressed the anti-hapten HTA response, whereas the covering of haptenic determinants was rather enhancing. On the other hand, a covering of haptenic determinants by anti-hapten antibodies in the second injection was greatly suppressive, while a covering of carrier determinants was ineffective. It was also found that the first injection of DNP-As on day 0 could be replaced by injection of the unconjugated carrier to prepare animals for the subsequent induction of the antihapten HTA response following a second single injection of the hapten-carrier conjugate. The results indicate that carrier determinants are first recognized by carrierspecific cells which interact with hapten-specific cells to induce formation of antihapten HTA.
Comparative Immunologic Studies on Fish and Invertebrate Collagens
Authors: Meigel W. , Pontz B. , Timpl R. , Hieber E. , Nordwig A. , Steffen C. , Kühn K.
Published: December 2022

Rabbit antisera prepared against collagens from carp and two invertebrate species were fractionated using an immunoadsorption technique. The two antibody fractions, one with and one without affinity for the rabbit collagen immunoadsorbent, were investigated for cross-reaction with various collagens derived from mammalian, fish and invertebrate species by hemagglutination and hemagglutination-inhibition studies. The antibody fractions without affinity for rabbit collagen, representing the main fractions, reacted only with the kind of collagen used for immunization or, to a smaller extent, with collagens from closely related species. The second type of antibody fractions (with affinity for rabbit collagen) exhibited cross-reaction with all the collagens investigated.
Local Immunologic Response to Immunization with Inactivated Newcastle Disease Virus
Authors: Zakay-Rones Zichriya , Levy R. , Spira G.
Published: December 2022

Chickens were immunized with Newcastle disease virus inactivated by β propriolactone. The antigen was administered by the intramuscular or intranasal route, and the appearance of HI antibodies in the serum and lungs was followed. Local secretory antibodies in the lung were found only as a consequence of intranasal immunization, whereas serum antibodies were found after administration of antigen by either route.
Spontaneous Renal Lesions and Glomerular Deposits of IgG and Complement in Guinea Pigs
Authors: Steblay Raymond W. , Rudofsky Ulrich
Published: December 2022

During recent studies on immunologically induced renal tubular disease (1), we noted a high incidence of spontaneous deposits of IgG and complement (C3) in glomeruli of untreated guinea pigs (2–4). We now describe the immunohistochemical, morphologic and functional status of kidneys of untreated guinea pigs of various ages.

Materials and Methods. Twelve female guinea pigs of the Albany strain3 were studied in each of the following age groups: near-term fetus, 3 weeks, 3, 6, 12, 18 and 24 months. Six female guinea pigs, 16 months of age, were surveyed from each of the following closed colonies:4 Rockefeller, Dunkin, Camm and Fort Detrick Hartleys, and English Short Hairs. Three strain 2 and six strain 13 female guinea pigs,5 one year of age, were studied. Three 24-hr quantitative urinary protein determinations (5) were made on all animals 6 months of age or older.
Erythrocyte-Bound C5 and C6 in Autoimmune Hemolytic Anemia
Authors: Kerr Robert O. , Dalmasso Agustin P. , Kaplan Manuel E.
Published: December 2022

In four patients with autoimmune hemolytic anemia, antiglobulin testing revealed, in addition to IgG, C3 and C4, erythrocyte-bound C5 and C6. In the presence of fresh serum, Rh-antiserum Ripley also caused non-lytic binding of C3, C4, C5 and C6 to normal type O, CDe red cells. These studies indicate that late-acting complement components (C5, C6) may be present on red cells of patients with immune hemolytic anemias without overt disruption of membrane integrity.
Species Distribution of J Chain
Authors: Weinheimer P. F. , Mestecky J. , Acton R. T.
Published: December 2022

A polypeptide chain, other than the heavy and light chains and the secretory component, has been observed in secretory immunoglobulin A (IgA) of both rabbit (1) and human (2) origin. This J chain, as it is called, is readily detectable as a fast-moving band on alkaline-urea disc electrophoresis gels of L-chain fractions from reduced and alkylated or S-sulfonated polymeric immunoglobulins (1, 2). It has been suggested that this component functions to join together the two 7S subunits of both 11S secretory and serum IgA (1). The presence of an analogous chain was recently described in studies involving human macroglobulinemic IgM (2) and in ovine (3) and porcine (Zikan, personal communication) secretory immunoglobulins. In the present investigation several other animal species representing key positions on the phylogenetic scale were examined in an attempt to establish whether or not this J chain is a common component of phylogenetically distinct polymeric immunoglobulin molecules.
The Lymphocytotoxic Reaction: The Mechanism of Rabbit Complement Action
Authors: Ferrone S. , Cooper N. R. , Pellegrino M. A. , Reisfeld R. A.
Published: December 2022

