bluuebunny/crossref_metadata_2025 huggingface dataset:
On the Transfer of DNP-BSA Hypersensitivity in Guinea Pigs by Means of Irradiated Donor Plasma. Evidence That the Cutaneous Reactivity Is Hapten-Specific and Mediated Via DNP Antibody
Authors: Richardson Walter P. , Paterson Philip Y.
Published: December 2022

Cutaneous sensitivity to 2,4-dinitrophenylated bovine serum albumin was transferred with plasma from sensitized, irradiated guinea pigs following the procedure of Dupuy and Good (1). Enhanced cutaneous reactivity was observed in recipients of plasma from irradiated (800 rads) donors as compared to plasma from nonirradiated donors. Cutaneous reactions of plasma recipients were greater at 3 to 4 hr than at 24 hr after skin testing and they exhibited clear-cut hapten, as opposed to carrier, specificity. The plasma used to transfer this sensitivity contained precipitating antibody which also was hapten-specific. These data demonstrate that the transferred sensitivity was “immediate” and due to circulating, precipitating DNP antibody producing a toxic antigen-antibody complex reaction of the Arthus type.
A Genetically Determined Antibody Response to Poly (l-Lysine) in Guinea Pigs
Authors: Levin Howard , Stollar B. David
Published: December 2022

The ability of poly(l-lysine) alone and poly(l-lysine) complexed to RNA to produce specific anti-poly(l-lysine) antibodies in strain 2 and strain 13 guinea pigs was tested with a sensitive complement fixation assay. The sera from all immunized strain 2 guinea pigs showed anti-poly(l-lysine) antibodies while none were detected in the sera from immunized strain 13 guinea pigs. The antibody nature of the reactive serum component was confirmed by ammonium sulfate precipitation, antiglobulin absorption and Sephadex G-200 gel filtration. The sera reacted specifically with poly(l-lysine) and not with poly (d-lysine), poly(d-arginine) or poly(l-ornithine). The results demonstrate that poly(l-lysine) itself, in adjuvant, is capable of eliciting specific anti-poly(l-lysine) antibodies and that this response is controlled by the poly(l-lysine) gene.
Evolutionary Studies of Rabbit Allotypes
Authors: Rodkey L. Scott , Hansen Ernil
Published: December 2022

Preparations of IgG F(ab′)2 fragments from domestic rabbits (NZW), hares (JR), and cottontail rabbits (Cot) were compared by indirect radioimmunoassay inhibition techniques for a comparison of the interrelatedness of the antigenic determinants reacting with anti-NZW allotype antisera.

JR F(ab′)2 molecules that were reactive with anti-NZW allotype antisera did not inhibit a reaction of NZW molecules with anti-allotype antisera. In another inhibition system, unlabeled JR molecules inhibited a reaction between I*JR molecules and anti-NZW allotype antisera better than did the homologous unlabeled NZW molecules. A third genus (Cot) was used for comparison and the results of the study suggested that the reactions observed could be explained best by the presence of different numbers of antigenic determinant sites shared between the three species that are identified by anti-NZW allotype antisera.

The present data appear to favor the presence of a limited number of germ line genes coding for variable regions of Ig molecules since the time for evolving numerous identical mutations in each of thousands of germ line genes is limited to less than 2 × 106 years in lagomorphs.
Immunochemical Studies on Dextrans
Authors: Torii Mitsuo , Sakakibara Keiko , Kabat Elvin A.
Published: December 2022

Human and rabbit antidextrans of α(1 → 2) specificity have been further characterized as to their combining sites based on oligosaccharide inhibition assays with three trisaccharides: 1) αdGlc(1 → 2)αdGlc(1 → 6)dGlc, 2) αdGlc·(1 → 6)αdGlc(1 → 2)dGlc and 3) αdGlc(1 → 2)[αdGlc(1 → 6)]dGlc. After absorption of α(1 → 6) specific antibody, the human antidextran sera contained antibodies complementary to trisaccharide 1 while the rabbit antisera contained antibodies complementary to both trisaccharides 1 and 2. With human antidextrans with non-α(1 → 2) specificity these trisaccharides were not the best inhibitors. The various rabbit antisera studied with dextrans of predominantly α(1 → 6) linkages showed combining site sizes with upper limits complementary to IM5, IM4 and IM3. The cross-reaction of horse antipneumococcal type XII antiserum was best inhibited by trisaccharide 1.
Transplantability and Antigenicity of Two Sublines of the TA3 Tumor
Authors: Sanford Barbara H. , Codington John F. , Jeanloz Roger W. , Palmer Peter D.
Published: December 2022

Loss of specificity in the strain A mammary ascites carcinoma subline TA3-Ha appears to be related to decreased immunogenicity rather than to increased immunoresistance. Allogeneic hosts could be induced to reject potentially lethal inocula of TA3-Ha cells by addition of cells from the more specific TA3-St subline or by presensitization with skin grafts from strain A mice. The nonspecific TA3-Ha subline absorbed markedly less activity from H-2 antisera than strain-specific TA3-St cells; however, the absorption capacity of nonspecific TA3-Ha cells for H-2 antisera was noticeably increased by lyophilization, suggesting decreased exposure of H-2 antigens at the cell surface of intact TA3-Ha cells. It is suggested that inaccessibility of histocompatibility antigens on the TA3-Ha cell may be related to presence of a unique sialoglycoprotein at the cell surface in this tumor subline. With the strain-specific TA3-St subline, lyophilization increased H-2k antigenicity while decreasing H-2d activity. The magnitude of these changes, however, was much less than that observed with either H-2d or H-2k antigenicity after lyophilization of TA3-Ha cells.
Role of the Thymus in Tolerance
Authors: Ha T.-Y. , Waksman B. H.
Published: December 2022

