bluuebunny/crossref_metadata_2025 huggingface dataset:
Chemical Modulation of Cell Surfaces by Sulfhydryl Compounds: Effect on C3b Receptors
Authors: Dierich Manfred P. , Ferrone Soldano , Pellegrino Michele A. , Reisfeld Ralph A.
Published: December 2022

C3b receptors on human erythrocytes, peripheral blood lymphocytes, and cultured lymphoid cells were shown to be sensitive to treatment with dithiothreitol, 2-mercaptoethanol, 2-amino-ethylisothiouronium bromide hydrobromide (AET), reduced glutathione, and l-cysteine (in order of decreasing effectiveness). Sensitivity was dependent on temperature, time, dose, and pH of the solution applied. Loss of C3b receptor activity became apparent at AET concentrations which caused an increase in erythrocytes' susceptibility to lysis. Raji cells which had lost C3b receptor activity due to AET treatment, started to re-express this receptor function after a lag period of 8 to 10 hr. AET treatment had no detectable effect on the expression of other surface markers, e.g., those detected on erythrocytes by concanavalin A and anti-AB blood grouping serum, or on cultured human lymphoid cells by HL-A alloantisera. However, changes of erythrocyte morphology could be seen in the scanning electron microscope. The sensitivity of C3b receptors to reducing agents suggests that disulfide bridges are important to this receptor's functioning. On the other hand, free sulfhydryl groups appear not to be involved because α-iodoacetamide did not alter the cell surface expression of the C3b complement receptor. Treatment of C3b receptor carrying cells with sulfhydryl compounds can be used as a tool to study the participation of the receptors in the cells' biologic functions.
Effects of Bacterial Products on Lymphocytes and Macrophages: Their Possible Role in Natural Resistance to Listeria Infection in Mice
Authors: Petit Jean-Claude , Unanue Emil R.
Published: December 2022

Fluids from Listeria monocytogenes cultures contain products that possess a number of biologic activities. The fluids contain a mitogenic substance for B lymphocytes. Thus, their injection into mice produces striking hyperplasia of the B cell zones of the spleen. Also, lymphocytes in vitro are stimulated to proliferate, regardless of whether they are first depleted of T lymphocytes by exposure to anti-θ serum plus complement. These fluids injected intraperitoneally activate macrophages as judged by morphologic criteria. Mice that receive the products develop a transitory state of nonspecific resistance to infection by live Listeria. The relationships between B cell mitogenicity and macrophage activation—and protection—were not established. This kind of reaction of lymphoid cells to Listeria products may represent an early nonspecific line of defense against bacterial invasion.
Alloantiserum-Mediated Suppression of Histocompatibility-Linked Ir Gene Controlled Immune Responses
Authors: Bluestein Harry G.
Published: December 2022

Alloantisera were produced in guinea pigs by immunizing one inbred strain with the lymphoid cells of another. The effect of these sera on the response of sensitized lymphocytes to antigen or mitogen was studied in vitro. Strain 13 anti-strain 2 alloantisera, when added to cultures at a 1:75 or greater dilution, specifically suppress responses to GA and to 2,4-dinitrophenyl (DNP)-GL, both of which are controlled by strain 2-linked immune response (IR) genes. Addition of the anti-strain 2 sera at higher concentrations diminishes the apparent specificity for strain 2-associated immune responses. for example, at a 1:30 dilution the anti-strain 2 sera suppress the response of sensitized Hartley lymphocytes to the mitogen phytohemagglutinin and to the antigen GT whose controlling immune response gene is linked to strain 13 histocompatibility antigens. When compared as a function of antiserum concentration, the ability of the anti-strain 2 alloantisera to suppress strain 2-associated immune responses in vitro closely parallels its cytotoxic capacity. Furthermore, the ability of the anti-strain 2 sera to suppress GA responses in vitro depends on the presence of strain 2 histocompatibility antigens on the responding lymphocytes. In contrast to its effective suppression of GA responses in sensitized lymphocytes bearing strain 2 histocompatibility antigens, the anti-strain 2 antisera has no effect on the response to GA in lymphocytes from responder Hartley guinea pigs that did not inherit the strain 2 histocompatibility locus. These studies indicate that the histocompatibility antigens on the lymphocyte membrane are the target through which the alloantisera mediate specific immunosuppression
Requirement of Thymus-Derived θ-Positive Lymphocytes for Rejection of DNA Virus (SV40) Tumors in Mice
Authors: Tevethia Satvir S. , Blasecki John W. , Waneck Gerald , Goldstein Allan L.
Published: December 2022

The nature of lymphocytes capable of inhibiting the growth of syngeneic papovavirus SV40 tumor cells in BALB/c was investigated by the tumor cell neutralization assay. Treatment of immune lymphocytes with anti-C3H (θ) AKR sera in the presence of complement eliminated the lymphocytes capable of inhibiting tumor cells in vivo. Treatment of immune lymphocytes with normal AKR sera and complement had no effect. Lymphocytes filtered through nylon wool column, a procedure known to remove B lymphocytes, retained their capacity to inhibit tumor cells in vivo. Thymectomized, irradiated, and reconstituted mice failed to develop lymphocytes sensitized to SV40 tumor-specific transplantation antigen (TSTA). These results suggest that cellular immune response to SV40 tumor cells in syngeneic mice is mediated by θ-positive thymus-derived lymphocytes.
Development of Antibodies to Viral and Tumor Antigens before Tumor Induction in Syrian Golden Hamsters Inoculated Intravenously at 3 Weeks of Age with SV40
Authors: Diamandopoulos George Th. , McLane Mary-Frances
Published: December 2022