The mechanism of action of rabbit complement (C) in the lymphocytotoxic reaction has been investigated in order to determine why it is the most effective source of C in this reaction, although it has a relatively low level of C as determined in the hemolytic test system. The studies presented here showed that the lymphocytotoxic reaction does not differ from the hemolytic reaction insofar as the requirement of C components is concerned. In addition, normal rabbit sera were found to contain a natural cytotoxic C-dependent antibody reactive with an antigen present on cultured human lymphoid cells and on peripheral lymphocytes. Removal of this antibody by specific absorption decreases the efficiency of rabbit serum as source of C in the lymphocytotoxic reaction. Therefore it is concluded that the superior effectiveness of rabbit serum as the source of C depends on the fact that it contributes to the cytotoxic reaction not only C components but also a natural antibody, which enhances the binding of C by HL-A antibody.
A Simple Method for Quantitation of Haptens Using “Indirect” Single Radial Immunodiffusion
Authors: Richter Wolfgang
Published: December 2022

Single radial immunodiffusion has been adapted to the quantitation of low molecular haptens, making use of the principle of hapten inhibition of precipitation. As model haptens, isomaltose oligosaccharides were quantitatively estimated in a precipitating antidextran protein-dextran conjugate system. The antidextrans had been raised in sheep by immunization with a bovine serum albumin-dextran conjugate. After preparation of the agar plates containing the antidextran, the wells were filled with the haptenic oligosaccharide solutions. The latter were allowed to diffuse into the agar and to bind to the antidextran for 30, 120, or 300 min, and then the hapten inhibition halos were visualized or “developed” by immersing the plates in a solution of an ovalbumin-dextran conjugate for 24 hr. Thereafter, the diameters of the transparent hapten inhibition halos, contrasting against the remaining opaque parts of the plate, were measured. A linear relationship between area of hapten inhibition and concentration of individual isomaltose oligosaccharides was obtained, when sufficient time for complete hapten diffusion was allowed before visualizing hapten inhibition zones. In the examples studied, the smallest amount of oligosaccharide hapten measurable was about 10 µg/ml. The potential application of this procedure for quantitation of other low molecular monovalent haptens is discussed.
Response of Canine Fetuses and Neonates to Antigenic Stimulation
Authors: Shifrine Moshé , Smith J. B. , Bulgin Marie S. , Bryant B. J. , Zee Yuan-Chang , Osburn B. I.
Published: December 2022

Canine fetuses of the same litter of beagles were injected with keyhole limpet hemocyanin (KLH), Mycoplasma gallisepticum, infectious bovine rhinotracheitis (IBR) virus or chicken erythrocytes. Three litters injected at gestation days 41, 47 or 51 produced no antibodies. Fetuses of a litter injected at gestation day 55 with 10 µg, 100 µg or 1000 µg of KLH and challenged with 1000 µg KLH 21 days after birth gave only a primary response. Fetuses of a litter infected with Brucella canis at gestation day 50 and killed 9 days later produced agglutinating antibodies. When 1-day-old pups were injected with KLH and challenged with the same antigen 40 days later, a secondary immune response was observed. When 1-day-old pups were injected with IBR virus, neutralizing antibodies were produced within 2 weeks. It was concluded that the canine fetus becomes competent for antibody production close to term.
The Isotopic Antiglobulin Assay: Application to the Study of Hybrid Cell Membranes
Authors: Rechsteiner M. C.
Published: December 2022

An isotopic antiglobulin assay was used to study the cell surface antigens of a hamster-mouse hybrid cell. Antisera were developed against the nuclei and whole cells of the parents and the hybrid. Whereas antisera to hamster and mouse whole cells cross-reacted extensively, antisera to nuclei were species-specific. The antinuclear sera were used to demonstrate the presence of hamster-specific and mousespecific cell surface antigens on the hybrid cell. The proportion of these antigens on the hybrid cell surface closely mirrored the number of chromosomes contributed by each parent. Antisera to hamster and mouse whole cells gave the same antiglobulin binding on the hybrid. Antiglobulin saturation of the hybrid cell surface by antiserum to either parent implied intimate mixing of hamster and mouse cell surface antigens. Further evidence for molecular intermixing was obtained by demonstrating that the binding of hamster-specific globulin to the hybrid cell surface prevented binding of mouse-specific globulin.
Purification of the First Component of Human Complement by Affinity Chromatography on Human γ Globulin Linked to Sepharose
Authors: Bing David H.
Published: December 2022