Thymocytes or spleen cells were transferred from normal adult Lewis rats that had received 100 mg bovine γ-globulin intraperitoneally to intact adult syngeneic recipients. The recipients, after challenge at 24 hr with antigen in complete adjuvant, showed inhibition of the Arthus and delayed hypersensitivity responses and a slight delay in the formation of hemagglutinating and hemolytic antibody but not anaphylactic antibody. The effect was specific and apparently was not due to transferred antigen (transfer of killed thymocytes or spleen cells had no effect); it was observed between 18 and 72 hr after the injection of bovine γ-globulin in donor animals.
Demonstration of Proliferation by Bone Marrow-Derived Lymphocytes of Guinea Pigs, Mice and Rabbits in Response to Mitogen Stimulation in Vitro
Authors: Elfenbein Gerald J. , Harrison Michael R. , Green Ira
Published: December 2022

We have shown by simultaneous complement receptor lymphocyte rosette and radioautographic techniques that guinea pig, mouse, and rabbit bone marrow-derived (B) cells proliferate in vitro in response to phytohemagglutinin, concanavalin A, pokeweed mitogen, and S. typhosa endotoxin lipopolysaccharide. PHA, Con A, and PWM are predominantly thymus-derived (T) cell mitogens in the guinea pig, mouse and rabbit. LPS is predominantly a B cell stimulant in the guinea pig but predominantly a T cell stimulant in the mouse.
Demonstration of Proliferation by Guinea Pig and Rabbit Bone Marrow-Derived Lymphocytes in Vitro in Response to Stimulation by Anti-Immunoglobulin Antisera
Authors: Elfenbein Gerald J. , Harrison Michael R. , Mage Rose G.
Published: December 2022

We have employed simultaneous complement receptor lymphocyte rosette and radioautographic techniques to investigate the proliferative response of guinea pig lymph node and rabbit peripheral blood bone marrow-drived (B) lymphocytes to stimulation with anti-immunoglobulin antisera. We have shown directly that all of the proliferative response due to anti-immunoglobulin antisera occurred in the complement receptor lymphocyte (B cell) subpopulation.
Studies on Brain Antigens
Authors: MacPherson Catherine F. C. , Yo Sui-Lan
Published: December 2022

Pretreatment of guinea pigs with a purified bovine spinal cord protein (β1-SCP) prevented them from developing experimental allergic encephalitis (EAE) when they were subsequently challenged with an excessive dose of bovine spinal cord in complete Freund's adjuvant. In contrast, guinea pigs treated with protein obtained from bovine liver or with protein extracted from the gray matter of bovine brain developed EAE similar in severity to the disease induced in animals which had not been pretreated. When used to challenge guinea pigs, β1-SCP was not encephalitogenic at a level of 300 µg. This was the total amount of β1-SCP used for the pretreatment of each animal. The protective activity of the β1-SCP is assumed to be mediated by anti-SCP immunoglobulin but at this stage of our research the mechanism of the protective action is not clear. The protective action may not be directed specifically against lymphocytes sensitized to the encephalitogenic protein of myelin because anti-β1-SCP antibody did not react in immunodiffusion experiments with myelin isolated by differential centrifugation.
Immunocompetent Cells of the Chicken. III Cooperation of Carrier Sensitized T Cells from Agammaglobulinemic Donors with Hapten Immune B Cells
Authors: Weinbaum F. I. , Gilmour D. G. , Thorbecke G. J.
Published: December 2022

Excellent adoptive anti-trinitrophenyl (TNP) antibody responses were obtained from cooperation between bovine serum albumin (BSA) sensitized spleen cells from agamma-globulinemic bursectomized chickens and TNP-Keyhole limpet hemocyanin immune spleen cells from normal chickens upon challenge with TNP-BSA in recipients. A contribution of anti-carrier antibody of bursal cell origin in this anti-hapten response could be ruled out since synergy was obtained in the absence of a detectable humoral response to BSA.
Retention of Lymphocyte Characteristics by Myelomas and θ+-Lymphomas: Sensitivity to Cortisol and Phytohemagglutinin
Authors: Ralph Peter
Published: December 2022

Seven θ+murine lymphomas in culture are very sensitive to killing by phytohemagglutinin; myeloma cell lines and a myeloid leukemia in culture are resistant. Pokeweed mitogen exerts an inhibitory effect both on lymphomas and myelomas, but only at high concentrations. Most of the lymphomas are inhibited by 10-6 M cortisol whereas myeloma cells are generally resistant to the steroid at 10-4 M. However, some IgA myelomas are as sensitive as lymphomas.
Physical Properties of Mouse Spleen Cells Involved in the Mixed Leukocyte Reaction and Graft-Vs-Host Disease
Authors: El-Arini M. O. , Osoba David
Published: December 2022