Three-week-old weanling male Syrian golden hamsters were inoculated i.v. with 108.5 TCID50 of oncogenic DNA simian virus 40 (SV40). All input virus was cleared from the blood stream within 48 hr. Antibody to SV40 V (virion) antigen appeared as early as the 1st postinoculation week, reached a peak by the 3rd week, and gradually decreased thereafter. Antibody to SV40 T (tumor) antigen was first detected by the 4th week. Its appearance preceded tumor induction by as many as 6 weeks for a tumor at the in situ stage, and 10 weeks for a palpable tumor. The results indicate that the development of antibody to viral and tumor antigens in hamsters inoculated i.v. at 3 weeks of age with high titer SV40 precedes rather than coincides with or follows the induction of tumors. The presence, therefore, of these antibodies in the serum of hosts in which the virus-cell interaction is of the transforming type, only signifies prior virus exposure, and can not be taken as evidence for the establishment of tumors in vivo.
Sequential Appearance of Three Different Anti-Fluorescein Combining Sites in Hyperimmunized Rabbits: Characterization by Circular Dichroism and Binding Studies
Authors: Gollogly John R. , Cathou Renata E.
Published: December 2022

Fluorescein (F1) bound in the combining site of anti-F1 antibodies exhibits extrinsic optical activity which is a function of the fine structure of the site. We have utilized this phenomenon to study structural changes in antibody combining sites as a function of time and repeated boosting. Four rabbits were immunized with F1-keyhole limpet hemocyanin (KLH) and successively boosted whenever the antibody production declined. Antibodies were purified from bleedings taken once or twice a week and characterized by the extrinsic circular dichroism (CD) of their hapten-antibody complexes. After the third boost, three characteristically different CD spectra were observed, each corresponding to a different combining site configuration.

Usually each individual bleed contained only one of the three antibody types and the particular type observed depended on the time after boosting: type I was present for the first 3 to 6 weeks of the antibody response, type II was present for a further 2 to 3 weeks and, finally, as the antibody production declined, only type III antibodies were found. This sequential response was repeated after each succeeding boost.

Each of the antibody types was functionally homogeneous with respect to its extrinsic CD properties. In contrast, binding data showed that they were somewhat heterogeneous. The induced CD bands of bound F1 originate in the xanthenone ring whereas the binding of F1 depends on the interactions of both the xanthenone and phenylcarboxylate moieties with the site. Therefore, these results suggested that the anti-F1 combining site consists of two distinct subsites. The functional homogeneity of each antibody type may be due to a common tertiary structure of the xanthenone-binding subsite whereas the heterogeneity indicated by the binding data may be due to heterogeneity within the phenylcarboxylate-binding subsite.

Possible mechanisms for the sequential appearance of these antibody types are also discussed.
Resistance of Sheep Erythrocytes to Immune Lysis by Treatment of the Cells with a Human Erythrocyte Extract: Studies on the Site of Inhibition
Authors: Hoffmann E. M. , Cheng W. C. , Tomeu E. J. , Renk C. M.
Published: December 2022

Sheep erythrocytes (E) were treated with an anti-complementary extract derived from human erythrocyte stromata (I-H) and then converted to the intermediate state . It was found that the treated decayed at about the same rate as control which had been generated from untreated sheep E. However, the extent of lysis of the treated intermediate was less than that of the control. Subsequent studies indicated that the same number of SAC was generated on treated E as was generated on control cells. Furthermore, the resulting EAC consumed essentially the same amount of C2 in both cases, but which had been formed from extract-treated sheep E consumed less C3 than did control (cells not treated with I-H). It was also found that I-H would inhibit lysis of but not when the intermediates were formed and then treated with the extract.

Preliminary purification was attempted with a combination of DEAE chromatography and gel filtration with Sepharose 4B. However, yields of active material were quite low. It was estimated that the factor, as isolated, had a very large molecular weight.
Cooperation and Cell-Mediated Cytotoxicity as Functions of two Subsets of T Cells
Authors: Dennert Gunther , Lennox Edwin S.
Published: December 2022

After stimulation of mice with alloantigens we compared the ability of their spleen cells to effect cell-mediated cytotoxicity and to assist in the triggering of bone marrow-derived lymphocytes. We did this using the DBA/2 mastocytoma, P815, as allogeneic immunogen on the one hand and as target for cytotoxicity and carrier for hapten on the other. Spleen cells from animals immunized with either low doses of irradiated or formaldehyde-treated P815 cells exhibit cooperating activity but not cytotoxic activity. In addition, we show that spleen cells from C57BL/6 mice immunized with P815 do contain cytotoxic thymus-derived (T) lymphocytes but are unable to trigger bone marrow-derived cells for a hapten response in vitro to hapten-coupled P815 antigen. That there may be some helper cells in cytotoxic spleens is not excluded, for we show that mixing cytotoxic spleens with those showing helper function blocks the expression of that helper function. Since we show that both functions, helping and killing, are sensitive to anti-θ serum, we conclude that they are carried out by two different subsets of T cells.
Interaction of Staphylococcal Protein a with Virus-IgG Complexes
Authors: Austin R. Marshall , Daniels Charles A.
Published: December 2022

