bluuebunny/crossref_metadata_2025 huggingface dataset:
T Cell Memory for the Cytotoxic Response to Hapten-Modified Target Cells
Authors: Koren Hillel S. , Wunderlich John R. , Inman John K.
Published: December 2022

This study describes the development of memory and cytotoxic murine T cells against syngeneic hapten N =[N-(3-nitro-4-hydroxy-5-iodophenyl-acetyl)-β-alanylglycylglycyl] associated antigen. Memory activity in this system had the following characteristics. a) In vitro challenged cells primed in vivo resulted in an augmented cytotoxic response compared to cells primed in vitro. b) The augmented cytotoxic response in vitro was antigen-specific for both target cells in the lytic reaction and stimulator cells in the secondary response. c) Memory activity was long lasting (at least 2 months). d) Memory cells were not cytotoxic. e) Memory activity as well as the cytotoxic cells generated in a secondary response in vitro were T cell dependent. These findings are consistent with the results of others who have investigated T cell dependent memory in other cell-mediated reactions.
The Effects of Age on the Immune Response to Type III Pneumococcal Polysaccharide (SIII) and Bacterial Lipopolysaccharide (LPS) in BALB/c, SJL/J, and C3H Mice
Authors: Smith A. Mason
Published: December 2022

Type III pneumococcal polysaccharide (SIII) and bacterial lipopolysaccharide (LPS) were used to evaluate B cell and T cell regulatory functions in BALB/c, SJL/J, and C3H mice of various ages. It was found that the BALB/c and C3H mice could mount high level plaque-forming cell (PFC) responses to SIII at various ages through 110 weeks whereas the levels of the SJL/J PFC responses had begun to decline by the age of 42 weeks through the age of 80 weeks. BALB/c mice were also capable of producing strong PFC responses to LPS at various ages through 110 weeks whereas the comparable SJL/J PFC responses to LPS had declined by 80 weeks of age. By using anti-lymphocyte serum (ALS) and low-dose paralysis to SIII, it was shown that suppressor T cell activity was apparently greater in young BALB/c mice than in older BALB/c mice. It was also found that paralysis to SIII in BALB/c mice was easier to achieve at an early age. SJL/J mice were found to have the necessary B cell activity to respond to SIII through 80 weeks of age and the PFC responses could be greatly enhanced by ALS. Implications of the roles of regulatory T cells in aging are discussed.
Production of IgG Antibodies and Enhanced Response of Nude Mice to DNP-AE-Dextran
Authors: Rüde Erwin , Wrede Jürgen , Gundelach Marie L.
Published: December 2022

DNP-AE-dextran, prepared by the binding of DNP-ε-aminocaproyl residues to aminoethylated dextran (predominantly α-1,6-linked), induced a T-independent anti-DNP antibody response in mice. However, certain differences were observed between the response to this antigen in normal and nude mice. Thus, the antibody titers of nu/nu mice from day 10 to 38 after immunization were significantly higher than those of nu/+ controls. Furthermore, DNP-AE-dextran induced a weak secondary response in nu/+ but not in nu/nu mice.

For both thymusless and normal mice the production of IgG in addition to IgM antibodies to DNP-AE-dextran could be established. The former included antibodies of the IgG1 subclass which were considered to be particularly thymus dependent (1). The higher response of nu/nu mice was reflected mainly in the increased production of IgG antibodies. Under the influence of a graft-vs-host reaction, a 10-fold increase in antibody titers to DNP-AE-dextran was observed, due entirely to an enhanced IgG response.
Isolation of Alternative Pathway C3 Convertase Containing Uncleaved B and Formed in the Presence of C3 Nephritic Factor (C3NeF)
Authors: Daha Mohammed R. , Fearon Douglas T. , Austen K. Frank
Published: December 2022

The capacity of serum from patients with membranoproliferative glomerulonephritis to interact with normal human serum to inactivate C3 was ascribed to the presence of C3 nephritic factor3 (C3NeF)4 (1). The action of C3NeF was magnesium ion dependent, operative in C4 deficient serum (2), and impaired in serum selectively deficient in B and D̄ (3) indicating that C3 cleavage occurred by the alternative pathway. Fluid phase interaction of C3, B, and D̄ in the presence of C3NeF permits recovery of C3 convertase with an estimated sedimentation rate of 10S containing at least C3̄ and B̄ (4). The present study shows that interaction of larger amounts of C3NeF with C3 and B in the absence of D̄ allows isolation of a 10S complex containing C3 and B̄, an uncleaved form of B, and manifesting C3-cleaving activity.

Materials and Methods. B (5), D̄ (6, 7), D̄ (7), and C3 (8, 9) were purified to homogeneity and quantitated as described.
Specific Antigen-Binding and Antibody-Secreting Lymphocytes Associated with the Erythrocyte Autoantibody Responses of NZB and Genetically Unrelated Mice
Authors: DeHeer David H. , Edgington Thomas S.
Published: December 2022

The receptor characteristics as well as incidence of antigen-binding lymphocytes (ABL) of B and T cell classes with membrane receptors specific for the exposed (X) and cryptic (HB) murine erythrocyte autoantigens were examined in NZB and nine control strains of mice. Whereas only NZB and NZB hybrid mice synthesize anti-X autoantibody pathogenetically implicated in the genetically determined autoimmune hemolytic anemia, the NZB as well as control strains synthesize the ubiquitous anti-HB anti-erythrocyte autoantibody. By utilizing immunocytoadherence assays, maximum numbers of specific ABL of both B and T lymphocyte classes were optimally demonstrated at erythrocyte:lymphocyte ratios of 20:1 and after lymphocyte fixation at 56°C for 20 min. Surface membrane receptor specificity was established by inhibition with semi-purified soluble X or HB autoantigen. Inhibition of immunocyto-adherence with class specific antisera to mouse immunoglobulins demonstrated that the receptors on both B and T cells were of IgM class. Specific receptors regenerated in vitro after trypsinization which excluded the role of cytophilic antibody in the immunocytoadherence reactions.

