bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on Cytotoxicity Generated in Human Mixed Lymphocyte Cultures
Authors: Seeley Janet K. , Masucci Giuseppe , Poros Anna , Klein Eva , Golub Sidney H.
Published: December 2022

In addition to the allospecific killing detected against Con A blasts, 5-day MLC (X vs Ym) cells have a cytotoxic effect against K562 and the autologous (X) lymphoblastoid cell line (auto LCL). This effect is designated as anomalous killing or, AK. Both the AK and allospecific killing could be generated with purified E-rosetting cells, indicating that the effectors were T cells. However, they appeared to belong to different populations, based on three criteria: 1) Upon secondary challenge with the original stimulating cells, allospecific killing displayed a classical memory response. In contrast, peak AK activity against both K562 and the auto LCL was weaker in the secondary, compared to the primary culture. 2) The AK and allo targets did not cross-compete in cold target inhibition tests. 3) AK and allocytotoxicity distributed differently after fractionation of the MLC cells on immobilized immune complexes (IC plates). AK effectors may be similar to natural killer (NK) cells, as they preferentially kill K562 and other NK-sensitive targets. However, the anti-K562 effect was not entirely the result of persisting NK cells because the activity could be generated from Fc receptor-negative, NK-negative precursors. Furthermore, in contrast to NK cells, the AK effector cells could not be efficiently depleted by attachment to IC plates, suggesting that NK and AK differ with respect to expression of Fc receptors. The relationship between these two effects, as well as the conditions involved in activating AK, requires further clarification.
Cyclophosphamide and Cortisone Acetate Inhibit Complement Biosynthesis by Guinea Pig Bronchoalveolar Macrophages
Authors: Pennington James E. , Matthews Wallace J. , Marino Joseph T. , Colten Harvey R.
Published: December 2022

To explore mechanisms of drug-induced alterations in local host defenses in the lung, the capacity of guinea pig alveolar macrophages to synthesize the second (C2) and fourth (C4) components of complement was studied following treatment with cyclophosphamide and cortisone acetate. Administration of either drug significantly inhibited biosynthesis of C2 and C4 within 1 day. After 1 week of treatment, local complement synthesis was inhibited approximately 80%, although serum levels of the corresponding proteins were normal.
Immunosuppression Induced by Avian Reticuloendotheliosis Virus: Mechanism of Induction of the Suppressor Cell
Authors: Rup Bonita J. , Spence Jean L. , Hoelzer Jacalyn D. , Lewis Robert B. , Carpenter Charles R. , Rubin Arnold S. , Bose Henry R.
Published: December 2022

Spleen cells from chickens infected with avian reticuloendotheliosis virus (REV) become cytostatically impaired in their ability to undergo phytohemagglutinin-P (PHA)-induced blastogenesis. This inhibition of T lymphocyte proliferation is mediated by a host-derived population of suppressor cells that are activated or induced during REV infection. Oncogenic preparations of REV consist of a replication-defective transforming virus and a nontransforming, replication-competent helper virus, designated reticuloendotheliosis-associated virus (REV-A). Infection of chickens with REV-A resulted in suppression of the PHA-induced responses of spleen cells from infected birds, and a population of suppressor cells was detected in their spleens. These suppressor cells could no longer be detected 3 weeks after REV-A infection. The loss of suppressor cell activity in the spleens of REV-A infected birds was correlated temporally with the loss of detectable virus in the serum of infected birds. The immunosuppression induced by REV-A was transient since PHA-induced lymphocyte responses of infected birds returned to normal by 5 weeks after infection. Cells transformed in vitro by the replication-defective transforming virus in the absence of its helper virus, which fail to produce either infectious or uninfectious virus particles, were capable of inducing immunosuppression and reticuloendotheliosis. Cell doses of both REV-transformed, nonvirus-producing cells and REV-transformed, nonvirus-producing cells and REV-transformed virus-producing cells that were effective in inducing immunosuppression also infuced lethal reticuloendotheliosis. Antiserum directed against the major envelope glycoprotein of REV-A lysed these cells in the presence of complement, indicating that the REV-transformed, nonvirus-producing cells contain either virion glycoproteins or their precursors on the surface membranes. Chick syncytial virus, duck infectious anemia virus, and spleen necrosis virus, other nontransforming members of the reticuloendotheliosis (RE) virus group, which share cross-reactive glycoproteins with REV, also induced immunosuppression in chickens within 6 days after infection. These results suggest that the viral glycoproteins or their precursors, expressed on the membranes of RE-group virus-infected cells, may be responsible for induction of a host-derived population of suppressor cells. The possibility that other viral or host specified polypeptides may be involved in the induction or activation of the suppressor cell during avian reticuloendotheliosis cannot be ruled out. The early induction of these suppressor cells appears to play an important role in the pathogenesis of avian reticuloendotheliosis.
Ultrastructural Demonstration of Calcium Loss from Local Regions of the Plasma Membrane of Surface-Stimulated Human Granulocytes
Authors: Hoffstein Sylvia T.
Published: December 2022