Protein A, an Fc-reacting molecule from Staphylococcus aureus Cowan I, neutralized infectious vaccinia virus-IgG (Vac-IgG) and herpes simplex virus-IgG (HSV-IgG) complexes. The amount of protein A-mediated neutralization was dependent on the concentration of anti-viral IgG and protein A incubated with the virus. Upon attachment of protein A to the HSV-IgG complex a portion of the virus formed an infectious HSV-IgG-protein A complex which could be neutralized by anti-protein A serum. In addition, attachment of protein A to HSV-IgG was found to inhibit complement-mediated neutralization. Heat-extracted Jensen's protein A caused less neutralization of HSV-IgG than the enzyme-extracted protein A-1. The experiments indicate that molecular size may play a major role in the mechanism by which protein A as well as other immunoglobulin-reacting substances neutralize virus-IgG complexes.
Antigen-Induced Trapping of Immunospecific Lymphocytes in the Spleen
Authors: Emeson Eugene E. , Thursh Donald R.
Published: December 2022

Tritium-labeled anti-chicken red blood cells (CRBC) and 14C-labeled anti-sheep red blood cells (SRBC) lymphocytes were adoptively transferred to three groups of syngeneic mice, previously challenged i.v. with CRBC, SRBC, or Brucella abortus (BA). Subgroups of each group were sacrificed at 24, 48, and 72 hr after challenge and the relative numbers of anti-CRBC to anti-SRBC lymphocytes present in the spleen, peripheral lymph nodes, and peripheral blood for each group of mice were inferred from the mean 3H/14C ratios of these tissues. The results of these studies clearly established that challenged spleens can selectively trap recirculating immunospecific lymphocytes. Selective trapping was maximum at 48 hr after challenge, but still persisted at a high level at 72 hr. An unexpected finding was the clear demonstration of a deficiency of immunospecific lymphocytes in the peripheral lymph nodes. This deficiency was maximum at 24 hr and then declined. The functional significance and implications of the selective trapping of immunospecific lymphocytes and the concomitant depletion of these cells in other tissues is discussed.
The Termination of Immunologic Unresponsiveness to the Cyanogen Bromide Fragments of Bovine Serum Albumin in Rabbits
Authors: Benjamin D. C. , Hershey Caroline W.
Published: December 2022

A specific unresponsive state to the cyanogen bromide fragments of bovine serum albumin (BSA) was terminated by injecting soluble native BSA. The quantity of antibody produced in response to primary injections of BSA was less than normal. Responses to booster injections were normal with a normal affinity for BSA. Antibody was produced to both sets of determinants on BSA (i.e., those to which the rabbits were unresponsive as well as those to which it was not). These results are discussed in relation to various proposed mechanisms of the maintenance and termination of the unresponsive state.
Clonal Nature of the Immune Response to Phosphorylcholine
Authors: Claflin J. Latham , Davie Joseph M.
Published: December 2022

The fine specificity of anti-phosphorylcholine antibodies was studied in different rodent species and 17 different inbred mouse strains. Measurements of the relative affinity and specificity of the antibodies were derived from a hemolytic plaque-inhibition assay with phosphorylcholine and two of its analogues, glycerophosphorylcholine and choline, as inhibitors. Within a given species, inhibition of plaque-forming cells was accomplished within a narrow range of ligand concentration and little individual variation was seen. Moreover, no changes in either the slopes or relative position of the inhibition curves occurred at different times after immunization. These findings suggest that a restriction in heterogeneity of the anti-phosphorylcholine antibodies exists in each of the rodent species. However, striking species-specific differences were observed in the binding characteristics of antibodies from different species of rodents.
Purification of Basophilic Leukocytes from Guinea Pig Blood and Bone Marrow
Authors: Dvorak Harold F. , Selvaggio Salvatore S. , Dvorak Ann M. , Colvin Robert B. , Lean Deborah B. , Rypysc Jane
Published: December 2022

Methods are presented for obtaining highly purified preparations of basophilic leukocytes from the blood (mean purity, 62%) and bone marrow (mean purity, 38%) of guinea pigs. The following steps are performed in sequence: harvest of blood or bone marrow cells from animals primed with antigen to develop a basophilia; phagocytosis of carbonyl iron particles by neutrophils and monocytes; sedimentation of erythrocytes with gelatin (in the case of peripheral blood); removal of phagocytic cells and residual-free iron particles by passage through a magnetic field; centrifugation on a newly developed 4.5 to 9.5% linear Ficoll density gradient; and removal of residual erythrocytes by Tris-NH4Cl (optional step for peripheral blood). The basophils isolated are viable, functional cells, and, depending on the source and degree of purity desired, may be obtained in quantities of 5 × 106 to 2 × 108 and in an overall yield of 25 to 100%.
Transplantation of a Marek's Disease Lymphoma in Syngeneic Chickens
Authors: Theis Gail A. , Schierman Louis W. , Mcbride Raymond A.
Published: December 2022

A lymphoma arising in an inbred B1/B1 chicken, after intramuscular injection of leukocytes from a Marek's disease virus (MDV)-infected chicken, has been transplanted serially in chickens of the same B genotype. The Marek's disease (MD) lymphoma grows progressively to the death of all injected syngeneic and hybrid B1/B2 chickens, although it is not transplantable to allogeneic B2/B2 chickens. Detection of alloantigens by specific alloantisera has confirmed the perpetuation of the original B1 phenotype in cells of the transplantable MD lymphoma. This tumor is considered, therefore, to be a transplantable tumor associated with MD. Studies in isolator-reared chickens have suggested that MDV remains in close association with cells of the transplant, since injection of MD lymphoma cells has resulted in appearance of MDV in feather follicles, development of anti-MDV antibodies, and transmission of MD to recipient chickens.
Formation of the C3-Cleaving Properdin Enzyme on Zymosan
Authors: Brade Volker , Nicholson Anne , Bitter-Suermann Dieter , Hadding Ulrich
Published: December 2022

