bluuebunny/crossref_metadata_2025 huggingface dataset:
Alloantigen Bound to Agarose Beads and Syngeneic Carrier Cells Are Capable of Stimulating Mouse Cytolytic T Lymphocytes in Vitro
Authors: Fast Loren D. , Fan David P.
Published: December 2022

Bead-bound antigen was prepared by coupling alloantigen covalently to agarose beads. Alloantigen-bearing syngeneic carrier cells were prepared by dilution of detergent solubilized alloantigen in the presence of syngeneic spleen cells. Both types of antigen were compared to spleen cells and reconstituted membrane fragments for the ability to stimulate cytolytic thymus-dependent lymphocytes in vitro. All these types of antigen could stimulate immune but not nonimmune spleen cells to form cytolytic T lymphocytes. The amount of lytic activity obtained with the bead-bound antigen was found to be only dependent upon the amount of H-2 antigen present in the culture and independent of the number of beads.
A Hapten-Specific Ir Gene
Authors: Pelkonen J. L. T. , Kaartinen M. , Karjalainen K. , Mäkelä O.
Published: December 2022

When mice of different strains were immunized with a conjugate of 2-phenyloxazolone (phOx) and chicken serum albumin (CSA), the antibody response was controlled by an Ir gene (Ir-phOx). H-2 alleles d and f were associated with a high response, k and a with an intermediate response, and allele b with a low response. The effect of the Ir gene was clear-cut in anti-carrier antibodies of the primary and the secondary response when the concentrations of anti-carrier antibodies varied between 1 and 350 µg/ml. Anti-hapten antibodies reached a ceiling of ca. 1000 µg/ml that was unaffected by the Ir gene. Before the ceiling was reached, antihapten antibodies were also subject to the control of the Ir-phOx gene.

When the same carrier CSA was coupled with other haptens, BOC-ABA-Tyr or NO2phOx, antibody responses were not under the control of the Ir-phOx gene. This gene is probably responsible for the differences that have been observed earlier in delayed hypersensitivity and antibody responses to skin painting by phOx.
Inhibition of the in Vitro Growth of Plasmodium Falciparum
Authors: Reese Robert T. , Motyl Mary R.
Published: December 2022

Sera from Aotus sp. monkeys (karyotypes II, III, and IV) which were immune to Plasmodium falciparum have been used to inhibit the in vitro growth of this human malaria parasite. Culture conditions used for the assays allowed 50- to 100-fold increases in the number of A+ erythrocytes infected in a 96-hr period in control cultures. Although normal monkey serum did not support growth as well as normal human serum, mixtures of normal monkey and human serum were found that did. Compared to such controls, as little as 3.5% immune monkey serum was found to cause approximately 56% inhibition in 4 days (2 replicative cycles). Purified globulin from immune monkeys inhibited 40% at 2 mg/ml and 75% at 7 mg/ml after a single replicative cycle. These data suggest that serum antibody is likely to play a major role in providing Aotus monkeys with protective immunity to P. falciparum.
Two New Recombinant H-2 Haplotypes, One of Which Juxtaposes Kb and Ik Alleles
Authors: Sachs David H. , Arn J. Scott , Hansen Ted H.
Published: December 2022

Two new recombinant H-2 haplotypes have been detected and established as congenic resistant lines on the C57BL/10 background. On the basis of serologic testing and immunoprecipitation analyses, the sublocus composition of the first recombinant haplotype, H-2bq1 (B10.MBR) has been shown to be KbIkSkDq, and that of the second recombinant, H-2sq3 (B10.SQR) to be KsIsSsDq. The occurrence of the Kb I-Ak juxtaposition after a recombination between H-2b and H-2m contrasts with the almost uniform failure to observe H-2b/H-2k recombinants in previous studies. This finding and the occurrence of a second recombination event involving the same chromosome soon after the first in our studies may imply that recombination within H-2 is not generally a random event. The B10.MBR line has proved useful in the production of specific anti-H-2Kb and anti-I-Ab antisera previously quite difficult to obtain without contamination by anti-Ia or anti-H-2K antibodies, respectively.
Nonspecific Suppressor Cells in Patients with Chronic Graft-Vs-Host Disease after Marrow Grafting
Authors: Tsoi Mang-So , Storb Rainer , Dobbs Sherrie , Kopecky Kenneth J. , Santos Erlinda , Weiden Paul L. , Thomas E. Donnall
Published: December 2022

Forty-four human long-term survivors after marrow transplantation for aplastic anemia or hematologic malignancy were studied for the presence of circulating nonspecific suppressor cells. Twenty-two of the patients were healthy and 22 had mild to moderately severe chronic graft-vs-host disease (GVHD). Patient mononuclear cells (of donor origin) were tested for their ability to suppress the responses of lymphocytes obtained from the respective marrow donors to alloantigens in mixed leukocyte culture (MLC) and/or to concanavalin A (Con A). Tests were carried out between 199 and 2393 (median 376) days after transplantation. Cells from only 1 of 22 patients without chronic GVHD showed suppression of donor cell blastogenesis responses. In contrast, cells from 11 of 22 patients with chronic GVHD showed more than 30% suppression of donor cell responses in MLC and/or to Con A. The finding of suppressor cells was not related to the time of testing after grafting nor to immunosuppressive therapy. Nonspecific suppressor activity was abrogated by irradiation with 1600 rads in vitro in five of six cases tested. Nonspecific suppressor cells may be one explanation for the severe combined immunodeficiency and the recurrent infectious complications characteristic of patients with chronic GVHD.
Mobility and Density of AgB, “Ia”, and Fc Receptors on the Surface of Lymphocytes from Young and Old Rats
Authors: Woda Bruce A. , Yguerabide Juan , Feldman Joseph D.
Published: December 2022

