bluuebunny/crossref_metadata_2025 huggingface dataset:
Idiotypic Determinants on the Monoclonal Immunoglobulins Associated with Papular Mucinosis
Authors: Wells J. Vivian , Fudenberg H. Hugh , Epstein William L.
Published: December 2022

Quantitative idiotypic studies were performed with monoclonal proteins from nine patients with the rare dermatologic disease papular mucinosis (PM). The PM serum protein is a basic “cathodal” IgG-1 monoclonal protein of λ light chain type and we postulated that these proteins might represent antibodies against acidic mucopolysaccharide components in the skin deposits or subcutaneous tissue of the PM patients. Specific anti-idiotypic antisera were prepared against each of four isolated PM proteins. The inhibition of the reaction between the 125I-labeled F(ab′)2 fragment of a PM protein and its homologous anti-idiotypic antiserum was tested with a series of proteins. In each case the homologous PM protein produced 98 to 100% inhibition whereas no inhibition was obtained with the other eight PM proteins, by a panel of 10 myeloma proteins of different classes, subclasses and light chain types, nor by fractions of normal IgG from PM patients or normal subjects. No inhibition was produced by a 2000-fold excess of normal IgG, suggesting that the idiotypic determinants of PM proteins are not represented in significant numbers of molecules in normal IgG. Further experiments for cross-reactivity between the four anti-idiotypic antisera found only slight cross-reactivity with one antiserum. Antisera against the γ chains of normal IgG from each of the four PM patients and from normal subjects consistently reacted with a small but significant percentage of the F(ab′)2 fragments of the PM proteins; the specificity was apparently not due to anti-light chain activity nor anti-human Fc activity and may have been directed against determinants in the variable region of the Fd portion.

There was no evidence for significant sharing of common idiotypic determinants by the PM proteins suggesting that, if these proteins were antibodies against acidic skin compounds, their specificities were directed against different compounds or against different determinants on the one compound.
Antibody Islet Forming-Cells: Localized Hemolysis Inhibition in Gel
Authors: Hege John S. , Epstein Wallace V.
Published: December 2022

Spleen cells from mice immunized with human γ globulin (HGG) and goat erythrocytes coated with human Forssman hemolysin were imbedded in agar and incubated with complement. Generalized hemolysis occurred revealing islets of intact erythrocytes representing single spleen cells releasing anti-HGG antibody. Back-ground hemolysis was increased and islets were more distinct when the goat erythrocytes were incubated with a carbodiimide reagent before coating with hemolysin.

Secondary HGG immunization yielded 50-fold more islets than did primary immunization. When background hemolysis was incomplete both islets and hemolytic plaques could be observed. Human IgM immunization yielded many more islets, but fewer plaques, than did IgG immunization. Evidence from this work and from the works of others suggests that islet-forming cells release antibody directed specifically against human immunoglobulin M, while plaque formation is the result of IgG antibody directed against human immunoglobulin G. In a system in which anti-globulin antibody modulates immune hemolysis, the immunoglobulin classes of both antiglobulin and hemolysin determine whether islets or plaques will occur.
Characterization of a Factor Required for the Differentiation of Myeloid and Lymphoid Cells in Vitro
Authors: Watson James , Prichard John
Published: December 2022

The properties of a factor which is secreted by a cloned tumor cell line (JLS-V5) have been examined. The factor has been assayed in two culture systems. The factor induced the formation of colonies containing granulocytes and macrophages from mouse bone marrow cells cultured in a semi-solid agar medium. The factor also stimulated a normal primary immune response against erythrocyte antigens in immunologically unresponsive mouse spleen cultures. These spleen cultures were prepared from unimmunized mice in medium containing a deficient fetal bovine serum. The factor has been purified 500-fold from the serum-free culture supernatants of JLS-V5 cells. When the factor was treated with a variety of chemical and enzymatic reagents and then fractionated by gel filtration, the biologic activity for each culture assay resulted from the same molecular weight fractions. Acrylamide gel electrophoresis indicates the factor has a m.w. of 70,000 to 80,000.
Production of Chemotactic Activity by Guinea Pig Immunoglobulins Following Activation of the C3 Complement Shunt Pathway
Authors: Sandberg A. L. , Snyderman R. , Frank M. M. , Osler A. G.
Published: December 2022

Guinea pig γ1 and γ2 immunoglobulins consume late C components (C3 to C9) when incubated with C4 deficient guinea pig serum. The γ2 antibodies produce greater hemolytic activity losses in normal than in C4 deficient guinea pig serum. In contrast, consumption of late component activity by the γ1 antibodies is similar in serum containing or lacking C4. Both types of immunoglobulins produce a serumderived factor chemotactic for rabbit polymorphonuclear leukocytes in either normal or C4 deficient serum. This factor has been identified as a cleavage product of C5.
Studies on Human Antibodies
Authors: Harisdangkul Valee , Kabat Elvin A.
Published: December 2022

The 1-(m-nitrophenyl) flavazoles (NPF1) of the isomaltose oligosaccharides obtained by condensation of o-phenylenediamine, m-nitrophenyl hydrazine sulfate with the isomaltose oligosaccharides were used to study their interaction with human antidextran by the quantitative hapten inhibition and fluorescence-quenching techniques. The NPF1 haptens inhibited in the dextran-antidextran system in whole serum in the same order as did the isomaltose oligosaccharides; that is, NPF1 of isomaltoheptaose (NPF1IM7) was the best inhibitor, inhibitory potency decreased progressively to the NPF1 of isomaltotetraose (NPF1IM4). Studies with antidextrans purified on a specific immunoadsorbent and eluted sequentially by isomaltotriose (IM3) and isomaltohexaose (IM6) showed that NPF1IM4 and NPF1IM5 inhibited the IM3 antidextran eluate better than the IM6 eluate, confirming the previous finding that antidextrans consist of subpopulations with different size combining sites.