During incubation of ZXd2, an insoluble intermediate of the properdin system, with properdin factors D and B a complex forms, designated ZX, which cleaves purified C3. Factor D is replaceable in this system by proteolytic enzymes, e.g., trypsin, pronase or plasmin. ZX formation with ZXd2, factor B and factor D or trypsin requires the presence of Mg++. ZX complexes formed either with factor D or with trypsin are identical with respect to their stability in the absence of divalent cations and with respect to the decay of the C3-cleaving enzymes. From the data obtained with proteolytic enzymes it is concluded that factor D acts as a protease in the formation of ZX. Sequential studies of ZX formation revealed that factor B binds reversibly to ZXd2 in the presence of Mg++. The resulting ZXd2 B complex was found to acquire enzymatic activity against C3 after treatment with factor D or trypsin. Our data suggest that in the ZXd2 system two steps are involved in ZX formation, namely binding of factor B to a receptor on ZXd2, probably C3b, which occurs only in the presence of Mg++, and subsequent proteolytic activation of this complex which is Mg++-independent.
Detection and Enumeration of Immunocytes Producing Different Classes of Vibriolysins after Primary and Secondary Immunization with Cholera Vaccine
Authors: Kateley John R. , Patel Chandu , Friedman Herman
Published: December 2022

The vibriolytic plaque-forming cell (PFC) assay was used to study the kinetics of the primary and secondary immune responses of mice and rabbits immunized with heat-killed cholera vibrios. Immunocytes releasing IgG antibody could be detected as readily as those immunocytes secreting IgM antibody in spleens of BALB/c mice and New Zealand White rabbits after a single injection of vaccine. Peak numbers of indirect (IgG) PFC were detected 3 to 4 days after the peak direct (IgM) PFC response (12 to 14 days). In contrast, only direct vibriolytic PFC were detected in spleens of NIH Albino mice during the primary response to cholera antigens. After a second injection of vaccine, IgM, IgG, and IgA PFC were detected in both mouse strains with peak responses for each immunocyte class occurring within the first week after booster injection. Heat-killed vibrios or a lipopolysaccharide (LPS) extract, but not cholera exotoxin or E. coli LPS, inhibited the vibriolytic response. Furthermore, viable cholera vibrios were a more sensitive indicator cell in the immunoassay than sheep erythrocytes sensitized with a cellular extract from heated vibrios.
Further Studies on the Immunologic Relationships among Measles, Distemper, and Rinderpest Viruses
Authors: Örvell Claes , Norrby Erling
Published: December 2022

The reaction between measles and canine distemper viruses prepared in Vero cells and acute, convalescent and hyperimmune sera against these viruses and rinderpest virus were studied in neutralization (NT), neutralization-enhancement (NE), complement-fixation (CF), immunofluorescence, and immunodiffusion tests. In addition, measles virus hemolysis-inhibition (HLI) and hemagglutination-inhibition (HI) tests were carried out with the different sera.

The nucleocapsid components of the three viruses could not be distinguished by the immunologic techniques employed. Sera against rinderpest virus contained HLI, HI, and NT antibodies against measles virus. In some sera HLI antibodies were present in marked excess over HI antibodies. NT antibodies against distemper virus were also found in these sera. Sera against distemper virus also displayed measles HLI antibody activity, but no measles virus-specific NT antibodies were detectable. Low titers of HI and NE antibodies were found in some sera. It is concluded that measles, distemper, and rinderpest virus share envelope antigens, which are more readily demonstrated in HLI than in HI tests with measles virus antigen.
The Effect of Lectins on Lysosomal Enzyme Release from Neutrophilic Leukocytes
Authors: Hawkins David
Published: December 2022

Exposure of neutrophilic leukocytes (PMN) to concanavalin A (Con A) before their reaction with immune complexes markedly inhibited the normal release of lysosomal constitutents. A similar but less striking inhibition of release was observed when zymosan-complement (ZC) was used as the immune reactant. Con A inhibition of release and binding of a Con A-ferritin conjugate to the PMN surface were specifically blocked by certain sugars. Three other lectins induced PMN agglutination but failed to inhibit immunologically triggered lysosomal enzyme release. Con A appears to interfere with immune reactant binding to specific receptors on the PMN surface.
Receptors for Antigen on Lymphoid Cells
Authors: Greeley Elizabeth , Berke Gideon , Scott David W.
Published: December 2022