The first component of human complement, C1̄, has been purified by affinity chromatography on a resin consisting of Sepharose covalently linked to human IgG. This resin selectively adsorbs at least 90% of the C1̄ detectable in a precipitate of human serum prepared at a final ionic strength of 0.03 and pH of 6.4. The C1̄ is eluted from the resin with 0.2 M 1, 4-diaminobutane. Virtually all the protein is recovered and the C1̄ has a high specific activity whether assayed with N-carbobenzoxy-L-Tyrosine-para-nitrophenyl ester or with EAC4, C2 and C-EDTA. Evidence that the protein obtained by this procedure is indeed C1̄ is based on the ability of C1̄ inhibitor to inhibit the enzymatic activity, of anti-C1s̄ antiserum to inhibit EAC4̄,2̄ formation and of the eluted protein to form the EAC1̄,4 intermediate. The procedure appears to be a simple reliable technique for preparing large quantities of human C1̄.
Bone Marrow-Derived Cells as Target Cells for Polynucleotide Adjuvants
Authors: Campbell P. A. , Kind P.
Published: December 2022

Mice were immunized with 107 sheep red blood cells (SRBC), with or without polyadenylic and polyuridylic acid adjuvant. Kinetics of the number of plaque-forming cells and cell units that could form hemolytic foci in recipient mice were studied. The adjuvant enhanced the number of plaque-forming cells on days 2 through 4 of the response but did not increase the number of hemolytic focus-forming units at any time. Since the limiting cell for the formation of hemolytic foci is probably the thymusderived cell, the adjuvant does not appear to act on cells of this type. The adjuvant appears to enhance the plaque-forming cell response at the level of the bone-marrow cell by a) increasing mitosis of bone marrow-derived cells, and/or b) by increasing the number of bone marrow-derived cells that interacted with thymus-derived cells.
Immunoglobulin E Antibodies in Guinea Pigs: Characterization of Monomeric and Polymeric Components
Authors: Dobson Colin , Rockey John H. , Soulsby E. J. L.
Published: December 2022

Immunization of Hartley strain guinea pigs with large doses (1 to 5 mg) of an ethylchloroformate polymerized Ascaris suum antigen, and immunization subsequently followed by infection with living second-stage larvae of the parasite, resulted in the production of both IgE (γE) and IgG1 (γG1) homocytotropic antibodies in high titer. Reaginic (PK) activity was assayed after a 6-day latent period and sedimented in sucrose density gradients at 8.3 ± 0.3S and 10.7 ± 0.3S. Both the 8S and 11S components were also demonstrated by antiglobulin enhancement of hemagglutination with specific anti-guinea pig IgE and anti-human IgE sera. The 11S reaginic antibody had a greater skin-sensitizing activity than the 8S component on the basis of the relative quantities detected following antiglobulin enhancement. The reaginic activity also was separated into different sized components by Sephadex G-200 gel filtration. Guinea pig PK activity was not absorbed by rabbit anti-guinea pig IgM (γM) or anti-IgG2/G1 (γG2/G1) sera. The IgG1 (γG1) (passive cutaneous anaphylatic (PCA) activity, assayed after an 18-hr latent period, was removed by the anti-IgG2/G1 (γG2/G1) serum. Reaginic activity was absorbed by rabbit anti-guinea pig IgE (γE) sera, and also by monospecific anti-human IgE (myeloma PS) serum. The anti-guinea pig IgE and anti-human IgE sera did not absorb guinea pig IgG1 PCA activity. Both monomeric and polymeric reaginic activities were absorbed by rabbit anti-guinea pig IgE serum. These experiments establish that guinea pig reaginic (IgE (γE)) antibody is the analogue of human reaginic antibody of the IgE class, and that the guinea pig and human IgE-globulins share common class-specific antigenic determinants.
The Relationship of Complement Consumption to Immune Fever
Authors: Mickenberg Ira D. , Snyderman Ralph , Root Richard K. , Mergenhagen Stephen E. , Wolff Sheldon M.
Published: December 2022

