bluuebunny/crossref_metadata_2025 huggingface dataset:
T Cell Requirement for Experimental Allergic Encephalomyelitis Induction in the Rat
Authors: Ortiz-Ortiz Librado , Nakamura Robert M. , Weigle William O.
Published: December 2022

The question of whether a cell-mediated or a humoral mechanism initiates EAE in rats sensitized with BP-CFA was investigated. The requirement of T cells for EAE induction was manifested when Tx, irradiated rats were reconstituted with normal lymphoid cells treated with ATS and then injected with BP-CFA. Neither EAE nor antibody was produced, indicating the T cell dependency of BP specific antibody production.

More precise information regarding the role of the T cell in the production of EAE was obtained by means of passive transfer of EAE with sensitized lymphocytes. Thus, transfer of lymphoid cells from rats previously sensitized to BP-CFA into Tx, irradiated rats elicited EAE and antibodies to BP. However, no EAE followed when the transferred cells were first depleted of T cells by treatment with ATS. Nevertheless, ATS pretreatment did not depress the levels of antibody to BP produced in the transfer recipients. The latter finding indicates that the cells from animals sensitized 9 days previously were already committed to the production of antibodies to BP. Therefore, a) T cells are absolutely necessary for induction of EAE and b) antibody detected by antigen-binding is not responsible for the pathogenesis of this disease.
Localized Deletion-Insertion Mutations: A Major Factor in the Evolution of Immunoglobulin Structural Variability
Authors: Potter Michael , Padlan Eduardo , Rudikoff Stuart
Published: December 2022

New and previously published amino acid sequences of mouse Vκ chains completed to Trp35 were examined. This sequence includes a framework loop from NH2 to Cys23 and an antigen contacting complementarity region L1 which lies between Cys23 and Trp35. There are many different sequence variations from NH2 to Cys23 among BALB/c mouse Vκ chains indicating multigenic control. Coordinately there is a remarkable structural heterogeneity of L1 size and sequence. Thus far chains with the same NH2-Cys23 sequence have similar sized L1, which suggests a close relationship between L1 size and characteristic NH2-Cys23 sequence. In this multigenic system selection for chains with size variations in L1 appears to have occurred. The mutational process that inserts or deletes triplets in this region is thought to be exploited in the derivation of new useful structural genes.
The Rat Basophilic Leukemia Cell Receptor for IgE
Authors: Kulczycki Anthony , McNearney Terry A. , Parker Charles W.
Published: December 2022

Rat basophilic leukemia cells were labeled either enzymically with 125I or biosynthetically by culture in the presence of 14C-glucosamine or 3H-amino-acids and then extracted with NP-40. IgE-anti-IgE precipitates insolubilized a radiolabeled macromolecule from these extracts largely or entirely absent in control IgG-anti-IgG precipitates. When specific precipitates were boiled in sodium dodecyl sulfate (SDS) and analyzed by polyacrylamide gel electrophoresis in the presence of SDS, most of the 14C or 125I radioactivity was in the area corresponding to an apparent m.w of 60,000 to 70,000 in 5.9% gels. In 10% and 12% gels, faster mobility was demonstrated indicating an atypical electrophoretic behavior often associated with glycoproteins and a presumptive m.w. of 50,000 or less. Since only IgE-containing precipitates localized lable in this region and since such precipitates from cells saturated with IgE prior to surface iodination failed to show this band, the labeled macromolecule appears to be the IgE receptor itself. Analysis of the acid hydrolysates of precipitated 14C radioactivity demonstrated that label was entirely in hexosamines and sialic acid. 125I and 14C labels in the receptor region were eliminated almost completely with pepsin and pronase and to a lesser extent with trypsin.
Biologic Properties of Nontoxic Derivatives of a Lipopolysaccharide from Escherichia Coli K235
Authors: McIntire Floyd C. , Hargie Martha P. , Schenck J. R. , Finley Richard A. , Sievert H. W. , Rietschel Ernst Th. , Rosenstreich David L.
Published: December 2022

Lipopolysaccharide (LPS)2 from Escherichia coli K235 was treated with o-phthalic anhydride to obtain a high degree of esterification of available hydroxyl groups, leaving a free carboxyl for each hydroxyl esterified (SPLPS). Although there was no demonstrable loss of fatty acids, this conversion of LPS to a polyanionic molecule altered dramatically the spectrum of biologic properties, most of which are normally attributed to the lipid A (LA) moiety. Mitogenicity for mouse B cells was decreased several hundred-fold; reaction with antibodies to LPS was abolished; pyrogenicity and toxicity were decreased by factors of 105 and 104; the ability to induce the Shwartzman reaction in rabbits was decreased 500-fold, and the ability to stimulate production of interferon in mice was decreased by more than 2 × 103. However, despite the loss of these properties, SPLPS retained the ability to act as an immunologic adjuvant. The nature of the anionic group is important, e.g., sodium succinyl-LPS (SuLPS) is 10-fold more pyrogenic and toxic than sodium phthalyl-LPS (SPLPS). Data on another LPS derivative, from which ester-linked fatty acid residues were removed before phthalylation, suggest that the ester-linked fatty acid groups in the lipid A moiety of SPLPS may not be necessary for its immunologic adjuvant effect.
In Vivo Activation of Properdin Factor B in Normotensive Bacteremic Individuals
Authors: Palestine Alan G. , Klemperer Martin R.
Published: December 2022

Activation of the alternative pathway of complement may be initiated by a variety of factors, including components of the bacterial cell wall (1, 2). It has been demonstrated in Gram-negative bacteremia leading to septic shock that C3 levels in serum are decreased (3). However, in bacteremia without shock, normal or increased C3 levels (3) and hemolytic complement activity (4) have been demonstrated.