We have investigated some of the physical properties of mouse spleen cells involved in the mixed leukocyte reaction (MLR) and in graft-vs-host disease (GVHD). When “transformed” cells were separated by velocity sedimentation they were found to be large cells sedimenting at 6.1 mm/hr. On density centrifugation, these cells are found in the light density region of the gradient (modal density, 1.055 g/cm3). In contrast, --positive cells and the precursors of “transformed” cells, i.e., cells capable of responding in the MLR and to phytohemagglutinin (PHA), are found in a higher density region (modal density, 1.071 g/cm3). Transplantation of cells from this region into appropriate recipients produces death from acute GVHD. However, cells from regions deficient in MLR- and PHA-responsive cells, but rich in hematopoietic colony-forming cells (modal density, 1.060 g/cm3), do not produce acute GVHD. Therefore, the density characteristics of ϑ-positive cells and of cells responding in the MLR and to PHA are similar to those of cells producing acute GVHD. This result is in keeping with the possibility that these classes of cells are either identical or subpopulations of a relatively homogeneous single population of cells.
Ontogeny of the Release and Reactivity to the Mediators of IgE-Induced Acute Allergic Reactions in the Rabbit: The Homologous PCA Reaction
Authors: Meng Aniko L. , Pinckard R. Neal , Halonen Marilyn
Published: December 2022

The qualitative and quantitative abilities of rabbits to elicit homologous passive cutaneous anaphylactic (PCA)2 reactions, and their reactivity to exogenously administered bradykinin, histamine, and compound 48/80, were studied as a function of age. Fourteen-day-old rabbits did not respond to PCA, histamine, or 48/80, whereas 74% responded to bradykinin. The lack of PCA reactivity could not be ascribed to lack of functional receptor sites for IgE antibody since skin mast cells and blood basophils of neonatal rabbits demonstrated the capacity for in vivo sensitization. Twenty-eight-day-old rabbits showed maximum incidence of response to PCA; however, PCA titrations demonstrated that adult levels of reactivity had not developed by this time. The incidence of PCA in 28-day-old rabbits was significantly greater than their response to either histamine or 48/80 (p < 0.0005). Forty-two days after birth the incidences of response to homologous PCA, histamine, 48/80, and bradykinin were no longer statistically different. These data indicate that the homologous PCA reaction in the young rabbit cannot be ascribed to the release of and reactivity to histamine alone, and that other mediators must be considered. Furthermore, the above observations suggest that the ontogeny of the homologous PCA reaction in the rabbit is likely due to a sequential maturation of factors responsible for the reactivity to mediators capable of inducing increased capillary permeability.
Protein a from Staphylococcus Aureus
Authors: Sjöholm Ingvar , Bjerkén Anders , Sjöquist John
Published: December 2022

Protein A from the cell walls of Staphylococcus aureus reacts with the Fc part of certain immunoglobulins from several species. Tetranitromethane has been used for the quantitative nitration of the four tyrosyl residues in protein A. The nitrotyrosines formed were subsequently reduced to the corresponding amino-tyrosyl groups with sodium dithionite.

After nitration, protein A lost almost completely its original reactivity towards the Fc fragments of IgG while amino-protein A retained about 40% of its activity, as estimated by quantitative single immunodiffusion tests against normal dog serum. The activity towards rabbit anti-protein A serum was not changed, while neither nitro protein A nor amino protein A agglutinated sensitized sheep red blood cells. The significance of the tyrosines in the active sites of protein A is discussed.
Prevention of Mif Activity by Agents Known to Increase Cellular Cyclic Amp
Authors: Koopman William J. , Gillis Marcia H. , David John R.
Published: December 2022

Agents known to increase intracellular levels of cyclic 3′5′ adenosine monophosphate (cyclic AMP), in a variety of tissues were examined for their effect on migration inhibitory factor (MIF)-induced inhibition of macrophage migration. β-Adrenergic agents (isoproterenol and epinephrine) and prostaglandins E1 and E2 consistently prevent MIF activity on macrophage migration. Aminophylline also prevents MIF activity. These observations suggest a possible role for cyclic AMP in modulating the biologic activity of MIF. The finding that dibutyryl cyclic AMP inhibits MIF activity supports this view.
Guinea Pig Lymphotoxin (LT)
Authors: Coyne John A. , Remold Heinz G. , Rosenberg Steven A. , David John R.
Published: December 2022

Guinea pig lymphotoxin (LT) produced by stimulation of sensitized lymph node cells, specifically by antigen or non-specifically by concanavalin A, was studied by methods previously used to characterize guinea pig migration-inhibitory factor. The results showed that LT induced by either antigen or mitogen was heat labile, eluted from Sephadex columns with peak activity in fractions containing molecules of 35,000 to 55,000 m.w., had an electrophoretic mobility at pH 9.1 similar to albumin and a buoyant density on CsCl similar to protein. It was destroyed by chymotrypsin but not by neuraminidase. These studies show that guinea pig LT can be readily differentiated from guinea pig migration-inhibitory factor. On the other hand, the LT produced by antigen stimulation has many features in common with LT produced by stimulation with concanavalin A.
Immunochemical Study of Group a Streptococcus Peptidoglycan Solubilized by 8 M Urea
Authors: Goldstein Israël , Caravano René , Parlebas Janine
Published: December 2022

Group A streptococcal cell walls, after extraction by formamide, may be partially solubilized in aqueous 8 M urea. This soluble peptidoglycan is not immunogenic, but contains material which precipitates strongly with group A streptococcus polysaccharide antisera as well as with antipeptidoglycan antisera. This immunologic reactivity is due to a free N-acetyl-glucosamine grouping. Coupling urea-soluble peptidoglycan with bovine serum albumin makes it immunogenic on the rabbit. Antibodies produced are directed to the peptidic moiety of peptidoglycan.
Studies on the Mechanism of Lymphocyte-Mediated Cytolysis
Authors: Plaut Marshall , Lichtenstein Lawrence M. , Gillespie Elizabeth , Henney Christopher S.
Published: December 2022