B lymphocyte ABL reactive with the X autoantigen were demonstrable in NZB, NZB hybrid, and control mice. Only in NZB and NZB hybrid mice, strains that uniformly synthesize anti-X autoantibody, were X ABL of T lymphocyte class demonstrated. The presence and incidence of T lymphocyte X ABL is compatible with the expression of a single dominant gene carried by the NZB strain. The incidence of B lymphocyte X ABL increased with age, suggesting proliferation of this cell population. HB ABL of both B and T lymphocyte classes were observed in all strains, concordant with the ubiquitous presence of humoral anti-HB autoantibodies. Differentiation of precursor B cells was evaluated by PFC assay of cells secreting specific autoantibodies. Anti-X PFC were observed only in NZB and NZB hybrid mice; and the observed frequency suggested that less than 3.5% of the specific ABL were differentiated for the secretion of anti-X autoantibody. Anti-HB PFC were observed in all strains and represented as high as 11.8% of specific ABL. Genetic determination of the anti-X anti-erythrocyte auto-antibody response does not prescribe the presence of precursors of the antibody-forming cell, but rather appears to influence regulation of the differentiation of these cells. These data suggest that circumvention of immunologic tolerance to this specific erythrocyte autoantigen may occur at the level of the T lymphocyte; or alternatively, that T lymphocytes as well as B lymphocytes, are induced to proliferate and differentiate in the NZB strain.
Replication of Measles Virus in Continuous Lymphoid Cell Lines
Authors: Gallagher Marie R. , Flanagan Thomas D.
Published: December 2022

The replication of measles virus was studied in a T cell-like line, MOLT-4, and a B cell-like line, Raji. Both cell lines supported measles virus replication. The infected lymphoid cells were assayed for infectious virus and expression of measles antigens at timed intervals up to 84 hr after inoculation. At time 0, after a 2-hr adsorption period at 4°C, both cell lines produced a maximum of 3% infectious centers and up to 22% antigen-positive cells. Eventually all cells in both populations became infected during the experimental period. When Raji and MOLT-4 cells were infected under similar conditions 100% infection of cells was achieved 12 hr earlier in Raji than in MOLT-4. Increasing multiplicity of infection (MOI) from 1 to 10 made no significant change in the number of infected cells during the early part of the cycle. Spread of virus was largely due to released virus and not cell to cell transmission. Since only a small proportion of the population adsorbed the virus at any one time, and eventually all cells were infected, it was concluded that the receptors for measles virus were expressed at some phase of the cell cycle.
The Host Antigen Phenomenon in Experimental Murine Schistosomiasis
Authors: Boyer Markley H. , Ketchum Dana G.
Published: December 2022

Adult Schistosoma mansoni grown in hamsters and transferred directly to the mesenteric vessels of C57BL10J mice previously immunized with hamster cells survive normally despite evidence of cytoxic and hemagglutinating antibodies directed against hamster cells. This finding indicates that species differences may play an important part in the manifestations of the host antigen phenomenon in experimental schistosomiasis.
Regulatory Substances Produced by Lymphocytes
Authors: Namba Yuziro , Waksman Byron H.
Published: December 2022

L cell mutant lines which were considerably more resistant to rat lymphotoxin (LT) than the original cell line were obtained by periodic additions of LT, partially purified from sensitized lymph node cell culture supernatants by DEAE-cellulose chromatography and Sephadex gel filtration. Addition of actinomycin D to cultures of these mutant cells abrogated resistance to LT, suggesting that resistance was not due to a loss of LT receptors but probably to increased activity of a repair mechanism. These mutant lines were also more resistant to the proliferation inhibitory effect of LT in low concentration and to that of diluted culture supernatants of lymph node cells stimulated with antigen (ovalbumin) than the original cell line, but they remained as sensitive to inhibitor of DNA synthesis (IDS) as the original line. The mutant lines also remained fully sensitive to both complement-dependent lysis by antibody and antibody-dependent cellular cytotoxicity, but showed increased resistance to the killing effect of rat lymph node cells sensitized with original L cells in vivo. These findings suggest that the lymphotoxic substance partially purified and characterized in this and in the previous paper may be an important mediator of T cell-mediated cytotoxicity.
IgM-Mediated, T Cell-Independent Suppression of Humoral Immunity
Authors: Ordal John , Smith Sean , Ness Dolly , Gershon Richard K. , Grumet F. Carl
Published: December 2022

The immunological unreactive state occurring in (T,G)-A-L nonresponder mice after secondary antigen challenge was investigated. Syngeneic IgM anti-(T,G)-A-L antibody-containing plasma, transferred at the time of the time of primary challenge, induced persistent suppression of autologous specific antibody production. Removal of plasma IgM with goat anti-µ antisera removed the ability of the plasma to suppress. The induction and maintenance of the suppressed state were not different in thymectomized or sham-thymectomized animals. Primed animals subjected to graft-vs-host reaction (GVHR) at the time of secondary challenge switched over to IgG production. Animals suppressed by passive antibody transfer reacted to GVHR, at the time of secondary challenge, with specific IgM but not IgG antibody production.

