bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on the Effect of Specific Antisera on the Metabolism of Cellular Antigens
Authors: Yu Alice , Cohen Edward P.
Published: December 2022

The metabolism of thymus leukemia (TL) antigens of ASL-1 leukemia cells (TL 1,2,3) as measured by specific activity (cpm/mg protein) of isolated TL antigens was investigated during antigenic modulation. TL antigens prelabeled with 3H-fucose or 125I disappeared more rapidly from ASL-1 cells undergoing modulation than from control nonmodulating cells. Although TL antigens were released into the culture medium the total quantity in the medium of modulating cells was the same as that released from control cells not undergoing modulation. The synthesis of TL antigens in cells undergoing modulation was indistinguishable from control cells. No difference in the incorporation of 3H-fucose into TL antigens was noted in cells incubated with TL antiserum and cells incubated with normal mouse serum.

The alteration in the metabolism of membrane-associated antigens after reaction with specific antisera was unique for TL antigens. Antibodies directed against antigens other than TL, also present in the TL 1,3 antiserum used for modulation, failed to induce changes in the metabolism of these non-TL antigens. Furthermore, no significant changes in the degradation of H-2a antigens were noted during modulation of TL antigens.

The synthesis of TL antigens by normal thymus and spleen cells was also studied. TL- A/J spleen cells or TL- C57BL/6 thymocytes failed to incorporate 3H-fucose into membrane components which were precipitable by TL 1,3 antiserum, whereas TL + A/J thymocytes were found to synthesize actively TL antigens.
Studies on the Induction and Expression of T Cell-Mediated Immunity
Authors: Bonavida Benjamin
Published: December 2022

Purified thymus-derived (T) cytotoxic lymphocytes were inhibited from mediating cytolysis of 51Cr-labeled target cells by alloantiserum directed against the target cell. Sensitization of BALB/c (H-2d) thymocytes was obtained in one way-mixed lymphocyte cultures against mitomycin C-treated C57BL/6 (H-2b) spleen lymphocytes and in vivo after injection into lethally irradiated C57BL/6 mice (educated T cells). EL-4 (H-2b) tumor cells were used as target cells. Several criteria for optimizing the blocking effect were investigated. Blocking was obtained with antibody-coated target cells but was absent when the lymphocytes were pretreated with the antiserum before admixing with the target cells. The blocking activity was dependent on the concentration of antiserum. It was estimated on a target cell basis that 50- to 200-fold increase in antibody concentration was required to mediate blocking than was necessary to induce lysis by complement. The blocking effect was observed with different ratios of effector to target cells (E:T) and was found to be independent of the concentration of effector cells present. Eluates obtained from antibody-coated target cells behaved very much like the original antiserum by reacting again with fresh target cells but not with the effector cells. When the cytotoxic assay was run for shorter incubation periods (90 min vs 180 min), more blocking activity was detected, and significant inhibition of cell-mediated cytotoxicity (CMC) was obtained with high dilutions of antiserum which were not inhibitory in longer incubation periods. Maximum blocking was obtained when the antiserum was present at the beginning of the assay; at 30 min or after, the addition of antiserum had no blocking effect. These results demonstrated that damage of target cells occurs rapidly after effector-target cell interaction long before the radioactive chromium marker is released.
Genetic Control of the Immune Response: the Effect of Non-H-2 Linked Genes on Antibody Production
Authors: Dorf Martin E. , Dunham Edward K. , Johnson Judith P. , Benacerraf Baruj
Published: December 2022

In mice genetic control of the ability to form an immune response to the random terpolymer of glu60, ala30, tyr10 (GAT) is regulated by an Ir gene which maps between the K and Ss-Slp region of the H-2 complex. Several responder mouse strains were immunized with limiting doses of GAT. All responder strains produced GAT antibody after immunization with 1 µg, but not with 0.1 µg GAT. Although all responder strains responded to the same limiting doses of antigen, the serum concentrations of anti-GAT antibodies varied considerably among strains. Thus A, A.By and (B10 × A)F1 mice demonstrated at least 10-fold higher GAT-binding capacities than B10 or B10.A mice. This dominant non-H-2 linked quantitative trait reflects differences in the concentration of specific antibody since anti-GAT antibodies from A and B10 mice were shown to have similar binding affinities, and inhibition studies with the copolymers GT and GA indicated that both the A and B10 antisera also had similar specificity. Measurements of antibody concentration have shown that sera from A strain mice contained 15-fold more specific antibody than sera from B10 animals. In addition, plaque assays demonstrated more antibody-forming cells in A strain animals.

Genetic analysis demonstrated that these strain differences were under multigenic control. One of the genetic components was shown to be linked closely with the heavy chain allotype loci.
Cellular Events in Tolerance
Authors: Scott David W.
Published: December 2022

Rats rendered tolerant to ultracentrifuged sheep γ-globulin (ShIg) make a poor anti-trinitrophenyl- (TNP) specific antibody response upon challenge with TNP-ShIg in adjuvant. This system has been utilized to investigate the specificity of carrier recognition in tolerance. Induction of tolerance to heterologous globulins (goat, bovine, horse, pig, and rabbit) produced varying degrees of cross-tolerance upon challenge with TNP-ShIg. The cross-reactivity was similar to that seen when ShIg-sensitized cells (immune) were stimulated in vitro to incorporate tritiated thymidine by heterologous globulins. However, rat antibody to ShIg showed more restricted cross-reactivity than either carrier cross-tolerance in vivo or cross-stimulation in vitro. This suggests that carrier recognition in both tolerance and cell-mediated immunity is similar and provides further evidence for broader reactivity in cell-mediated (T cell) than in humoral (B cell) immunity.
Antigenic Studies of J Chain
Authors: Meinke Geraldine C. , Spiegelberg Hans L.
Published: December 2022