Most cell-associated calcium exists as bound rather than free ionized calcium. Since calcium and other cations are important regulators of polymorphonuclear leukocyte (PMN) function, the localization of bound calcium in resting PMN and its redistribution after exposure to a variety of immune stimuli was investigated by means of ultrastructural cytochemistry. Potassium pyroantimonate was used to precipitate cations in situ in some experiments; in others, cells were loaded with exogenous calcium during fixation to localize binding sites. Both methods resulted in the appearance of discrete electrondense deposits along the plasmalemma of resting cells. The deposits could in both cases be removed with 3 mM EGTA as well as EDTA. Additional pyroantimonate was precipitated primarily in the heterochromatin regions of the nucleus, and in azurophile granules, but not in the cell sap. Preincubation of PMN with particulate stimuli, such as opsonized zymosan, immune complex precipitates, or Con A covalently bound to Sepharose beads, resulted in the rapid (less than 10 sec) segmental loss of pyroantimonate precipitable cations from regions of plasma membrane that were in close apposition to the bound particle but not from more distant areas of membrane. The distribution of precipitates in the remainder of the cell, including regions of plasmalemma not in contact with particles, was unaltered. Exposure of the entire cell surface to soluble stimuli such as Con A, the chemotactic peptide F-met-leu-phe, or the calcium ionophore A23187, resulted in the circumferential loss of pyroantimonate precipitable calcium from the entire plasma membrane but not from any intracellular location. The data suggest that cations are associated with the plasma membrane of resting PMN in concentrations sufficiently high to precipitate pyroantimonate and that these membrane-associated cations are released from sites of receptor-ligand interaction.
Non-H-2 Linked Control of Low Versus High Responses of Antigen-Induced Lymph Node Cell Proliferation: Possible Role for Antigen-Presenting Cells
Authors: Suzuki Koji , Garrison Fathman C. , Tomasi Thomas B.
Published: December 2022

Quantitative differences in the magnitude of antigen-induced proliferative responses of sensitized lymph node cells between low (C3H/Anf or C3H/Cr) and high (C3H/Hej or CBA/j) responder H-2k mice have been observed 1 to 2 weeks after in vivo sensitization with antigen (OVA, PPD, and GAT). We have shown that antigen presentation is less effective in the sensitized lymph node cell populations from low responder mice compared with those from high responder mice, suggesting that the number and/or functional status of antigen-presenting cells in the regional lymph nodes may be a key factor in determining the magnitude of antigen-induced proliferative responses. These data are consistent with the hypothesis that cell traffic after sensitization plays an important role in determining the immune responsiveness of lymph nodes.
The Immune Response of Lewis Rats to Peptide 68–88 of Guinea Pig Myelin Basic Protein
Authors: Jen Chou C.-H. , Fritz Robert B. , Chou Frank C.-H. , Kibler Robert F.
Published: December 2022

A series of peptides, produced from peptide 68–88 by selective enzyme cleavage, were used to define the amino acid sequences required for the induction of experimental allergic encephalomyelitis (EAE), an in vitro lymphocyte proliferative response (LPR), and serum antibody by peptide 68–88 in the Lewis rat. Here we present data that indicate that the T cell determinants for induction of EAE, an LPR, and helper function in the production of antibody are located in the same region of the molecule and that a minimum of 13 amino acids are involved; i.e., residues 71–85.
Modulation of the Host Response in Human Schistosomiasis
Authors: Ottesen Eric A.
Published: December 2022

Previous studies have demonstrated that peripheral blood mononuclear cells (PBMC) from patients with chronic schistosomiasis are unable to generate strong in vitro proliferative responses to parasite antigens but that responses to mitogens and nonschistosome antigens are normal. In the present study PBMC from eight patients with schistosomiasis mansoni and five normal individuals were fractionated with nylon wool columns to remove adherent cells. Whereas the removal of such cells regularly diminished the proliferative responses of normals to both antigens and mitogens, in patients with chronic schistosomiasis, removal of adherent cells significantly enhanced the responsiveness to parasite antigens (p < 0.05). The enhancement was specific for schistosome antigens (derived from the cercarial, adult, and egg states of the parasite) and did not extend to nonparasite antigens or mitogens. The suppression found originally in the unfractionated PBMC could be consistently reinduced in vitro by the addition of adherent to nonadherent cells of these patients.

Studies on a single patient evaluated during both acute and chronic stages of S. mansoni infection showed the absence of adherent suppressor cells early in infection, but they showed development of such cells by 20 months. Thus, our observations demonstrate clearly that adherent suppressor cells are a regular constituent of the PBMC of patients with chronic schistosomiasis and that the development of such cells may be partially responsible for the progressive modulation of specific lymphocyte responsiveness to parasite antigens seen in these patients as their acute infections advance to more chronic stages.
M467: A Murine IgA Myeloma Protein that Binds A Bacterial Protein
Authors: Mason Smith A. , Miller Jeffery S. , Whitehead Dan S.
Published: December 2022

We have studied the binding of M467, an IgA murine myeloma protein, to flagellin from seven species of Salmonella. It was found that M467 was reacting with antigenic determinants that were common to all the flagellins studied. These determinants were not related to serotypic antigens. Electronmicrographs of unreduced M467 showed a variety of polymeric species bound to flagella in a manner than could produce immobilization as well as agglutination and precipitation through cross-linking of antigenic determinants. Immunodiffusion in agar gel revealed that M467 was recognizing more than one group of peptide determinants on the flagellins studied. Passive hemagglutination inhibition and a solid phase radioimmunoassay provided evidence that there were differences in binding avidities between M467 and the various Salmonella flagellins studied. It was concluded that M467 is binding more than one specific group of antigenic peptide determinants on flagellin molecules. Flagellin from four of the seven species of Salmonella studied were deficient in one or more of these determinants.
Effects of Human Leukocyte Dialysates Containing Transfer Factor in the Direct Leukocyte Migration Inhibition (LMI) Assay
Authors: Borkowsky William , Lawrence H. S.
Published: December 2022

We report on an evaluation of the direct leukocyte migration inhibition (LMI) test as a potential in vitro assay of Transfer Factor activity. Crude dialysates of human leukocyte extracts containing Transfer Factor (DLE) were observed to cause both antigen-dependent and antigen-independent effects on the migration of nonimmune leukocyte populations.