Transfused normal spleen cells partially abrogated suppression only when (suppressed) hosts had been lethally irradiated. Spleen cells from antigen-plus-antibody suppressed donors, upon transfer to previously normal, syngeneic hosts, were less immunocompetent than spleen cells from untreated donors. These data are consistent with a model of IgM mediated, T cell-independent persistent suppression of humoral immunity.
Neutrophil Receptors for IgG and Complement: Their Roles in the Attachment and Ingestion Phases of Phagocytosis
Authors: Scribner Duncan J. , Fahrney David
Published: December 2022

The functional roles of IgG and C3b in phagocytosis by human peripheral neutrophils were investigated. Phagocytosis of Staphylococcus aureus in the presence of human serum was severely depressed by heat inactivation of serum at 56°C for 30 min. Experiments with varying particle: leukocyte ratios in the presence of complement-inactivated sera showed that particle-bound C3b can mediate a 10-fold enhancement of the overall phagocytic rate. When sheep erythrocytes were sensitized with either IgG or IgM, only the former were bound to and readily internalized by neutrophils. Erythrocytes sensitized with both IgM and C3b were bound but not internalized. Furthermore, the presence of Fc fragments during incubation of S. aureus or latex beads with neutrophils in the presence of IgG or fresh serum affected a total inhibition of internalization but did not significantly alter adherence. Quantitative data regarding IgG sensitization indicated that bound C3b results in at least a 3-fold decrease in the amount of sensitizing IgG required for 50% maximal phagocytic response by neutrophils. On the basis of the above results, it is argued that particle-bound C3b functions primarily in the adherence phase and that bound IgG serves as a trigger for the internalization phase of phagocytosis.
Effect of Cross-Immunization on Monotypic Antibody Responses to Herpes Simplex Virus Types 1 and 2
Authors: Smith Jerry W. , Glorioso Joseph C.
Published: December 2022

New Zealand White rabbits were immunized subcutaneously with partially purified UV-inactivated preparations of herpes simplex virus (HSV-1 or HSV-2) in complete Freund's adjuvant. After the initial immunizations, designated groups of animals received additional amounts of either HSV-1 or HSV-2 at 35-day intervals. Sera were absorbed with lysates of cells infected with heterotypic virus and the residual monotypic antibodies were detected by 51Cr-release assay using HSV-infected target cells. A positive correlation was found between the ratio of neutralizing antibodies to HSV-1 and HSV-2 (II/I index) and the content of monotypic antibodies. Results showed that production of monotypic antibodies to HSV-1 and HSV-2, under the conditions employed, was independent of previous exposure to HSV.
Effects of Sodium Periodate Modification of Lymphocytes on the Sensitization and Lytic Phases of T Cell-Mediated Lympholysis
Authors: Schmitt-Verhulst Anne-Marie , Shearer Gene M.
Published: December 2022

Sensitization of mouse splenic lymphocytes in vitro with sodium periodate-treated autologous spleen cells stimulated a one-way mixed lymphocyte reaction and led to the generation of thymus-derived cytotoxic effector cells. These effectors were capable of lysing in 4 hr periodate-treated syngeneic and, to a lesser extent, periodate-treated allogeneic target cells. These results suggest that sensitization by periodate-treated autologous cells could result either from a specific reaction to modified self components or from a nonspecific mitogenic stimulation. Effector cells generated by allogeneic sensitization were detected on periodate-modified targets, irrespective of the H-2 antigens expressed by the targets. The effects of periodate modification on both stimulator and target cells were reversible by sodium borohydride, suggesting that the biologic effects of sodium periodate are dependent on the formation of a free aldehyde group on cell surface glycoproteins. Pretreatment of stimulator cells with neuroaminidase prevented the effect of periodate treatment, suggesting that the sensitization involves oxidized sialic acid residues. During the 4-hour 51Cr-release assay periodate-treated targets could be used to detect cytotoxic effector cells of any specificity. Fresh spleen cells and lymphocytes cultured for 5 days without antigen or in the presence of lipopolysaccharide did not lyse periodate-treated targets. An increasing level of cytotoxicity was detected on periodate-treated targets when the effector cells were generated, respectively, by stimulation with concanavalin A, by sensitization with periodate-modified autologous cells, or by sensitization with unmodified allogeneic stimulator cells. Although the lysis of periodate-treated targets is itself nonspecific, effector cell specificity could be determined by selective blocking of the lytic phase with cells syngeneic to the stimulators. These results indicate that a nonspecific interaction can occur between lymphocytes and periodate-treated target cells, but that this interaction leads to lysis only when the lymphocytes were activated to become cytotoxic effectors.
Antibodies Specific to Isoniazid and Isonicotinic Acid
Authors: Tse Kam S. , Vijay Hari , Attallah Nicholas , Sehon Alec H.
Published: December 2022