Analysis of spleen cell populations from old Lewis rats (>24 months) and from young Lewis rats (3 to 4 months) in a fluorescence-activated cell sorter indicated that with aging there is a loss of brightly stained Ia and Fc receptor- (FcR) positive cells. The density of AgB, Ia, and FcR was diminished on the surface of cells from old rats. The rate of capping of all three membrane proteins was slower on cells from old rats. Colchicine treatment allows capping of AgB with a single ligand only in young rats. Fluorescence photobleach recovery experiments (FPR) show that the fluidity of the lymphocyte membrane from old rats is diminished and the lateral diffusion of AgB is decreased. The colchicine and FPR experiments suggest that the changes in capping in old rats are due to, in part, alterations in membrane fluidity and cytoskeletal function.
Macrophages/Monocytes Require Cell-to-Cell Contact in Order to Regulate the Growth of a Murine Lymphoma Cell Line
Authors: Hewlett Guy , Opitz Hans-Georg , Flad Hans D. , Schlumberger H. Dieter
Published: December 2022

The mechanism of the macrophage-regulated proliferation of the murine lymphoma cell line FIO 30 has been further investigated. It appears that the macrophage is alone in its ability to support FIO 30 growth; a macrophage-like cell line, however, is unable to provide the stimulus required by the FIO 30 cells for their proliferation. Investigations into the nature of this stimulus indicate that the serum factor pro-MaSF and a macrophage cell surface component act synergistically to support the growth of the FIO 30 cell, but only when the two cell types are in close promixity. MaSF has also been further characterized and is shown to be closely associated with serum albumin.
Macrophage Stimulation by Bacterial Lipopolysaccharides
Authors: Doe William F. , Henson P. M.
Published: December 2022

Peritoneal macrophages from the endotoxin-unresponsive C3H/HeJ substrain of mice were entirely refractory to activation in vitro by protein-free LPS, a defect that was not overcome by co-culture of spleen cells from the responder C3H/St substrain with LPS resistant C3H/HeJ macrophages. The defect in responsiveness appears confined to the lipid A activation signal since C3H/HeJ macrophages were fully activated after in vitro treatment by lipid A protein (LAP)—LPS complex, isolated LAP, and BCG. Moreover, after exposure to allogeneic tumor cells in vivo, C3H/HeJ macrophages were cytotoxic for tumor target cells in vitro. By contrast, macrophages from the responder C3H/St strain were fully activated by protein-free LPS to become cytolytic for tumor cells in vitro. C3H/HeJ macrophages, therefore, exhibit a highly selective defect characterized by unresponsiveness to the lipid A activation signal of protein-free LPS and resistance to the toxic effects of high concentrations of LPS that were lethal to the responder C3H/St strain.
Specific Helper T Cells Permit Differentiation of Thymic Anti-Self-Trinitrophenyl Cytotoxic Precursor Cells
Authors: Cooley Margaret A. , Schmitt-Verhulst Anne-Marie
Published: December 2022

Thymocytes, in contrast to peripheral T cells, were unable to generate significant numbers of anti-self-TNBS cytotoxic T cells in primary in vitro cultures. The addition to such cultures of syngeneic 1000 rad irradiated lymph node or spleen cells from mice sensitized 6 or 7 days previously with trinitrochlorobenzene (TNCB) permitted the development of an anti-self-TNBS cytotoxic T cell response. This effect was shown to be T-dependent since it was largely abolished by anti-Thy 1 plus complement and was present in nylon-wool column purified T cells.

The lymph node cells act as specific helper cells in that a) they must be restimulated in vitro by the priming antigen and b) they do not take part in the lytic phase of the response. The specificity of the helper effect is further indicated a) by the lack of help from nonsensitized lymph node cells and b) by absence of an enhanced helper effect for thymic anti-allogeneic or anti-allogeneic-TNBS responses in the presence of specific T helper cells for the anti-self-TNBS response.

Analysis of the thymocyte subpopulation able to provide precursor cells was approached by separating thymocytes into a “medullary-like” minor, peanut-lectin-agglutinable (PNA-) fraction and a “cortical-like” major, peanut-lectin-agglutinable (PNA+) fraction. It was found that the PNA- fraction was enriched in thymocytes able to mount an autonomous anti-self-TNBS cytotoxic response, whereas the PNA+ population was totally unable to generate anti-self-TNBS effector cells in the absence of helper cells. The presence of helper cells from TNCB-painted mice permitted differentiation of anti-self-TNBS cytotoxic precursor cells from the PNA+ thymocyte cultures and enhanced the level of cytotoxic cells generated in PNA- thymocyte cultures.
Inhibition of Specific Immune Responses by Feeding Protein Antigens
Authors: Hanson Donald G. , Vaz Nelson M. , Maia Luiz C. S. , Lynch James M.
Published: December 2022

Single and multiple intragastric feedings of ovalbumin (OVA) to high-responder BDF1 mice resulted in virtually complete unresponsiveness to later parenteral immunization with OVA in Al(OH)3 adjuvant, as indicated by antibody titers in Farr assays and in passive cutaneous anaphylaxis (PCA) assays for IgE antibodies. Repeated feedings of partially tolerizing doses of antigen had a cumulative inhibitory effect on subsequent responsiveness. Slight increases in primary antigen binding were detected in the serum of antigen-fed mice not given parenteral immunization, although rates of clearance of radiolabeled OVA from the blood were similar in normal and tolerant animals. Serum recovered from tolerant mice 7 or 14 days after feeding did not inhibit antibody responses of normal recipients. Responsiveness of normal or OVA-primed syngeneic spleen cells was substantially reduced upon transfer into recipients that were previously fed OVA, when compared with transfer into saline-fed recipients. The differences in responsiveness of lymphocytes transferred into OVA- and saline-fed mice were completely eliminated by irradiation (850 R) of the recipient animals either 7 or 20 days after feeding and immediately before the transfer.