All NPF1 of isomaltose oligosaccharides caused specific fluorescence quenching of the purified antidextrans which could be reversed specifically by addition of unlabeled IM6. Since relatively large amounts of nonspecific attenuation also occurred with γ globulin devoid of antidextran activity or with L-tryptophan solution, absolute values of maximum quenching (Qmax) could not be obtained, but the relative value of Qmax for each NPF1 was estimated from Scatchard plots by using various values of Qmax and taking the one which gave an extrapolated value of two combining sites since purified antidextrans are IgG and have two combining sites/molecule. NPF1IM3 with the NPF1 group closest to the immunodominant non-reducing end of IM3, which would be closest to the antibody-combining site, gave the largest Qmax and NPF1IM7 with the NPF1 group farthest from the immunodominant end gave the smallest Qmax. Differences in Qmax values were seen for the various antidextran fractions eluted by methyl α-D-glucoside, IM3 and IM6. Average intrinsic association constants, K0, of the purified antidextrans obtained from the Scatchard plots ranged from 5 to 8 × 104 for NPF1IM7 and from 1 to 1.3 × 104 for NPF1IM3. NPF1IM3 and NPF1IM4 gave smaller K0 values with the IM6 antidextran eluate than with the IM3 and methyl α-D-glucoside eluates, providing further evidence that small haptens are bound less effectively in combining sites of larger size.
A Protein in Normal Nurse Shark Serum which Reacts Specifically with Fructosans
Authors: Harisdangkul Valee , Kabat Elvin A. , McDonough Robert J. , Sigel M. Michael
Published: December 2022

The purified fructosan-specific protein (FSP) from normal nurse shark serum showed several bands in acrylamide gel electrophoresis, pH 9.3. Three purified preparations from three individual sharks appeared to have the same banding pattern but two pools from four shark sera showed a different band distribution. One preparation of FSP appeared as a single symmetrical peak on analytical ultracentrifugation with an s020,w of 10.64; the other showed two bands. The molecular weight calculated from sedimentation velocity, viscosity measurement and partial specific volume is 280,500. The protein contains four subunits linked noncovalently. Amino acid analysis showed large amounts of aspartic acid, glutamic acid, serine, threonine and cystine. No carbohydrate was detected. The circular dichroism spectrum of the purified FSP showed a negative peak at 220 to 222.5 nm. In the near ultraviolet, three positive bands were seen at 289, 280 and 268 nm. When sugars which bind specifically to FSP were added to the protein solution, increased ellipticities were seen between 260 to 285 nm with no change below 240 nm.
Electron Microscopic Localization of Rabbit Immunoglobulin Allotype b4 on Blood Lymphocytes by an Indirect Ferritin Immune Complex Labeling Technique
Authors: An Teisa , Miyai Katsuma , Sell Stewart
Published: December 2022

Immunoglobulin allotypic determinant b4 may be detected on the surface of rabbit peripheral blood lymphocytes by electron microscopy. Soluble complexes of ferritin and rabbit antiferritin of a given allotype were incubated with lymphocytes coated with anti-allotype antibody to form labeled lymphocytes. Seventy to 80% of the lymphocytes examined demonstrated specific focal ferritin-immune complex labeling scattered over the membrane surface. This technique should provide a sensitive and relatively simple method for the ultra structural detection of other immunoglobulin determinants.
Immunoglobulin-Containing Cells in Lungs of Hamsters Infected with Mycoplasma Pneumoniae
Authors: Fernald Gerald W. , Clyde Wallace A. , Bienenstock John
Published: December 2022

Immunoglobulin production in lungs of hamsters experimentally infected with M. pneumoniae was studied with the indirect immunofluorescent technique. Only IgA was detected in normal controls as single plasmacytoid cells in the bronchial lamina propria. Varying numbers of cells infiltrating the peribronchial spaces during the infection were fluorescence-positive for IgM, IgA, 7Sγ1, and 7Sγ2 globulins. IgM appeared early and was followed by 7Sγ globulin production during the period of maximum pneumonia at 2 weeks. IgA-cells increased, but were the least affected, during the infectious process indicating that in this model of local immunization a unique local immunoglobulin response occurs. Non-immunoglobulin stainable lymphocytes increased in number during the period of infection, tending to surround small blood vessels. Upon reinfection of previously infected, immune animals, a perivascular-peribronchial lymphoid reaction was observed on the 3rd day. Very few of these cells were fluorescence-positive. The histology of these recall lesions suggested that delayed hypersensitivity is a major component of acquired immunity to M. pneumoniae.
Delayed Hypersensitivity to Polysaccharides from Nocardia
Authors: Ortiz-Ortiz L. , Bojalil L. F. , Contreras M. F.
Published: December 2022