An antiserum to human J chain was prepared by injection of purified J chain in complete Freund's adjuvant into the popliteal lymph nodes of rabbits. The antiserum was used to screen urine and serum samples of patients with multiple myeloma and macroglobulinemia for the presence of J chain, and to determine the extent of cross-reaction between human and other mammalian J chains. Free J chain was detected by gel diffusion immunoelectrophoretic analysis in the urines but not the sera of two of 100 patients tested. The urinary J chain cross-reacted extensively with isolated J chain and had a similar electrophoretic mobility. The J chains of various species cross-reacted extensively with human J chain suggesting a great deal of structural homology among these chains. The most extensive cross-reaction was observed between man and monkey, whereas dog, mouse, and guinea pig J chains were antigenically deficient when compared to human J chain.
Blastogenesis of Autologous, Allogeneic, and Syngeneic (Identical Twins) Lymphocytes in Response to Lymphokines Generated in Dinitrochlorobenzene-Sensitive Human Leukocyte Cultures
Authors: Levis William R. , Whalen John J. , Miller A. Edgar
Published: December 2022

Dinitrochlorobenzene (DNCB) coupled to peripheral blood erythrocytes or leukocytes forms a particulate complex (DNCB-antigen) capable of inducing lymphocyte transformation when added to peripheral blood leukocyte cultures prepared from human subjects specifically sensitized to DNCB. Biologically active soluble factor(s) are released into the cell-free supernatant of sensitive leukocyte cultures when stimulated by DNCB-antigen, but are not released in unstimulated control cultures. The active soluble supernatant factor(s) induced blastogenesis and DNA synthesis when added to secondary target leukocyte cultures prepared from subjects not sensitized to DNCB. Under the conditions employed in these experiments, maximal DNA synthesis was induced in secondary target leukocyte cultures when supernatants of primary sensitive cultures were collected 24 to 72 hr after stimulation and decreased rapidly thereafter. Increasing amounts of the active soluble factor(s) induced almost proportional increases in DNA synthesis in target leukocyte cultures, illustrating the quantitative nature of the blastogenic assay. Allogeneic and syngeneic leukocyte cultures from subjects not sensitized to DNCB showed levels of DNA synthesis comparable to autologous sensitive leukocyte cultures in response to the soluble factor(s). Thus, the soluble factor(s) which are specifically released after the induction of allergic contact dermatitis to DNCB appear to be lymphokine(s) which express blastogenic activity independent of the genetic origin or antigenic sensitivity of the target leukocyte population.
Lymphokine Activity in Vivo in Relation to Circulating Monocyte Levels and Delayed Skin Reactivity
Authors: Yoshida Takeshi , Cohen Stanley
Published: December 2022

The i.v. injection of migration inhibition factor- (MIF)containing supernatants from cultures of sensitized lymphocytes incubated with specific antigen results in a marked reduction in the number of monocytes in the peripheral blood of recipient guinea pigs. This treatment results also in suppression of delayed-type skin reactions to unrelated antigens in such recipients.

When actively immunized guinea pigs were studied, it was found that detectable macrophage migration inhibitory activity appeared in their sera shortly after i.v. challenge with antigen. This activity was maximal at 6 to 12 hr, which was somewhat later than the reduction in peripheral blood monocytes induced by i.v. injection of either specific antigen or exogenous MIF-containing fluids.

These results provide proof of the ability of lymphokines to participate in in vivo reactions. Neither the presence of MIF in the antigen-activated supernatants used for suppression of circulating monocytes and skin reactivity nor the appearance of MIF activity in serum of actively immunized and antigen-challenged animals implies that MIF itself is the responsible agent of the observed biologic activities although it is clearly a strong candidate for such a role. Regardless of which lymphokine(s) proves to be the mediator of the suppressive effects described here, these observations suggest the possiblity of modifying the expression of immunologic reactivity by such substances in appropriate clinical states.
The Enhancement of MIF Activity by Inhibition of Macrophage Associated Esterases
Authors: Remold Heinz G.
Published: December 2022

Macrophages pretreated with diisopropyl fluorophosphate (DFP) or incubated with soybean trypsin inhibitor demonstrate enhanced inhibition of migration caused by migration inhibition factor (MIF). This enhancing effect of DFP is time, dose, and temperature dependent. Incubation of MIF in serum treated with DFP had no effect. In addition, hydrolysis products of DFP were inactive. These findings strongly suggest that a macrophage-associated serine esterase, which is present in an active form before the addition of MIF and which counteracts MIF, is involved in the MIF-macrophage interaction. Its significance as a control mechanism in the macrophage-mediator response is discussed.
Priming in Recipient Mice Unable to Generate a Primary Antibody Response: Kinetics of Priming for an IgM Response
Authors: Groves David L. , Niblack Gary D. , Christian Elizabeth S.
Published: December 2022