The results indicated that the state of tolerance after ingestion of OVA is actively maintained, but did not support the hypothesized mediation of this tolerance by orally induced antibodies or serum factors that have been associated with unresponsiveness to ingested antigens in similar experimental systems.
Mechanisms of C-Type Viral Leukemogenesis
Authors: Lee John C. , Ihle James N.
Published: December 2022

Various inbred strains of mice respond immunologically to genetically transmitted ecotropic C-type viruses. Part of this response is T cell blastogenesis with type specificity for the viral envelope glycoprotein gp71. Of those nonviremic, nonleukemic strains, and F1 crosses examined, in which virus expression occurs early in life, gp71-specific blastogenic T cells were detected within the first 2 months of age and temporally preceded the development of a humoral immune response. However, in the viremic, highly leukemic strain of AKR mice, gp71-specific T cell blastogenesis in vitro was readily detectable throughout the preleukemic phase, the first 5 months of age. In appropriate F1 crosses and backcrosses, the persistent in vitro blastogenic response segregated with viremia and leukemia. These data suggest that in vivo T cell stimulation by endogenous viral gp71, caused by viremia, may contribute to virus-induced leukemogenesis in mice.
Translocation of Dimeric IgA through Neoplastic Colon Cells in Vitro
Authors: Nagura Hiroshi , Nakane Paul K. , Brown William R.
Published: December 2022

We studied the translocation of dimeric IgA across epithelium, using neoplastic human colon cells in culture as a source of epithelial cells, and immunoelectronmicroscopy with peroxidase-labeled antigens and antibodies. The cells had some of the ultrastructural characteristics of normal, mature epithelial cells, i.e., polarity, desmosomal junctions, and secretory component on their basal and lateral plasma membranes. Horseradish peroxidase-labeled dimeric IgA, exposed to the cells at 0°C, bound selectively to secretory component on the cell surfaces. At 37°C, the bound dimeric IgA was taken into the cells by endocytosis and transported apically through the cytoplasm in vesicles. After 30 min, IgA was discharged across the apical surface. Neither colchicine (10-4 M) nor cytochalasin B (10-5 M) interfered with binding or endocytosis of dimeric IgA, but colchicine inhibited intracellular transport of the IgA-containing vesicles.

These experiments demonstrated that dimeric IgA can be transported through living intestinal epithelial cells in vitro. The transport includes 1) specific binding of IgA dimers to secretory component on plasma membranes, 2) endocytosis of IgA in vesicles, 3) transcytoplasmic transport of the IgA-containing vesicles by a process involving microtubules, and 4) discharge of IgA at the apical surfaces.
Cell-Mediated Immunity in Interstitial Nephritis
Authors: Neilson Eric G. , Phillips S. Michael
Published: December 2022

The present studies investigate the effector role of lymphocytes in guinea pigs with an interstitial nephritis. Several observations were made relative to a number of functions expressed by these cells. The results of adoptive cell migration studies suggest that a subpopulation of T cells in nephritic animals traffic renotropically to either normal or damaged kidneys on transfer. Similar lymphocytes were also tested in vitro for direct effector function by utilizing target kidney cell monolayers in a cell-mediated cytotoxicity assay. The kinetics of the observed cytotoxic response were studied over the life span of nephritic animals. Optimal target-cell lysis occurred 12 to 17 days after sensitization, simultaneous with the onset of active histopathologic changes. The cytotoxicity was stoichiometrically titratable and relatively specific for fetal kidney tissue. In addition, cells from the spleen or lymph nodes of diseased animals effectively suppressed this cytotoxic response. These findings demonstrate that a diverse population of T lymphocytes are both capable of damaging the renal interstitium as well as modulating effector-cell functions on a regional basis with the immune system.
Purification and Properties of Human B Cell-Activating Factor
Authors: Wood David D.
Published: December 2022

Human monocytes, when appropriately stimulated in vitro, release into the culture medium a factor (BAF) that stimulates the IgM response of T-depleted murine splenocytes to heterologous erythrocytes. The behavior of this factor on gel filtration, isoelectric focusing, ion exchange chromatography, and isopycnic centrifugation was studied. BAF appears to be a molecule of 15,000 daltons, pI 6.5, 1.33 g/ml with low solubility at low ionic strength. It is stable to acid, mild heating, and long-term storage. Activity is lost in alkali or by boiling. Papain may reduce BAF activity slightly, whereas trypsin and chymotrypsin have no significant effect. These properties are similar to those of other monokines reported to have a similar m.w.
Chemotactic Factor-Induced Generation of Superoxide Radicals by Human Neutrophils: Evidence for the Role of Sodium
Authors: Simchowitz Louis , Spilberg Isaias
Published: December 2022

The role of sodium ion in superoxide (O-2) generation by human peripheral neutrophils was investigated. Cells were activated by exposure to the synthetic tripeptide, N-formyl-methionyl-leucyl-phenylalanine (FMLP), and O-2 release was assessed by ferricytochrome c reduction after 5 min of incubation at 37°C in the presence of FMLP 4 × 10-8 M. In the absence of monovalent cations (isotonic glucose), negligible O-2 generation occurred.