The inhibition of thymus-derived lymphocyte-mediated cytolysis by histamine has been studied in a cytolytic system with immune C57BL/6 effector cells and DBA/2 mastocytoma target cells. Histamine inhibited cytolysis in a reversible manner, and stimulated intracellular cyclic AMP levels in the effector cell. Histamine inhibition was specifically antagonized by burimamide but not by other antihistamines, such as diphenhydramine. The specificity of this antagonism suggests that the histamine receptor on effector thymus-derived cells is of the histamine2 type only.
Proliferation of Stimulated Human Peripheral Blood Lymphocytes: Preferential Incorporation of Concanavalin a by Stimulated Cells and Mitogenic Activity
Authors: Pauli Richard M. , DeSalle Louis , Higgins Patrick , Henderson Earl , Norin Allen , Strauss Bernard
Published: December 2022

Binding of concanavalin A (conA) to human peripheral blood lymphocytes or to lymphoblasts is inhibited by α-D-mannopyranosides or α-D-glucopyranosides. After 1 hr incubation of cells with con A over 80% of the bound con A can be washed off with methyl-α-D-mannoside; after 24 hr only about 25% of the cell associated con A can be removed by competitor. This sequestration is temperature dependent. Cells which have been treated with con A for 20 hr still have as many combining sites for freshly added con A as do non-pretreated cells, even though about 75% of the previously added con A remains associated with the cells. Con A stimulated cells can be separated from non-stimulated cells on Ficoll gradients. Irreversibly bound con A is preferentially accumulated in the stimulated cells. However, this irreversibly bound con A does not stimulate. We suggest that irreversibly bound con A is inside the cell and that internalization is a result of the early activation of energy generating reactions by the mitogenic stimulus. The stimulatory effect depends on reactions at the cell surface.
Synthesis, Assembly and Secretion of γ-Globulin by Mouse Myeloma Cells
Authors: Baumal Reuben , Scharff Matthew D.
Published: December 2022

Normal lymphoid cells obtained from hyperimmunized BALB/c mice and cells from most but not all IgG-producing mouse myeloma tumors and cell lines synthesized more light (L) than heavy (H) chains. While most myeloma tumors secreted their excess L chains, two tumors, MOPC 21 and MOPC 173, degraded the excess L chains and did not secrete them. A third group of tumors produced the same number of H and L chains. In the case of the MPC-11 tumor, excess L chains were synthesized by cloned tumors and cell lines.
A Sensitive Microassay System for Lymphocyte-Mediated Cytotoxicity Induced by PHA and PPD
Authors: Amino Nobuyuki , Degroot Leslie J.
Published: December 2022

A sensitive in vitro microassay for lymphocyte cytotoxicity and lymphotoxin (LT) modified from the method of Takasugi and Klein has been characterized in detail. The cytotoxicity assay (nonspecific target cells + activator + lymphocytes) was performed by using phytohemagglutinin (PHA) and purified protein derivative (PPD). Human skin fibroblasts were not useful as target cells because of insensitivity, but L cells (mouse fibroblasts, subline 5b) proved satisfactory. The fractional survival rate of target cells incubated with activated lymphocytes, relative to incubation with non-activated lymphocytes, was calculated. This discounted nonspecific lymphocyte toxicity.

The sensitivity of four variations of the microassay systems was compared and a “mixture assay” (preincubation of L cells and lymphocytes before addition to the microplate) was found to be most sensitive. Lymphocytes at a concentration of 0.02 × 106/ml (activated by PHA) and 1.0 × 106/ml (activated by PPD) showed nearly 100% target cell destruction. This assay was approximately 50 times more sensitive than the standard incubation of lymphocytes on L cell micromonolayer for 2 days.

Cytotoxicity of the assay performed with direct contact of lymphocytes and target cells was constantly higher than cytotoxicity of the culture supernatant. However, cytotoxicity was strongly correlated in the two, indicating that LT or LT-like materials might be the mediator in this system.

The population of PPD-sensitive lymphocytes in the peripheral blood of a tuberculin skin test positive donor was calculated to be 2.0%.
A Latex Agglutination Test for Measuring Antibodies to Streptococcal Mucopeptides
Authors: Heymer Berno , Schachenmary Walter , Bültmann Burkhard , Spanel Reinhard , Haferkamp Otto , Schmidt Willard C.
Published: December 2022

The preparation and application of mucopeptide (MP)-coated latex particles for determining antibody to streptococcal group A MP is described. Anti-MP hyperimmune sera, normal animal and human sera, and commercial antisera to various antigens were tested. MP-latex agglutination proved to be both specific and highly sensitive, permitting the detection of low antibody levels in 20 to 40% of normal sera; these antibodies were not demonstrable by means of other serologic techniques. Data confirming the specificity of the reaction are presented. No cross-agglutinations were observed with the exception of commercial anti-C reactive protein (CRP) serum. This latter phenomenon may be due to traces of MP present in the preparations used to produce anti-CRP serum.