Rabbit antisera to isoniazid (INH) and its major metabolite, isonicotinic acid (INA), were prepared by immunization with conjugates of these compounds with human serum albumin. The antisera were rendered hapten-specific by exhaustive absorption with the immunizing carrier. Purified anti-hapten antibodies were also isolated with appropriate immunosorbents. As demonstrated by inhibition of the quantitative precipitin curves and of precipitating immune complexes in immunodiffusion tests, the antibodies to the two haptens reacted with either INH or INA, and also with isonicotinamide (INC); these three related molecules share the isonicotinyl group. The relative effectiveness of inhibition by free hapten of precipitating immune complexes consisting of either anti-INH or anti-INA antibodies and the related hapten-protein conjugates was INH > INC > INA.
In Vitro Hemolysis of Autologous Erythrocytes Caused by Immune Murine Spleen Cells and Spherules of the Fungus Coccidioides Immitis
Authors: Danley David L. , Pappagianis Demosthenes , Benjamini Eliezer
Published: December 2022

This communication describes an in vitro system wherein mouse erythrocytes are lysed in the presence of spherules of the fungus Coccidioides immitis and spleen cells from syngeneic mice immunized with a variety of antigens. The antigens include: tobacco mosaic virus in complete Freund's adjuvant (CFA), CFA alone, separate components of CFA, sheep erythrocytes, and allogeneic tumor. Spleen cells from mice sublethally infected with C. immitis are also capable of participating in this response. The lytic phenomenon, which does not require complement, is dependent upon the number of spleen cells per culture, the number of spherules per culture, the time of culture incubation, the amount of antigen injected into the animal and the time after immunization at which spleen cells are recovered. Live spherules or spherules killed with heat, with dimethylsulfoxide, or with formalin were effective participants, together with immune spleen cells, in the lytic reaction.
Mechanisms of Resistance Against Experimental Trypanosoma Cruzi Infection: The Importance of Antibodies and Antibody-Forming Capacity in the Biozzi High and Low Responder Mice
Authors: Kierszenbaum Pelipe , Howard James G.
Published: December 2022

The role of antibodies and the host's antibody-forming capacity in resistance to Trypanosoma cruzi infection has been investigated in the Biozzi high and low responder lines of mice. Ab/L animals with low antibody-forming capacity were found to be more susceptible to i.p. infection with trypomastigotes than high-responders (Ab/H), whereas non-selected Swiss albino mice showed an intermediate level of susceptibility. The correlation between antibody-forming potentiality and susceptibility was consistently observed with both the Y and Tulahuén strains of the parasite, which differ in their preferential tissue tropism as well as virulence. A similar divergence in susceptibility was observed after subcutaneous infection with the Y strain which produces a more fulminating disease. Although Ab/L mice neither produced a significant antibody response to the parasite nor responded to prophylactic immunization with killed T. cruzi epimastigotes, they could be effectively protected by passive transfer of immune plasma.
An Enzyme-Linked Procedure for the Detection and Estimation of Surface Receptors on Cells
Authors: Cameron Deborah J. , Erlanger Bernard F.
Published: December 2022

An enzyme immunoassay procedure has been developed for the visualization and estimation of lymphocyte surface receptors. β-Galactosidase (β-gal'ase) was covalently linked to sheep anti-rabbit immunoglobulin (SARIg), to ovalbumin (OA), and to adenosine (A). Exposure of rabbit peripheral lymphocytes to SARIg-β-gal'ase and subsequent incubation with the fluorogenic substrate fluorescein-β-digalactopyranoside allowed visualization of B cells by fluorescence microscopy. Treatment with each of the above β-gal'ase conjugates and incubation with another fluorogenic substrate, 4-methyl-β-D-umbelliferylgalactopyranoside, made possible the estimation of the various receptors by spectro fluorimetry. Procedures used for enrichment of T and B cell populations could be monitored. Among the findings was that immunoglobulin-bearing cells (presumably B cells) formed rosettes with sheep erythrocytes. The quantitative procedure was used to study lymphocyte population changes during immunization with an A-O A conjugate. An increase in the binding of A-β-gal'ase, SARIg-β-gal'ase, and OA-β-gal'ase by peripheral lymphocytes occurred repeatedly 6 to 8 days after immunization but lasted only a few days despite a continued high circulating antibody titer. These results are consistent with the possibility that immunization induces migration of lymphocytes into the circulation from bone marrow and/or thymus.
Increased Iodine Uptake by Obese Strain Thyroid Glands Transplanted to Normal Chick Embryos
Authors: Sundick Roy S. , Wick Georg
Published: December 2022

The Obese strain (OS) of chickens spontaneously develops autoimmune thyroiditis. Since a defect or abnormality of the thyroid gland may be involved in this disease experiments were performed to compare the iodine uptake of OS with normal thyroid glands. To minimize the interaction of the OS immune system with the thyroid gland during these studies, thyroids were removed from 16-day-old embryos and transferred to the chorioallantoic membrane (CAM) of 9-day-old normal White Leghorn (NWL) embryos. NWL thyroid glands were transferred to the same CAM. Six days later the 20-hr 131I uptake of the transplants was determined. Twenty OS thyroid lobes had a mean 131I uptake of 2960 ± 740 cpm, whereas the NWL thyroids incorporated significantly less iodine (890 ± 160 cpm; p < 0.025). These results, along with a previous report suggest that an abnormality of the thyroid gland might be an important factor in the development of autoimmune thyroiditis.
Specific Removal of Bovine Serum Albumin (BSA) Antibodies in Vivo by Extracorporeal Circulation over BSA Immobilized on Nylon Microcapsules
Authors: Terman David S. , Tavel Tracie , Petty Donald , Tavel Arnold , Harbeck Ronald , Buffaloe George , Carr Ronald
Published: December 2022