The enhanced production of direct plaque-forming cells (DPFC) to a second dose of sheep erythrocytes (Srbc) has been demonstrated in irradiated mice restored with a limiting number of unprimed spleen cells. The limiting number of spleen cells was such that only a proportion of the recipients could respond to a primary dose of Srbc given at the time of spleen cell transfer. The most important finding was that 100% of such mice could respond to a second dose of Srbc given 1 week after cell transfer and the magnitude of the response was 18- to 36-fold greater than that of similar mice receiving only the first dose of antigen. The development of enhanced DPFC responsiveness was shown to require the initial dose of antigen and both its induction and elicitation were antigen-specific. Since enhanced responsiveness occurred in a population of mice not all of which could make a primary DPFC response, these results have been interpreted to indicate that priming occurs in a cell population not directly involved in initiating the primary antibody response.
Participation of Different Cell Populations in Antigen- and Mitogen-Induced Lymphocyte Proliferation. Evidence That Blastogenic Factor-Sensitive Lymphocytes Are Inactivated by 5-Bromo-2′-Deoxyuridine
Authors: Lazda Velta A. , Baram Peter
Published: December 2022

The nature of the subpopulations of lymphocytes which respond by blast transformation to stimulation by specific antigens and phytohemagglutinin (PHA) was investigated. Leukocytes from rhesus monkeys sensitized to keyhole limpet hemocyanin (KLH) and purified protein derivative from Mycobacterium tuberculosis culture filtrates (PPD) were stimulated with KLH or PPD in culture. The replacement of medium and the readdition of the same antigen on day 4 did not initiate a second round of DNA synthesis. The addition of a second antigen, however, markedly increased the 3H-thymidine (3H-T) incorporation within 1 day. However, no additive stimulatory effect occurred after simultaneous stimulation with KLH and PPD, possibly indicating competition for a limited cell population.

The lymphocyte proliferative response to KLH and PPD stimulation could be inhibited as much as 95% by treatment from days 2 to 4 with 5 × 10-6 M 5-bromo-2′-deoxyuridine (BrdU) and exposure to visible light. Such treatment also markedly reduced the expected response of the cultures to restimulation with a second antigen, suggesting that the observed response to both antigens was dependent upon the same BrdU-sensitive cells. BrdU-light inactivated cultures lost most of their capacity to respond to blastogenic factor. These results were interpreted as indicating that: 1) a large proportion of the BrdU-sensitive cells are blastogenic factor-reactive and 2) distinct subpopulation of cells is not recruited by blastogenic factor produced by specific antigen-sensitive lymphocytes.

Peak incorporation of 3H-T in antigen-pretreated cultures, when stimulated with PHA, occurred later than in cultures stimulated by a second antigen, suggesting that PHA was stimulating a new population of cells. Moreover, an additive stimulatory effect occurred when PHA and KLH or PHA and PPD were added to cultures simultaneously. BrdU-light treatment of antigen-stimulated lymphocytes had little effect on subsequent stimulation by PHA. Antigen and PHA stimulation, therefore, may involve different cell populations.
Antigen Binding T and B Lymphocytes
Authors: Hämmerling Günter J. , McDevitt Hugh O.
Published: December 2022

The present investigation describes inhibition by anti-Ig and anti-H-2 sera of 125I-(T,G)-A--L-binding T and B cells from murine lymph nodes. It was found that T-ABC from unimmunized and immunized mice are partially blocked by anti-IgM sera and not by anti-IgG sera. Furthermore, specific anti-H-2 sera block only T-ABC, but not B-ABC. The possible role of cytophilic IgM in the inhibition of T-ABC by anti-Ig, and the meaning of inhibition by anti-H-2 are discussed.
Clonal Nature of the Immune Response to Phosphorylcholine
Authors: Claflin J. Latham , Lieberman Rose , Davie Joseph M.
Published: December 2022

The clonal nature of the immune response to phosphorylcholine (PC) was studied at the level of the antibody-producing cell (PFC) with the highly sensitive parameters of idiotypes and binding characteristics as specific markers. As a basis for comparison, the characteristics of the anti-PC IgM antibody produced in different inbred strains of mice were related in PFC inhibition assays with those exhibited by four different PC-binding myeloma proteins, HOPC 8, McPC 603, MOPC 167, and MOPC 511. Each of these myelomas possesses distinctly different idiotypic determinants and binding affinities for PC, glycerophosphorylcholine (GPC) and choline (C). The induced anti-PC antibody in each of the strains tested was solely of the IgM class and bound PC τ; GPC τ; C. In BALB/c and A, but not CBA, no affinity maturation occurred during the immune response which indicates a restriction in the affinity heterogeneity. As determined from the PFC inhibition profiles, BALB/c and A strain anti-PC antibody had the same binding characteristics as HOPC 8 and that of BALB/c possessed the idiotypic determinants of this myeloma protein. As shown in a previous paper, the receptors on PC-specific antigen-binding cells (ABC) possessed the identical characteristics of class, specificity, and idiotypes as shown by the secreted antibody. These results, therefore, provide evidence for a direct link between the antibody secreted by PFC and the receptor molecules on ABC.
In Vitro Studies of Histamine Release from Rabbit Leukocytes by Divalent Haptens
Authors: Magro Albert M. , Alexander Alice
Published: December 2022

The in vitro histamine release by washed leukocytes from rabbits immunized to produce reagin specific for the benzyl penicillolyl (BPO) and the 2,4 dinitrophenyl (DNP) haptenic groups was studied. The animals were immunized to produce reagin specific for one of the haptenic groups (DNP or BPO) and for both haptenic groups (DNP and BPO). The divalent and univalent haptens used to initiate histamine release from the leukocytes were bis-DNP-octamethylenediamine (di-DNP), DNP-n-octylamine (uniDNP), bis-BPO-octamethylenediamine (diBPO), BPO-n-octylamine (uniBPO) and N-DNP,N-BPO, octamethylenediamine (mixed BPO-DNP).