A method was developed for examining the binding specificities of plaque-forming cells (PFC) which were immune to either goat red blood cell surface antigens or to the trinitrophenyl (TNP) haptenic group. Immune rat lymph node cells were plated on poly-l-lysine fixed red blood cell (RBC) monolayers which functioned dually as immunoadsorbents and as plaque indicators of both adherent and nonadherent PFC populations. Anti-TNP PFC demonstrated marked specific adherence to TNP-GRBC. The adherence was optimal following a 30-min adsorption phase and was essentially independent of the adsorption temperature in the 4° to 37°C range. Specific adherence of anti-GRBC PFC to GRBC monolayers at 4°C was not detectable, but did occur at adsorption temperatures of 25° or 37°C. Temperature differences in specific adsorption in the two systems were not due simply to differences in the density of antigenic determinants on the adsorbing monolayers. Specific adherence of anti-GRBC PFC at 4°C was able to occur in the presence of DEAE-dextran, indicating the importance of surface charge effects. This immunoadsorbent-indicator system provides a means of studying the recognition events occurring at the PFC surface.
Regulatory Role of T Cells in IgG Antibody Formation and Immune Memory to Type III Pneumococcal Polysaccharide
Authors: Braley-Mullen Helen
Published: December 2022

Mice immunized with Type III pneumococcal polysaccharide (S3) produce only IgM antibodies and there is no evidence for immune memory after a second challenge with the antigen. However, when mice undergoing an allogeneic effect are immunized with S3, IgG antibody is produced by some, but not all, strains of mice. Mice also produce IgG antibody specific for S3 when they are immunized twice with a thymus (T) dependent form of antigen, S3 coupled to sheep erythrocytes (S3-SRBC), and these mice develop S3-specific immune memory. Since the allogeneic effect and immunization with S3-SRBC would be expected to result in activation of alloantigen-specific or carrier-specific T cells, the results suggest that T cells play an important regulatory role in IgG antibody formation and immune memory for a presumably T-independent antigen. IgG antibody was also produced when mice were immunized with S3 and treated with anti-lymphocyte serum (ALS). Since ALS presumably removes suppressor T cells it is suggested that S3-specific “helper” T cells may exist but that suppressor cells normally prevent them from being expressed when mice are immunized with S3 alone.
Adjuvanticity of Synthetic Polynucleotides for Induction of Experimental Allergic Encephalomyelitis in Guinea Pigs
Authors: Paterson Philip Y. , Drobish Diane G.
Published: December 2022

Complexes of polyadenylic acid and polyuridylic acid and complexes of polyinosinic acid and polycytidylic acid in incomplete Freund's adjuvant provide the prerequisite adjuvanticity for induction of experimental allergic encephalomyelitis in guinea pigs sensitized to spinal cord homogenate. The experimental allergic encephalomyelitis-adjuvanticity of these polynucleotide complexes resides in the pyrimidine component of each complex, i.e., polyuridylic acid and polycytidylic acid. These findings are discussed in the light of contemporary views linking viruses and nucleic acids to autoimmune disease of animals and man.
The Effect of Measles Virus Infection on T and B Lymphocytes in the Mouse
Authors: McFarland Henry F.
Published: December 2022

The effect of measles virus infection on T and B lymphocytes in the mouse has been studied with a system of cell cooperation for production of an anti-hapten response. The effect of virus was studied by infecting helper cell or antibody-forming precursor cell donors with measles virus at various times before cell transfer into recipient mice and measuring the anti-hapten response after challenge with carrier-hapten conjugates. These studies indicate that measles virus produces a significant reduction in the anti-hapten response. This reduction in antibody formation is caused predominately by measles virus suppression of T helper cell function and possibly by a lesser effect on B cell function. The suppression of T cells is transient and corresponds to the time during which measles antigen can be demonstrated in the mouse spleens. Although infectious virus cannot be recovered from the lymphocytes, the suppression of T cells could only be demonstrated after inoculation with a virus strain capable of replicating in the mouse. These findings suggest that the virus may be present in a non-infectious or defective form. The effect of measles virus in T cells does not appear to be due to a depletion of T cells. The possible mechanisms for the suppressive effect are discussed.
The Molecular Dimensions of Mitogenic Factor from Guinea Pig Lymph Node Cells
Authors: Gately Celia L. , Gately Maurice K. , Mayer Manfred M.
Published: December 2022

The molecular dimensions of a mitogenic factor from guinea pig lymph node cells were investigated by means of gel filtration, sucrose density gradient ultracentrifugation, and CsCl isopycnic ultracentrifugation. The mitogenic factor was produced by stimulation of immune lymph node cells from inbred guinea pigs, strain 2 or 13, with the specific antigen, ovalbumin; and mitogenic activity was assayed by measuring incorporation of tritiated thymidine by target lymph node cells from syngeneic animals not immune to ovalbumin. The D20,w of the mitogenic factor was estimated to be (9.9 ± 0.2) × 10-7 cm2/sec by gel filtration on Sephadex G-100, and the apparent S20,w was estimated to be 2.4 ± 0.2 S by sucrose density gradient ultracentrifugation. The buoyant density was determined to be 1.324 ± 0.006 g/ml by CsCl isopycnic ultracentrifugation, and from this value the partial specific volume of mitogenic factor was estimated to be 0.71. From these data the molecular weight of the mitogenic factor was calculated to be 20,000, and the frictional ratio was calculated to be 1.2.
Complement Activation by Interaction of Polyanions and Polycations
Authors: Rent Rosemarie , Ertel Nancy , Eisenstein Reuben , Gewurz Henry
Published: December 2022