Studies were undertaken to determine whether BSA, once immobilized on activated nylon microspheres, would be capable of specifically removing circulating BSA antibody in vitro and in vivo in an extracorporeal circulation system in dogs. Nylon microspheres were prepared and, after gentle hydrolysis and glutaraldehyde treatment, demonstrated a retention of up to 34.5 mg of BSA. In vitro studies showed that BSA immobilized on microspheres removed a significant percentage of BSA-binding activity. For in vivo studies, an extracorporeal circulation system was established and mongrel dogs were then injected with anti-BSA and anti-HSA antibodies. After an equilibration period, BSA microspheres were introduced into the extracorporeal circulation system. After the insertion of BSA microcapsules, serum exhibited a sharp reduction in BSA binding over the next 15 min, with a more gradual diminution over the ensuing 60 to 90 min. There was no significant reduction in anti-HSA binding over the same time frame. This effect could not be attributed to release of BSA from the microspheres since no 125I-BSA was detected in the serum or organs of the dogs at the conclusion of the experiments. After extracorporeal circulation over nylon microspheres, there were only minimal changes in the host's hematocrit or leukocyte counts and no significant thrombotic material or cellular debris was recoverable on the capsules. These data suggest that antigen immobilized on nylon microspheres may specifically withdraw circulating antibodies in vivo with minimal release of its antigenic material and little alteration in the host's hematologic status.
Role of T Lymphocytes in the Humoral Immune Response
Authors: Warren Robert W. , Murphy Suzanne , Davie Joseph M.
Published: December 2022

The effect of activated T lymphocytes (ATC) on the avidity distribution of PFC in the secondary response was studied in normal mice. The total PFC response was not significantly changed for either direct or indirect PFC by administration of ATC before secondary antigen challenge. However, marked suppression occurred of indirect PFC that secreted high avidity antibody; no suppression was seen of high avidity direct PFC. At the same time, significant stimulation was seen of relative and absolute frequencies of indirect PFC that secreted middle and low avidity antibody. These effects were dependent on Thy 1-bearing, nylon nonadherent cells which demonstrated carrier specificity. In further characterization of these effects, it was found that increasing the number of ATC transferred produced progressive loss of high avidity PFC and compensatory increase in lower avidity PFC. Moreover, in these experiments, suppression of the high avidity response was inducible with the administration of ATC 5 weeks before to 3 days after the secondary immunization. Thus, it is likely that the avidity-modifying effects are dependent on T lymphocytes which influence the late stages of B lymphocyte maturation.
The Effects of Antigen Dose and Adjuvant on the Antibody Response; Amplification of Restricted B Cell Clones
Authors: Civin Curt I. , Levine Herbert B. , Williamson Alan R. , Schlossman Stuart F.
Published: December 2022

Anti-DNP antibody production in inbred strain 2 guinea pigs immunized with chemically defined DNP-oligolysines was enhanced by increasing the antigen dose or employing more potent adjuvants. However, the increase in antibody titer was not accompanied by an increase in antibody heterogeneity. Only a small number of anti-DNP antibody-producing B lymphocyte clones were triggered to produce antibody, regardless of the antigen dose or the adjuvant used for immunization or the resulting specific antibody titer. This restricted response occurred despite the capacity of these animals to synthesize a large number of distinct anti-DNP antibodies and of DNP-oligolysines to bind all these antibodies. These observations are inconsistent with the hypothesis that adjuvant and increased antigen dose enhance antibody titer by recruitment of more diverse precursor B cells (clonal recruitment), but support the hypothesis that adjuvant and increased antigen dose act by enhancing the expansion of a restricted few B lymphocyte clones (clonal expansion).
The C5b-9 Complex: Subunit Composition of the Classical and Alternative Pathway-Generated Complex
Authors: Podack Eckhard R. , Kolb William P. , Muller-Eberhard Hans J.
Published: December 2022

The membrane attack complex of complement (C5b-9) is identical in composition regardless of which pathway of activation was instrumental in its formation. Band V protein was consitently a subunit of the soluble complex. Since band V protein is not required for complement-dependent cytolysis, it probably represents a membrane site equivalent in serum of the nascent C5b-9 complex.
Enhancement of DNA Synthesis and cAMP Content of Mouse Thymocytes by Mediator(s) Derived from Adherent Cells
Authors: Oppenheim Joost J. , Shneyour Amir , Kook Abraham I.
Published: December 2022

Supernatants of adherent mouse peritoneal exudate cells or human mononuclear cells were used as the source of lymphocyte activation factor (LAF). LAF was found to potentiate the effect of mitogens such as PHA and Con A on DNA synthesis by mouse thymocytes. However, LAF also was capable of reducing vigorous thymocyte reactions to Con A. Thus, LAF usually enhanced the effect of PHA on DNA synthesis by BALB/c thymocytes to a relatively greater degree than that of Con A. This change in the ratio of Con A to PHA response of thymocytes suggests that LAF can serve as a regulator of thymocyte DNA synthesis. Moreover, in the presence of LAF, allogeneic thymocytes developed the ability to have bidirectional mixed thymocyte reactions. Exposure to LAF not only improved the ability of parental thymocytes to act as responder cells, but, in addition, led to increased stimulatory activity of F1 thymocytes, presumably by promoting the differentiation of stimulator cells.