There was a progressive increase in the magnitude of FMLP-induced O-2 generation with increasing Na+ concentration up to 90 mM, where the response was noted to plateau. Varying the K+ concentration (1 to 10 mM) had no effect on the amount of O-2 produced in the presence of Na+ 140 mM.

FMLP also stimulated 22Na+ and 45Ca2+ uptake by the cells in a dose- and time-dependent fashion. FMLP-induced 22Na+ uptake appeared to be independent of the external Ca2+ concentration (0 to 4 mM). In contrast, there was a progressive decrease in the magnitude of the FMLP-induced increase in 45Ca2+ uptake as the Na+ concentration was reduced by replacement with choline+ or glucose. These studies support a requirement for Na+ in FMLP-induced O-2 generation and suggest that a Na+ influx may underlie the nature of this requirement. The data are also consistent with the hypothesis that a Na+ influx may precede the Ca2+ influx in the FMLP-induced activation sequence.
T Cell Regulation of Polyclonal B Cell Responsiveness
Authors: Goodman Michael G. , Weigle William O.
Published: December 2022

In addition to the x-linked B cell maturation deficit previously reported in CBA/N mice, a functional T cell defect has now been observed. T lymphocyte regulation of the polyclonal PFC response was studied within the context of this x-linked immunodeficiency model. The ability of 1) B cells from (CBA/N × CBA/CaJ)F1, male mice to respond to nonspecific T cell helper signals and 2) T cells from NCF1 male mice to provide such signals was investigated under in vitro conditions by using bacterial lipopolysaccharide (LPS) as the polyclonal activator. B lymphocytes from both male and female NCF1 mice were receptive to T cell help rendered by NCF1 female T cells. Male T cells, however, were unable to augment polyclonal B cell responses of either NCF1 male or female B cells to LPS. Treatment with ATS + C reduced the polyclonal response of female but not male spleen cells to LPS. This deficit could not be overcome by the use of greater numbers of NCF1 male T cells. The observation that this deficiency in T cell regulation is not due to active suppression suggests that the results may be attributable to an intrinsic T cell defect.
Idiotypic Analysis of Anti-GAT Antibodies
Authors: Ju Shyr-Te , Pierres Michel , Germain Ronald N. , Benacerraf Baruj , Dorf Martin E.
Published: December 2022

Previous studies demonstrated that anti-GAT antibodies of all inbred strains of mice share a common idiotype, termed CGAT idiotype. The CGAT idiotype is present on a major fraction of anti-GAT antibodies and is induced by the “GT”-related antigenic determinants on GAT molecules. The present paper describes another common idiotype of anti-GAT antibodies, termed GA-1 idiotype, the identification of which is based on a heterologous idiotypic interaction between guinea pig anti-idiotypic antiserum made against pooled specifically purified D1.LP anti-GAT antibodies and a monoclonal hybridoma anti-GAT antibody from the F9-102.2 cell line. The homologous anti-F9-102.2 idiotypic antiserum detected private idiotypic specificities.

GA-1 and CGAT are distinct idiotypes induced upon GAT immunization. Unlike CGAT idiotype, GA-1 idiotype is expressed on a minor fraction of the anti-GAT antibodies and “GA”-related antigenic determinants are responsible for the inducation of GA-1 idiotype. Thus, anti-GA, anti-GAT and anti-GLA40 antisera from 21 different inbred strains of mice expressed GA-1 idiotype. However, both the CE/J anti-GAT and anti-GA antisera lack detectable levels of GA-1 idiotype.
Experimental Malaria in the CBA/N Mouse
Authors: Jayawardena Anil N. , Janeway Charles A. , Kemp John D.
Published: December 2022

CBA/N mice carry an X-linked, recessive gene, which results in the absence of a B cell subset, and is expressed primarily as an inability to respond to a certain class of thymus-independent antigens. We have examined the responses of these mice to the malaria parasite Plasmodium yoelii and found that primary infections induced by this parasite are more severe and last longer in mice with the X-linked defect than in normal controls. The decreased resistance of the defective mice is associated with a striking deficiency in their IgM antibody response. After recovery from a primary infection, defective mice resist reinfection with the homologous parasite as well as normal mice. Although as resistant as normal controls, B cells from defective mice transfer considerably less immunity to naive recipients than B cells from normal animals. Hence, two modes of thymus-dependent protective immunity may contribute to the host response to P. yoelii. Control of an acute primary infection appears to involve a thymus-dependent antibody response that CBA/N mice are deficient in. Resistance to reinfection may be mediated primarily by a different mechanism.
Different Responses of B Lymphocyte Colony-Forming Cells in Bone Marrow and Spleen of Mice Injected with Bacterial Cell-Wall Components
Authors: Staber Fritz Georg , Layton Judith E. , Tarcsay Lajos
Published: December 2022

The responses of B lymphocyte colony-forming cells (BL-CFC) and hematopoietic colony-forming cells (HP-CFC) to three structurally unrelated chemically highly purified bacterial cell-wall components (BCWC) were studied in vivo. The total femoral BL-CFC content in C57BL/6 mice was found to be decreased approximately 70-fold 6 days after injection of lipid A and approximately 40-fold after injection of the outer membrane lipoprotein of Escherichia coli. Murein caused a much smaller decrease. In contrast to the marrow, the total splenic BL-CFC content did not change significantly after injection of any of the three BCWC. The response pattern of HP-CFC differed completely from that of BL-CFC. Although in the marrow only slight fluctuations were observed, the total spleen content of HP-CFC increased markedly after injection of either lipid A or lipoprotein but not after murein injection.