The ability of purified polysaccharides, isolated from either Nocardia asteroides or Nocardia brasiliensis, to elicit a delayed-type hypersensitivity reaction was studied. From the results of chemical and physical analysis done on the purified carbohydrates obtained from these microorganisms, it was shown that these materials were devoid of demonstrable protein, RNA and DNA materials. Both polysaccharides, separately, elicited a delayed-type hypersensitivity reaction when they were skin tested in either N. asteroides- or N. brasiliensis-sensitized guinea pigs. The skin reaction, however, was usually larger in homologous sensitized animals. In addition, they also elicited an Arthus-type reaction. The presence of a common polysaccharide among them was shown by passive cutaneous anaphylatic (PCA) and double diffusion assays. Meanwhile, the carbohydrates did not induce any inflammatory reaction in nonsensitized animals.
The Effect of Complement Depletion on Neutrophil Migration in Acute Immunologic Arthritis
Authors: DeShazo Claude V. , McGrade Maureen T. , Henson Peter M. , Cochrane Charles G.
Published: December 2022

The influence of complement components upon the movement of neutrophils and development of immunologic injury was tested in immune complex injury of rabbit synovial tissue. Suspensions of purified neutrophils were infused in the joint space of rabbits depleted of neutrophils which were prepared for a reversed passive Arthus reaction in the synovial tissues. The neutrophils migrated through the tissues to the site of antigen-antibody-complement deposition in the walls of blood vessels. Injury followed the accumulation of neutrophils in the vessel walls and was assessed by leakage of radiolabeled proteins from the circulation into the joint space, deposition of carbon black from the circulation into the area of the injured vessel and diapedesis of red blood cells. Animals genetically deficient in C6 or depleted of C3 with cobra venom factor showed a retarded rate of neutrophil migration through the synovial tissues to the site of antigen-antibody complex. Injury was delayed and diminished in the rabbits deficient in complement. The results indicate the presence of complement dependent chemotaxis in vivo. They also demonstrate the reconstitution of immunologic injury in neutrophil depleted animals by the replacement of purified neutrophils.
Antibody Cytophilic for Lymphocytes
Authors: Modabber F. , Coons A. H.
Published: December 2022

The ability of cell suspensions from spleen to bind antibody raised against β-galactosidase of Escherichia coli was tested in vitro. To exclude macrophage binding most macrophages were removed by their adherence property from the spleen cell suspensions, and the lymphocyte-rich fraction was used to test for cytophilic antibody. Hundreds of 0.2-µl droplets containing 30 to 40 cells were made in oil chambers; the number of such cells was chosen so that macrophages would be statistically excluded from one-third of the total drops. The antigen-binding activity of cells treated with antiserum in those droplets containing no macrophages was calculated, and lymphocytes from spleen were shown to bind cytophilic antibody.

In addition, thymocytes were also able to bind antibody and the binding as shown to be inhibited by exposing the cells to normal mouse serum. This indicated that the receptors for cytophilic binding on thymus cells could be preempted by molecules present in normal serum. These observations are consistent with the assumption that cytophilic antibody plays a role in lymphocyte-antigen interactions and may thereby influence the regulation of the immune response.
Dichotomy and Improved Specificity of Immunosuppressive Effect of Anti-Thymocyte Globulin
Authors: Judd K. P. , Bryant C. J. , Trentin J. J.
Published: December 2022

Cytotoxicity of horse anti-mouse thymocyte globulin (HAMTG) against mouse thymus was completely removed by multiple absorptions with mouse thymus but not by absorptions with kidney or spleen. Cytotoxicity against spleen was completely removed by multiple absorption with spleen but not by absorption with kidney or thymus. Absorptions with thymus resulted in complete removal of immunosuppressive potency as judged by duration of survival of H-2 incompatible murine cardiac allografts, but absorptions with kidney or spleen did not significantly affect the capacity to prolong cardiac allograft survival. HAMTG treatment significantly reduced hemolytic plaque-forming cells (PFC), rosette-forming cells (RFC) and hemagglutinating antibody (HA) titers in mice immunized with sheep red blood cells (SRBC). Absorptions with either spleen or thymus, but not kidney, removed this immuno-suppressive activity against PFC, RFC and HA. These studies suggest that HAMTG, in addition to containing antibodies specific for thymus, also contains antibodies which are cross-reactive with spleen and kidney. The thymus specific antibodies are of major importance in determining the ability to promote cardiac allograft survival. Antibodies either specific for or cross-reactive between thymus and spleen are important for suppression of the spleen dependent PFC, RFC and HA response to SRBC. The fact that exhaustive absorption of HAMTG with spleen completely removed the ability to suppress PFC, RFC and HA without reducing the ability to suppress allograft rejection suggests that a different population of “T” cells is necessary for allograft rejection than for humoral antibody formation.
Antibody Response to Escherichia Coli Lipopolysaccharide and Type III Pneumococcal Polysaccharide by Congenitally Thymusless (Nude) Mice
Authors: Manning Judith K. , Reed Norman D. , Jutila John W.
Published: December 2022

The concept of thymus-dependency of immune responses arose from studies of neonatally thymectomized animals and thymectomized, irradiated, bone marrow-reconstituted adult animals. In addition to an impaired cellular response, these animals exhibit a depressed humoral response to some antigens, such as sheep erythrocytes (SRBC), T-2 bacteriophage and certain serum proteins, but produce a normal humoral response to other antigens, such as ferritin, Salmonella flagellar antigen and MS-2 bacteriophage (1). These results have led to a classification of antigens as either thymus-dependent or thymus-independent.