These indications that LAF affected differentiation were investigated further by studying its effect on the cAMP content of thymocytes. LAF stimulated significant immediate but transient elevations of intracellular cAMP and adenylate cyclase activity in thymocyte membranes. In contrast, the mitogens themselves failed to elevate or to influence the effect of LAF on the content of intracellular cAMP of thymocytes. Furthermore, the potentiating effect of LAF on mitogen-induced thymocyte DNA synthesis at times was enhanced by exogenous cGMP, carbachol, or imidazole. These findings suggest that LAF, through its stimulation of cAMP levels in thymocytes may in turn promote thymocytes to differentiate sufficiently to become competent to proliferate in response to mitogens.
Interaction Between Lymphocytes and Inflammatory Exudate Cells
Authors: Nakamura Shuji , Yoshinaga Masaru , Hayashi Hideo
Published: December 2022

Mouse polymorphonuclear leukocytes (PMN) were harvested from the peritoneal cavity stimulated with sodium caseinate. The cells were cultivated in vitro and the supernatant of these cultures (SUP) was tested for enhancing potency on DNA synthesis by syngeneic thymocytes stimulated with phytohemagglutinin (PHA). The enhancing potency of the SUP was markedly influenced by duration of the donor cultures of PMN, population density of the cultures, and protein concentrations in the medium, respectively.

The enhancing factor in the SUP was found to be non-dialysable, heat-labile, and stable in the pH ranging between 3 and 9 but labile in the pH below 2 or above 10; its m.w. was approximately 19,000 when measured by gel-filtration on Sephadex G-75. The factor had a proteolytic activity on 3H-acetyl hemoglobin (3HHb) at neutral pH (7.2). Both the proteolytic activity and thymocyte-helping potency of the SUP were similarly abolished by adding protease inhibitor (Trasylol) in soluble form, or by passing through a column of the inhibitor insolubilized. It was thus assumed that the enhancing effect of PMN on thymocyte response was associated with a neutral protease released from the cells: it may be termed a lymphocyte-helping protease (LHP).
Studies on the Maturation of Immune Responsiveness in the Mouse
Authors: Landahl Carol A. , Chakravarty Ashim , Sulman Miriam , Kubai Louis , Auerbach Robert
Published: December 2022

Experiments were designed to investigate the role of the spleen in the development of the murine immune system. By using mice splenectomized within 24 hr of birth, as well as mice with a hereditary, congenital absence of the spleen, the primary immune response to sheep erythrocytes was examined. The immunocompetence of lymph node cells from spleenless or control mice was assessed in vitro, in organ and in cell suspension cultures, and in vivo, by transfer into lethally irradiated syngeneic recipients followed by antigenic stimulation. The immunologic capacities of thymus and bone marrow cells were similarly tested by injection separately or in combination into irradiated syngeneic mice. Lymph node cells from spleenless animals appeared fully competent both in vitro and in transfer experiments. Neither neonatal splenectomy nor congenital absence of the spleen significantly reduced the capacity of bone marrow or thymus cells to participate in the immune response to sheep erythrocytes.
Detection of Antibodies to Bacterial Cell Wall Peptidoglycan in Human Sera
Authors: Heymer Berno , Schleifer Karl-Heinz , Read Stanley , Zabriskie John B. , Krause Richard M.
Published: December 2022

A radioimmunoassay has been developed for the measurement of antibodies to peptidoglycan in human sera including patients with rheumatic fever and juvenile rheumatoid arthritis. The assay is based on the percentage of binding of the hapten 125I-L-Ala-γ-D-Glu-L-Lys-D-Ala-D-Ala, the major peptide determinant of peptidoglycan. Because of differences in the avidity of the antibodies in different sera, the amount of antibody was expressed as pentapeptide hapten-binding capacity (pentapeptide-HBC in ng/ml of serum).

Fourteen out of 105 normal blood donors had a pentapeptide-HBC value ≥ 75 ng/ml serum. Values in healthy children 5 to 18 years of age were ≤ 50 ng/ml. Sixty-eight percent of the individuals with rheumatic fever had values ≥ 75 ng/ml, an indication that streptococcal infections can stimulate an immune response to peptidoglycan. Thirty-five percent of the patients with juvenile rheumatoid arthritis had values ≥ 75 ng/ml. Such a finding points to a possible association between bacterial infections and juvenile rheumatoid arthritis.
Induction of Specific Suppressor T Cells in Vitro
Authors: Eardley Diane D. , Gershon Richard K.
Published: December 2022

We describe conditions for generating sheep red blood cell-specific suppressor T cells in Mishell-Dutton cultures. The production of specific suppressor cells is favored by increasing antigen dose in the initial culture but can be produced by transferring more cells when lower doses of antigen are used. Transfer of small numbers of cells cultured with low doses of antigen leads to a specific helper effect. Transfer of large numbers of educated cells leads to nonspecific suppression.

Suppression can be effected by the effluent cells from nylon wool columns which do not make detectable PFC. A fraction of these cells become resistant to treatment with anti-T cell sera and complement after culture. The suppressor cells are radiation sensitive and must be able to synthesize protein to suppress. They take 2 to 3 days of education to reach maximum suppressive efficiency and will not suppress cultures if added 2 to 3 days after culture initiation. Their production is favored by the absence of mercaptoethanol, suggesting that the observed suppression is not “too much help”. The ability to generate specific suppressor cells in vitro should be of great benefit in determining the factors that regulate their appearance in vivo.
Suppressor Cells in Spleens from “Nude” Mice: Their Effect on the Mitogenic Response of B Lymphocytes
Authors: Smith J. Bruce , Eaton Gordon J.
Published: December 2022