The decreased incidence of marrow BL-CFC was found to be maximal 4 to 6 days after injection, and normal levels of marrow BL-CFC were detectable again 2 weeks after injection. Inhibitory factors produced by marrow cells, especially prostaglandin E, appear not to be responsible for the decreased B lymphocyte colony-forming capacity of post-lipid A or post-lipoprotein marrow. The frequency of immunoglobulin-positive cells was also decreased in the marrow of mice injected with BCWC, but to an extent far too low to account completely for the decreased incidence of BL-CFC.
The Requirement of Ia-Positive Accessory Cells for the Induction of Hapten-Reactive Cytotoxic T Lymphocytes in vitro
Authors: Yamashita Uki , Hamaoka Toshiyuki
Published: December 2022

The antigen-presenting mechanism to hapten-reactive cytotoxic T cells was studied by using fractionated responder T cells, hapten-modified syngeneic stimulator cells, and accessory cells. When the effect of various treatments on the stimulatory activity by spleen cells in the induction of cytotoxic T cells was studied by coculturing TNP-modified spleen cells and nylon wool column-purified T cells, the stimulatory activity was associated with the spleen cell population adhering to plastic dishes, Sephadex G-10, and nylon wool, and was sensitive to anti-Ia serum and complement treatment. The activity was resistant to anti-Th B serum and anti-Thy 1 serum and complement treatment. In order to figure out the role of Ia antigens for induction of cytotoxic T cells, Ia-negative tumor cells were used as stimulator cells with TNP-modification, and the effect of addition of spleen cells containing Ia-positive cells to the system was studied. The addition of accessory cells restored the induction of TNP-reactive cytotoxic T cell activity under conditions in which the TNP-modified tumor cells alone could not induce cytotoxic T cells to stimulate nylon wool column-purified T cells. In this system, not only syngeneic but also allogeneic spleen cells had the accessory activity. Anti-Ia serum could inhibit the induction of cytotoxic T cells when added continuously to the culture, and this inhibition was specific for the accessory cells added, but not for the responder T cells or stimulator cells. Thus, Ia-positive accessory cells seem to play a crucial role in the induction of cytotoxic T cells in vitro. The possible role of Ia-positive accessory cells is discussed in the context of cellular interactions in the antigen-recognition mechanism of cytotoxic T cells.
Subpopulations of Human T Lymphocytes
Authors: Gupta Sudhir
Published: December 2022

Purified peripheral blood T lymphocytes were incubated with inducers of cyclic nucleotides and examined for the numbers of T cells with receptors for IgM (Tµ), IgG (Tγ), or IgA (Tα). Isoproterenol and theophylline, agents known to increase cAMP levels at 10-3 to 10-6 M concentrations, significantly decreased the number of Tµ cells but had no effect on Tγ or Tα cell numbers. This effect of isoproterenol could be completely blocked by a β-adrenergic blocking agent, propranolol. Receptors for IgM on Tµ cells regenerated after treatment with theophylline when cells were washed and further incubated at 37°C over a period of 12 to 24 hr in the absence of theophylline. Phenylephrine, at 10-3 to 10-6 M concentrations, significantly increased the numbers of Tµ cells but had no effect on Tγ or Tα cell numbers. The effect of phenylephrine could be completely blocked with an α-adrenergic blocking agent, phentolamine. The significance of the results are discussed.
Induction and Specificity of Antisera from Mice Immunized with Syngeneic UV-Induced Tumors
Authors: DeLuca Dominick , Kripke Margaret L. , Marchalonis John J.
Published: December 2022

Sera from mice inoculated with syngeneic fibrosarcomas induced by ultraviolet light (UV) were tested to determine if antibodies specific for the immunizing tumor were present. The assay used for this determination involved indirect immunofluorescence binding of anti-sera to tissue culture cells derived from the appropriate UV-induced tumor. The degree of binding of a given antiserum was determined by either fluorescence microscopy or quantitative cytofluorometry.

Antisera specific for the syngeneic UV-induced tumor used for immunization were produced. Binding by such antisera was tumor specific and was not due to the presence of contaminating antibodies directed against AKR murine leukemia viruses (MuLV). Only two instances of cross-reactivity were observed, and these involved allogeneic UV-induced tumors. These sera did not react with embryonic fibroblasts, tail skin epidermal cell lines, methylcholanthrene (MCA)-induced fibrosarcomas, or spontaneous fibrosarcomas. On the other hand, anti-UV-tumor sera, which also contained antibodies that react with MuLV, were cross-reactive with a number of tumors. Upon removal of anti-MuLV antibodies by appropriate adsorptions, the cross-reactive activity of these sera was lost, but the specific activity against the immunizing UV-induced tumor remained. We conclude that syngeneic mice challenged with UV-induced tumors can make a tumor-specific humoral immune response to individual tumors.
Regulation of Antibody Response in Different Immunoglobulin Classes
Authors: Kishimoto Tadamitsu , Hirai Yoshikatsu , Nakanishi Kenji , Azuma Ichiro , Nagamatsu Atsuo , Yamamura Yuichi
Published: December 2022

The suppressive effect of antigen-conjugated muramylpeptides or 6-O-mycoloyl muramylpeptides selectively on the induction of IgE antibody response was demonstrated. Preadministration of DNP-mycoloyl muramylpeptides completely inhibited the induction of the anti-DNP IgE antibody response with DNP-ovalbumin (DNP-OA). The selective suppression of the IgE response was due to the induction of DNP-specific suppressor T cells by DNP-mycoloyl muramylpeptides, and the suppressor cells were shown to be radiosensitive. Preadministration of OA-conjugated muramylpeptides partially inhibited the primary and secondary induction of an anti-OA IgE antibody response. The suppressor effect was also due to the induction of OA-specific suppressor T cells. Application of allergen-conjugated muramylpeptides as therapeutic agents in human allergic diseases was suggested.
Target-Effector Interaction in the Natural Killer Cell System
Authors: Roder John C. , Klein Margareta
Published: December 2022