There is some controversy as to whether the concept of a thymus-independent immune response which is based on the responses of thymectomized animals is valid. First, it is possible that the immune response to some antigens matures relatively early in ontogeny and appropriate thymus-derived antigen-reactive cells are seeded out from the thymus before its removal at birth.
The Role of Cyclic 3′,5′ Adenosine Monophosphate in the Specific Cytolytic Activity of Lymphocytes
Authors: Henney Christopher S. , Bourne Henry R. , Lichtenstein Lawrence M.
Published: December 2022

The in vitro destruction (51Cr release) of DBA/2 mastocytoma cells by lymphocytes from immune C57BL mice has been used as a system to study the relationship between lymphocyte cyclic 3′,5′-adenosine monophosphate (cAMP) levels and cytolytic activity. It was established that there is a direct correlation between increased intracellular levels of cAMP and the inhibition of cytolytic activity of a lymphocyte population.

Treatment of lymphocytes with adenyl cyclase stimulating agents (histamine, isoproterenol and prostaglandins E1 and E2) caused increases in cAMP levels and inhibited cytolytic activity. The time course of the increase in cAMP content following exposure to drug paralleled closely the time course of inhibition of cytolytic activity. The effect of isoproterenol on both cAMP content and cytolytic activity was short lived; the prostaglandins produced longer lasting effects on both systems.

Theophylline, which inhibits the enzymatic degradation of cAMP also inhibited cytolysis. This inhibitory effect was at least additive to that produced by isoproterenol.

The specificity of the relationship between intracellular cAMP levels and cytolytic activity was shown by the following observations. 1) Cyclic AMP and dibutyryl cyclic AMP inhibited lymphocyte activity. 5′-AMP at comparable concentrations did not. 2) The prostaglandins PGE1 and PGE2 both increased cyclic AMP content and consistantly inhibited lymphocyte-mediated cytolysis. Prostaglandin PGF 1α was inactive in both systems. 3) Propanolol, a β-adrenergic blocking agent, specifically prevented the effect of isoproterenol, a β-adrenergic agonist, on both cyclic AMP and cytolytic activity, but did not prevent the effect of the prostaglandins or histamine in either system.

These results suggest that cAMP occupies a central modulatory role in the expression of cytolytic activity by lymphocytes.
Regulation of Homocytotropic Antibody Formation in the Rat
Authors: Tada Tomio , Okumura Ko , Taniguchi Masaru
Published: December 2022

The effect of neonatal high dose of and preimmunization with hapten or carrier determinants on the subsequent homocytotropic antibody (HTA) response to a hapten-carrier conjugate was examined in the rat. Animals given neonatal high dose of Ascaris suum extract (As) could not produce anti-hapten HTA following immunization with dinitrophenylated Ascaris extract (DNP-As) and Bordetella pertussis vaccine, while those treated with DNP-bovine serum albumin (BSA) produced high titers of HTA against DNP-As. Preimmunization of animals with As in complete Freund's adjuvant also caused inhibition of anti-hapten HTA response to DNP-As, while that with DNP-BSA had no influence. Passive transfer of thymocytes from donors preimmunized with As in complete Freund's adjuvant restored the ability of tolerant recipients to produce HTA against DNP-As. These results indicate that neonatal high dose of carrier molecules induced tolerance in carrier-specific “helper cells,” while the hyperimmunization with the carrier caused proliferation of “regulator cells,” both resulting in the inhibition of HTA formation against the hapten on the same carrier. It is also suggested that the regulator cells and helper cells having opposite effects may be identical.
Specific Heterologous Enhancement of Immune Responses
Authors: Rubin Arnold S. , Coons Albert H.
Published: December 2022

A soluble factor which enhanced the plaque-forming antibody response against sheep erythrocytes was found in tissue culture fluid of spleen cell cultures from mice injected with tetanus toxoid 1 to 2 months earlier to which 1 ng of toxoid was added on the 2nd day of a 5-day period of incubation. Such enhancement was demonstrated in normal spleen cell suspensions to which sheep erythrocytes were added at the beginning, and the supernatant on day 2. In such cultures, enhancing factor was generated and appeared in the supernatant only after the addition of the specific antigen used for priming. Also, at certain concentrations, such supernatants produced inhibition of the response and hence contained a soluble inhibitory factor. The enhancing factor is both non-dialyzable and heat stable (56°C, 30 min).
Antigenic Specificity of Anti-Erythrocyte Autoantibody Responses by NZB Mice: Identification and Partial Characterization of Two Erythrocyte Surface Autoantigens
Authors: Linder Ewert , Edgington Thomas S.
Published: December 2022

Two independent but coexistent spontaneous anti-erythrocyte autoantibody responses by NZB mice are described. The two specific murine erythrocyte surface antigens uniquely associated with these autoantibody responses, and thus referred to as autoantigens, are identified and partially characterized. Both appear to represent species-specific antigens and are not recognizably segregated on a genetic basis within six strains of mice. One response is directed against an exposed erythrocyte surface determinant herein referred to as the X antigen. This response may be of primary significance in the pathogenesis of the autoimmune hemolytic anemia of NZB mice. The second type of autoantibody response is directed to a site, unexposed on the normal murine erythrocyte surface but exposed by proteolysis with bromelin, and is referred to as the HB antigen. An alternative mechanism for HB antigen exposure and binding of anti-HB autoantibody appears to be mediated by in vivo erythrocyte surface modification, possibly secondary to the binding of anti-X autoantibody in vivo. Specific assays are described for each of these generically independent autoantibodies and erythrocyte surface antigens. The possible implication of these autoantigens in the immune complex disease of the NZB strain is also suggested.
The Purification and Immunochemical Characterization of a New Specific Antigen, Differentially Expressed on the Surface Membranes of Mouse Lymphoid Cells
Authors: Von Fellenberg Roland , Guggisberg Elisabeth
Published: December 2022