Interactions of heparin and protamine in fresh human serum, in amounts far below those required for complement depletion by either agent alone, were found to induce virtually complete depletion of total hemolytic complement activity. This depletion was dependent on time, temperature, pH, divalent cations, and serum concentration. The predominant complement component hemolytic activity depleted was C1; under appropriate reaction conditions C4 and C2 were depleted as well. Equivalent amounts of heparin along induced lesser but substantial depletion of C1, potentiation of C4 and C2, and minimal depletion of C3-9, whereas equivalent amounts of protamine had no effect upon complement component activities. We conclude that interaction of heparin with protamine, like interaction of antibody with antigen, markedly enhances its ability to interact with the first component of complement and activate the classical complement pathway. It is suggested that complement activation by interactions between certain polyanions and polycations, like interactions between antigens and antibodies, may have a role in the initiation of inflammatory reactions.
Carrier-Determined Tolerance in Vitro
Authors: Galanaud Pierre , Aldo-Benson Marlene , Borel Yves
Published: December 2022

A primary immune response to normal BDF1 spleen cells was obtained in vitro to the T-independent antigen TNP-T4 coliphage. This anti-TNP response was suppressed by exposing the spleen cells for 6 hr to TNP bound either to isologous or heterologous γ-globulin. The suppression was hapten specific. In contrast, TNP-albumin conjugates did not induce tolerance in vitro.
Interaction of IgE with Rat Basophilic Leukemia Cells
Authors: Carson Dennis A. , Kulczycki Anthony , Metzger Henry
Published: December 2022

Rat basophilic leukemia cells which bind rat IgE-release particulate material as they lose viability. This material appears to contain the surface receptors for IgE in their native form. Up to 80% of the receptor activity has been recovered in a fraction which contains only 4 to 5% of the total cellular protein.
Use of Radioimmunoassays to Determine the Concentration of Streptococcal Group-Specific Antibodies in Rabbit Antisera
Authors: Bernstein David , Klapper David G. , Krause Richard M.
Published: December 2022

A radioimmunoassay was developed for the quantitation of antibodies to the Group A, A-variant, and C carbohydrates in rabbit streptococcal antisera. The assay employs radiolabeled purified Group carbohydrate which has been tyrosylated to allow for incorporation of 125I. This assay has an advantage over the quantitative precipitin test because it measures non-precipitating antibody in addition to precipitating antibody. Furthermore, 7S anti-IgG in rabbit antisera give a falsely elevated value for the antibody concentration in the quantitative precipitin test. This did not occur with the radioimmunoassay. The assay described is reproducible, sensitive, and uses little antisera.
Identification and Some Properties of Rat Secretory Component
Authors: Vaerman Jean-Pierre , Heremans Joseph F. , Bazin Hervé , Beckers Andrée
Published: December 2022

An antiserum, prepared against partially reduced and alkylated rat milk SIgA, was shown to contain antibodies reacting with a rat milk protein of α2 mobility, which possessed antigenic determinants common with SIgA, eluted from Sephadex G-200 at the same position as mammalian secretory components, and could be released from rat SIgA by mild reduction and alkylation. This protein, called rat secretory component, was also detected in saliva, tears and urine. Rat free secretory component (FSC) possessed antigenic determinants which were inaccessible in rat SIgA, as found in other species. In vitro, ra FSC combined with rat serum polymeric IgA or IgM, but not with monomeric IgA or IgG. These data emphasize the many similarities between the human and rat SIgA systems
Niridazole
Authors: Mahmoud Adel A. F. , Mandel Mark A. , Warren Kenneth S. , Webster Leslie T.
Published: December 2022

Niridazole, a human anthelminthic drug, is shown to be a potent long-acting suppressor of delayed hypersensitivity. Much lower doses than used in the therapy of helminth infections suppressed granuloma formation around Schistosoma mansoni eggs for periods up to 32 days and inhibited delayed footpad swelling in mice previously sensitized with eggs. Marked prolongation of skin allograft survival in mice across the H-2 area was produced. The drug is administered orally, has been used clinically on a massive scale, and appears to be relatively free of the cytotoxic effects of most immunosuppressive agents.
Adoptive Transfer of Experimental Allergic Encephalomyelitis (EAE): Prevention of Successful Transfer by Treatment of Donors with Myelin Basic Protein
Authors: Driscoll Bernard F. , Kies Marian W. , Alvord Ellsworth C.
Published: December 2022

After onset of experimental allergic encephalomyelitis (EAE), guinea pigs can be effectively treated by injection with myelin basic protein (BP). In order to localize the site of action of BP, cells from sensitized donors treated with BP one, two, three, or four times after disease onset have been transferred to normal recipients. One injection of BP has no effect on ability of cells to transfer EAE. Two injections partially inhibit transfer. After the third and fourth injections the sensitized cells lose their capacity to transfer EAE. The therapeutic effect of BP previously demonstrated in actively sensitized guinea pigs must involve the specifically sensitized cells rather than the target organ.
Alloimmune Cytotoxic T Cells: Evidence that They Recognize Serologically Defined Antigens and Bear Clonally Restricted Receptors
Authors: Bevan Michael J.
Published: December 2022

Murine T cells immunized with allogeneic cells exert a cytotoxic effect on appropriate target cells, as measured by 51Cr release. Lysis of the 51Cr-labeled target can be inhibited by addition to the assay of unlabeled cells which bear target antigens. With variants of lymphoid tumors which express different amounts of serologically defined H-2 antigens as competitors in the assay, a positive correlation is shown to exist between the amount of serologically defined antigen and the degree of inhibition of T cell-mediated lysis. The immunizing small lymphocytes are shown to be very poor inhibitors of 51Cr release from target cells which are more susceptible to T cell-mediated lysis. This is interpreted as showing that small lymphocytes are bound very poorly by cytotoxic T cells. With H-2 cross-over strains which carry only the H-2K or H-2D antigens which a cytotoxic population is directed against, it is shown that, 1) approximately equal amounts of cytotoxicity are directed against the K and D antigens, and 2) separate cytotoxic T cells recognize the K and D antigens. Thus, by this criterion, cytotoxic T cells are monospecific and therefore do not acquire their antigen-specific receptors cytophilically.
Blast Transformation of Rabbit B-Derived Lymphocytes by a Mitogen Extracted from Nocardia
Authors: Bona C. , Chedid L. , Damais C. , Ciorbaru R. , Shek P. N. , Dubiski S. , Cinader B.
Published: December 2022