Latex agglutination should prove useful for establishing the occurrence of anti-MP antibody and for elucidating the role of anti-MP antibody in the various toxic or inflammatory reactions induced by MP in vitro and in vivo.
Hyperacute Rejection of Skin Allografts in the Mouse by the Administration of Alloantibody and Rabbit Complement
Authors: Koene Robert A. P. , Gerlag Paul G. G. , Hagemann Jacq. F. H. M. , van Haelst Urbaan J. G. , Wijdeveld Paul G. A. B.
Published: December 2022

B6AF1 recipients of B10.D2 mouse skin grafts were treated with anti-lymphocyte serum to prolong graft survival. On day 10 after transplantation, when the grafts were well healed in, the hosts received a single intravenous injection of B6AF1 anti-B10.D2 serum (= antiserum to H-2.31) together with rabbit complement. This treatment was followed by hyperacute rejection of the grafts within 24 to 48 hr. Guinea pig complement was ineffective if used instead of rabbit complement. if, in the same model, graft survival was prolonged by treating the hosts with antiserum to H-2.31 (enhancement), hyperacute rejection could be produced on day 10 by the injection of rabbit complement alone. At this time circulating antibody to H-2.31 was demonstrated in the serum of the enhanced mice. The results suggest that mouse skin allografts are completely sensitive to the cytotoxic action of alloantibody in vivo provided an effective species of complement is present at the same time.
Application of Electroimmunodiffusion and Crossed Electroimmunodiffusion to the Comparative Serology of a Microorganism (Histoplasma Capsulatum)
Authors: Sweet George H. , Wilson Donna E. , Gerber Jay D.
Published: December 2022

Electroimmunodiffusion and crossed electroimmunodiffusion were evaluated and compared to immunodiffusion with respect to their resolution capabilities and their abilities to detect antigenic relationships, both qualitative and quantitative, among yeast-phase growth filtrates of eight strains of Histoplasma capsulatum. Electroimmunodiffusion was judged superior to immunodiffusion and crossed electroimmunodiffusion in all respects. A comparison was also made between burro and rabbit antisera and the former was found to resolve a larger number of antigens. By electroimmunodiffusion considerable antigenic variation among strains was detected and correlations were noted between the antigenic properties and length of time in culture and geographic origin of the strains.
Delayed Hypersensitivity to Beryllium Compounds
Authors: Marx James J. , Burrell Robert
Published: December 2022

Assessment of delayed hypersensitivity to beryllium compounds was made by using in vivo and in vitro techniques in sensitized and normal guinea pigs. Hartley strain guinea pigs could be sensitized by repeated intradermal injections over a 6-week period, but not by single injections of beryllium intraperitoneally or intramuscularly. Maximum subtoxic doses of BeSO4 (5 µg) and BeF2 (2.5 µg) were determined for the intradermal skin test in normal guinea pigs. When these doses of salts were used to challenge sensitized animals, classical delayed hypersensitive skin reactions ensued. When normal and sensitized guinea pigs were challenged with an insoluble form of beryllium (BeO), the reactions obtained were considered typical of classical beryllium granulomata in only the sensitized animals.

Soluble mediators of cellular hypersensitivity were assayed as in vitro correlates of delayed hypersensitivity. There was a 74% positive correlation of migration-inhibitory factor (MIF) and skin test results in sensitized animals. This sensitivity to beryllium was transferable by cells when assessed by both in vivo and in vitro techniques in the passively sensitized recipients. Lymphocytotoxin was also demonstrable in cell suspensions from sensitized animals when exposed to beryllium, but not in unsensitized controls.

Patients with diagnosed chronic pulmonary berylliosis were assessed for sensitivity by in vitro inhibition of peripheral blood leukocyte migration and showed MIF production when exposed to BeSO4 in vitro. An unexplained inhibition of leukocyte migration was observed when peripheral leukocytes from these patients were exposed in vitro to tuberculin purified protein derivative (PPD), even though the patients were all skin test-negative before steroid therapy. Peripheral blood leukocytes from patients with chronic lung disease other than berylliosis and no history of exposure to beryllium were MIF-negative to BeSO4, but positive to PPD.

The available data support the hypothesis that reactivity to beryllium compounds is due to classical delayed hypersensitivity, and the techniques that were used may be applicable as diagnostic tools.
Homocytotropic Autoantibodies in Rabbits Subjected to Cryosurgery
Authors: Riera Clelia M. , Faillaci María G. , Yantorno Carlos
Published: December 2022

Homocytotropic antibodies can be induced in rabbits by parasite infection (1, 2) or by immunization with heterologous antigens (3, 4). The presence of such antibodies in primary responder and hyperimmune animals was recently demonstrated (5, 6).

This study reports the induction of homocytotropic autoantibodies in male rabbits subjected to injury of the accessory glands by in vivo freezing. It is known that such treatment is able to evoke a specific hemagglutinating autoantibody response of the 19S and 7S type (7, 8).

Materials and Methods. Thirty-five adult rabbits, cross-bred from various generations of New Zealand and Flemish Giant were used. Rabbits were subjected to one or several freezings of the accessory glands following a procedure previously described (8). Bleedings were performed approximately at 3, 7, 10, 15, 20, and 25 days after each cryogenic treatment.