It has been reported recently that properdin, C3, C5, C6, and C9 levels are lower in patients with bacteremia who develop shock than in normal individuals and normotensive bacteremic patients (5). Factor B3 levels in the above study were in general lower for patients in shock. However, factor B levels in all patients with bacteremia did not differ significantly from controls.

Circulating conversion products of factor B have been found in serum from two patients with arthritis after intestinal bypass surgery (6). Activation in these patients was postulated to be caused by an immunoglobulin cryoprecipitate.
Lymphocytic Choriomeningitis Virus-Induced Immunosuppression: A Virus-Induced Macrophage Defect
Authors: Jacobs Robert P. , Cole Gerald A.
Published: December 2022

Primary immunizing infections with LCM virus result in a transient depression of the in vitro proliferative responses of splenic lymphocytes to mitogens specifically reactive with T cells or B cells. This depression of lymphocyte function is the result of a virus-induced defect in an adherent, phagocytic cell population required for in vitro lymphocyte activation. Depressed responses persist for about 1 week after virus clearance and can be corrected by the addition of normal PEM or 2-ME to infected spleen cell cultures. Although the precise nature of this defect remains unclear, it is postulated that it is due to a productive infection of macrophages and their precursors that renders them dysfunctional. Secondary LCM virus infections do not result in depressed in vitro responses to mitogens, presumably because of rapid virus clearance and limited numbers of infected cells.

Primary infections of immunologically immature mice, mice rendered functionally athymic, or mice treated with nonspecific immunosuppressive agents result in LCM virus persistence. Shortly after infection, these animals show a similar depression of immunologic reactivity that returns to normal as the virus carrier state becomes established. Despite virus persistence, few PEM from established LCM virus carrier mice contain viral antigens and these cells function normally. Thus, LCM virus-induced immunosuppression appears to reflect a subtle cytopathic effect of LCM virus replication that is not mediated by the virus-specific cell-mediated immune mechanisms responsible for acute LCM virus disease.
Colchicine-Sensitive Structures and Lymphocyte Activation
Authors: Greene Warner C. , Parker Christine M. , Parker Charles W.
Published: December 2022

The possible modulatory role of microtubules or closely related colchicine-sensitive structures in the response of human lymphocytes to mitogenic lectins was investigated. Colchicine (0.1 to 10 µM) and vinblastine (0.1 to 10 µM) inhibited early [14C]-aminoisobutyric acid and late [3H]-thymidine uptake in phytohemagglutinin- and concanavalin A-stimulated human lymphocytes but failed to alter 45Ca uptake. Lumicolchicine, an inactive congener of colchicine, was ineffective in all three systems. Both microtubular agents accentuated and prolonged the early cyclic AMP response to lectin. Little or no alteration in cyclic AMP levels was seen with colchicine or vinblastine alone or in combination with PGE1 (10 µM) or epinephrine (1 µM) suggesting that the effect on cyclic AMP metabolism is largely selective for lectin stimulation. Neither microtubular agent altered 125I-concanavalin A binding. Since the inhibition of DNA synthesis was maximal only when the inhibitory agents were present throughout the culture period and early aminoisobutyric acid uptake is affected, it appears that these agents are acting on an early event, or events, in the activation sequence. Although the mechanism of the inhibition is not known, the effect of colchicine and vinblastine in prolonging the cyclic AMP response to lectin may be involved. Alternatively, alternations in microtubule assembly may exert effects on membrane architecture interfering with propagation of the stimulus from the membrane to the cell interior.
Inhibition of Normal Mouse Lymphocyte Mitogen Responses by Xenogeneic or Allogeneic Antibodies to the MuLV Glycoprotein gp71
Authors: Lee John C. , Ihle James N.
Published: December 2022

Xenogeneic and allogeneic antisera to the major envelope glycoprotein (gp71) of murine leukemia viruses (MuLV) inhibited the mitogenic response of normal mouse splenic lymphocytes to phytohemagglutinin (PHA) and lipopolysaccharide (LPS). This inhibition was specific for gp71 as demonstrated by the inability of xenogeneic antisera to other viral glycoproteins or structural proteins to inhibit and by the ability of purified antigens to block specifically the inhibitory effect. The ability of antisera to gp71 to inhibit LPS responses, however, is highly dependent on the strain and age of mouse spleen cells used and appears correlated with the expression of endogenous viruses. Moreover, the preferential inhibition of LPS responses suggests that this expression may be predominately B cell specific. The results suggest that the inhibitory effect is mediated via antibody binding to lymphocytes and that expression of viral envelope antigens on the cell surface which bind immunoglobulins can block or interfere with the binding or uptake of mitogens. A variety of natural mouse immune sera and “tumor” sera, having antibodies directed against gp71, can similarly inhibit mitogen responses; and this inhibition can be specifically blocked with MuLV or gp71.
The Formation of Stable E-Rosettes by Human T Lymphocytes Activated in Mixed Lymphocyte Reactions
Authors: Galili Uri , Schlesinger Michael
Published: December 2022

The aim of the present study was to determine whether activation of human T-lymphocytes affects their interaction with sheep red blood cells (SRBC). Less than 3% of the E-rosettes formed by freshly isolated peripheral blood lymphocytes (PBL) and SRBC are stable and do not disintegrate after incubation at 37°C. In contrast, about 30% of PBL kept in culture for 5 days in the presence of mitomycin C-treated allogeneic lymphocytes were found to form stable E-rosettes. Whereas no rosettes were formed by freshly isolated PBL incubated with human red blood cells at 24°C, 15% of the cells recovered from mixed lymphocyte reactions (MLR) formed such rosettes. When responder PBL were maintained in culture in the absence of allogeneic stimuli the proportion of cells forming stable E-rosettes depended on the serum present in the medium. Less than 5% of the responder cells kept in medium containing human serum or in serum-free medium formed stable E-rosettes, whereas 18% of the cells maintained in medium containing fetal calf serum formed stable E-rosettes.