We find that only those DLE preparations that are obtained from donors with intense cutaneous DTH reactions to antigen have the capacity to cause antigen-specific effects consistently, and DLE obtained from donors with moderate or absent cutaneous reactivity are generally not active in this regard. Additionally, this antigen-specific activity of DLE is dose dependent. These two variables are consonant with similar requirements for successful in vivo transfer of cutaneous DTH.

The pulsing of nonimmune migrating cell populations with immune DLE for 30 to 60 min is an additional requirement for maximum effects to be achieved. When these experimental conditions are met, there is a strong suggestion that antigen-specific inhibition of migration can be conferred by DLE on nonimmune cell populations, which is concordant with the pattern of cutaneous DTH reactions expressed by the donor of DLE. The data suggest further that the component(s) of DLE responsible for this activity resides in a >3500 m.w. dialysis fraction and it does not behave like a superantigen or a chemotactant in this assay system.

When immune DLE is incubated with immune leukocyte populations, the anticipated inhibition of migration in the presence of antigen is not augmented but is abrogated instead. The antigen specificity of such suppressor-like activity is currently under investigation.

We conclude that the LMI test appears to provide a suitable in vitro assay to investigate further the nature and mechanism of action of DLE and its components and provide a means of separating the moiety or moieties responsible for antigen-specific effects before in vivo testing.
Promonocytes Have the Functional Characteristics of Natural Killer Cells
Authors: Lohmann-Matthes Marie-Luise , Domzig Wolfgang , Roder John
Published: December 2022

Promonocytes isolated from a 5-day-old L-fibroblast-conditioned liquid bone marrow culture show strong NK cell cytotoxicity. They kill YAC target cells in a short-term 125IUDR-release assay whereas P815 targets are unaffected. This NK-like cytotoxicity is enhanced in the presence of interferon preparations. Morphologically, these promonocytes resemble a medium size lymphocyte with a high nucleus to cytoplasma ratio, they are non-adherent, nonphagocytic, and negative in nonspecific esterase staining. Promonocytes are precursor cells from macrophages, which have not yet developed the typical macrophage criteria. Within 24 to 48 hr they mature to adherent macrophages. We have shown previously, that the same promonocytes have the capacity to perform K cell killing of antibody-coated tumor target cells. The cytotoxic effector functions of promonocytes are abolished when the cells are treated with the alloantimacrophage serum Mø 1.2 plus rabbit C. The relationship or similarity between K cells, NK cells and promonocytes is discussed.
Role of C3 in the regulation of a splenic PFC response in rabbits.
Authors: Romball C G , Ulevitch R J , Weigle W O
Published: December 2022

The effects of in vivo C3 depletion on the immune response were examined in rabbits by assaying for splenic PFC after immunizing normal or cobra venom factor-treated animals with aggregated human gamma-globulin. The response to this T-dependent antigen has previously been shown to be regulated such that several cycles of PFC appear following a single intravenous injection of antigen. C3 depletion had no effect on the first peak of PFC (appearing 5 days after injection), but resulted in depression of the second peak of PFC (day 13). In rabbits depleted of C3, antigen localization in splenic germinal centers was markedly decreased. Delaying C3 depletion until after antigen localization had occurred resulted in no depression of the second peak of PFC. These results suggest that one mechanism by which C3 affects immune responses in vivo is via its role in influencing the persistence of antigen. In the absence of C3, no significant localization of antigen occurs, resulting in interference with the cyclical production of antibody.
Generation of H-2-reactive T cell lines that bear the 5936 idiotype(s).
Authors: Rubin B , Golstein P , Nordfang O , Hertel-Wulff B
Published: December 2022

The present experiments showed 1) that it was possible to produce mouse T cell lines against MHC determinants with a relatively high success rate by stimulation of purified T cells with allogeneic cells in the presence of irradiated syngeneic spleen cells; 2) that these lines could be led to react against selected H-2 specificities; 3) that only T cell lines established from Ig-1b allotype mice contained 5936-Id+ T cells (5936-Idiotypes are defined by an antiserum against B6 anti-CBA IgG produced in rabbit no 5936, which was tolerant to mouse gamma-globulin); and 4) that antigenic determinants coded by IAk genes induce the 5936-Idiotype(s). The latter data are in accordance with the 5936-idiotype characteristics of primary MLC T blasts. All T cell lines contained both specific MLC-responding cells and cytolytic cells. However, studies on the functional capacity of 5936-Id+ T cells from both primary MLC and the T cell lines showed that neither MLC-responding cells nor cytolytic cells directed against H-2Kk, IAk, or H-2Dk were 5936-Id+. Thus, 5936-Id+ T cells may be regulator cells induced by IAk antigens.
Genetic control of the immune response of mice to highly dinitrophenylated human gamma-globulin (DNP56HGG).
Authors: Newton R C , Warner C M
Published: December 2022