The data indicate that 1) reagin of both specificities is on the majority of the basophils of rabbits immunized to both DNP and BPO; 2) the reagin on the basophil surface is not grouped by specificity; 3) internal bridging between two Fab pieces of a single reaginic antibody molecule is probably not a basic mechanism by which histamine is released; and 4) the hapten must bridge the membrane bound reaginic antibody to another point away from the antibody molecule in order for the hapten specific release of histamine to be initiated.
Evaluation of C3 Receptors on Lymphoid Cells with Different Complement Sources
Authors: Dierich Manfred P. , Pellegrino Michele A. , Ferrone Soldano , Reisfeld Ralph A.
Published: December 2022

Receptor activities for cell-bound degradation products of the third complement component, C3b and C3d, were demonstrated on cells from nine cultured human lymphoid lines and spleen lymphocytes from eight strains of mice. These receptor activities were found with rosette formation and immune adherence assays by utilizing sensitized sheep erythrocytes coated with C3 derived from human, mouse, guinea pig, and rabbit serum. Some cultured human lymphoid cells displayed receptor activities for both C3b and C3d, others showed only the one for C3d, and two cell lines reacted negatively. All murine cells examined showed receptor activities for murine C3d and to a lesser extent for human C3d together with receptor activity for guinea pig C3b; however, human C3b-carrying cells could not be attached to murine lymphocytes. Sensitized sheep erythrocytes coated with C1-C5 of rabbit serum deficient in C6 were found to react much like erythrocytes coated with human C3b and thus provided a useful reagent for the detection of C3b receptor activities on human lymphoid cells. There was no correlation between the expression of C3b and C3d receptors and the H-2 or HL-A phenotype or the antigenic determinants of β2-microglobulin which suggests that these complement receptors have unique chemical structures representing additional and useful cell surface markers.
Saturation of the Reticuloendothelial System with Soluble Immune Complexes
Authors: Haakenstad Alan O. , Mannik Mart
Published: December 2022

The clearance kinetics of passively administered soluble human serum albumin (HSA)antiHSA complexes were studied in mice at varied doses. Complexes with a molecular composition exceeding Ag2Ab2 (τ 11S complexes) were preferentially cleared by the hepatic reticuloendothelial system. As the dose of administered complexes was increased to levels known to induce glomerulonephritis, however, saturation of the reticuloendothelial system occurred. Three lines of evidence supported this conclusion: 1) The proportion of injected complexes cleared with the fastest half-life became less as the dose was increased. 2) Saturation was demonstrated in the first-order reaction that describes the clearance of τ 11 S complexes (τ Ag2Ab2). 3) The specific hepatic uptake of complexes approached a maximum as the dose was increased. Due to saturation of the reticuloendothelial system, the τ 11 S complexes persisted in the circulation and thereby the likelihood of their non-hepatic tissue deposition was enhanced. The possible contribution of saturation of the reticuloendothelial system to the pathogenesis of glomerulonephritis in experimental serum sickness is discussed.
Studies on Antibody Induction in Vitro
Authors: Schaefer Annette E. , Fishman Marvin , Adler Frank L.
Published: December 2022

The elimination of cells with specific receptors for the solubilized T2 antigen (S-T2) was accomplished by passage of spleen cells from unimmunized rabbits through antigen-coated glass bead columns. This procedure resulted in a population of lymphoid cells incapable of antibody-production in response to S-T2 in vitro. Such a population, devoid of antigen-specific cells, was tested for its ability to produce antibody in response to two fractions of RNA extracted from peritoneal exudate cells exposed to the T2 phage and then fractionated on methylated BSA Kieselguhr columns. The unresponsiveness of the cells filtered through antigencoated glass beads to S-T2 extends to the MAK III fraction containing RNA-antigen complexes, but not the MAK II fraction containing informational RNA. Therefore, the response to the informational RNA is independent of cells possessing specific antigen recognition.
Leukemia Virus-Induced Immunosuppression
Authors: Mortensen Richard F. , Ceglowski Walter S. , Friedman Herman
Published: December 2022

Measurements of lymphocyte mediated cytolysis of allogeneic target cells (C57BL/6 EL4) by sensitized BALB/c spleen cells revealed a rapid depression of this in vitro correlate of cellular immunity after infection with Friend leukemia virus (FLV). Immunosuppression occurred with infection either before or after alloantigen sensitization. Cell-mediated cytotoxicity by sensitized lymphocytes from the leukemia resistant C57BL/6 strain for DBA/2 mastocytoma cells was unimpaired when FLV was administered after immunization. However, a significant, but transient, depression of the response was observed when C57BL/6 mice were infected 4 days before immunization, but not at 8 or more days before immunization. A kinetic analysis of the cytolysis indicated that there was a decreased number of cytolytic effectors present in the spleens of the FLV-infected BALB/c mice rather than a diminution of the lytic efficiency of the cytolytic cells. The cytolytic activity carried out by the remaining effectors still occurred via a single interaction between a target cell and a sensitized lymphocyte as in the noninfected mice. The lowered cytotoxicity by BALB/c lymphocytes was proportional to the infecting dose of FLV and could not be accounted for by a simple dilution of effector cells with neoplastic cells. The number of cytolytic lymphocytes decreased logarithmically with time after FLV infection. These results demonstrate that FLV is capable of interfering with a T cell (Θ-bearing) dependent event as well as the B cell function of antibody formation. The depression of cell-mediated immunity by infection with FLV after immunization clearly contrasts with the absence of depression of humoral antibody formation when FLV is administered after antigen.
Interaction of C-Reactive Protein Complexes with the Complement System
Authors: Kaplan Melvin H. , Volanakis John E.
Published: December 2022