The proportion of cells forming stable E-rosettes increased before any increase in DNA synthesis was detectable in MLR. Indeed, a high proportion of cells forming stable E-rosettes appeared in MLR taking place in serum-free medium, without any accompanying increase of DNA synthesis.

Depletion of cells forming EAC′-rosettes from responder PBL increased the proportion of cells forming stable E-rosettes in MLR. Exposure of the cells recovered in MLR to specific anti-T sera inhibited the formation of both stable and regular E-rosettes. Exposure of the cells recovered in MLR to anti-Ig serum had no effect on the formation of regular rosettes. Anti-Ig serum strongly inhibited the formation of stable E-rosettes by cells grown in medium containing human serum, but had no effect on the formation of stable E-rosettes by cells grown in either serum-free medium or in serum containing fetal calf serum.

It is concluded that activated human T lymphocytes are characterized by their capacity to form stable E-rosettes, resistant to incubation at 37°C, and by their capacity to acquire an immunoglobulin coat, possibly by binding immunoglobulin molecules present in their environment.
Thymic Function in NZB Mice
Authors: Bach Marie-Anne , Niaudet Patrick
Published: December 2022

Administration of a circulating thymic factor isolated from normal pig blood prevented the development of the exaggerated production of anti-polyvinyl pyrrolidone (PVP) antibody in young NZB mice. However, treatment was ineffective if initiated after the 4th week of life at a time when endogenous serum thymic factor (TF) normally disappears in these mice. These data suggest that circulating TF is necessary for the survival of short-lived suppressor T cells normally implicated in the regulation of the production of antibodies against PVP, a thymus-independent antigen. In older NZB mice, TF treatment increased paradoxically anti-PVP antibody production, which suggests that “amplifier” T cell activity could also be under TF influence.
Interaction of C-Reactive Protein with Lymphocytes and Monocytes: Complement-Dependent Adherence and Phagocytosis
Authors: Mortensen Richard F. , Osmand Alexander P. , Lint Thomas F. , Gewurz Henry
Published: December 2022

The serum constituent C-reactive protein (CRP), which activates the classical complement (C) pathway when rε-acting with its substrates, was examined for its ability to mediate reactions of opsonic adherence and phagocytosis. Erythrocytes coated with C-polysaccharide (CPS) and reacted with CRP (E. CPS-CRP) failed to adhere to B cells and displayed only minimal adherence to monocytes. However, upon the addition of absorbed C or purified C components these cells were found to possess the cleavage products C4b and C3b, which in turn resulted in attachment of these cells to both human B lymphocytes and peripheral blood monocytes. E. CPS-CRP treated with C in the absence of antibody were readily phagocytosized by glass-adherent human monocytes. The phagocytosis of E. CPS-CRP-C was not only mediated by CRP but also required the presence of CRP on the surface of the red cells. The extent of ingestion was proportional to the amount of CRP on the red cell intermediate and was reduced by blocking monocyte receptors with aggregated human γ-globulin (HGG) at concentrations which did not impair the uptake of other particles. The mediation by CRP of reactions of opsonic adherence and phagocytosis as outlined in these studies points to a significant role for CRP in reactions of host defense and inflammation.
Studies on Human Brain-Thymus Cross-Reactive Antigens
Authors: Maznina T. P. , Kushner S. G.
Published: December 2022

Antigens shared by human brain and thymocytes and by human and mouse tissues were studied with rabbit anti-human thymocyte antiserum (RAHT). It was found that cytotoxicity of RAHT serum against mouse thymus cells was not absorbed by mouse liver or bone marrow cells. Human brain and thymus cells completely absorbed the anti-thymocyte activity from this antiserum. It was suggested that human brain had antigenic determinants identical or very similar to those found on human thymocytes. Activity of RAHT antiserum against mouse thymus cells was completely removed by an absorption of mouse brain and thymocytes. These results demonstrated that there were shared antigenic determinants between human and mouse tissues.
C4 Synthesis in C4-Deficient Guinea Pig Radiation Chimeras: Restoration of the Classic Complement Pathway
Authors: Webster Robert O. , Rudofsky Ulrich H. , Pickering Richard J.
Published: December 2022

Bone marrow transplants from normal Albany strain guinea pigs established a functional classical pathway of complement (C) in C4-deficient (C4D) guinea pigs. Seventeen days after transplant the Albany → C4D chimeras had detectable C4 and total hemolytic C activities. Maximum C4 levels (2 to 8% of normal) were reached by day 73 and restored total C to 40% of normal. Classical pathway function persisted for about 150 days and, thereafter, declined to undetectable levels by day 385.