Semen was obtained from normal animals by means of the artificial vagina (9).
Supressor Cells in the Graft vs Host Reaction
Authors: Hardin John A. , Chused Thomas M. , Steinberg Alfred D.
Published: December 2022

The importance of cellular interaction in the regulation of the immune response has become apparent in recent years. The first example of such interaction to be observed was synergy between bone marrow and thymus cells in the antibody response to sheep erythrocytes (1). Subsequently, two subpopulations of thymicderived (T)1 cells have been shown to cooperate positively in cellular immune responses such as graft-vs-host disease (GVH) (2, 3) and allograft rejection (4). A negative or inhibitory interaction has recently been described in humoral immunity (5–8) mediated by “regulator” or “suppressor” T cells which interact with bone marrow-derived (B) cells to reduce the antibody response. The cell-mediated GVH reaction is reduced in the presence of antigenic competition (9). Evidence has been presented to implicate suppressor T cells or a product thereof as the cause of antigenic competition (10, 11). These and other observations (12) suggest that the GVH reaction might be inhibited by suppressor T cells.
The Responsiveness of Tolerant Lymphocyte Populations after Transfer to Normal Recipients
Authors: McGregor D. D.
Published: December 2022

Thoracic duct lymphocytes taken from Lewis rats exhibiting tolerance of F1 hybrid skin grafts can recirculate from blood to lymph in recipients syngeneic with the skin donors. They do so in a functionally active condition as evidenced by the ability of recirculating donor cells to cause a graft-vs-host reaction in recipient rats belonging to a second F1 hybrid cross. This phenomenon was exploited in experiments designed to detect tolerant lymphocytes within cell populations that were either unresponsive or only feebly responsive to BN alloantigens. The results demonstrate that lymphocyte populations from tolerant donors retain approximately the same level of unresponsiveness when housed for up to 2 weeks in normal, non-tolerant recipients. Failure of the transferred cells to regain an appreciable level of responsiveness in these circumstances implies that tolerance to major histocompatibility antigens is a stable property of circulating lymphocytes, and lends support to the concept that tolerance induction involves either the selective deletion or permanent inactivation of antigen reactive cells.
Heart Autoantibodies in Mice: Genetic, Bacterial, and Age Determinants
Authors: Schwab John H. , Brown Roger R.
Published: December 2022

IgM autoantibody specific for heart sarcolemma can be induced in the mouse by immunization with protoplast membranes from group A streptococci. An IgM autoantibody of similar specificity develops “spontaneously” with increasing age in non-immunized normal mice and reaches a very high incidence in old mice of certain inbred strains. Comparison of strains of A-congenic mice indicates that the production of this “spontaneous” autoantibody is associated with the H-2 locus. The genetic control is apparently dominant since the F1 hybrid of high and low incidence strains is also of high incidence. This antibody is independent of sex. IgM antisarcolemma antibody can be absorbed from both immunized and “spontaneous” sera with protoplast membranes from group A streptococci but not with group D membranes or group A cell walls. Antibody against group A streptococcal membranes, measured directly by membrane agglutination, is present in the serum of all old normal mice tested. These agglutinin titers do not correlate with heart autoantibody. The autoantibodies give no reaction with membranes in liver or kidney and much less reaction with skeletal muscle compared to heart. An IgM antinuclear autoantibody which gives a distinctive diffuse pattern of staining by indirect immunofluoresence is also induced by immunization with group A streptococcal protoplast membranes.
Regulation of Antibody Response in Vitro
Authors: Kishimoto Tadamitsu , Ishizaka Kimishige
Published: December 2022

Rabbits were primed with DNP-coupled Ascaris antigen (DNP-Asc) precipitated with aluminum hydroxide gel, received a supplemental immunization with bovine γ globulin (BGG) or ragweed Fr. D (Rag), and their mesenteric lymph node cells were stimulated in vitro with DNP-Asc or DNP-secondary carrier conjugate (DNP-BGG or DNP-Rag). IgM, IgG, and IgE antibodies were all formed in vitro, when the cells were stimulated with DNP-Asc. Stimulation of the same cells with DNP-secondary carrier also resulted in the formation of anti-DNP-antibodies of the three immunoglobulin classes, when donors of lymphocytes received the carrier with alum. Carrier-specific cells generated by the supplemental immunization with the same carrier in complete Freund's adjuvant (CFA) showed helper function for IgM and IgG antibody response but not for IgE antibody response. Similarly, lymph node cells from the rabbits primed with DNP-Rag and receiving supplemental immunization with Asc in alum responded to DNP-Asc to form IgM, IgG and IgE antibodies. In this system, supplemental immunization with CFA was occasionally effective for IgE antibody response to DNP-Asc, however, alum was a more effective adjuvant than CFA to prepare a carrier-specific cell population having helper function for IgE antibody response. The helper function of the carrier specific cell population for IgE antibody formation did not have any relationship with either the helper function of the same cell population for IgG antibody response or with the size of the carrier-specific cell population estimated by 3H-thymidine uptake. Evidence was obtained that lack of IgE antibody response to DNP-secondary carrier with the lymphocytes from the CFA group was not due to an increase of precursors of carrier-specific antibody-forming B cells. The results indicated that carrier-specific helper cells for the IgE antibody response were different from those for IgG/IgM antibody response.
Extraction and Purification of Soluble HL-A Antigens from Exhausted Media of Human Lymphoid Cell Lines
Authors: Pellegrino Michele A. , Pellegrino Anna , Ferrone Soldano , Kahan Barry D. , Reisfeld Ralph A.
Published: December 2022

HL-A alloantigens could be isolated in soluble form by 3 M KCl extraction of membranous debris present in the exhausted, cellfree media of several cultured human lymphoid cell lines. These antigens were highly specific, possessed good immunologic potency, and could be obtained in high yield from this source material. Purification of HL-A antigens could be readily achieved by preparative acrylamide gel electrophoresis resulting in the isolation of an electrophorectically homogeneous moiety with a 20-fold increase in serologic reactivity. Yield and immunologic potency of HL-A antigens from exhausted culture media did not vary during different phases of the cell growth cycle in contrast to whole lymphoid cells where optimal yields were achieved in the late log or resting phase. The condition of the cell membrane, cell metabolism, and membrane turnover seemed to have an appreciable influence on the yield and immunologic potency of soluble antigens that can be extracted with 3 M KCl from cultured human lymphoid cells.
Technical Aspects of the Rosette Tests Used to Detect Human Complement Receptor (B) and Sheep Erythrocyte-Binding (T) Lymphocytes
Authors: Mendes Nelson F. , Tolnai Marina E. A. , Silveira Nelson P. A. , Gilbertsen Richard B. , Metzgar Richard S.
Published: December 2022