A lymphoid-cell specific surface membrane antigen of the mouse has been purified over 200-fold by differential solubilization, ion exchange chromatography and gel filtration. The serologic activity corresponded to a fraction of 67,000 molecular weight. Two main protein bands could be detected in acrylamide gel electrophoresis together with a very faint third band. Double gel diffusion revealed a single strong band of precipitation, whereas no immunoprecipitation could be detected in conventional—or crossing-over immunoelectrophoresis. This discrepancy is discussed. The antigen was strongly expressed on cells having an organ distribution similar to bone marrow-derived cells and slightly expressed on a subpopulation of cells which was similarly abundant in all lymphoid organs tested. During ontogeny the antigen was strongly expressed on thymocytes with a transient maximum between days 4 and 8 after gestation.
The Biologic Significance of Fc Receptor on Mouse B-Lymphocytes
Authors: Paraskevas F. , Orr K. B. , Anderson E. D. , Lee S-T. , Israels L. G.
Published: December 2022

Evidence exists that antigen is trapped in lymphoid follicles complexed with antibody (1–6) and that antigen-antibody complexes (Ag-Ab) prepared in vitro localize in these same areas (7). The localization of the complexes depends on the presence of the Fc of the antibody since complexes of the antigen with Fab fragment do not localize in the follicles (8). These studies consider that the binding of Ag-Ab complexes and Fc fragment occurs by an Fc receptor present on reticular cells.

We have previously described the presence of a receptor for antibody Fc on the surface of mouse spleen lymphocytes (9–11). This receptor could bind isolated Fc fragments as well as the Fc of antibody in Ag-Ab complexes. The binding was independent of complement.

In the present communication we present some evidence for the possible biologic role of this receptor during the primary response and during the interaction of rabbit anti-mouse thymocyte serum (ALS) with lymphocytes both in vitro and in vivo.
In Vitro Morphine Binding by Sera from Morphine-Treated Rabbits
Authors: Ringle David A. , Herndon Betty L.
Published: December 2022

Results of a number of studies on the development of tolerance to morphine suggest that at least some aspects of this tolerance are immunologic-like in character (for reviews see Friedler (1) and Cochin (2)). Evidence that the morphine configuration is immunogenic is provided by Ryan et al. (3) who recently reported the apparent presence of antibody against morphine in the sera of approximately 40% of heroin addicts studied. Preliminary findings from our laboratory, in studies reported here, have indicated that in the rabbit prolonged exposure to morphine is associated with an increase in serum binding of 14C-labeled morphine, both after withdrawal from morphine and after a brief re-exposure to morphine.

Two albino New Zealand female rabbits were given multiple doses of morphine sulfate or morphine hydrochloride (Merck, USP) administered subcutaneously in three courses of injections spaced 1 and 2 months apart.
Effect of Spleen Cells on Cytotoxicity by Immune Lymph Node Cells: Cell-Mediated Immune Suppression (? Enhancement) in Vitro
Authors: Klein William J.
Published: December 2022

The reactivity of spleen cells (SP) from C57BL mice immunized with BP8 ascites tumor was assessed in an in vitro assay system for cell-mediated immunity. In this system it has been shown previously that immune lymph node cells (ILNC) obtained 5 to 7 days after subcutaneous immunization are capable of killing the tumor cells. The activity of these ILNC could be blocked completely by microliter amounts of hyperimmune sera possessing enhancing activity in vivo.

Normal C47BL SP, normal C3H SP, SP from C3H mice bearing the ascites tumor, and SP from C57BL mice immunized with Freund's adjuvant were all incapable of killing the tumor cells in culture. SP obtained from C57BL mice 5 to 7 days post-immunization were inactive. Hypotonic treatment did not modify these results. Lymphocyte-mediated cytotoxicity (LCT) was observed with SP obtained 9 days or longer after subcutaneous immunization. In contrast, more direct stimulation of the spleen with intravenous immunization produced evidence of immune reactivity in SP obtained 5 days after immunization. None of the various SP capable of LCT were able to effect 100% cytotoxicity of the tumor cells.

Most significantly, SP obtained 5 to 7 days after subcutaneous immunization were able to block completely the LCT of ILNC obtained from the same animals. These results suggest that a portion of the SP population is producing cells capable of LCT while another part of the population consists of cells which antagonize this effect. This antagonistic sub-population may act by interfering with ILNC-tumor cell contact or by elaborating substances which act on the ILNC. These substances could be “enhancing antibodies” or non-antibody molecules.
Regulation of Antibody Response in Vitro
Authors: Ishizaka Kimishige , Kishimoto Tadamitsu
Published: December 2022

Rabbits were immunized by intraperitoneal injections of DNP-Ascaris antigen precipitated with aluminum hydroxide gel to form reaginic antibodies. Mesenteric lymph node cells from the primed rabbits were stimulated in vitro by incubating with homologous antigen for 24 hr and then cultured for 6 days in the absence of antigen. Measurements of IgG and reaginic antibodies in culture fluids showed that the lymph node cells from the rabbits which formed reaginic antibodies after primary immunization produced IgG antibodies. In vitro reaginic antibody formation was generally observed when donors of lymph node cells showed both primary and secondary reaginic antibody responses after the primary and booster injections. The titer of reaginic antibody in the culture fluid, as determined by the PCA reaction, did not correlate with the concentration of IgG antibodies in the fluid, or with the reaginic antibody titer of the serum of donors at the time of sacrifice. The molecular size of reaginic antibody in the culture fluids was intermediate between those of IgM and IgG antibodies. The skin-sensitizing activity of the antibody was completely lost by treatment with 0.01 M dithiothreitol followed by alkylation and slightly diminished after heating at 56°C for 2 hr. Such properties of the reaginic antibodies in the culture fluid were comparable to those of IgE antibodies in the serum.