Dibutyryl cAMP (dB-cAMP) and the cAMP elevating agents, prostaglandin E1, theophylline, and histamine markedly suppressed NK cytolytic function in a dose-and rate-dependent manner. The inhibition was rapidly induced and persisted in the presence of the drugs. Separate pretreatment of targets and highly purified NK cells, isolated by a target binding and velocity sedimentation technique, revealed that PGE1 and dB-cAMP acted at the level of the effector cell in a short-term cytolytic assay. In contrast to the inhibitory effects of cAMP elevating agents, dB-cGMP and carbamylcholine caused a small but significant acceleration in the rate of lysis and could compete with inhibitory doses of dB-cAMP to reduce the level of suppression thereby suggesting that the cAMP-cGMP ratio might be important in NK-mediated lysis. Insulin had no effect on NK activity, whereas T cell-mediated cytolysis was augmented by insulin and cGMP if the effector cells were taken early after alloimmunization but not later. Neither cAMP- nor cGMP-elevating agents affected the frequency of NK-target cell conjugates. These results are compatible with the hypothesis that cyclic nucleotides may be involved in triggering the lytic event within NK cells.
Slow Reacting Substance (SRS) from Ionophore A23187-Stimulated Human Leukemic Basophils
Authors: Yecies Lewis D. , Wedner H. James , Parker Charles W.
Published: December 2022

SRS was generated from human leukemic basophils upon stimulation with ionophore A23187. Radiolabel from [14C]-AA was incorporated into SRS with continued comigration of radioactivity and bioactivity through several chromatographic systems including DEAE-cellulose, silicic acid, and RP-HPLC. Human basophilic leukemia SRS displayed physiochemical properties similar to those of rat basophilic leukemia cell SRS.
Identification and Separation of Thy-1 Positive Mouse Spleen Cells Active in Natural Cytotoxicity and Antibody-Dependent Cell-Mediated Cytotoxicity
Authors: Mattes M. Jules , Sharrow Susan O. , Herberman Ronald B. , Holden Howard T.
Published: December 2022

The expression of the Thy-1 antigen on mouse spleen cells responsible for NK activity and ADCC was investigated by using a monoclonal IgM anti-Thy-1.2 antibody. Both C-mediated cytotoxicity and the fluorescence-activated cell sorter were used to fractionate cells. The effector cells were found to be heterogeneous in their expression of Thy-1. Effector cells from nude BALB/c mice were predominantly Thy-1 positive; some of the NK cells in CBA spleens appeared to be Thy-1 positive, but at least one-third of the lytic activity was due to Thy-1 negative cells. The effects of treatments on NK cytotoxicity and ADCC were very similar, supporting the hypothesis that the same cells mediate both activities.
Mechanism of Cell-Mediated Cytotoxicity at the Single Cell Level
Authors: Grimm Elizabeth , Bonavida Benjamin
Published: December 2022

A single cell method has been developed for directly measuring the frequency of cytotoxic T lymphocytes (CTL). The method is based on the ability of CTL to bind corresponding target cells and to form CTL-target conjugates visible by microscopy. Conjugates are incubated at 37°C in agarose to prevent CTL recycling. Single target lysis is assessed by trypan blue uptake. Target cell lysis was evident as early as 15 min and reached a plateau at 2 hr with an average of 60 to 80% of the conjugated target cells lysed. In a population of nylonwool nonadherent BALB/c peritoneal exudate T lymphocytes sensitized to C57BL/6 alloantigens, the frequency of CTL was estimated to range from 12 to 24% of the total T lymphocytes. The frequency of CTL and the kinetics of lysis were a function of the state of immunity, the method of sensitization, and the target cell employed. The single cell method described here has several features, namely, its independence of CTL recycling, the elimination of the variability in 51Cr release by target cells, and its prevision for a direct estimate of CTL frequency. Furthermore, the method is simple, reproducible, highly quantitative with very low numbers of CTL, and adaptable to non-T cell-mediated cytotoxic systems.
The biologic effects of allogeneic effect factor on T lymphocytes. I. The mitogenic activity and the autonomous induction of cytotoxic T lymphocytes by AEF.
Authors: Altman A , Cardenas J M , Bechtold T E , Katz D H
Published: December 2022

Studies presented herein illustrate the capacity of the soluble mediator, allogeneic effect factor (AEF), which is derived from histoincompatible cell interactions, to induce the in vitro differentiation of normal murine splenic lymphocytes into mature cytotoxic cells capable of exerting activity on H-2-identical target cells. This process requires the presence of T lymphocytes during the sensitization phase, and the lytic activity on tumor cells is mediated by cytotoxic T lymphocytes (CTL). The capacity of AEF to induce differentiation of such CTL does not require the presence of stimulating target cells in the sensitization phase. The induction of CTL requires the presence of AEF at the initiation of culture, although exposure to AEF as brief as 1 hr is sufficient to induce fresh spleen cells to differentiate into CTL during the subsequent 5 days in culture. In addition to its ability to induce CTL, AEF is highly mitogenic for T lymphocytes. However, the mitogenic and the CTL-inducing activities of AEF can be experimentally dissociated, indicating that different subpopulations of T lymphocytes may be involved in the response to AEF. In contrast to similar soluble helper factors derived from allogeneic cell interactions, AEF appears to be unique in its ability to autonomously induce a primary CTL response in vitro.
The protein-A plaque assay: a new system for the detection of cells secreting a given idiotype.
Authors: Hammarström L , Smith C I , Pettersson D , Mellstedt H , Holm G
Published: December 2022