In contrast, Albany guinea pigs transplanted with C4D marrow maintained normal C4 levels throughout the experiment, suggesting that the C4-producing cells are radioresistant and long-lived. Unlike unmanipulated C4D animals, Albany → C4D chimeras were unable to produce antibodies to guinea pig C4 when immunized with normal guinea pig serum. These experiments suggest that bone marrow cell progeny produce C4 in vivo.
Synthetic Glycolipid Adjuvants
Authors: Behling Ulrich H. , Campbell Barbara , Chang Chung-Ming , Rumpf Christopher , Nowotny Alois
Published: December 2022

In the course of the organic synthesis of model compounds similar in some features to the lipid moiety of endotoxic lipopolysaccharide (LPS), N-acylated-D-glucosamine derivatives were prepared. One of these, N-palmitoyl-D-glucosamine, has been previously found to be mitogenic for athymic nude mouse B cells. This and other N-acylated homologs were tested for adjuvant activity in the immune response to human γ-globulin (HGG) and sheep red blood cells (SRBC) in mice. Comparable or superior enhancement of the immune response was obtained for these glycolipids when compared to LPS in assays measuring anti-SRBC or HGG hemagglutinin titers. In the determination of hemolytic plaque formation, considerable adjuvant effect was shown by the lauroyl derivative, and less but still significant enhancement was achieved by the N-palmitoyl-d-glucosamine. In the rosette formation assay, in addition to the above two glycolipids, N-oleyl-D-glucosamine showed good adjuvant effect. In the latter two assays, the LPS was a superior adjuvant as compared to the synthetic glycolipids.

The radiation protective effect of some of the better synthetic adjuvants was also investigated in mice. It was found that although LPS was more effective in this assay, the N-myristoyl-D-glucosamine and N-decanoyl-D-glucosamine compounds gave a definite protection, since up to 40% of the lethally irradiated (700 R) mice survived.
Leukocyte Migration Inhibitory Factor (LMIF) Production in Unidirectional Mixed Lymphocyte Cultures
Authors: Górski Andrzej J. , Dupont Bo , Hansen John A. , Good Robert A.
Published: December 2022

Puromycin treatment of lymphocytes was used to develop a one-way test for leukocyte migration inhibitory factor (LMIF) production in the mixed lymphocyte culture (MLC) reaction. Lymphocytes incubated with this protein synthesis inhibitor induced a vigorous mediator production by nontreated allogeneic cells, being themselves unable to respond to stimulator cells. When puromycin-treated cells were stimulated with the mitogens PHA, ConA, or PWM, overall protein and DNA synthesis were significantly decreased with concomitant abolishment of LMIF production. Viability of stimulator lymphocytes was found to be essential for generation of the mediator in MLC reaction.
Structure and Function of Immunoglobulin Domains
Authors: McNabb Trudy , Koh T. Y. , Dorrington K. J. , Painter R. H.
Published: December 2022

Fc receptors have been shown to be present in human placental tissue with properties distinct from those on macrophages and lymphocytes. A single class of receptor was observed with an intrinsic affinity of 4 × 106 M-1 for human IgG1. The order of affinity for IgG subclasses was IgG1 = IgG3 τ; IgG4 τ; IgG2. IgA and IgM were not bound. Fc from IgG1 bound with the same affinity as the whole molecule and to the same number of receptor sites, 2 × 1012/mg placental protein. IgG1 was no longer bound after mild reduction and alkylation whereas the binding of Fc was unaffected by this treatment. Neither Cγ2 nor Cγ3, the two domains which comprise the Fc region of IgG1, bound to the placental receptor. This implies that this Fc receptor is unlike those found on most cell surfaces and that placental binding is an exception to the theory that each domain has evolved to perform independent functions.
The Complement Profile in Babesiosis
Authors: Chapman William E. , Ward Peter A.
Published: December 2022

Rats have been infected with Babesia rodhaini, a protozoan agent that induces immune complex disease. Assays for complement have revealed depletion of C2, C3, C4, C5, and whole complement in the course of infection. No evidence of depletion in the alternative (properdin) pathway was found. These findings are consistent with the conclusion that the classical complement pathway is activated during the course of the protozoan infection.
Studies on the Specificity of in Vitro Induced Lymphocytotoxicity to SV40-Transformed Fibroblasts
Authors: Warnatz H. , Krapf F.
Published: December 2022

Cytotoxic effector lymphocytes were induced by in vitro immunization of lymph node and spleen cells from AKR-mice (H-2k) and from BALB/c-mice (H-2d) to syngeneic SV40-transformed fibroblasts. The T cell-dependent cytotoxicity was specific for target cells expressing the same H2-specificity as the immunizing cells. Nontransformed fibroblasts as stimulator cells did not induce efficient cytotoxicity to transformed or nontransformed target cells. Incubation with phytohemagglutinin during the sensitization period modified the specificity of the T cell-mediated lysis of syngeneic SV40-transformed fibroblasts: allogeneic as well as syngeneic target cells were destroyed by these effector cells. However, the polyclonal stimulant activates preferentially cytotoxicity to H2-matched target cells. The in vitro generation of cytotoxic effector cells was restricted to living SV40-transformed fibroblasts as immunizing cells; it was not possible to immunize lymphocytes in the presence of membrane proteins prepared from the SV40-transformed cells. The cytotoxicity of the in vitro immunized lymphocytes was inhibited by incubation with membrane protein preparations from syngeneic or allogeneic SV40-transformed fibroblasts.
Effects of C-Reactive Protein on Platelet Function
Authors: Fiedel Barry A. , Gewurz Henry
Published: December 2022