The immune response to highly dinitrophenylated human gamma-globulin (DNP56HGG) was tested in inbred strains of mice. Significant differences in the anti-DNP response among inbred strains were found, including the magnitude of serum antibody and the location of plaque-forming cells (spleen or lymph nodes). The strain differences persisted when the dose and adjuvant were changed. The genetic control of the anti-DNP response to DNP56HGG was investigated. The analysis of the response of congenic and F1 hybrid mice to DNP56HGG suggests that at least two genes are involved in the control of the anti-DNP response. The two genes are demonstrated by complementation in the F1 generation, and show no correlation with H-2 haplotype or IgG2a allotype. A third gene may be implicated by differences in response observed between male and female mice.
Dominance of an immune response by secondary cells: quantitation by allotype analysis.
Authors: Brown A R , DeWitt C L , Bosma M J , Nisonoff A
Published: December 2022

Dominance of an immune response by secondary cells was demonstrated in a cell transfer system utilizing allotype-congenic mice. Spleen cells from BALB/c mice immunized with TNP-KLH were adoptively transferred into sublethally irradiation (200 R) congenic partners of a different allotype (C.B.-17 mice). Subsequent to the cell transfer, the recipients were challenged with TNP-KLH and bled periodically. Anti-TNP antibodies were isolated and analyzed for their content of donor and host IgG allotypes. Through day 58 after the adoptive transfer, 80% or more of the IgG antibody measured was of donor allotype. By day 110, most but not all recipients made IgG antibody principally of host allotype. In contrast, control C.B-17 recipients, which received nonimmune BALB/c spleen cells but otherwise treated as above, produced IgG antibodies almost entirely of host allotype. Total anti-TNP synthesis was reduced markedly in recipients of immune BALB/c cells so that there was a decrease of about 95% in the production of anti-TNP antibodies of host allotype, as compared with controls that received nonimmune cells. The results are discussed in terms of dominance of an immune response by secondary B cells.
Mechanisms of regulation of cell-mediated immunity. VI. Antigen density dependence of the induction of genetically restricted suppressor cells.
Authors: Pierres A , Bromberg J S , Sy M S , Benacerraf B , Greene M I
Published: December 2022

We have investigated the relationship of antigen density on cell surfaces to the induction of genetically restricted suppressor T cells (Ts). It was found that Ts able to suppress the development of hapten-specific contact sensitivity were induced by the i.v. inoculation of trinitrobenzene sulfonate (TNBS) hapten-coupled cells. The Ts induced by this technique were found to be gentically unrestricted in terms of generation or expression when 10 mM TNBS were used to prepare the hapten-coupled cells. Transferable suppression could be obtained by 10 mM coupled allogeneic cells or, alternatively, TNP-coupled Ia- syngeneic erythrocytes or even H-2-negative syngeneic tumor cells. However, 1 mM TNBS-derivatized hapten-coupled cells induced Ts that were able to suppress only recipients syngeneic with the tolerogen. The significance of these findings to our understanding of the induction and expression of Ts and the triggering signals that are necessary for their activation are discussed.
Induction of prostaglandin E release from macrophages by colchicine.
Authors: Gemsa D , Kramer W , Brenner M , Till G , Resch K
Published: December 2022

Rat peritoneal macrophages released high amounts of prostaglandin E (PGE) when treated in vitro with 10(-7) to 10(-4) M colchicine. PGE production occurred after a lag period of 4 hr and proceeded at a constant rate for more than 24 hr. Lymphocytes could not be stimulated to PGE release by colchicine. Disaggregation of microtubules appeared to be an essential event, since lumicolchicine was inactive and addition of heavy water (D2O) abolished colchicine-induced PGE formation. Cytochalasin B (5 microgram/ ml) did not interfere with PGE production by colchicine during the initial 12 hr, but thereafter it gave rise to an activity capable of degrading or converting newly synthesized PGE. Although details of the mechanisms by which colchicine in association with disrupted microtubules may induce PGE release remain unclear, these observations suggest that components of the cytoskeleton may efficiently influence the biosynthesis of prostaglandins.
Cytolytic T lymphocyte precursors reactive against mutant Kb alloantigens are as frequent as those reactive against a whole foreign haplotype.
Authors: Widmer M B , MacDonald H R
Published: December 2022

Minimal frequencies of CTL-P in C57BL/6 spleen responding to H-2 mutational or whole haplotype differences were estimated by using a limiting dilution MLC technique. The frequency of CTL-P reacting against cells from each of three different H-2Kb mutant strains, carrying H-2 haplotypes ba, bg2, or bh, was determined to be within the range of frequencies of CTL-P directed against cells from strains carrying the independently derived H-2d and H-2k haplotypes.
Radioimmunoassay for the P2 protein of bovine peripheral nerve myelin.
Authors: Sarvas H O , Milek D J , Weise M J , Carnow T B , Fudenberg H H , Brostoff S W
Published: December 2022

The P2 protein of bovine nerve root myelin was radiolabeled with 125I in homogeneous solution by the chloramine-T method and purified by gel filtration on Sephadex G-75 to obtain monomeric 125I-P2. Antigen-antibody complexes were isolated by the silica gel, double antibody, or polyethylene glycol techniques by using rabbit antibody to P2. As little as 1 ng/ml P2 could be detected. The RIA was used to measure the P2 content in nerve tissue and isolated myelin. The presence of P2 in spinal cord as well as in the peripheral nervous system was confirmed. Peptides isolated by CNBr digestion of the P2 protein were tested in the RIA. CN-1, comprising 80 to 90 residues from the interior of the molecule displayed complete immunologic cross-reactivity with intact P2. Neither CN-2, representing 18 amino acids from the COOH terminal, nor CN-3, representing 20 amino acids from the NH2 terminal, showed cross-reactivity. Since the major determinant for experimental allergic neuritis in the rabbit is located in peptide CN-2, our present data suggest that the major neuritogen and the major determinant(s) for humoral antibody response in this species may be at different locations within the P2 molecule.
Concentration of IgG1 and IgG2a allotypes in serum of nude and normal allotype-congenic mice.
Authors: Bosma G C , Owen J , Easton G , Marshall G , Dewitt C , Bosma M J
Published: December 2022