With a modification of the protein-A plaque assay, cells secreting a given idiotype could be detected. Different anti-idiotypic antisera raised against the M-component of macroglobulinemia Waldenström patients were used. The antibodies did not cross-react with cell clones of other patients or normal controls. Spontaneous plaques, without prior cultivation, were mainly shown to be of IgM class, the majority being idiotype specific, in blood and bone marrow lymphocytes from these patients. This is in contrast to the predominance of Ig or IgA found in normal blood donors. A low but recognizable stimulation of the malignant clone may be observed when cells were stimulated by certain polyclonal B cell activators.
IgG B memory cell subpopulations: differences in susceptibility to stimulation by TI-1 and TI-2 antigens.
Authors: Tittle T V , Rittenberg M B
Published: December 2022

Spleen cells from mice primed with the thymus dependent (TD) antigen trinitrophenyl keyhole limpet hemocyanin several months earlier can be stimulated in vitro to produce an IgG anti-hapten response to TD as well as thymus independent (TI) forms of the hapten. Selective killing of TD or TI-2 responding B cells can be accomplished with the corresponding antigen by bromouridine deoxyribose (BUdR) and light treatment without affecting the other population. In contrast, we show here that selective killing with TI-1 antigens does not occur. Rather, the TI-1 antigens, TNP-Brucella abortus or TNP-lipopolysaccharide, eliminate both TD and TI-2 responding IgG memory B cells. All TNP-responding B cells are similarly eliminated if cultures are challenged simultaneously with TD and TI-2 antigens before BUdR and light but not when they are challenged with either a TD or TI-2 antigen separately. We conclude that IgG memory B cell precursors stimulated to produce anti-TNP by TD or TI-2 forms of the hapten are defined by only two functionally distinct subpopulations and that TI-1 antigens can stimulate both of these populations at least to divide.
Analysis of cytotoxic effector cell populations by kinetic and monolayer adsorption techniques.
Authors: Lynch D H , DeWitt C W
Published: December 2022

Cytolytically active lymphocytes were generated by immunizations between rat strains and evaluated by both kinetic and monolayer adsorption techniques. Kinetic analysis yields two values (Ka and Vmax), which are characteristic of interactions between individual effector and target cell populations. The Ka values this obtained correlate with the quantitative level of adsorption of CTL onto cellular monolayers, and we conclude that Ka is a measure of the binding avidity of effector and target cells.
The interaction of C5 with C3b in free solution: a sufficient condition for cleavage by a fluid phase C3/C5 convertase.
Authors: Isenman D E , Podack E R , Cooper N R
Published: December 2022

We have measured the interactions of C3b with C5 in free solution under conditions that favor detecting weak binding interactions (high C3b and low C5 concentrations and low ionic strength). When a mixture of 125I-C5 (2 X 10(-8) M) and unlabeled C3b (3.8 x 10(-5) M) was ultracentrifuged in a sucrose gradient, virtually all of the C5 sedimented to the position of a 13 to 14S complex. In contrast, a sedimentation rate for C5 of 9S was obtained in the absence of C3b. The ability to bind C5 was observed to be a property of C3b since native C3 was unable to bind C5. It was also found that beta 1H by itself could inhibit the binding of C5 to cell-bound C3b. From inhibition studies, we estimate that the association constant for the C3b-C5 interaction is on the order of 2 x 10(6) M-1 in a low ionic strength buffer (mu = 0.06) and 5-fold weaker at physiologic ionic strength. C5 bound to C3b in free solution was cleaved by nephritic factor-stabilized fluid phase C3bB. C5 activation did not occur on omitting C3b. We conclude that the ability to bind C5 is a property of C3b molecules whether surface bound or in free solution and that when C5 is bound in either fashion, it can be cleaved by a fluid phase C3/C5 convertase.
Characterization of lymphocyte responsiveness in early experimental syphilis. II. Nature of cellular infiltration and Treponema pallidum distribution in testicular lesions.
Authors: Lukehart S A , Baker-Zander S A , Lloyd R M , Sell S
Published: December 2022

The nature of the cellular infiltration and the distribution of Treponema pallidum during rabbit testicular infection were examined by immunofluorescence and light microscopy. Low numbers of treponemes are demonstrable in the perivascular regions on day 3 post-infection. On days 6, 10, and 13, large numbers of organisms are found in the interstitial spaces. The treponemes do not appear to invade the walls or lumina of the seminiferous tubules, although tubular atrophy is obvious. On days 17, 24 and 31, treponemes are no longer identifiable by immunofluorescence in infected testicles. The cellular infiltration, which is apparent on day 6, reaches its peak on day 13, corresponding to the amount of swelling observed grossly. The infiltrate is primarily lymphocytic, but macrophages are also observed during the peak cellular response. These cells are located, as are the treponemes, in the interstitial spaces. The lymphocytes are demonstrated by specific immunofluorescence to be predominantly T cells. The peak T cell infiltration in the infected testicle is followed rapidly by the disappearance of the organisms from that organ. Thus, it is postulated that infiltration by specifically sensitized T cells results in the clearance of large numbers of T. pallidum from infected tissues.
Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens.
Authors: Ozato K , Mayer N , Sachs D H
Published: December 2022