The reaction of C-reactive protein (CRP)-positive sera and crystalline CRP with pneumococcal C-polysaccharide at equivalence induced consumption of human complement and C3 conversion in the absence of detectable antibody. Sera with negative or trace reaction for CRP most often gave no or minimal consumption of complement on addition of C-polysaccharide. Analysis of complement components consumed in the reaction of CRP-positive sera with C-polysaccharide, in the absence of detectable antibody, demonstrated consumption of 80% or more of total hemolytic complement, C1, C4, and C2, and 42 to 66% consumption of C3-9. Quantitative estimates of the minimal amount of CRP in acute phase sera which permitted fixation of complement closely approximated the minimal value obtained with crystalline CRP. All normal human sera tested permitted consumption of complement by CRP complexes, including sera from infants 5 to 7 months of age with physiologic hypogammaglobulinemia and sera previously absorbed with pneumococcal cell walls to remove any presumptive trace of antibody to C-polysaccharide. Both the CRP-C-polysaccharide precipitation and complement consumption reactions were 100% inhibited by phosphoryl choline but were not at all inhibited by N-acetyl galactosamine, the major determinant of antibody specificity. CRP reacted with phosphoryl choline of the choline phosphatides, lecithin and sphingomyelin, by complement fixation and by flocculation. Immunodiffusion analysis of the washed complexes obtained by reaction of CRP-positive sera with C-polysaccharide and with lecithin demonstrated strong lines of precipitation for CRP, C1q, and C3, whereas immunoglobulins were absent or detectable only in faint traces.

These combined data have permitted the conclusion that complement consumption was attributable to CRP complexes and that it proceeded via the classical pathway independent of the presence of specific antibody or immunoglobulin. The specificity of CRP for choline phosphatides and its capacity to activate the complement system are considered in relation to the possible functions of CRP in the inflammatory process.
Acquired Immunity in Opossum (Didelphis Virginiana) Embryos
Authors: Rowlands David T. , Blakeslee Dennis , Angala Epifania
Published: December 2022

Opossum pouch young were immunized with bacteriophage (f2, øx-174, and T4), haptens (2,4 dinitrophenyl (DNP), fluorescein conjugates (FTC), and 1-dimethylaminophthalene (DNS)), or proteins (ribonuclease (RNAase), sperm whole myoglobin, and lysozyme). The haptens were placed on three carriers (bovine serum albumin, hemocyanin, or bovine γ-globulin. The sera obtained from animals at sacrifice 2 weeks after immunization were studied by bacteriophage neutralization with either the intact organisms or T4 coupled to one of the haptens or to one of the proteins. When the levels of antibodies were compared with the age of the pouch young, a distinct hierarchy of responsiveness was observed. The order of appearance of responsiveness was: bacteriophage f2, DNP, bacteriophage øx-174, FTC, bacteriophage T4, and RNAase. This hierarchy could not be explained as a difference in sensitivity of the assay, since the assays were of similar sensitivity in each case.
Physical and Immunologic Properties of Pneumococcal Capsular Polysaccharide Produced during Human Infection
Authors: Coonrod J. Donald
Published: December 2022

A mean molecular weight of ≥ 100,000 was found for type-specific pneumococcal capsular antigens (PCA) present in sera, bullous fluid, and pleural or cerebrospinal fluids from 11 patients with pneumococcal infection and for type-specific purified pneumococcal polysaccharide vaccines (PPP). The molecular weight of serum PCA was not decreased by trypsinization or incubation of the PCA in serum at 37°C. PCA present in the urine of individuals with pneumococcal infection was markedly smaller than PCA present in the other body fluids. Urine PCA had only partial immunologic identity with PPP and reacted differently from PPP or PCA in other body fluids in immunoelectrophoresis. These differences could not be attributed to degradation of PCA by urine per se. The mean molecular weight of circulating PCA and its immunoreactivity in gel diffusion tests did not change with time. PCA present in the urine during periods of prolonged antigenemia was persistently different from the serum PCA in gel diffusion tests and had a low molecular weight. Urine PCA from different patients infected with the same pneumococcal type were immunologically identical in double diffusion tests.

Low molecular weight PCA in the urine of individuals with pneumococcal infection may be derived from eigher selective renal excretion of low molecular weight PCA produced by the infecting pneumococci or from in vivo degradation of large molecular weight PCA produced during infection.
The IgM Antibody Site
Authors: Oriol Rafael , Rousset Monique
Published: December 2022

Rats immunized with DNP-Salmonella and DNP-BGG were bled at different times ranging from 5 days to 8 months. The 19S and 7S antibody fractions were separated by gel filtration and their binding affinities were measured with tritiated ε-DNP-Lys. Five days after the first injection, both 7S and 19S fractions were heterogeneous and their average K0 was low. The binding affinity of the 7S fraction increased progressively with time. The 19S fraction showed a different pattern of evolution. Two restricted populations of sites evolved from the starting heterogeneous, but normally distributed family. The K0 of the high affinity population of 19S antibodies followed the same maturation process as the corresponding 7S antibodies, whereas the K0 of the low affinity population of 19S antibodies remained fairly constant throughout the whole experiment.
The Binding of α-1 Antitrypsin to Human IgA
Authors: Tomasi Thomas B. , Hauptman Stephen P.
Published: December 2022