It was recently demonstrated that C-reactive protein (CRP)4 inhibits the response of human platelets to heat-aggregated human γ-globulin and thrombin and that this inhibition is characterized by a dose-dependent reduction in aggregation, activation of platelet factor 3 (PF3), and release of β-glucuronidase. In the present experiments, CRP was found also to inhibit the ability of washed human platelets to aggregate in response to poly-l-lysine (PLL); in these experiments, the magnitude of the inhibitory effect was dependent upon the m.w. of PLL used as the stimulating agent, and was more effective with low (15,000 daltons) than with high (400,000 daltons) m.w. polymers. CRP similarly inhibited ADP- and epinephrine-stimulated platelet aggregation in platelet-rich plasma (PRP), and this was characterized by relatively minimal suppression of the primary wave with marked suppression of the secondary ADP-mediated wave of aggregation. CRP also inhibited the platelet aggregation induced by collagen in PRP, although it had no effect upon the adherence of platelets to collagen. Finally, CRP inhibited the activation of PF3 and the release of serotonin during stimulation of platelets with ADP, and this inhibition was temporally related to the onset of the secondary wave of aggregation. These experiments extend the platelet reactivities inhibited by CRP, show that CRP expresses its inhibitory capacity in platelet-rich plasma as well as upon isolated platelets, raise the possibility that CRP exercises its effects by inhibiting or interfering with the release and/or utilization of endogenous platelet ADP, and support the concept that CRP plays an important role in the control of platelet responsiveness to a variety of stimuli during acute inflammatory reactions.
Age-Associated Decline in the in Vitro Development of Cytotoxic Lymphocytes in NZB Mice
Authors: Hirano Toshio , Nordin Albert A.
Published: December 2022

The development of cytotoxic lymphocytes (CL) in in vitro mixed lymphocyte culture (MLC) was investigated in young (14 weeks), middle (40 weeks), and aged (80 weeks) NZB mice. Cytotoxic activity against H-2b alloantigens was measured by using the 51Cr release assay. The antigen dose to elicit the optimum development of CL in vitro was the same for all ages of NZB mice, but the level of the development of CL was consistently low and could be delayed by up to 24 hr in aged mice. This decline in the development of CL was not related to the increase of the frequency of autoantibody against red blood cells nor to the decrease of the frequency of θ-positive cells in aged NZB mice. Aged (83 weeks) DBA/2 mice showed a similar decline in the development of CL. This decline of T cell function in aged NZB mice might be related to a physiologic aging process rather than to autoimmune disease.
Leukocyte-Derived Complement Inhibitor
Authors: Bernard Alain , Walter William , Teshima Hideki , Boumsell Laurence , Good Robert A. , Day Noorbibi K.
Published: December 2022

E, pretreated with leukocyte cultures supernatant (ES), binds C1̄ through C1q; ES and EIgM that bind the same amount of C1̄ as measured in a hemolytic assay have the same uptake of 125I-C1q; ESC1q and EIgMC1q, carrying the same number of molecules of C1q per cell, have the same uptake of C1r and C1s; soluble immune complexes prevent the binding of C1̄ and C1q to ES. The activity of C1̄ bound to ES is impaired; ESC1̄ can react with C4 but not with C2. The C4 turnover and the C1̄ INH turnover by ESC1̄ are reduced so that ES-bound C1̄ is protected from destruction by C1̄ INH. These modifications are fully reversed when C1̄ is transferred from ES to EA: C1̄ recovers its ability to react with C2, and to react fully with C4 and C1̄ INH. Thus the C1s activity can be modulated inside the C1̄ molecular complex upon binding of C1q to a lymphocyte product. In addition, the 125I-C1q uptake is proportional to the amount of IgM hemolysin used to sensitize E; it has, however, an exponential relationship to the amount of IgG or S used to sensitize E. The ratio of 125I-C1q uptake to whole C1̄ uptake measured in a hemolytic assay is lower than 2. This indicates that one molecule of IgM is sufficient to bind one molecule of C1q on E, that several molecules of IgG or S are required to bind one molecule of C1q, and that one molecule of C1q is sufficient to create a lytic site on E.
Cross-Reactions between 2,4-Dinitrophenyl and Menadione (Vitamin K3) and the General Problem of Antibody Specificity
Authors: Johnston Marilyn F. M. , Eisen Herman N.
Published: December 2022

Early and late antisera to 2,4-dinitrophenyl (DNP) and to menadione (K3)4 were compared for reactivity with diverse DNP and K3 ligands. Late antisera were generally more reactive (higher affinity) than early antibodies. Nevertheless, the early antibodies had higher affinity for homologous ligands than late antibodies had for heterologous ligands. In all but one of eight antibody subsets, homologous ligands were bound with higher affinity than heterologous ligands; the exception, a small (ca. 5%) and possibly heteroclitic subset from late anti-DNP sera, had higher affinity for K3-butyrate than for DNP-lysine. Early antisera appeared to be more specific than late antisera by one criterion (the precipitin reaction), but less specific by a more fundamental criterion (K0/Kx, the ratio of intrinsic affinities for homologous and cross-reacting ligands). By several methods (precipitation, binding to heterologous immunoadsorbants) 25 to 75% of the antibodies in both anti-DNP and anti-K3 sera were cross-reactive; even more cross-reactivity was evident by equilibrium dialysis, where it appeared that virtually all anti-DNP antibodies can bind K3 and that virtually all anti-K3 can bind DNP. This extreme level of cross-reactivity is in accord with molecular models, which, as shown previously, demonstrated some structural similarity between DNP and K3. Because their cross-reaction is not as “strange” as was once thought, it seems unwarranted to regard this cross-reaction as support for the view that antibodies in general are multispecific.
Demonstration of Human Nephritogenic Tubular Antigen in the Serum and Organs by Radioimmunoassay
Authors: Miyakawa Yuzo , Kitamura Kiyoshi , Shibata Seiichi , Naruse Takuji
Published: December 2022