Nocardia water soluble mitogen (NWSM) is known to stimulate mouse and rabbit lymphoid cells and to act selectively on murine B-derived lymphocytes. In this paper, evidence is presented that NWSM is also a mitogen for rabbit bursal equivalent cells and does not bring about blast transformation of thymus-derived rabbit cells. Pretreatment of rabbit spleen lymphocytes with antibody directed against rabbit thymus lymphocyte antigen (anti-RTLA serum) and with complement did not affect the strong increase of thymidine incorporation which follows stimulation with NWSM. The mitogen-induced polyclonal activation of antibody-forming cells and resulted in the presence of 30% of cells with the ultrastructural characteristics of plasmocytes. These observations led to the conclusion that NWSM is a mitogen for a B-derived lymphocyte of the rabbit.
Stimulation of a T-Independent Primary Anti-Hapten Response in Vitro by TNP-Lipopolysaccharide (TNP-LPS)
Authors: Jacobs Diane M. , Morrison David C.
Published: December 2022

The trinitrophenyl hapten (TNP) has been covalently conjugated to bacterial lipopolysaccharides (LPS) to give TNP-LPS. The site of attachment has been suggested to be in the core polysaccharide and lipid A region of the molecule and approximately 2.4 hapten molecules are bound per monomer LPS molecule. The TNP-LPS has been demonstrated to be immunogenic in vitro at very low concentration. This antigen has further been shown to initiate a T-independent TNP-PFC response. The immunogenicity of TNP-LPS is abrogated by mild alkaline hydrolysis, suggesting a requirement for intact lipid A in the initiation of an immune response at the very low concentrations of antigen used.
Rosette Formation by Canine Peripheral Blood Lymphocytes
Authors: Bowles Charles A. , White Gordon S. , Lucas Diane
Published: December 2022

The surface membrane characteristics of canine peripheral blood lymphocytes were investigated using erythrocyte (E) and erythrocyte antibody complement (EAC) rosette assays and immunofluorescent staining techniques. Canine thymus-derived lymphocytes formed nonimmune E rosettes with human and guinea pig erythrocytes at approximately the same percentages (mean percentage 36.3 and 32.4, respectively). Rosettes did not form with erythrocytes from seven other animal species. Cell surface immunoglobulins were demonstrable in a small percentage (4%) of the E rosette-forming lymphocyte population. Fragments of human erythrocytes inhibited E rosette formation by intact human red cells, but did not result in a significant decrease in rosette formation by intact guinea pig erythrocytes; likewise, guinea pig fragments had no inhibitory effect on rosette formation by human erythrocytes, demonstrating that separate receptors were required for the two red cell types. EAC rosette formation was not affected by addition of intact or fragmented human and guinea pig erythrocytes.

Canine bone marrow-derived lymphocytes were characterized by immunoglobulin on the cell surface and EAC rosette formation. Serial tests of lymphocytes from one dog revealed a wide variation in percentage of cells forming E and EAC rosettes. A close correlation was observed between the immunoglobulin-bearing cells (mean percentage 46.6) and those forming EAC rosettes (mean percentage 49.3).
The Primary Structure of a Human Lambda II Chain
Authors: Köhler Heinz , Rudofsky Susanna , Kluskens Larry
Published: December 2022

The human myeloma protein Boh (γ2, λ) was isolated and completely reduced and aminoethylated. The light chain was obtained by chromatography on Sephadex G-100 in 4 M guanidine HCl. The amino-terminal sequence on the blocked light chain could be determined by automatic sequence degradation after PCAase treatment. Twenty-one peptides were isolated from a tryptic digest and 12 peptides from a chymotryptic digest. The sequence determination on these peptides was performed by automatic sequencing methods. The light chain of Boh protein belongs to the λ II subgroup. Unique substitutions have been found at position 8 (Arg) and position 62 (Tyr). Furthermore, the Boh light chain has six cysteine residues, the additional (sixth) cysteine being adjacent to the invariable intrachain-S-S linking cysteine at position 91. Sequence comparison of λ II proteins reveals a high degree of homology emphasizing the biologic significance of the hypervariable region sequences.
Immune Specific Production of Interferon by Human T Cells in Combined Macrophage-Lymphocyte Cultures in Response to Herpes Simplex Antigen
Authors: Valle Martti J. , Bobrove Arthur M. , Strober Samuel , Merigan Thomas C.
Published: December 2022