The conditions for rosette formation between normal human thymus-derived (T) lymphocytes and unsensitized sheep erythrocytes (E) and between bone marrow-derived (B) lymphocytes and sheep erythrocytes sensitized with rabbit antibody and mouse complement (EAC) have been examined in detail in this collaborative study. Mean percentage values for E and EAC obtained at Duke University were 55.4 and 13.3, and at the Escola Paulista de Medicina were 30.4 and 26.2. Eighty-nine to 98% of thymus cells formed E rosettes. Another method for detecting complement receptor lymphocytes utilizing human erythrocytes sensitized with antibody and complement (HEAC) was compared with the EAC indicator. At Duke University the mean percentage value for HEAC was 14.4 and at the Escola Paulista de Medicina that value was 18.3. The variation in rosette percentages when the same normal donor was tested on different days and the effect of storage of blood and/or lymphocytes on rosette formation were also examined. Rosette formation between T lymphocytes and E was found to be temperature dependent, with maximum values occurring between 10°C and 25°C with no rosette formation occurring at O°C or 37°C. Some thymocytes, however, were capable of forming E rosettes at 37°C. When EAC peripheral blood lymphocyte rosettes were determined below 37°C, some T lymphocytes were able to form rosettes with E receptors on the EAC indicator cells. Use of HEAC instead of EAC eliminated the interference of E rosette formation with the latter indicator. When centrifugation was compared with rotation in the procedures to detect complement receptor lymphocytes, it was found that fewer B cells were detected when the centrifugation procedure was omitted. This paper further demonstrates the use of rosette formation and gradient centrifugation for the specific depletion of B or T cells from peripheral blood lymphocytes.
A Neutrophil Immobilizing Factor Derived from Human Leukocytes
Authors: Goetzl Edward J. , Gigli Irma , Wasserman Stephen , Austen K. Frank
Published: December 2022

The neutrophil-immobilizing factor (NIF) derived from human leukocytes inhibited the random migration and chemotactic response of purified human neutrophils and eosinophils but not of monocytes. Concentrations of NIF sufficient to suppress mobility did not diminish phagocytic capacity or adherence to plastic Petri dishes of human neutrophils; the increased activity of the hexose monophosphate shunt associated with either phagocytosis or direct interaction with chemotactically active concentrations of kallikrein or C5a was also not suppressed. It is postulated that the in vivo role of NIF may relate to its ability to concentrate neutrophils in inflammatory foci without depressing their phagocytic potential or metabolic activation.
Antigen Stimulation of 3H Thymidine Incorporation by Subpopulations of Human Peripheral Blood Cells
Authors: Eisen Seth A. , Lyle Leon R. , Parker Charles W.
Published: December 2022

Using human peripheral blood lymphocytes at various stages of purification we have re-examined the question of whether foreign proteins and hapten-protein conjugates to which the cell donor has no known previous immunization can stimulate in vitro 3H thymidine incorporation. Responses were examined in dextran-sedimented mixed leukocytes, purified lymphocytes isolated by isopycnic centrifugation in a Ficoll-Hypaque gradient (FH cells), and FH cells that had been further purified by passage through a short nylon wool column (nylon purified cells). The dextran sedimented, FH, and nylon purified cells were cultured in autologous serum with various antigens for 5 days and the increase in 3H thymidine incorporation was measured. In mixed leukocytes and FH cells all of the antigens examined (bovine serum albumin, bovine γ-globulin, Candida albicans, histoplasmin, trinitrophenyl-bovine serum albumin, dinitrophenyl-bovine serum albumin, and fluorescein-bovine γ-globulin) produced a significant increase in 3H thymidine uptake. Appropriate controls indicated that the response could not be ascribed to endotoxin. Serum protein and fungal antigens stimulated nylon purified cells as well or better than the leukocyte or FH populations. Contrariwise, the three hapten-protein conjugates inhibited 3H thymidine incorporation by nylon-purified cells. Although there are several alternate possibilities, we favor the interpretation that the in vitro response to serum protein and fungal antigens represents a secondary immune phenomenon in which few if any monocytes are required, whereas stimulation by hapten-protein conjugates involves a primary immune (monocyte dependent) response. However, the possibility that the hapten-protein stimulation is a secondary response involving cells sensitized to cross-reacting antigens is by no means excluded. The inhibition of nylon-purified cells by hapten-protein conjugates may conceivably represent an in vitro correlate of tolerance.
Genetic Control of the Immune Response in Guinea Pigs to the Known Sequence Polymer (Tyr-Glu-Ala-Gly)n
Authors: Maurer Paul H. , Odstrchel Gerald , Merryman Carmen F.
Published: December 2022

The immune response in guinea pigs against the known sequence polymer, (tyr-glu-ala-gly)n has been studied. Hartley guinea pigs and inbred strain 13 guinea pigs react against the polymer (cellular and humoral responses) but strain 2 guinea pigs do not. Breeding studies have established that the response is controlled by a single dominant gene and that the locus involved is also linked to the major histocompatibility locus of the species.
Failure to Obtain Histamine Release from Rat Mast Cells Exposed to Human Allergic Serum and Specific Antigen or IgE Myeloma Protein and Anti-IgE
Authors: Plaut Marshall , Lichtenstein Lawrence M. , Bloch Kurt J.
Published: December 2022

Attempts were made to induce histamine release from rat mast cells exposed directly to serum from allergic patients plus specific antigen (one-stage procedure) or from mast cells initially exposed to such serum and subsequently challenged with antigen (two-stage procedure). Varying conditions of incubation, type of buffer and pH were tested; negative results were obtained. Similar attempts were made using IgE myeloma protein and rabbit antibody to IgE to induce histamine release from rat mast cells; these experiments were also unsuccessful.