Nephritogenic tubular epithelial antigen (Tub-Ag), which had been found in immune complexes deposited along the glomerular capillary walls of some patients with idiopathic membranous glomerulonephritis, was solubilized from renal tubuli of humans by Pronase digestion. Soluble Tub-Ag was then purified by gel filtration and ion exchange chromatography on DEAE-Sephadex. Three DEAE fractions, designated as 0.07 M, 0.13 M, and 0.23 M fractions, raised antibodies in rabbits, and fluoresceinated antibodies against any of these fractions reacted exclusively with luminal layer of proximal tubular epithelia. Among three fluoresceinated antibodies, however, only the one directed to 0.07 M fraction bound with immune complexes which were deposited along the glomerular capillary walls of the patients with Tub-Ag-mediated membranous glomerulonephritis. Purified Tub-Ag (DEAE 0.07 M fraction) was physicochemically homogenous with a S20 value of 8.2. Utilizing 125I-labeled Tub-Ag, a sensitive and quantitative radioimmunoassay of Tub-Ag was developed. Tub-Ag was demonstrated to occur naturally in serum and urine, as well as in all the organs tested including kidney, intestine, liver, spleen, stomach, heart, and lung. Tub-Ag was detected even in the sera of anephric patients on maintenance hemodialysis (60.8 ± 7.8 radioimmunoassay units/ml), although at slightly lower levels than in those of normal individuals (69.9 ± 10.6 units/ml). The size of serum Tub-Ag was identical to that of molecules bearing Tub-Ag activity solubilized by Pronase from all the organs tested, indicating that circulating Tub-Ag was maintained mainly by organs other than kidneys.
Differences in the Age-Dependent Release of a Low Molecular Weight Suppressor (LMWS) and Stimulators by Normal and NZB/W Lymphoid Organs
Authors: Ranney David F. , Steinberg Alfred D.
Published: December 2022

The age-dependent release of soluble suppressors and stimulators of DNA synthesis by cultured thymocytes and spleen cells from C57BL/6 and BALB/c mice was compared with their release by NZB/W lymphoid organs. Spleen cells from the normal strains released high levels of suppressor early in life and gradually decreasing quantities with age. NZB/W spleen cells exhibited an early deficiency followed by a later excess in the production of suppressor. These differences were quantitated by dose-response studies. Thymocytes from the normal strains released stimulatory factors throughout life. In contrast, NZB/W thymocytes stopped releasing stimulatory activity and began to produce suppressor after 2½ to 4 months of life. This abnormal elaboration of suppressor by thymocytes occurred 2 months before its reappearance in the autologous spleen cell supernatant. Both the early and late-appearing NZB/W suppressors were of low molecular weight (<1000). This activity was designed as low molecular weight suppressor (LMWS). Its aberrant production by the lymphoid organs of NZB/W mice correlated well with their reported functional immunologic abnormalities. The following items were discussed: the production of LMWS by adherent spleen cells, its relationship to previously described regulators, its partial purification and initial chemical characterization, and exclusion of the naturally occurring inhibitors of lymphocyte activation, cortisol, corticosterone, cold thymidine, and cyclic AMP as the active molecule.
Macrophage-Lymphocyte Interaction
Authors: Greineder Dirk K. , Shevach Ethan M. , Rosenthal Alan S.
Published: December 2022

Inhibition by anti-Ia sera of guinea pig T lymphocyte proliferation induced by allogeneic macrophages (MLR) and NaIO4 or neuraminidase-galactose oxidase-treated macrophages has been investigated in order to identify the target cell upon which the antisera act. Anti-2 and anti-13 alloantisera were found to inhibit both MLR and aldehyde-induced T cell reactivity when directed against the specificity of the stimulatory macrophage. Little or no inhibition was observed when these antisera were directed against the T lymphocyte specificity when cultures were harvested at the time of peak proliferation. In addition, anti-2 serum was found to inhibit macrophage-lymphocyte rosette formation at 20 hr between neuraminidase-galactose oxidasetreated strain 2 macrophages and strain 13 lymphocytes. These findings demonstrate that inhibition of T cell proliferation can be produced by anti-Ia sera directed against the macrophage and raise the possibility that Ir gene products may function in part at the level of the macrophage.
Mechanism of the Growth-Promoting Effect of Serum Albumin on Concanavalin A-Activated Lymphocytes: Protective Effect of the Plasma Proteins
Authors: Polet Herman , Spieker-Polet Helga
Published: December 2022

The concentration of concanavalin A (Con A) required for optimal growth of lymphocytes is 10 to 20 times greater in the presence of plasma than in the presence of serum albumin (BSA). This shift in Con A requirement is mainly caused by Cohn fractions III and IV, which bind and probably remove free Con A. Lymphocytes, once they are activated by Con A, lose all proliferative activity unless protected by certain proteins. Cohn fraction VI and β-lactoglobulin were found to be most effective in protecting the cells. The protective proteins do not promote growth; only BSA promotes growth of Con A-activated lymphocytes. The mechanism of the growth-promoting effect of BSA of Con A-activated lymphocytes was investigated. The continuous presence of BSA in the culture medium during and after Con A activation is required for optimal lymphocyte growth. Temporary replacement of BSA by any of the protective proteins markedly reduces, whereas protein-free medium abolishes, growth. The mechanism of the growth-promoting effect of BSA is not concerned with regulating the uptake of Con A by lymphocytes. Red blood cells, crenated by washing in protein-free medium, revert immediately to their normal globular shape by the addition of BSA or FAF BSA, whereas the protective proteins fail to do so, indicating a direct effect of BSA on the external surface of the erythrocyte membrane and presumably also of the lymphocyte. The hypothesis is proposed that BSA promotes growth by conveying total structural integrity to the cell surface membrane of Con A-treated lymphocytes.
Mixed Lymphocyte Reactions: Augmentation by Drugs That Inhibit Stimulator Cell Protein Synthesis
Authors: Metzler Charles M. , Kostyk Thomas G. , Gershon Richard K.
Published: December 2022