BALB/c (C) and C57BL/6 (B6) allotype--congenic strains of mice (C, C.B, B6, B6.C) were made congenitally anthymic by the introduction of the mutant nude gene (nu). The resulting nude mice (C.nu, C.B.nu, B6.nu and B6.C.nu) were found to produce highly variable quantities of IgG1 and IgG2a. Whether these mice showed high, normal, or low quantities of IgG1 or IgG2a relative to their normal counterparts depended on their age and genetic background. In all nude mice, IgG1 production appeared more sensitive to the loss of T cells than did IgG2a production. An extreme example of this was the deficiency of IgG1 but not IgG2a in adult B6.nu and B6.C.nu mice. Substantial quantities of both IgG1 and IgG2a were found in adult C.nu and C.B.nu mice.
Cytotoxic T cells generated in the autologous mixed lymphocyte reaction. I. Primary autologous mixed lymphocyte reaction.
Authors: Tomonari K
Published: December 2022

In the course of the culture of an autologous mixed lymphocyte reaction (AMLR), T cells proliferated in response to autologous non-T cells, and differentiated to cytotoxic T cells (AMLR killers). DNA synthesis was necessary to generate AMLR killers, as the elimination of autoreactive proliferating cells with BUdR and UV light completely abrogated AMLR killer cytolysis. Amlr killers lysed various lymphoid cell lines, including autologous B cell lines, autologous or allogeneic mitogen blasts stimulated by Con A, PHA, or pokeweed mitogen, variious nonlymphoid cell lines derived from human, mouse, or rat, and weakly normal autologous or allogeneic non-T cells. KMT-17, methylcholanthrene-induced rat fibrosarcoma, was the only resistant cell line to have been tested. AMLR killers had characteristics similar to NK cells, Major histocompatibility antigens were not the target antigens for AMLR killers. AMLR killers distinguished the blasts stimulated by alloantigens as self from the blasts stimulated by mitogens as non-self.
Hybridoma autoantibodies to DNA.
Authors: Andrzejewski C , Stollar B D , Lalor T M , Schwartz R S
Published: December 2022

MRL/1 mice develop a severe form of systemic lupus erythematosus and spontaneously produce high levels of autoantibodies to DNA. Spleen cells from these animals were used as fusion partners to prepare hybridomas that produce autoantibodies to DNA. Since exogenous immunization was not employed, the hybridoma antibodies were derived from populations of B cells that spontaneously produced autoantibodies. Each hybridoma autoantibody had a distinctive binding specificity with S-DNA, N-DNA, or oligonucleotides.
Therapy of leukemia by nonimmune syngeneic spleen cells.
Authors: Cheever M A , Greenberg P D , Fefer A
Published: December 2022

In previous studies mice bearing an established transplanted syngeneic leukemia were cured by the adoptive transfer of immune lymphoid cells inoculated as an adjunct to chemotherapy, whereas nonimmune cells had no effect. By employing a smaller initial tumor inoculum to enhance the sensitivity of the chemoimmunotherapy model, the current study demonstrated that cells from normal nonimmune donors can also eradicate tumor. Thus, adult BALB/c mice given a lethal dose (1 x 10(4)) of LSTRA on day 0 were treated on day 5 with cyclophosphamide (CY) alone or CY plus nonimmune cells. Untreated mice all died by day 10. Cy alone prolonged median survival time (MST) to day 20, but all mice died. Treatment with CY plus 1 x 10(7) nonimmune cells prolonged the MST to day 26 and cured 16 of 48 mice. The effector cells, present in the spleens of normal and nude mice, were nonadherent lymphocytes resistant to anti Thy + C. These results are consistent with this effector cell in adoptive chemoimmunotherapy being an NK cell. However, they are distinct from classical NK cells since they are not directly cytotoxic in vitro in a short term CRA and do not neutralize tumor in vivo in a Winn assay.
Analysis of the heterogeneity of the mouse H-2K, D, and L gene products.
Authors: Krakauer T , Hansen T H , Camerini-Otero R D , Sachs D H
Published: December 2022

The detection and partial characterization of the major histocompatibility antigens encoded by the three H-2 loci, K, D, and L were performed by analyzing immunoprecipitates from detergent-solubilized radiolabeled cells on two-dimensional gels. Each locus was found to code for a population of proteins heterogeneous in both charge and m.w. Only part of the heterogeneity was shown to be due to sialylation as demonstrated by shifts in mobility after neuraminidase treatment of antigen preparations. The 2-D gel analysis of the H-2K, H-2D, and H-2L antigens of the d haplotype shows that each constitutes a distinct series of products.
Multiple myeloma: an immunologic profile. Cytotoxic and suppressive effects of the EA rosette-forming cell.
Authors: Paglieroni T , Mackenzie M R
Published: December 2022