It was found that a removal of most of the adherent cells in lymph node cell suspension before antigen stimulation in vitro did not have a significant effect on the formation of IgG, IgM and IgE antibodies in the culture. An addition of monospecific anti-IgG with antigen for in vitro stimulation suppressed in vitro IgG and IgM antibody formation but not the IgE antibody formation.
Determination of Relative Affinity and Heterogeneity of Mouse Anti-DNP Antibodies by a Plaque-Inhibition Technique
Authors: Miller Gary W. , Segre Diego
Published: December 2022

A plaque-inhibition technique was used to study the response of mice to the dinitrophenyl (DNP) determinant at the cellular level. Sheep erythrocytes were sensitized for use in the plaque assay by exposing them to dinitrophenylated antisheep hemolysin of low hemolytic efficiency. DNP-lysine was added to the agar to inhibit specifically indirect plaques formed by mouse spleen cells producing anti-DNP antibodies. The average relative affinity of these antibodies was defined as the reciprocal of the molar concentration of DNP-lysine producing 50% plaque inhibition. The average relative affinity increased by a factor of 130 between 1 and 8 weeks after initial immunization. A highly significant correlation, apparently linear, was found between the average relative affinity and the average association constant of the serum antibody as measured by a modified Farr technique. The slopes of the plaque-inhibition curves in the vicinity of 50% plaque inhibition were assumed to be inversely proportional to the degree of heterogeneity of the antibodies with respect to affinity. There was a highly significant positive correlation between affinity and heterogeneity.
Reaginic Antibody Formation in the Mouse
Authors: Ishizaka Kimishige , Okudaira Hirokazu
Published: December 2022

DBA/1 mice were immunized by an intraperitoneal injection of 0.2 to 1.0 µg of ovalbumin precipitated with aluminum hydroxide gel and the antibody response was observed. Both reaginic and γ1 antibodies were detected in the serum 10 days after the immunization and the antibody formation persisted for more than 1 month. The animals showed a secondary reaginic antibody response upon booster injection. An intravenous injection of 80 µg N of mouse anti-ovalbumin IgG antibody into the immunized mice within 24 hr after primary immunization prevented both reaginic and γ1 antibody responses for a period of over 3 weeks, during which time passive antibody was detectable in the serum. Evidence was obtained that an essentially normal immune response followed after the passive antibody was catabolized. Both reaginic and γ1 antibodies were detected in the serum 40 days after immunization. An increased interval between immunization and passive antibody resulted in progressively decreasing efficiency of inhibition of the antibody response. Passive administration of IgG antibody did not terminate pre-existing reaginic antibody formation nor suppress secondary antibody response.
The Isolation and Some Physiochemical and Biologic Properties of the Type III Antigen of Group B Streptococci
Authors: Russell H. , Norcross N. L.
Published: December 2022

The type III antigen of Lancefield's group B streptococci is known to be responsible for the virulence of the organism. Some of the chemical, physical and immunologic properties of the antigen were studied. The antigen was extracted from the bacteria by hot HCl extractions. It was further purified by treatment with pepsin, trypsin, and by fractionation on a DEAE-cellulose, ion-exchange, chromatographic column.

The type III antigen was characterized by hydrolysis and chromatography, nitrogen, phosphorus, amino acid and carbohydrate content. It is a polysaccharide composed of 38.9% galactose, 22.8% glucosamine and 17.8% glucose. The latter sugars were present at a ratio of 2:1:1 µmols/mg, respectively. The antigen also contains 3.1% uronic acid, 0.8% muramic acid and less than 1% each of alanine, aspartic acid, glutamic acid, serine and threonine. The antigen has a sedimentation coefficient of 1.67S and a m.w. of 15,561. The partial specific volume was 0.62 g/ml. Immunoelectrophoresis of the antigen resulted in migration toward the cathode. The cathodal migration explains the separation which was obtained on the DEAE-cellulose column.

Glucuronic acid appears to be one of the immunodominant components of the antigen. It offers 34% inhibition of the quantitative precipitin reaction between the type III antigen and type III antisera.
Presidential Address
Authors: Dixon Frank J.
Published: December 2022

Understanding the pathogenesis of immunologic disease, i.e., the way in which immunologic events cause injury and/or malfunction, demands several kinds of information. First, one must identify the particular antigen(s) which initiates and serves as the target of the pathogenic immune response. This is often a difficult task, since many of these antigens are substances normally present or at least commonly present in the absence of an immune response or disease. This fact suggests that transient events such as incidental infections or exposures to environmental agents may have an adjuvant effect which increases immunogenicity of potential antigens or in other ways terminates the tolerance barrier. Second, one must know a number of things about the host which are important in determining the self-infliction of immunologic injury. It appears that the magnitude and perhaps balance, i.e., cellular vs humoral aspects, of the immune response, are in part genetically predetermined and therefore may predispose to immunologic disease.
Genetic Control of the Immune Response of Inbred Guinea Pigs to 2,4-Dinitrophenyl Guinea Pig Albumin. Frequency of Antigen-Binding Lymphocytes and Avidity of Antibodies Secreted by Plaque-Forming Cells
Authors: Davie Joseph M. , Paul William E. , Green Ira
Published: December 2022

Strain 2 guinea pigs immunized with 1 µg of 2,4-dinitrophenyl guinea pig albumin (DNP-GPA) containing an average of six DNP groups/molecule have no detectable antibody-secreting cells (direct or indirect plaque-forming cells) up to 28 days after immunization. As previously demonstrated, these animals lack an immune response (IR) gene, linked to the gene complex controlling strain 13 histocompatibility antigens, which controls the response to DNP-GPA.