The administration of cobra venom factor or soluble human serum albumin (HSA)-anti-HSA complexes in antigen excess to rabbits resulted in a lowering of total hemolytic complement and C3 titers which persisted more than 24 hr. When such animals were challenged with complexes made up in 10 times antigen excess, the consumption of complement, febrile and thrombocytopenic responses were significantly diminished from those seen in untreated controls, and the duration of post-challenge leukopenia was shortened. These observations suggest a role for complement consumption, particularly of components C3-9, in the manifestation of these responses. Pyrogenic tolerance to complexes could not be explained by their more rapid removal from the circulation or a failure of the thermoregulatory centers to respond to preformed leukocytic pyrogen, suggesting that impairment in complement consumption either by depletion of C3 or interference with the activation of C1 might be responsible for this state. In contrast to these findings using complexes in 10 times antigen excess, similar prior treatment of immunized rabbits given HSA or of normal rabbits that received a mixture of soluble and insoluble complexes in 3 times antibody excess failed to suppress febrile or hematologic responses. If the formation of antigen-antibody complexes is essential for the production of these responses in immunized animals challenged with specific antigen, it appears that the large, rapidly-cleared complexes formed near equivalence are most likely involved in producing these biologic activities. Whereas some of the properties of these complexes make them more able to fix complement in vitro, it seems unlikely that complement-mediated mechanisms are significantly involved in the pathogenesis of the in vivo responses measured in these investigations.
Membrane Immunoglobulins of Lymphocytes in NZB Mice
Authors: Bhoopalam Nirmala , Yakulis Vincent J. , Costea Nicolas , Heller Paul
Published: December 2022

Mammalian lymphocytes of spleen, marrow, lymph nodes and peripheral blood have been found to have membrane sites with the structural characteristics of immunoglobulin which have been considered to be receptors for antigens and appear to be indicative of the capacity of these cells to synthesize immunoglobulins (1–7). We have recently found (8) that the proportion of these receptor-carrying lymphocytes (RCL) diminishes in BALB/c mice during the development of plasmacytoma. We were interested to determine if there was also a diminution in the percentage of RCL in other diseases associated with immunologic deficiency, e.g., in NZB mice during the development of the characteristic autoimmune disease.

NZB and CF1 mice were divided into three groups according to age: older than 6 months, less than 3 months, and a small group of newborns. Old mice of the BALB/c, SWR and C57B1 strains were also studied. Heparinized blood was obtained from each mouse by puncture of the retro-orbital venous sinus.
The Immunologic Release of Constituents from Neutrophil Leukocytes
Authors: Henson Peter M.
Published: December 2022

The reaction of rabbit neutrophils with immune complexes induced release of constituents to the external medium. The release which accompanied adherence of neutrophils to immune complexes bound to a nonphagocytosable surface resulted from a direct extrusion of granules to the exterior of the cell. Alkaline phosphatase from the secondary granule was released before constituents of the primary granule and remained at the site of excretion, adherent to the external neutrophil membrane.

In contrast, while phagocytosis of particulate complexes stimulated release of materials from the primary granules, alkaline phosphatase was released in only small amounts. A possible mechanism of enzyme release during phagocytosis of large particles was demonstrated to occur by two steps. Degranulation into the phagocytic vacuole was followed by opening of the vacuole during uptake of additional particles. Neutrophils were not lysed during either release process.
Anthrax Indirect Microhemagglutination Test
Authors: Buchanan Thomas M. , Feeley James C. , Hayes Peggy S. , Brachman Philip S.
Published: December 2022

A new application of the indirect microhemagglutination (IMH) test has been developed for anthrax. The antigen was prepared by DEAE cellulose, Dowex 1-X1, and Sephadex G-50 column chromatography of a culture filtrate of the avirulent, non-proteolytic, non-encapsulated Vollum strain of Bacillus anthracis grown anaerobically in a chemically defined liquid medium. Tanned, sensitized sheep red blood cells (SRBC) were tested with complement-inactivated SRBC-absorbed serum samples from 72 anthrax patients, 91 persons who had been vaccinated against anthrax, and 103 controls. The IMH test detected antibodies in 93% of patients, 98% of vaccinees, and none of the controls. Run-to-run reproducibility was 76% within one twofold dilution, and 93% within two twofold dilutions. The IMH test proved more sensitive and less time-consuming than the currently used agar-gel precipitation inhibition (AGPI) test.