Inhibiting the protein synthesis of F1 stimulator cells in a mixed lymphocyte reaction (MLR) can enhance the DNA synthetic response and cytotoxic activity of the responder parental lymphocytes, independent of the number of stimulator cells used. Thus stimulator lymphocytes act not only as a source of antigen but play an additional regulatory role in the MLR.
B Lymphocyte Alloantigen Specificities Present on Cultured Lymphoblastoid Cell Lines
Authors: Honig R. , Billing R. , Matsuyama M. , Terasaki P. I.
Published: December 2022

Thirty-three of 34 cultured human lymphoblastoid B cell lines have been shown to express four out of five polymorphic non-HLA specificities expressed by normal B lymphocytes. The specificities were detected by 32 alloantisera produced by absorption with pooled platelets to remove HLA activity and selected from over 400 pregnancy sera. One B group (B2) which was expressed by 23% of a panel of normal B lymphocytes was not found on any of the cultured lines tested. Four T cell lines tested and myeloid line (K562) did not react with any of the B cell alloantisera.
Quantitative Analysis of the Proliferative Activity Induced in Murine Thymocytes by Concanavalin A
Authors: Gerhart Stanley , Mills Gordon , Monticone Vicky , Paetkau Verner
Published: December 2022

A quantitative analysis of the proliferative response induced in murine thymocytes by concanavalin A (Con A) is described. Exogenous 3H-thymidine labels 35 to 40% of the newly incorporated TMP residues under optimal conditions. The density label 5-bromo-2′-deoxuridine (BrUdR) does not affect DNA metabolism in this system. With this nucleoside, it is shown that newly synthesized DNA is the result of semi-conservative replication, not repair. Double labeling of DNA provides a monitor for cells traversing the cell cycle (S phase to subsequent S phase). The average cycle time is 12.5 hr, and the shortest cell cycle time is 10 hr. The growing fraction of active cells is about two-thirds. The data show that different subpopulations of thymocytes begin proliferating after various times in culture. Once effectively stimulated by Con A, some of the cells can traverse the cell cycle at least twice more after the mitogen is removed.
Reversal of Cell Surface Abnormalities of T Lymphocytes in Hodgkin's Disease after in Vitro Incubation in Fetal Sera
Authors: Fuks Zvi , Strober Samuel , King Donna P. , Kaplan Henry S.
Published: December 2022

The capacity of peripheral blood lymphocytes from patients with untreated Hodgkin's disease to form E rosettes with sheep erythrocytes and to respond in vitro to PHA stimulation were found to be profoundly impaired. In 49% of the patients, the percentage of E rosette-forming cells (E-RFC) was more than two standard deviations below the mean for normal donors. Overnight incubation of the peripheral blood lymphocytes from these patients in culture media containing 20% fetal calf serum was followed by restoration of the percentage of E-RFC up to normal levels. Similar results have been observed after incubation in fetal human serum, but not in adult human AB serum or adult bovine serum. Incubation of peripheral blood lymphocytes from untreated patients in 20% fetal calf serum also resulted in a remarkable restoration of their capacity to respond normally to PHA. Possible mechanisms involved in these reversible cell surface and in vitro lymphocyte function abnormalities in Hodgkin's disease are discussed.
Nonspecific Inhibition of Tumor Growth in Vivo by Admixed Allogeneic Tumor-Sensitized Lymphoid Cells and Identical Inactivated Allogeneic Tumor Cells
Authors: Sendo F. , Nakayama M. , Gotohda E. , Kobayashi H.
Published: December 2022

With the in vivo tumor neutralization test (Winn test), growth of a transplanted tumor (KMT-17) from Wistar-King-Aptekman rats was inhibited by allogeneic tumor (AH-66 from Donryu rats)-sensitized syngeneic lymphoid cells admixed with mitomycin C (MMC)-treated AH-66 cells. The observed tumor inhibition may be immunologically nonspecific, since no cross-antigens were detected by membrane immunofluorescence on the surfaces of KMT-17 and AH-66 cells. Close contact among KMT-17, AH-66-sensitized lymphoid cells and MMC-treated AH-66 cells was required for the inhibition of KMT-17 growth. AH-66 cells pretreated with formalin or ultrasonication lost tumor inhibitory activity when they were admixed with AH-66-sensitized lymphoid cells, and only MMC-treatment effectively preserved the tumor inhibitory activity of AH-66 cells. The sensitized spleen cells, draining lymph node, or peripheral blood cells inhibited tumor growth when they were admixed with MMC-treated AH-66 cells, whereas nucleated cells from bone marrow, thymus, or distal lymph node did not. Growths of KMT-17 were inhibited by admixed sensitized spleen cells and MMC-treated AH-66 even when pre-irradiated rats were used as recipients.
Generation of a Neutrophil Chemotactic Agent by Spermatozoa: Role of Complement and Regulation by Seminal Plasma Factors
Authors: Clark Robert A. , Klebanoff Seymour J.
Published: December 2022

The interactions of polymorphonuclear leukocytes, spermatozoa, seminal plasma, and serum have been examined in an in vitro leukocyte chemotaxis system. Bovine or human spermatozoa have no neutrophil chemotactic activity alone, but generate potent activity when incubated with serum. Generation of this activity requires magnesium, but not calcium ions and is blocked by pre-heating the serum. Chemotactic activity is generated normally in serum deficient in immunoglobulins or C4, but is not produced in serum deficient in C3. Thus, spermatozoa activate complement via the alternative pathway. The chemotactically active agent has been identified as the low m.w. C5 cleavage product (C5a) on the basis of heat stability, gel filtration characteristics, and specific inhibition by antiserum to C5. The spermatozoal constituents responsible for complement activation are largely heat stable, resistant to diisopropylfluorophosphate and on gel filtration compise a heterogeneous group of large m.w. materials.