Complex formation between IgA and several serum proteins including albumin (1), βlipoproteins (2), haptoglobins (3), anti-hemophilic globulin (4) and in one case, to α1 glycoprotein (3) has been previously noted. However, with the exception of albumin, the binding of these proteins to IgA myeloma proteins has not been regularly observed. It has been reported (3) that 53% of IgA myeloma proteins as well as 74% of monoclonal IgM proteins complex with albumin. Subsequently, a thorough study by Mannik (5) demonstrated that the binding of albumin by both IgA and IgM proteins involved covalent disulfide bonds. Presumably, the binding site is located in the heavy chain and could involve the free SH group thought to be present in the α-chain (6), although this has not been directly demonstrated.

During the course of a study attempting to localize the site of albumin binding to IgA proteins, an antiserum to albumin was employed which also contained antibodies to α1 antitrypsin (α1AT).2
Mechanism of Allograft Rejection in New Zealand Mice
Authors: Gelfand Michael C. , Parker Leroy M. , Steinberg Alfred D.
Published: December 2022

Aging New Zealand mice develop impaired cellular immune functions including delayed rejection of primary skin allografts, a model of cellular immunity. In the first paper in this series it was found that old NZB/W mice are deficient in a recirculating T cell subpopulation which is present in young NZB/W mice. This T cell synergizes with a second T cell subpopulation present in both young and old NZB/W mice to effect prompt allograft rejection. The present study investigates the mechanism by which recirculating T cells cooperating in skin allograft rejection are eliminated as New Zealand mice age. It was found that a host factor diverts lymphoid cells from the recirculating pool. This host factor was found to have specificity for an antigen common to thymocyte and brain suggesting that it was similar to antisera to the Thy 1 (θ) antigen; the naturally occurring antithymocyte antibody (NTA) present in New Zealand mice has the same type of specificity. It appears that NTA leads to a reduction in recirculating T cells as New Zealand mice age resulting in impaired cellular immunity.
The Relationship between PF/DIL and Activated Human Hageman Factor
Authors: Johnston Alan R. , Cochrane Charles G. , Revak Susan D.
Published: December 2022

Permeability factor of dilution (PF/dil) activity, prepared by dilution of human plasma in contact with glass, was removed by insolubilized antibody to human Hageman factor. Insolubilized normal globulin or antibodies to prekallikrein failed to remove PF/dil activity. Dilutions of Fletcher trait plasma (a prekallikrein deficiency) failed to generate PF/dil activity but addition of purified prekallikrein reconstituted the capacity to generate PF/dil activity. These data support the conclusion that PF/dil and activated Hageman factor are identical and that generation of PF/dil (HFa) requires fluid phase activation with kallikrein.
The Splenic Suppressor Cell
Authors: Folch Hugo , Waksman Byron H.
Published: December 2022

The present study confirms and extends the finding that DA rat spleen cells include a subpopulation of adherent cells which act at high doses of phytohemagglutinin (PHA) or concanavalin H (ConA) to suppress DNA synthesis by other cells. Suppressor activity is lacking in spleens of doubly thymus-deprived rats. Mitomycin C treatment does not reduce the suppressor activity of normal whole spleen. It is inferred that the suppressor cell is a T lymphocyte or depends for its action on the presence of T lymphocytes and that its action does not require cell proliferation. DNA synthesis is much more affected by suppression at high mitogen levels than RNA or protein synthesis. A parallel is drawn between the separate regulation of DNA synthesis and other macromolecular synthesis in lymphocytes and so-called “pleiotypic” effects in other types of cells.

Spleen cells of young rats give a relatively low response (3H-thymidine uptake) to optimal or supraoptimal doses of PHA, which is markedly increased by removal of adherent cells on glass wool and is not affected by addition of purified macrophages. In contrast spleen cells of older animals give a high response, which falls when adherent cells are removed but is restored to the original level by addition of macrophages. It is concluded that older animals' spleens differ from those of younger rats by a variable loss of suppressor cells and an increasing macrophage dependence of PHA-responsive cells. The former may account for the reported increase in autoantibody formation with age. Reactivity of cells of a given age was not affected by culturing them in young or old rat serum pools, respectively.

Stress or injection of hydrocortisone acetate result in loss of suppressor cell adhesiveness to glass wool and increased macrophage dependence of PHA-responsive cells at 5 days. At 2 to 3 weeks there is a return to normal levels of adhesiveness and macrophage dependence, accompanied by a marked increase of suppressor activity. The 5-day changes are thought to reflect direct effects of steroid on the properties of T lymphocyte membranes, and the later changes altered levels of thymus hormone or possibly a surge of “post-thymic cells” into the peripheral lymphoid organs.
Role of Cyclic Adenosine 3′,5′-Monophosphate in Lymphocyte Mitogenesis
Authors: DeRubertis Frederick R. , Zenser Terry V. , Adler William H. , Hudson Thomas
Published: December 2022