A cell population recovered after velocity sedimentation fractionation of “nude” but not haired mouse spleen cells suppressed the response of spleen cells from both “nude” and haired mice to the B cell mitogen, LPS. The suppressor activity of these cells was abrogated by treatment with anti-ϑ antiserum and complement but not by treatment with the same antiserum preabsorbed with mouse brain. It is possible that these suppressor cells come from the pool of T cell precursors known to be present in “nude” bone marrow and spleen, and that they do not require thymic influence in order to perform the suppressor function.
Interaction of Soluble C3 Fragments with Guinea Pig Lymphocytes. Comparison of Effects of C3a, C3b, C3c, and C3d on Lymphokine Production and Lymphocyte Proliferation
Authors: Koopman W. J. , Sandberg A. L. , Wahl S. M. , Mergenhagen S. E.
Published: December 2022

C3a, C3b, C3c, and C3d were generated from purified guinea pig C3 by trypsin treatment. These fragments were characterized immunochemically and functionally by rosette inhibition. C3b is capable of binding to both C3b and C3d receptors on lymphocytes whereas C3d binds only to C3d receptors. C3b stimulates guinea pig spleen cells to elaborate a macrophage chemotactic factor which is similar in m.w. to that generated in response to PHA or LPS and is antigenically unrelated to C3 or C5. In contrast, neither C3a, C3c, or C3d stimulate guinea pig lymphocytes. Neither C3 nor any of its major fragments induce cellular proliferation. These data are compatible with the hypothesis that C3b triggers spleen cells to release a macrophage chemotactic factor by cross-linking C3b and C3d receptors.
Antibody-Dependent, Lymphoid Cell-Independent Plaques
Authors: Hanna Edgar E.
Published: December 2022

The report by Muchmore et al. (1) of artifactual sheep erythrocyte (SE) hemolytic plaques occurring in their assays of human lymphocyte cultures confirms and extends our observations to a human system. We reported (2–4) that background anti-SE antibody contained in the normal rabbit serum, a supplement of the medium for our rabbit spleen cell cultures, generated numerous lymphoid cell-independent plaques (LCIP). These artifactual plaques were generated by complement-mediated local lysis of the preformed anti-SE antibody-sensitized agglutinates of SE which formed after addition of SE as antigen to our cultures. Muchmore et al. have reached the same conclusions on the origin of artifactual plaques in their cultures of human lymphocytes (1). The source of preformed anti-SE antibody in their cultures was human plasma used as a supplement in their culture medium (1). Another type of artifactual plaque formation was observed by Roseman and co-workers (5), who suggested that passive carry over of antibody on several particulate vehicles may predispose PFC assays to “pseudo-plaques”.
Characterization of Effector Lymphocytes Associated with Immunity to Murine Sarcoma Virus (MSV) Induced Tumors
Authors: Plata Fernando , MacDonald H. Robson , Engers Howard D.
Published: December 2022

Cytolytic T lymphocytes (CTL) were generated in secondary mixed leukocyte-tumor cell cultures (MLTC) with syngeneic RB1-5 tumor cells as stimulating cells and with responding spleen cells from regressor mice that had rejected a murine sarcoma virus (MSV)-induced tumor. CTL precursor cells were found to be exclusively of thymic origin and non-T cells were apparently not required for CTL generation. When the size variations of CTL from syngeneic MLTC were analyzed by velocity sedimentation it appeared that a transition from small precursor cells to larger effector cells occurred during the first 5 days in culture; this change in cell size was then followed by a shift toward small-sized cells. Furthermore, the CTL generated in syngeneic MLTC in the MSV tumor immune system were compared with those CTL obtained in allogeneic mixed leukocyte cultures (MLC) and were shown to exhibit fundamental similarities.
Measurement of H-2 and Non-H-2 Antigens in the Mouse with the Footpad Swelling Test
Authors: Kon Neal D. , Klein Paul A.
Published: December 2022

The footpad swelling (FPS) test for delayed-type hypersensitivity in the mouse was evaluated for its ability to measure histocompatibility (H) antigens present on intact spleen cells or solubilized from a methylcholanthrene-(MCA) induced fibrosarcoma.

Mice were immunized to H-antigens (H-2 or non-H-2) with skin allografts and challenged 14 days later with test antigens. Spleen cell-bound and solubilized H-antigens elicited strong and specific FPS reactions in allograftimmunized but not in unimmunized mice. The test could distinguish H-2 from non-H-2 antigens as well as D from KI region specificities of the H-2 gene complex.

These experiments suggest that the FPS test is a reliable in vivo method for measuring both cell-bound and solubilized H-antigens in the mouse.
Immunogen Structure and Cell Selection
Authors: Trump Gary N.
Published: December 2022

The cellular requirements for the in vitro response to DNP-O-Bio-Gel-P were determined. Results of these experiments indicate that: 1) splenic adherent cells are required for generation of anti-hapten PFC to the solid phase immunogen; and 2) cultured nude-mouse spleen cells have far less capacity to respond to the immunogen than do cells from wild type mice. These experiments suggest that both T and B cells are required for responsiveness to the moderately haptenated form of DNP-O-Bio-Gel-P.
Lymphokine-Like Factors Produced by Human Lymphoid Cell Lines with B or T Cell Surface Markers
Authors: Yoshida Takeshi , Kuratsuji Tadatoshi , Takada Akikazu , Takada Yumiko , Minowada Jun , Cohen Stanley
Published: December 2022

In the present series of experiments we examined the ability of eight continuous cell lines derived from human lymphocytes to release lymphokines into their culture media. We have demonstrated that all could produce a macrophage migration inhibitory factor and a neutrophil chemotactic factor. This ability was independent of the B or T cell origin of the cell lines. As a further extension of previous reports, we report that the migration inhibitory effect does not seem to be due to cytotoxic effects as judged by trypan blue exclusion studies, and more important by the demonstration that the inhibition is reversible.