IgE myeloma protein did not prepare rat skin for reverse passive cutaneous anaphylaxis mediated by anti-IgE. IgE myeloma protein which is capable, at low concentration, of inhibiting the sensitization of human basophils by IgE antibodies, failed to inhibit sensitization of rat mast cells by rat IgE antibodies both in vivo and in vitro despite its use in very high concentration. While some inhibition of the IgGa antibody-mediated release of histamine was obtained with IgE myeloma protein, large amounts of this protein were required, and some inhibition was achieved with human IgG as well, making it difficult to evaluate the inhibitory effect. The findings suggest that human IgE does not compete with rat IgE for receptors on rat peritoneal mast cells.
Thymus Function in Spontaneous Lymphoid Leukemia
Authors: Nagaya Hiroshi
Published: December 2022

More than 90% of AKR mice died of spontaneous lymphoid leukemia before the age of 12 months. However, the mice did not develop leukemia until the age of 6 months suggesting that a latent period or aging was necessary for the development of leukemia. Moreover, the incidence of leukemia was reduced to nil when the mice were thymectomized before the age of 6 months.

For studying the thymic influence on leukemogenesis, 1- or 5- to 6-month-old AKR mice were thymectomized and received syngeneic 5- to 6- or 1-month-old thymus grafts. When 1-month-old mice were thymectomized and grafted with thymus glands from 5- to 6-month-old mice, 13 of 21 (62%) died of leukemia before the recipient mice reached the age of 6 months. Only 3 of 17 control mice thymectomized at the age of 1 month and grafted with 5- to 6-month-old spleens died of leukemia. When 5- to 6-month-old mice were thymectomized and grafted with thymus glands from 1-month-old mice, none of 19 mice developed leukemia for at least 5 months or until the recipient mice were 11 months old. The latent period for leukemogenesis appears to be required by the reticular epithelial cells of the thymus grafts rather than by the lymphoid cells of host origin migrating from bone marrow to thymus.
Selective Loss of H-2 Antigenic Reactivity after Chemical Modification
Authors: Pancake Samuel J. , Nathenson Stanley G.
Published: December 2022

Murine H-2 alloantigen glycoproteins were subjected to chemical modification by different modifying reagents and the effects of such modification on antigenic reactivity were evaluated. Modification of tyrosine residues using N-acetylimidazole or tetranitromethane was followed by loss of antigenic reactivity of all H-2 glycoproteins with reagents thought to be specific for amino groups, i.e., reductive alkylation and acetamidination, revealed a clear dissimilar susceptibility to inactivation. For example, for the H-2d alloantigens, specificity H-2.4 was almost totally lost and H-2.31 was almost completely retained. For the H-2b alloantigens, H-2.33 was almost completely inactivated, whereas H-2.2 and H-2.5 were affected to a much lesser degree.

The reductive methylation procedure was found to change about 90% of the lysine residues to dimethylysine. The alteration of these amino groups with a direct effect on the antibody binding properties of some of the antigenic determinants on the antigen provides additional support for the view that protein structure determines antigenic H-2 reactivity.
Transient Appearance of Cells Secreting Antibody of Different Specificities after Immunization of Mice with Trinitrophenylated Erythrocytes
Authors: Couderc Jacques , Bleux Christian , Birrien Jean-Louis , Liacopoulos Panayotis
Published: December 2022

A sensitive variant of the method detecting hemolytic plaque-forming cells (PFC) was used to study the possibility of production of antibody of two different specificities by single cells. 2,4,6-Trinitrophenyl (TNP), conjugated pigeon erythrocytes (PRBC) were given in a single immunizing injection to mice, and their spleen cells were plated on days 3, 4, and 5 after this injection with both PRBC and TNP-sheep erythrocytes (SRBC). A significant number of PFC lysing both erythrocytes (3.24 × 106 cells) was found on the 3rd day after immunization. On the 4th day, out of 13.39 × 106 cells examined in all experiments, 6.260 (468 × 106 cells) monospecific PFC and 137 (10.2 × 106 spleen cells) bispecific PFC were found, whereas on the 5th day, only 0.74 × 106 cells were bispecific. Studies on cross-reactivity between PRBC and SRBC or TNP-SRBC showed that only 0.11 to 0.2 × 106 cells produce cross-reacting antibody. Furthermore, inhibition studies with TNP-bovine albumin or TNP-ε-aminocaproic acid showed that these inhibitors did not influence anti-pigeon PFC at all, but sharply reduced the number of anti-TNP-SRBC PFC and provoked a disappearance of bispecific PFC. On the other hand, short term incubation with puromycin also caused disappearance of bispecific PFC. The same proportion of double PFC was found after immunization of mice with native PRBC and SRBC. It can be inferred, therefore, that shortly (on the 3rd and 4th day) after primary immunization with two unrelated antigens, a low but significant percentage of cells simultaneously produce and secrete antibody of two different specificities.