Nonimmunized non-responder strain 2 animals and non-responder offspring of a backcross between strain 2 and (2 × 13)F1 guinea pigs have normal numbers of lymphocytes capable of binding 125I-DNP-GPA to their surface, indicating that these animals have no quantitative deficit in precursors of anti-DNP antibody-forming cells. Moreover, when strain 2 guinea pigs are immunized with 100 µg of DNP-GPA, they produce anti-DNP antibodies containing a fraction of high affinity molecules similar to that present in similarly immunized strain 13 animals. This indicates that strain 2 animals have no deficit among precursor cells bearing high affinity receptors. Thus, the IR gene controlling the immune response to DNP6GPA does not control the frequency of precursor cells bearing receptors which bind DNP-GPA.
Alloimmune Hemolytic Plaque-Forming Cells: Kinetics of IgM and IgG Alloantibody Releasing Cells during a Primary and Secondary Immune Response of Chickens
Authors: Bacon Larry D. , Schierman Louis W. , McBride Raymond A.
Published: December 2022

The kinetics of alloimmune hemolytic plaque-forming cells found in spleens of chickens after primary and secondary immunization was investigated. Rabbit anti-chicken immunoglobulin was employed for plaque development since chicken alloantibodies are non-hemolytic. Plaques formed by cells secreting IgM and IgG were developed separately by using rabbit antisera specific for each class of immunoglobulin.

The kinetics as well as numbers of cells which produced antibodies specific for B blood group alloantigens during both the primary and secondary response were similar to what has been found for the mouse heteroimmune system. Certain advantages of the chicken alloimmune system for enumeration of hemolytic plaque-forming cells are discussed.
The Effect of Mycobacterial RNA on the Primary Antibody Response of Mice to Bovine γ Globulin
Authors: Youmans Guy P. , Youmans Anne S.
Published: December 2022

Mycobacterial RNA-protein preparations, obtained from viable cells of the attenuated H37Ra strain, will stimulate the production of antibody to bovine γ globulin in mice in a manner and intensity comparable to that of the synthetic polynucleotides poly A:U or poly I:C. In this respect, though, the mycobacterial RNA was less effective than Escherichia coli endotoxin or the double-stranded coliphage Mu-9 ds-RNA. The adjuvant activity of mycobacterial RNA was reduced by treatment with ribonuclease or with trypsin. The possible role of mycobacterial RNA in the adjuvant activity of intact mycobacterial cells is discussed.
Myelin Basic Proteins of Mammalian and Submammalian Vertebrates: Encephalitogenic Activities in Guinea Pigs and Rats
Authors: Martenson Russell E. , Deibler Gladys E. , Kies Marian W. , Levine Seymour , Alvord Ellsworth C.
Published: December 2022

Central nervous system myelin basic proteins of various vertebrates, mammalian and submammalian, have been examined for encephalitogenic activity in guinea pigs and Lewis rats. Myelin basic proteins from mammalian, avian (chicken), reptilian (turtle) and amphibian (frog) species were encephalitogenic in rats at a level of 50 µg; those of fish (carp, shark) were completely inactive. None of the myelin basic proteins from the submammalian species was encephalitogenic in guinea pigs at levels as high at 250 µg. Guinea pigs responded equally well to 5 µg of all mammalian myelin basic proteins tested except the smaller of the two rat proteins, which was completely inactive at this level. In contrast, rats responded well to guinea pig and both the larger and the smaller rat proteins, but not as well to proteins of the other mammals (bovine, rabbit and human) at a 10-µg level. The different patterns of reactivity of guinea pigs and rats to mammalian and submammalian myelin basic proteins suggest that the two species recognize different amino acid sequences in the basic protein molecule as being encephalitogenic determinants. Additional studies carried out in highly susceptible strains of rats showed that whole spinal cord tissue of salamanders, as well as frog, is encephalitogenic, but that carp spinal cord is not. Evidence of rapid autolytic changes in spinal cord tissue from cold-blooded animals together with the observed heterogeneity of fish basic proteins indicate the need for caution in interpretation of negative results with carp and shark proteins.
Secretory Newcastle Disease Virus Antibodies from Chicken Respiratory Tract
Authors: Zakay-Rones Z. , Levy R. , Spira G.
Published: December 2022

The nature of the antibodies extracted from lungs of chickens immunized intranasally by Newcastle disease virus (NDV) was analyzed by chromatography on Sephadex G-200, DEAE cellulose, as well as by electrophoresis on acrylamide gel. Specific local antibody activity was always located at the same region as serum antibodies with a molecular weight of 160,000 to 180,000. By immunodiffusion and immunoelectrophoresis this antibody was shown to be antigenically identical to humoral IgG serum antibodies. Concentrations of hexosamine were 1.55% for serum and 1.35% for lung antibodies. The S values as determined by analytical ultracentrifugation were 6.9 and 7.1 for local and humoral antibodies, respectively. It was concluded, therefore, that local anti-NDV antibodies belong to the IgG type of immunoglobulin, and that this immunoglobulin is responsible for the systemic as well as for the local immunity in the chickens.
Chemical Carcinogenesis and Humoral Antibody Synthesis
Authors: Brai Melchiorre , Patrucco Aida , Osler Abraham G.
Published: December 2022