For assessment of the role of cyclic adenosine 3′,5′-monophosphate (cAMP) in lymphocyte mitogenesis, adenylate cyclase activity, cAMP content, 3H-thymidine (3H-THY) incorporation, and blast transformation were examined in lymphocytes incubated with mitogenic agents, and with prostaglandins, cholera toxin, or theophylline. When tested over a wide dose range with mouse spleen cells, the mitogens phytohemagglutinin-P (PHA-P), staphylococcal enterotoxin B (SEB), concanavalin-A (CON-A), and endotoxin did not stimulate adenylate cyclase activity or increase cellular cAMP content. Similarly, in human lymphocytes, neither SEB, CON-A, nor mitogenic doses of PHA-P altered cellular cAMP levels. A rapid increase in human lymphocyte cAMP was noted in response to high doses of PHA-P, but such doses resulted in little induction of 3H-THY incorporation compared to lower doses that were ineffective in elevating cAMP. Further, PGE1, cholera toxin, and theophylline, agents which all significantly increased lymphocyte cAMP content, uniformly suppressed both basal and mitogen-induced 3H-THY incorporation and blast transformation in a dose-related manner.

The inhibitory effects of PGE1, cholera toxin, and theophylline on mitogen induced 3H-THY incorporation were time-dependent. After a 36-hr exposure of mouse spleen cells to CON-A increases in cellular cAMP no longer inhibited subsequent DNA synthesis, perhaps owing to the fact that those metabolic events influenced by cAMP were already completed in the majority of mitogen-activated cells by 36 hr. Moreover, washing mouse spleen cells which had been concomitantly incubated with CON-A and with PGE1 or theophylline for periods up to 36 hr abolished the inhibitory actions of the latter agents on 3H-THY incorporation. It seems possible from these results that elevated cellular cAMP levels may only delay, by a reversible process, the flow of mitogen-activated lymphocytes through their growth cycle and that lymphocytes remain susceptible to such inhibition for only a finite period after stimulation with mitogens.
Cytotoxic Activity of Lymphocytes
Authors: Lucas Zoltan J. , Walker Sharyn M.
Published: December 2022

A simple, quantitatively reproducible assay of lymphocyte-mediated cytotoxicity which duplicates results achieved by microscopically counting residual target cells or by 51Cr release has been developed. The method is based on quantitation of the intracellular 86Rb incorporated by target cells in monolayer culture after exposure to sensitized lymphocytes. Factors of paramount importance in detecting cytotoxicity are degree of sensitization of the lymphocytes, antigenic density of the target cell, and use of optimal standardized geometric conditions for the assay. Elimination of plastic adherent cells is essential to demonstrate immunologic specificity of the attacking cells. Nonspecific destruction of syngeneic monolayers is avoided by this procedure.
Antigenic Composition, Not the Degree of Polymerization, Determines the Requirement for Thymus-Derived Cells in Immune Responses to Salmonella Flagellar Proteins
Authors: Langman Rodney E. , Armstrong W. Douglas , Diener Erwin
Published: December 2022

Two flagellar proteins, prepared from either SW1338 or SL167 strains of Salmonella, have a differential requirement for thymus-derived (T) cells in producing a primary IgM response in mice. By using two methods for T cell depletion, the SW1338 flagellar protein, in both monomeric and polymeric forms, induced an IgM response that was essentially the same as that found in normal mice. In contrast, the SL167-derived antigen failed to initiate the appearance of antibody-forming cells in T cell-deprived mice. These findings add a further dimension to the consideration of mechanisms which underlie the collaboration between T and B cells, namely the role of epitope composition. In view of a failure to demonstrate differences between monomeric and polymeric forms of the antigen, it is argued that the arrangement of epitopes is not an essential requirement for T cell-independent IgM responses in vivo.
Comparison of Cross-Reactivities between Albumins of Various Species at the Level of Antibody and Helper T Cells—Studies in Mice
Authors: Weinbaum Fredric I. , Butchko Gregory M. , Lerman Stephen , Thorbecke G. Jeanette , Nisonoff Alfred
Published: December 2022

A study was made in LAF1 mice of the cross-reactivity between bovine serum albumin (BSA) and albumins from a wide variety of species with respect to their interaction with anti-BSA and reactivity with BSA-sensitized helper T cells. Inhibition studies and direct binding of 125I-albumins to anti-BSA were used to determine humoral cross-reactivity. Challenge by various trinitrophenyl- (TNP)-albumins of BSA-immune helper cells and TNP-keyhole limpet hemocyanin immune spleen cells in an adoptive anti-TNP plaque-forming cell response was used as a measure of cross-reactivity at the T cell level.

In most cases, cross-reactivities at both the humoral and T cell levels correlated fairly well. Goat and sheep albumin were highly cross-reactive, guinea pig, dog, and rabbit albumin gave medium-range reactivity, and rat and chicken albumin were essentially negative. Two major exceptions were observed both in the direction of greater T cell reactivity (pig albumin) and in the direction of greater humoral reactivity (horse albumin).