Bovine seminal plasma has no neutrophil chemotactic activity alone; considerable activity, however, is generated by incubating low, but not high concentrations of seminal plasma with serum. The chemotactic agent produced in serum is C5a. Bovine seminal plasma also contains potent heat labile inhibitors of chemotaxis. Two distinct inhibitors differing in m.w. (50 to 100,000 and 15,000) and heat stability have been isolated. The inhibitors appear to act directly on the neutrophil rather than on the chemotactic factor. Human seminal plasma exhibits slight chemotactic activity alone but does not generate significant additional activity on incubation with serum. In contrast to the inhibitory activity of bovine seminal plasma, the human material contains heat-stable, nondialyzable factors which enhance the neutrophil chemotactic response.

These studies document a specific mechanism for the directed migration of polymorphonuclear leukocytes toward spermatozoa. The findings may provide an explanation for the observed in vivo leukocyte accumulation in the female genital tract in association with spermatozoa and seminal plasma. The biologic significance of this phenomenon relates to potential leukocyte-spermatozoa interaction which may influence the likelihood of fertilization.
Autologous Human Cellular Cytotoxicity Induced by Mitogenic and Nonmitogenic Lectins
Authors: MacDermott Richard P. , Nash Geoffrey S. , Saint John G. , Clark Eloise A. , Zaras Alexander G. , Boldt David
Published: December 2022

Autologous systems are of great interest in the study of cellular cytotoxicity. By eliminating considerations of previous allogeneic sensitization, an autologous system allows straight forward study of effector mechanisms. The induction of lymphocytotoxicity by a variety of plant lectins toward xenogeneic, allogeneic, or syngeneic target cells has been described by Holm and Perlmann (1–4) and other investigators (5–7). Although early studies of lectin-induced lymphocytotoxicity demonstrated a close correlation between the degree of mitogenic stimulation of the lymphocytes and subsequent cytotoxicity (2), recent work has suggested that the phenomena of mitogenesis and of cytotoxicity may be independent of one another (5, 7). The ability to fractionate phytohemagglutinin into its major components (8), and the existence of other plant lectins that are nonmitogenic, provide an opportunity to study human autologous cellular cytotoxicity induced by lectins, with particular attention directed to the relationship between cytotoxicity and mitogenicity.
Thyroid Antibodies in Spontaneous Autoimmune Thyroiditis in the Buffalo Rat
Authors: Noble Bernice , Yoshida Takeshi , Rose Noel R. , Bigazzi Pierluigi E.
Published: December 2022

Thyroid antibodies in the sera of BUF rats are closely correlated with spontaneous thyroiditis; their detection may facilitate the study of this animal model of organspecific autoimmunity. In a group of 115 retired BUF breeders (females older than 1 year), 26% had mononuclear cell infiltration of the thyroid and high titers of thyroid antibodies detectable by indirect immunofluorescence (IF) and chromic chloride passive hemagglutination (CCH). In contrast, low-titered thyroid antibodies were present in 9% of the rats that had normal thyroids.

Sequential studies performed on a group of 76 neonatally thymectomized BUF rats showed that at 2 months 24% had high titers of thyroid antibodies detectable by IF and 8% by CCH and at 3 months these percentages increased to 27% by IF and 25% by CCH. When the rats were sacrificed at 4 months, at a time when spontaneous disease is not seen in untreated animals, 26% were found to have mononuclear cell infiltration of their thyroids. Approximately 75% of these rats with thyroiditis had been positive for thyroid antibodies at 2 months and 90% at 3 months. At sacrifice all of these animals had high-titered antibodies to thyroid antigens. In contrast, low-titered thyroid antibodies were present in 36% of the animals without thyroiditis.

Intravenous injection of BUF thymus cells into neonatally thymectomized rats failed to reduce the incidence of thyroiditis and thyroid antibodies. Approximately 33% of these animals had both thyroid infiltration and serum antibodies, whereas 19% of those with normal thyroids had low-titered thyroid antibodies.

Titers of circulating thyroid antibodies were closely correlated with the initial and intermediate stages of thyroiditis, i.e., animals with less infiltration had the lowest titers, whereas animals with intermediate levels of infiltration had high antibody titers. On the other hand, rats with a high degree of thyroid infiltration had relatively lower titers of thyroid antibodies. Direct IF of infiltrated thyroids revealed the presence of rat immunoglobulins in these organs, suggesting a possible direct or indirect role of autoantibodies in the pathogenesis of the disease.

We attempted to detect delayed hypersensitivity by skin testing with thyroid antigens and observing reactions at 4, 24, and 48 hr. All of 123 rats were negative, 20% of which had thyroiditis and thyroid antibodies. No serum MIF activity was detected in rats with thyroiditis and those with normal thyroids. The absence of delayed hypersensitivity reactions in these experiments provides further support for the contention that spontaneous thyroiditis in the BUF rat may be antibody mediated.