Hybridoma cell lines secreting antibodies to mouse H-2 or Ia antigens have been generated by fusing mouse immune lymphocytes with appropriate myeloma lines. Among the 11 established clones reported here, nine produce anti-H-2 antibodies and two produce anti-Ia antibodies. The specificities and cross-reactions of these monoclonal antibodies have been studied in detail. One hybridoma antibody reacted only to Kk antigens without any detectable cross-reactions, thus suggesting reaction to a private specificity of the Kk molecule. All other anti-H-2 hybridoma antibodies appeared to detect public specificities as defined either by reactions with products of more than one H-2 locus or with different alleles at one or more loci. The two anti-Ia antibodies both reacted with I-E/C products, but exhibited different cross-reactivity patterns. Strain distribution analyses so far indicate that the public specificities detected by these monoclonal antibodies are considerably different from those that had been established by traditional serology. Since public specificities defined by the hybridoma antibodies must by definition represent cross-reactions, these findings may have important implications relating to the structure and evolution of MHC gene products.
Contact hypersensitivity, humoral immunity, and specific unresponsiveness can be induced in Syrian hamsters with simple haptens.
Authors: Streilein J W , Sullivan S , Thompson S
Published: December 2022

Several inbred strains of Syrian hamsters have been immunized by skin painting with highly reactive haptens. Classical contact hypersensitivity has been assayed by the ear swelling response: the response is hapten-specific, exaggerated after re-immunization with the original hapten, and can be transferred adoptively to naive syngeneic hamsters with viable lymphoid cells, suggesting that contact hypersensitivity is mediated in hamsters by T lymphocytes. Moreover, skin painting with hapten induces significant serum titers of anti-hapten antibodies, indicating that antigen-specific B lymphocytes are also activated. Hamsters can be rendered unresponsive to these same haptens by conventional methods: 1) i.v. inoculation of the soluble hapten sulfonate or 2) inoculation of hapten-derivatized syngeneic lymphoid cells. Hamsters treated with these "tolerizing" maneuvers develop profound hapten-specific unresponsiveness that can be adoptively transferred to naive recipients with living lymphoid cells. "Unresponsive" animals, however, make strong anti-hapten antibody responses that rival the humoral immune responses found after skin sensitization. The data suggest that an active process is involved in the induction and maintenance of the unresponsive state, but responsibility can not be assigned firmly to putative suppressor T cells or to an antibody-mediated B suppressor modality.
The role of the spleen in tumor growth kinetics of the murine B cell leukemia (BCL1).
Authors: Slavin S , Morecki S , Weiss L
Published: December 2022

BCL1 is a transplantable B cell leukemia resembling human chronic lymphocytic leukemia-lymphoma maintained by cell passage in BALB/c mice. After BCL1 inoculation (10(7) cells), all mice developed extreme B lymphocytosis in the blood (less than or equal to 440,000 lymphocytes/mm3) and marked splenomegaly (50 times normal nucleated cell numbers). BCL1 infiltrated the spleen before peripheral leukemia was overt (3 days vs 28 days, respectively). BCL1 development in splenectomized mice was characterized by a delayed onset of leukemia (greater than 20,000 cells/ mm3 at 59 vs 28 days in intact mice), doubled median survival (102 vs 53 days, respectively), and reduced peak level of leukemic counts in the blood (125,000 vs 440,000 cells/mm3). Early splenectomy at different time intervals, ranging between 1 hr to 3 days after BCL1 inoculation, significantly delayed onset of the disease and prolonged survival, indicating that homing to the spleen occurred as early as 1 hr after inoculation. Splenectomy at 7 days still delayed onset of leukemia but did not affect survival. No significant effect on BCL1 kinetics was noticed when splenectomy was done on day 21. All splenectomized mice showed significantly lower peripheral blood counts as compared to intact mice (98,000/mm3 vs 440,000/mm3, respectively). The data show that the spleen plays a major role in the pathogenesis and prognosis of BCL1.
Cellular events in tolerance. VIII. Analysis of the proliferative capacity of antigen-binding cells isolated from normal or tolerant spleens.
Authors: Venkataraman M , Scott D W
Published: December 2022

Previous studies in our laboratory have demonstrated that hapten-binding lymphocytes (ABC) can be isolated from the spleens of mice rendered tolerant to the hapten. These cells are reduced in PFC precursor frequency when stimulated with either specific antigen or mitogen (Reference 6). In the present studies, we have combined antigen-specific cell isolation and an analysis of thymidine uptake by purified B cells to determine whether hapten-binding B cells in tolerant mice can be stimulated to proliferate in response to specific antigen. Compared with normal ABC, "tolerant" ABC showed a significantly reduced thymidine incorporation responsiveness in vitro. This suggests that unresponsiveness in adult mice can result in an impairment of clonal proliferation. Hence, these "tolerant" ABC appear to be functionally though not physically, deleted.
Induction and suppression of immunoglobulin synthesis in cultures of human lymphocytes: effects of pokeweed mitogen and Staphylococcus aureus Cowan I.
Authors: Pryjma J , Muñoz J , Galbraith R M , Fudenberg H H , Virella G
Published: December 2022

The synthesis and secretion of immunoglobulins by human peripheral blood mononuclear cells in cultures stimulated with pokeweed mitogen or Staphylococcus aureus Cowan I were evaluated by enumeration of cells containing cytoplasmic immunoglobulins and cells actively secreting immunoglobulins, and by quantitation of immunoglobulins released into culture supernatants. The two mitogens caused comparable stimulation of immunoglobulin production; however, in contrast to pokeweed mitogen, S. aureus was active in cultures depleted of T lymphocytes, and its stimulatory effects were resistant to the influence of suppressor T cells generated by co-stimulation with concanavalin A or by preincubation without mitogenic stimulus. These results indicate distinct pathways of induction and suppression of immunoglobulin synthesis for these two polyclonal B cell activators, and suggest that stimulation by S. aureus is less thymus dependent than that induced by pokeweed mitogen.