Human peripheral blood lymphocytes, highly enriched for T cells, were obtained by passing gravity-sedimented leukocytes through nylon wool columns. The eluted cells were cultured with autologous macrophages and the mixture was studied for its capacity to produce interferon in vitro in response to stimulation with herpes simplex virus antigen. The interferon produced by the combined macrophage-lymphocyte cultures was shown to depend upon the presence of T cells; elimination of these cells by treatment with an anti-T cell serum plus complement greatly diminished the amount of interferon produced. The memory for the immune-specific release of interferon also appeared to be carried by the T lymphocytes rather than the glass-adherent macrophages. Furthermore, the results suggest that under our conditions of culture immune-specific interferon originates from T cells.
Rabbit Peyer's Patches, Appendix, and Popliteal Lymph Node B Lymphocytes: A Comparative Analysis of Their Membrane Immunoglobulin Components and Plasma Cell Precursor Potential
Authors: Craig Susan W. , Cebra John J.
Published: December 2022

We have characterized and compared the differentiative potential of B lymphocytes obtained from rabbit appendix (APP), Peyer's patches (PP), and popliteal lymph nodes (PLN) by immunofluorescence analysis of membrane immunoglobulin (Ig) components, by radioiodination of membrane components followed by isolation and identification of cell surface Ig, and by cell transfer studies. As assessed by cell transfer, lymphoid cells from PP and APP appear to be highly enriched sources of IgA plasma cell precursors. In addition, these populations of lymphocytes have a small but significant potential to give rise to IgM and IgG plasma cells. In contrast, as shown previously (1), lymphocytes from PLN have a comparatively small potential to generate IgA plasma cells. The ability of cells from PLN and PP to give rise to plasma cells is correlated with the presence of 35 to 38% of lymphocytes from both sources having membrane light chain determinants detectable by immunofluorescent staining. When the cells were stained directly for µ chain determinants, about 33% of PLN cells and 28% of PP cells were positive. These latter percentages were shown to reflect cells bearing both endogenously synthesized b4 light chain and µ heavy chain. By indirect immunofluorescence, α and γ chain determinants were also detected on PP and PLN cells; however, these Ig did not appear to be products of the cells which bore them as they were not regenerated after removal of Pronase. It was further demonstrated that purified rabbit copro-sIgA and colostral sIgA are cytophilic for PP and PLN cells. The results are discussed with respect to the origin of IgA plasma cells and the postulated bursal role of mammalian gut-associated lymphoid tissue.
Strain Differences in Ease of Tolerance Induction to Bovine γ-Globulin: Dependence on Macrophage Function
Authors: Lukić Miodrag L. , Cowing Carol , Leskowitz Sidney
Published: December 2022

DBA/2 mice are highly susceptible while BALB/c mice are resistant to the induction of tolerance with ultracentrifuged BGG. As measured by the clearance of a test dose of 125I-BGG, 2 mg ultracentrifuged BGG is sufficient to induce complete tolerance in DBA/2 mice, while BALB/c mice exhibit a clearance rate similar to immune controls.

BALB/c mice pretreated with carrageenan (a macrophage toxic agent) lose their resistance to tolerance induction with 2 mg ultracentrifuged BGG, while treatment with cobra venom factor does not alter their resistance to tolerance induction.

Stimulation of the reticuloendothelial system with BCG produces a reduction in the susceptibility to tolerance of DBA/2 mice. This effect is transitory, with the maximum effect 3 weeks after BCG infection.

Biologic filtration of BGG through BALB/c mice results in a preparation which is highly tolerogenic for recipient mice and this ability to filter the immunogenic portion of BGG is radioresistant.

The results provide evidence for the concept that the differential susceptibility to the induction of tolerance is related to a macrophage function in the inductive phase of immunity.
Dimeric and Monomeric Forms of HL-A Antigens Solubilized by Detergent
Authors: Cresswell Peter , Dawson Jeffrey R.
Published: December 2022

HL-A antigens solubilized by papain contain two non-covalently linked subunits, a 34,000 m.w. glycoprotein and an 11,000 m.w. protein (1–3), estimated by sodium dodecyl sulfate (SDS)2 gel electrophoresis. The glycoprotein exhibits differences in electrophoretic mobility depending on HL-A specificity (3) and probably bears the antigenic determinant (4). The small protein, when assessed by immunologic and electrophoretic techniques, is identical to β2-microglobulin (4–7), a protein found in serum and in elevated amounts in nephrotic urine, which is significantly homologous to the constant region domains of IgG (8, 9). Numerous studies with immunofluorescent techniques have recently shown that β2-microglobulin is also associated with HL-A antigens on the lymphocyte membrane (10–13), and partially purified HL-A antigens solubilized by detergents contain β2-microglobulin (7) together with a glycoprotein of molecular weight 43,000 to 44,000 (14). In this report, we present evidence that the detergent-solubilized HL-A-β2-microglobulin complex, both labeled with radioactive precursors and in large-scale unlabeled preparations, exists in part in a dimeric form.
Studies on the Inhibition of C5̄6̄-Induced Lysis (Reactive Lysis)
Authors: Baker Patricia J. , Lint Thomas F. , McLeod Bruce C. , Behrends Carole L. , Gewurz Henry
Published: December 2022

Multiple polyanions and polycations were tested for their ability to influence formation of from C, C7, and sheep erythrocytes. Six of 11 polyanions tested, including polyanethol sulfonate, heparin, and dextran sulfate, inhibited this reaction. By contrast, polycations (five of seven tested), including polybrene, protamine, and polyornithine, potentiated formation of EC. The inhibition was similar to that previously described for anionic serum factors termed C-INH, while the potentiation seemed to involve neutralization of serum C-INH. Thus, this step of the complement attack mechanism seems amenable to modulation by certain polyelectrolytes, and may thereby be susceptible to pharmacologic manipulation.