Methylcholanthrene in an amount sufficient to induce tumors in 100% of injected mice failed to influence humoral antibody responses as estimated by activity or weight measurements. Mice injected with the carcinogen before or after immunization with sheep erythrocytes and human serum albumin responded at the same levels during the primary and secondary phases of antibody synthesis as did the controls. The early immune response in tumor-bearing mice was also indistinguishable from that of normal animals, despite the presence of marked splenomegaly in the former group.
The Carbohydrate Composition of Immunoglobulins from Diverse Species of Vertebrates
Authors: Action R. T. , Niedermeier W. , Weinheimer P. F. , Clem L. W. , Leslie G. A. , Bennett J. C.
Published: December 2022

Immunoglobulins and their respective heavy (H) and light (L) polypeptide chains from species representing the major classes of vertebrates were analyzed for their carbohydrate composition by gas chromatography of the alditol acetate derivatives of the monosaccharides released by acid hydrolysis. Mannose, galactose, glucosamine and sialic acid were present in the immunoglobulins from all the species investigated. Fucose however was absent from the immunoglobulins of two species of animals studied. Most of the carbohydrate was found associated with the H chains. The carbohydrate composition was felt to be heterogeneous in that the quantities of fucose and sialic acid found often accounted for less than one residue per H chain. Whereas species representative of the mammals and birds had mannose to galactose ratios greater than one, the ratio of these sugars to each other in the immunoglobulins from animals below the birds was on the order of one.
Chicken Immunoglobulin Resembling γA
Authors: Bienenstock John , Perey D. Y. E. , Gauldie J. , Underdown B. J.
Published: December 2022

The immunoglobulin classes recognized in the chicken are γG and γM. Because of an interest in this laboratory in the mechanisms of local mucosal immunity and the possible experimental manipulations of chicken central lymphoid tissue, it was felt that a redefinition of chicken immunoglobulins, particularly with reference to a possible γA class, should be undertaken before a satisfactory exploration of mucosal immunity in the chicken could occur. Accordingly, we immunized a rabbit against chicken γG Fab fragments prepared according to the methods of Tenenhouse and Deutsch (1). We further prepared antisera in goats to chicken γM and γG heavy chains as described previously (2). The anti-chicken γG was monospecific for the γ chain, and did not react with Fab fragments. The anti-γM antiserum was rendered monospecific by absorption with newly hatched chicken serum. Both antisera gave single precipitin arcs in immunodiffusion experiments against normal chicken serum.
Synthesis of β2-Microglobulin by Stimulated Lymphocytes
Authors: Bernier George M. , Fanger Michael W.
Published: December 2022

β2-microglobulin is a human serum protein of low molecular weight and unknown function (1). Normally present in serum in low concentrations (2 µg/ml), it is degraded by the kidney (2). In the presence of renal failure serum concentrations as high as 70 µg/ml may occur (3, 4). Patients with defective renal tubular function excrete increased amounts of β2-microglobulin in their urine (4), suggesting that tubular reabsorption is an important step in the catabolism of this protein. In its dependence upon the kidney for degradation, β2-microglobulin is similar to immunoglobulin light chains (5), although microglobulin is not antigenically related to any known immunoglobulin.

Recently, Smithies and Poulik have reported the partial amino acid sequence of β2-microglobulin (6). They found significant homology between β2-microglobulin and two adjacent loops (C2 and C3) of the heavy chain of human γG globulin. This finding suggested that β2-microglobulin might be synthesized in lymphoid tissue and prompted us to investigate whether this protein was produced by lymphocytes during in vitro culture.
Appendix and γM Antibody Formation. V. Appendix and Thymus Cell Synergism in the Direct and Indirect Plaque-Forming Cell Responses to Sheep Erythrocytes in the Rabbit
Authors: Ozer H. , Waksman B. H.
Published: December 2022

Thymectomized, lethally irradiated rabbits were restored to normal or near-normal immune responsiveness against sheep erythrocytes by a combination of autologous or allogeneic thymocytes and appendix cells.
The Immune Response to Sheep Erythrocytes in the Lizard Calotes Versicolor
Authors: Kanakambika P. , Muthukkaruppan VR.
Published: December 2022

The immune response of the lizard, Calotes versicolor, to sheep red blood cells has been characterized in terms of the appearance of circulating antibodies as well as hemolysin-releasing plaque-forming cells (PFC). PFC appear in the spleen by the end of the 1st week after immunization and reach a maximum number at 14 days. The maximum level of serum antibody response is reached during the 3rd week. After a second immunization the rise in PFC and serum antibody is more rapid with a shorter latent period but without a concomitant increase in antibody level. PFC can be detected only in the spleen and blood of immunized lizards but not in other tissues such as the thymus, bone marrow, cloacal complex, lung, liver or kidney.

Histologic changes occurring in the spleens of immunized lizards are also described. After immunization, there appear to be a greater abundance of mitotic figures, increased vascularity within the splenic pulp, and a reduction in the relative number of lymphocytes.