The results do not permit any definite conclusion as to the nature of the T cell receptor. They suggest, however, that such receptors may have discriminatory capabilities comparable to those of immunoglobulin.
Functional Heterogeneity among the T-Derived Lymphocytes of the Mouse
Authors: Kappler John W. , Hunter Philippa C. , Jacobs Diane , Lord Edith
Published: December 2022

A number of T cell functions were followed in mice after thymectomy in adult life. It was found that T cells involved in these functions could be divided roughly into two subpopulations: one whose activity was relatively long-lived (longer than 20 weeks) and one, relatively short-lived (on the order of a few weeks). In the long-lived class were those T cells involved in the helper function in the primary IgM and IgG responses to SRBC in vivo and in vitro. Priming for the secondary demonstration of helper activity or delayed hypersensitivity specific for SRBC seemed to involve both classes, although the contribution from the short-lived class was greater. These findings are consistent with models for T cell heterogeneity in which a short-lived population of T cells contains precursors distinct from a second long-lived population containing memory cells. Both classes can apparently contribute to a pool of activated effector cells in a process driven by antigenic stimulation.
Marsupial Immunoglobulins: An Immunoglobulin Molecule Resembling Eutherian IgA in Serum and Secretions of Setonix Brachyurus (Quokka)
Authors: Bell Robin G. , Stephens Christopher J. , Turner Keven J.
Published: December 2022

An immunoglobulin class present in several external secretions and serum of the marsupial Setonix brachyurus (Quokka) is described. This immunoglobulin possesses antibody activity and shares light chain determinants with other serum immunoglobulin classes but has heavy chain antigenic determinants which are exclusive to this class. In milk the molecular weight exceeds 200,000 and in serum a bi-modal distribution suggests two separate molecular weight forms, one in excess of 200,000 and another of approximately 150,000. Local challenge of the mammary gland in lactating quokkas induced an antibody response in milk which was restricted to this class. The immunoglobulin was designated IgA by virtue of its biologic and physicochemical characteristics which were analogous to those of eutherian IgA.
Immunogenic Properties of Modified Antigen E
Authors: Ishizaka Kimishige , Kishimoto Tadamitsu , Delespesse Guy , King Te Piao
Published: December 2022

Denaturation of antigen E in 8 M urea or by reduction-carboxymethylation treatment resulted in loss of antigenic determinants in native molecules. The denatured antigen preparations, as well as the α and β polypeptide chains isolated from the antigen upon its dissociation failed to combine with rabbit or human IgG antibody against the native antigen, or to induce erythema wheal reactions in ragweed-sensitive individuals. The denatured antigens and the polypeptide chains, however, enhanced anti-DNP antibody response of DNP-ragweed (DNP-Rag)-primed rabbit lymph node cells to DNP keyhole limpet hemocyanin (KLH). When the primed lymph node cells were treated with DNP-KLH for 24 hr, and then cultured in the presence of one of these modified antigens, both IgG and IgE anti-DNP antibodies were formed in vitro. The activity of the modified antigens to stimulate carrier-specific helper cells was comparable to that of native antigen. All of the modified antigens induced DNA synthesis of peripheral lymphocytes of ragweed-sensitive patients but not the lymphocytes of normal individuals. The results indicated that denatured antigen E as well as α and β polypeptide chains possess determinants specific for T cell receptor.
The Lack of Cellular Cooperation in the Immune Response to E. Coli 0127
Authors: Poe Wanda J. , Michael J. Gabriel
Published: December 2022

Cellular cooperation in the immune response to Escherichia coli 0127 was investigated. Spleen cell cultures prepared from BDF1 and congenitally athymic “nude” mice responded in vitro to E. coli bacteria, and removal of adherent cells from the spleen cell populations had no effect on the response to the bacterial antigen.

Nude mice immunized with sheep erythrocytes (SRBC) responded poorly to the SRBC antigen, but the injection of 7.5 × 107 thymocytes with the SRBC antigen increased the response 10-fold. In contrast, nude mice immunized with E. coli responded well to the bacterial antigen, and the injection of thymocytes did not alter this response. These results support the view that the immune response to E. coli 0127 is independent of thymus-derived cells, and demonstrate that the response ia also independent of adherent cells.
Lysis of Human Erythrocytes Due to Activation of the Alternate Complement Pathway by Nephritic Factor (C3NeF)
Authors: Arroyave Carlos M. , Vallota Enrique H. , Müller-Eberhard Hans J.
Published: December 2022

Nephritic factor (C3NeF) occurring in the serum of patients with hypocomplementemic chronic glomerulonephritis is known to activate complement via the alternate pathway. Addition of isolated C3NeF to normal serum resulted in assembly of the membrane attack mechanism of complement, the stable 22.5S C5b-9 complex.

In the presence of normal serum C3NeF caused lysis of erythrocytes from patients with paroxysmal nocturnal hemoglobinuria or of glutathione-treated normal human erythrocytes. The extent of cell lysis was proportional to the amount of C3NeF employed. Sera from patients with hypocomplementemic chronic nephritis containing C3NeF were shown to be hemolytically inactive because of lack of C3. Cell lysis induced by C3NeF ensued upon addition of isolated C3 to nephritic sera. The results show that C3NeF is capable of initiating cell injury through activation of the alternate pathway, and that the impairment of this pathway in C3NeF-containing sera is due to C3 deficiency. Lysis of glutathione-treated erythrocytes initiated by C3NeF may be used as an alternative assay for C3NeF.
Carbohydrate Inhibition Studies of the Naturally Occurring Human Antibody to Neuraminidase-Treated Human Lymphocytes
Authors: Rogentine G. Nicholas , Plocinik Barbara A.
Published: December 2022

Normal human sera, including autologous sera, contain complement-dependent lymphocytotoxic antibodies to Vibrio cholera neuraminidase-treated human peripheral blood lymphocytes and human lymphoid cells maintained in long-term tissue culture, but not to untreated cells. Carbohydrate hapten inhibition studies showed that these antibodies were inhibited by oligosaccharides containing β-d-galactosyl nonreducing end residues, indicating that the antigenic determinant is at least in part carbohydrate. There is evidence for both antigen and antibody heterogeneity.