Lymphoid cells bearing receptors for human IgG anti-Rh antibody but not otherwise identifiable as T or B cells were isolated from the peripheral blood of human multiple myeloma patients. These cells do not demonstrate surface immunoglobulin, do not secrete immunoglobulin in culture after stimulation with pokeweed mitogen, and do not possess receptors for the third component of complement. They do not form rosettes with sheep erythrocytes, and do not respond to known T cell mitogens. These cells are not phagocytic and do not possess demonstrable nonspecific esterase. Thus, they must be considered at present, members of the "third population" lymphoid cells. These cells are active in the cytotoxic response by myeloma peripheral blood lymphocytes to myeloma plasma cells accounting for approximately 60% of such activity. They are also responsible for part of the suppression of pokeweed mitogen-stimulated immunoglobulin production, and at equivalent suppressor to target cell ratios are more active than immunoregulatory monocytes. Thus, these cells play an important, not previously widely appreciated role in the pathophysiology of the human myeloma patient.
Detection and characterization of membrane antigens of Toxoplasma gondii.
Authors: Handman E , Goding J W , Remington J S
Published: December 2022

Toxoplasma gondii tachyzoites were surface radioiodinated by the lactoperoxidase technique, and the solubilized membrane proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis. Four major labeled proteins with apparent m.w. of 43,000, 35,000, 27,000, and 14,000 were detected. None of the radioiodinated proteins bound to concanavalin A-Sepharose. When a panel of eight different fluorescein-conjugated lectins was used in an attempt to characterize further the nature of the cell membrane, none of the lectins bound to intact tachyzoites. Two-dimensional polyacrylamide gel electrophoresis did not reveal any significant differences among three different strains of Toxoplasma. Each of the radioiodinated surface proteins was precipitable by sera from mice chronically infected with the same strain as well as by a series of sera from mice infected with other strains. Sera from humans with acute Toxoplasma infection showed more variability in that some precipitated all labeled proteins whereas others precipitated only two or three of them. Monoclonal antibodies (2G11 and 3E6) prepared by hybridization of spleen cells from Toxoplasma-immune mice with myeloma cells consistently precipitated both the solubilized 35,000 and 14,000 dalton proteins, whereas 1E3 precipitated the 43,000-dalton protein and 1E11 the 27,000-dalton protein.
Natural regulatory cells in murine bone marrow: inhibition of in vitro proliferative and cytotoxic responses to alloantigens.
Authors: Dorshkind K , Klimpel G R , Rosse C
Published: December 2022

A lymphocyte-enriched fraction of murine bone marrow (BML), obtained by sucrose density fractionation, contains natural regulatory cells that can profoundly suppress the proliferative and cytotoxic response of syngeneic lymph node cells to irradiated alloantigens in a mixed lymphocyte culture (MLC). A close correlation exists between the inhibition of alloantigen-induced proliferation and the generation of cytotoxic effectors. The suppression of proliferation is dependent on the dose of BML added to the cultures but is not due to cell crowding, since red blood cells, thymocytes, and irradiated splenocytes, all syngeneic to the lymph node responders, do not inhibit proliferation to the degree observed with BML. The addition of BML to cultures does not cause the maximum proliferative response to change from the usual day 5 peak, indicating that there is no change in culture kinetics. The release of nonradioactive thymidine by BML cannot explain the suppression. The target of suppression is maximally affected during the first 24 hr of culture, since adding BML to MLC later than this resulted in negligible inhibition of proliferation. Thus, the natural regulatory cell-mediated suppression reflects inhibition of "early" events in the proliferative response to alloantigens.
Genetic control of the immune response to ferredoxin: linkage and mapping of T cell proliferation and antibody production genes to the MHC of mice.
Authors: Sikora L K , Levy J G
Published: December 2022

The genetics of the immune response in the mouse were studied by using the antigenically simple, stable, naturally occurring protein ferredoxin (Fd) from Clostridium pasteurianum. The immune status of mice primed and boosted with Fd was assessed by using two parameters of immunity: T cell proliferation and serum antibody production with the ELISA method. In both assay systems, the response has been shown to be H-2 linked: k, b, and s haplotypes respond to Fd, and H-2d mice are nonresponders. It is apparent that different immunoregulatory events modulate the response in the responder strains; these factors become evident in the recombinant analysis of the response and to date an immunoregulatory gene(s) has been mapped to at least the K/I-A subregions. F1 analysis demonstrated a gene dose-dependent response of the strains studied.
Lymphoid procoagulant activity and mitogenesis in the C3H/HeJ mouse: discordant response to lipopolysaccharide stimulation.
Authors: Levy G A , Edgington T S
Published: December 2022

Lymphoid cells from the C3H/HeJ mouse respond abnormally to bacterial lipopolysaccharide (LPS) in a number of in vitro functions. Abnormalities in both macrophage and lymphocyte populations have been suggested. In the present study, splenic mononuclear cells were isolated from C3H/HeJ, C3H/St, A/J, and BALB/c mice, and analyzed for mitogenesis and induction of procoagulant activity (PCA) by LPS prepared by either phenol or butanol extraction methods. All strains responded vigorously with resultant increased 3H thymidine uptake to Con A and PHA, and all but the C3H/HeJ responded well to butanol or phenol extracted LPS. However, the C3H/HeJ was unresponsive to phenol-extracted LPS and exhibited a poor but significant response to butanol-extracted LPS. In contrast, lymphoid cells from all strains, including the C3H/HeJ, exhibited a marked and equal increase of PCA in response to both phenol- and butanol-extracted LPS by using both splenic and peripheral blood lymphoid cells. The results obtained indicate that although the C3H/HeJ mouse may be unresponsive to a number of LPS-stimulated functions, there is no associated defect in PCA generation, a lymphocyte collaboration-dependent event.
Histocompatibility-2 system in wild mice. X. Frequencies of H-2 and Ia antigens in wild mice from Europe and Africa.
Authors: Götze D , Nadeau J , Wakeland E K , Berry R J , Bonhomme F , Egorov I K , Hjorth J P , Hoogstraal H , Vives J , Winking H , Klein J
Published: December 2022