The demonstration that neutrophil chemotactic activity is removable by treatment with a previously described and characterized anti-lymphokine antiserum suggests that this factor may be chemically similar to some of the lymphokines (migration inhibitory factor or chemotactic factor for macrophages) produced by antigen-activated guinea pig lymphocyte cultures. The failure to effect similar removal or inactivation of the macrophage migration inhibitory factor may simply reflect greater interspecies heterogeneity in this factor, or more interestingly, that this does not correspond to “classic” MIF.

These results extend previous observations by defining further the capacity of lymphoid cell lines in continuous culture to produce mediators and by characterizing their relationship to conventional lymphokines produced by antigen or mitogen-activated lymphocytes.
Specificity of Allogeneic and Xenogeneic Cell Recognition in the Fetal Lamb
Authors: Asantila Tuula , Toivanen Paavo
Published: December 2022

Cells prepared from liver, thymus, and spleen of fetal lambs at different stages of gestation were confronted with allogeneic and xenogeneic cells in MLC. Specific elimination of the responding cells with BUdR and UV light together with a subsequent restimulation was used to study the specificity of the reaction. The response of fetal liver cells was not based on the existence of specifically recognizing cellular subpopulations; the response was concluded to be due either to stimulatory products released by the stimulating cells or to the multipotentiality of the responding cells. Specifically recognizing cells first appeared in the thymus at 58 days postconception and in the spleen at 70 days.

In the response of sheep lymphocytes against allogeneic and xenogeneic (mouse, human) cells, a cross-reactivity occurred. Fetal lamb lymphocytes were also capable of recognizing intraspecies differences on the xenogeneic cells. This capacity developed simultaneously with the specific recognition of allogeneic cells. No clear difference was observed in the reactivity of fetal thymus cells and spleen cells when compared to that of adult peripheral blood lymphocytes. These findings indicate that immunologically specific recognition of foreign cells is created in the sheep during the early intrauterine development.
Control of Fc and C3 Receptors on Myeloid Leukemic Cells
Authors: Lotem Joseph , Sachs Leo
Published: December 2022

An experimental system has been developed to study in cloned lines of cells the control of Fc and C3 receptors by different compounds. The cells used were clones of mouse myeloid leukemic cells and the compounds used were the protein MGI2 (macrophage and granulocyte inducer) in serum from mice injected with bacterial endotoxin and the steroid inducer (SI) dexamethasone. Eight clones were isolated which could be divided into three groups. One group (MGI+SI+) was induced to form EA and EAC rosettes by MGI and only EAC rosettes by SI, the second group (MGI-SI+) was not inducible by MGI but was induced by SI to form EA or EA and EAC rosettes, and the third group (MGI-SI-) was not inducible for EA or EAC by MGI or SI. There were two types of MGI+SI+ clones, one type (D+) could be induced by MGI to differentiate to mature macrophages and granulocytes, and the other type (D-) could not be induced to differentiate to mature cells. The MGI-SI+ and MGI-SI- clones were all D-. The results indicate that there are different cellular sites for MGI and SI and that induction of EA and EAC rosettes did not seem to be mediated by cyclic AMP.

Experiments with specifically bound 3H-BSA-anti-BSA complexes have indicated that there was an increase in the amount of 3H-BSA-anti-BSA bound per rosette-forming cell following induction by MGI or SI, and there were differences in the amount of 3H-BSA-anti-BSA bound per rosette-forming cell in different clones. These clones also showed differences in the shape of the curve for the number of EA rosette-forming cells obtained with erythrocytes coated with decreasing concentrations of antibody. The results suggest that such curves and those obtained with EAC rosettes can be used to determine the relative abundance of EA and EAC receptors on rosette-forming cells. EA rosettes on the myeloid leukemic cells, like those on normal macrophages and granulocytes, were specifically inhibited by IgG2α and by the Fc but not the Fab fragment of IgG. The EAC rosettes were inhibited by destroying the C3 component of complement. The different clones maintained their specific properties for at least 6 months in culture. The present system should, therefore, also be useful for studies on the genetic control of the regulation of Fc and C3 receptors.
Recruitment of Effector Lymphocytes by Initiator Lymphocytes
Authors: Livnat Shmuel , Cohen Irun R.
Published: December 2022

In previous studies, we have shown that initiator T lymphocytes (ITL) sensitized in vitro recruit effector T lymphocytes in vivo. When ITL were sensitized against fibroblast antigens in vitro and injected into footpads of syngeneic recipients, they induced enlargement of the draining popliteal lymph node (PLN) and the development there of specific effector lymphocytes of recipient origin. To study the basis of this lymph node response in recruitment, we injected 51Cr-labeled spleen cells i.v. into recipients of sensitized ITL and found that the labeled circulating lymphocytes were trapped in the reacting PLN. The trapping depended on surface properties of the labeled circulating lymphocytes, as revealed by various enzymatic treatments. The trapping process was radiosensitive, both on the part of the trapped lymphocytes and the lymph nodetrapping mechanism. Thus, sensitized ITL injected into the hind footpads migrate to the PLN and induce the trapping of circulating recruitable lymphocytes, which either differentiate into or regulate the differentiation of effector T lymphocytes.