In this study, data are presented on serologic H-2 typing of 320 wild mice collected in several parts of Europe and Egypt. The sample was typed for 39 class I (K, D) and 16 class II (Ia) antigens. The phenotype frequencies of class I and class II antigens showed high variability in their distribution among different areas, ranging from absence or presence in low frequency in one area to presence in about one-half of the mice in another area. The average phenotypic frequency of class I private antigens was 6.3%; at least 60% of class I alleles (blanks) could not be identified with the available reagents. The data suggest that there might be more than 100 alleles for each class I locus, H-2K and H-2D. The average phenotypic frequencies of Ia-1 private antigens was 10.8%. About 75% of Ia-1 alleles (blanks) could not be identified. The number of Ia-1 alleles was estimated to be in the range of 20 to 50.
Inherited 7S immunoglobulin deficiency in chickens: presence of suppressor T cells that suppress synthesis of 7S immunoglobulin but not IgM.
Authors: Chanh T C , Benedict A A , Tam L Q , Abplanalp H , Gershwin M E
Published: December 2022

University of California (UCD) line 140 chickens, previously described to have an inherited 7S Ig deficiency and dysgammaglobulinemia associated with early bursal defects, and autoimmune phenomena were examined for the presence of suppressor cells. Pokeweed mitogen (PWM) stimulated UCD 140 peripheral blood lymphocytes (PBL) to synthesize IgM but not 7S Ig. In co-culture, UCD 140 PBL specifically inhibited 7S Ig but not IgM synthesis of PWM-stimulated PBL from normal birds. Normal 7S Ig synthesis usually was suppressed 40 to 80%, although PBL from some normal birds were not suppressible. Adoptive transfer of UCD 140 spleen cells to normal chicks resulted in early and transient decreased serum 7S Ig levels. Homology at the B locus (major histocompatibility complex) was not required for suppression. Higher levels of suppressor cells were found in UCD 140 than in normal birds as early as 13 days of age. The suppressor cells were found to be T cells as indicated by inactivation with anti-T antiserum but not with anti-B antiserum, and by suppression associated with T cell fractions obtained by nylon wool fractionation. UCD 140 B cells, obtained by nylon wool fractionation, were stimulated with PWM to synthesize 7S Ig when provided with normal T cells. These findings are discussed with particular reference to early bursal anomalies.
Aggregated human gamma-globulin-induced proliferation and polyclonal activation of murine B lymphocytes.
Authors: Morgan E L , Weigle W O
Published: December 2022

Murine splenic B lymphocytes are stimulated to proliferate and undergo polyclonal activation in the presence of heat-aggregated human gamma-globulin (AHGG). Splenic macrophages are required to generate the proliferative and polyclonal antibody responses. The polyclonal response as opposed to the proliferative response is dependent upon the presence of T lymphocytes. The stimulatory molecule responsible for the AHGG-induced proliferative and polyclonal activation is derived from incubation of AHGG with splenic macrophages. The biologically active moiety is derived from the Fc portion of the HGG molecule and is approximately 14,000 m.w. The method of aggregation is critical for the generation of stimulatory HGG preparations. Heat aggregation is the only method produced a biologically active preparation
Idiopathic paraproteinemia. III. Increased frequency of paraproteinemia in thymectomized aging C57BL/KaLwRij and CBA/BrARij mice.
Authors: Radl J , de Glopper E , van den Berg P , van Zwieten M J
Published: December 2022

The influence of thymectomy on the appearance of idiopathic paraproteinemia (IP) during aging was investigated in mice of the C57BL/KaLwRij and the CBA/BrARij strains, which under normal conditions develop IP in high and low frequency, respectively. Compared with sham-thymectomized mice, C57BL mice thymectomized at a young adult age showed a markedly increased frequency and an earlier onset of IP during aging; this was even more pronounced in neonatally thymectomized mice. A similar effect of thymectomy was also observed in mice of the CBA strain. Restriction in the heterogeneity of the serum immunoglobulins and the appearance of transient homogeneous Ig components was another frequent finding and this often preceded the appearance of IP in mice of both strains. Thymectomy did not substantially influence either the incidence of paraproteinemias due to a B cell malignancy or the isotype distribution among the paraproteins produced. The results are compatible with the hypothesis that IP develops in three stages as a consequence of an age-related immunnodeficiency that primarily affects the T immune system.
Ia antigen turnover. II. The kinetics of biosynthesis and release of Ia alpha- and beta-chains by murine spleen cells in culture.
Authors: Emerson S G , Murphy D B , Cone R E
Published: December 2022

The kinetics of biosynthesis, release, and turnover of Ia antigens were determined by 3H-leucine labeling, immunoprecipitation and SDS-PAGE analysis. Continuous incorporation and pulse-chase experiments demonstrated that the half-life of the total cell Ia pool was 8 hr, with radiolabeled Ia antigens released from the cell within 4 hr of culture initiation. Both Ia alpha- and beta-chains were released by the cells. I-A antigen turnover was more rapid than I-E antigen turnover. However, detailed comparison of I-A and I-E alpha- and beta-chain biosynthesis demonstrated that the E beta (Ae) chains were synthesized more rapidly than E alpha chains, at rates similar to A alpha- and A beta-chains. Once assembled, however, E alpha- and E beta (Ae)- chains were turned over at the same rate. This kinetic anomaly may reflect post-translational association of polypeptides coded by separate genes into complete I-E antigens.
Cleavage of C4b by C3b inactivator: production of a nicked form of C4b, C4b', as an intermediate cleavage product of C4b by C3b inactivator.
Authors: Nagasawa S , Ichihara C , Stroud R M
Published: December 2022

We have investigated the mechanism of cleavage of C4b into C4c and C4d by the C3b inactivator (C3bINA) and revealed the formation of a nicked form of C4b as an intermediate cleavage product. The cleavage of C4b by the C3bINA was a two-step reaction. The first cleavage occurred on the alpha-chain (89,000 daltons) yielding two fragments, 73,000 daltons and 16,000 daltons. These fragments were bound to each other or to the beta or gamma chain through disulfide linkages. Therefore, an altered form of C4b, C4b', consisting of four disulfide-linked polypeptide chains with the same m.w. as C4b, was produced as an intermediate cleavage product. Subsequently, the second cleavage occurred on the alpha-chain fragment of 73,000 daltons to produce the two generally recognized fragments, C4c and C4d. In both cleavage reactions, a high m.w. cofactor protein, C4bC3bINACo, which is the same protein described as the C4 binding protein, was required, suggesting that both of the proteolytic processes are catalyzed by the C3bINA.
Differential binding of fluorescein-labeled lectins to mouse thymocytes: subsets revealed by flow microfluorometry.
Authors: Fowlkes B J , Waxdal M J , Sharrow S O , Thomas C A , Asofsky R , Mathieson B J
Published: December 2022

Fluorescein-labeled lectins bound to mouse thymocytes were analyzed by flow microfluorometry. This technique has identified several lectins that bind differentially to thymocyte subsets. The most complex fluorescence distributions were obtained using lectins with nominal specificities for galactose or N-acetylglucosamine. Inhibition of binding by sugars confirmed that the fluoresceinated lectins were bound to cells at their carbohydrate binding site. Simultaneous analyses of lectin fluorescence and forward light scatter intensity showed that cell subpopulations of different sizes can exhibit marked differences in the level of binding such that the amount of lectin bound per cell is often independent of cell size. A minor population of dull or unstained cells, delineated by several of these lectins, correlates with the subpopulation of medium-sized thymocytes resistant to in vivo cortisone treatment.
Immune response to phosphorylcholine. VII. Functional evidence for three separate B cell subpopulations responding to TI and TD PC-antigens.
Authors: Fung J , Köhler H
Published: December 2022

A T-independent PC antigen, the C-polysaccharide, was used to induce a primary response to PC in splenic foci containing precursors taken from normal, unprimed BALB/c mice. The precursor frequency for the TI response was compared with the frequency for a T-dependent PC antigen, PPC-TGG-Hy. Idiotype analysis of the precursors for TD and TI responses indicate that the TD responses are more diverse with respect to clonal dominance of idiotype, TEPC-15, than that of TI responses. If splenic fragments were doubly immunized with both TI and TD antigens, the combined PC-specific precursor frequency was superadditive. TD-antigen-induced responses were found resistant to tolerogen and anti-idiotype suppression, whereas TI responses were extremely sensitive to both manipulations. Since under limiting dilution conditions precursors sort out independently, the superadditive response seen in dual immunized fragments can only be interpreted as evidence for an additional subpopulation of B cells that responds to the combined signals of TD and TI antigens. This postulated third B cell population is also extremely sensitive to tolerogen and anti-idiotype suppression.
Human T cell antigens defined by monoclonal antibodies: the 65,000-dalton antigen of T cells (T65) is also found on chronic lymphocytic leukemia cells bearing surface immunoglobulin.
Authors: Royston I , Majda J A , Baird S M , Meserve B L , Griffiths J C
Published: December 2022

Peripheral blood lymphocytes from 15 patients with chronic lymphocytic leukemia (CLL), four patients with lymphosarcoma cell leukemia (LCL), four patients with hairy cell leukemia, and three patients with a monoclonal gammopathy (two with IgM, one with IgA) were tested for reactivity with the anti-T cell monoclonal antibody, T101. By immunofluorescent staining, all of the patients had circulating monoclonal surface Ig+ (sIg+) lymphocytes except for three CLL patients whose leukemia cells were sIg-. The leukemia cells of all of the sIg+ CLL cases were reactive with T101 antibody by indirect immunofluorescence; however, the abnormal cells in all of the remaining cases were unreactive. Reactivity of sIg+ CLL cells with T101 was confirmed by a radioactive binding assay, absorption analysis, and complement-dependent cytotoxicity. Moreover, a 65,000-dalton protein (T65), similar to that found on T cells, was precipitated by T101 antibody from the surface of sIg+ CLL cells. The fluorescent staining of sIg+ CLL cells by T101 antibody was weak as was the staining of the sIg. This was in contrast to the LCL cells, which had intense staining sIg and absence of staining with T101 antibody. These data demonstrate the existence of two major subtypes of CLL that have phenotypes sIg+ and T101+ and sIg-T101-. The implication of the finding of dual T and B markers on the major type of CLL, but not other B cell malignancies is discussed.