bluuebunny/crossref_metadata_2025 huggingface dataset:
Una pieza de la Real Fábrica de Artillería de Sevilla en Londres. El “Cadiz Memorial” de Saint James Park
Authors: Baena Gallé José Manuel
Published: January 2023

En Londres existe un monumento creado a partir de una pieza de artillería española originaria de la Real Fundición de Sevilla. Es una de las consecuencias de la guerra de la Independencia y también muestra de las relaciones de España y el Reino Unido en aquellos momentos. Con este obús se realizará una escultura pública de gran interés iconográfico y con enormes consecuencias tanto artísticas como políticas. Se trata del “Cadiz Memorial” en el Saint James Park londinense.
Le lieu-dit des « Trois-Piliers » : un ensemble à caractère monumental du iiie s. apr. J.-C. dans la périphérie nord-est de Reims/Durocortorum ?
Authors: Rollet Philippe , Jouhet Émilie
Published: December 2022

La fouille du 59 rue Belin dans le quartier des « Trois-Piliers » a permis de mettre au jour une grande salle semi-enterrée à la manière d’un cryptoportique. Longue d’au moins 57 m et large de 7,80 m, elle est aménagée d’une série de piliers le long de son mur oriental et bordée à l’extérieur d’un large égout. Construit au début du iiie s., cet édifice est partiellement détruit par un incendie à la fin du deuxième tiers du iiie s. Sa réoccupation intervient rapidement, vers les années 260, avec la construction en son sein de deux puits massifs cuvelés avec des blocs de grand appareil en remploi qui attestent, par les restes de sculpture qu’ils ont conservés, la proximité d’un vaste monument, probablement un temple. Si plusieurs publications mentionnent les fouilles de la rue Belin, qui remontent tout de même à la fin des années 1990, en faisant notamment référence aux éléments lapidaires issus des puits ou à la couche d’incendie particulièrement étendue qui scelle les vestiges de la grande salle, le présent article expose l’ensemble des données archéologiques ayant trait à l’édifice (plan, composantes architecturales, techniques de construction, etc.), tout en le replaçant dans son environnement archéologique, avant d’en proposer l’interprétation.
Regulation of the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Barthold David R. , Prescott Benjamin , Stashak Philip W. , Amsbaugh Diana F. , Baker Phillip J.
Published: January 2023

A modification of the technique of localized hemolysis-in-gel was used, in conjunction with class-specific anti-γ-globulin sera, for detecting IgG and IgA plaque-forming cells (PFC) in mice immunized with Type III pneumococcal polysaccharide (SSS-III). Small, but significant, numbers of IgG and IgA PFC could be detected, and the numbers increased at least 10-fold after treatment with antilymphocyte serum (ALS). The results of plaque-inhibition studies affirmed that such PFC represent cells making antibody having the same class-specificity as the antiserum used for their detection. These findings suggest that the magnitude of not only the IgM, but also the IgG and IgA, antibody response to SSS-III is governed by the activities of regulatory thymus-derived cells having opposing functions and that ALS-induced enhancement cannot be attributed to a reversal or alteration of antibody-mediated feedback inhibition. Furthermore, the kinetics observed for the development of the IgM, IgG, and IgA antibody response to SSS-III suggest that clones of precursor cells, restricted mainly to the synthesis of one class of immunoglobulin, exist before immunization and increase independently in size upon antigenic stimulation.
In Vitro Immune Stimulation-Inhibition to Spontaneous Canine Tumors of Various Histologic Types
Authors: Fidler Isaiah J. , Brodey Robert S. , Bech-Nielsen Steen
Published: January 2023

Utilizing dogs with progressively growing tumors we are reporting on the in vitro reactivity of their lymph node or peripheral blood lymphocytes to their own tumor cells. Tumors, lymph nodes, and blood were collected aseptically during surgical treatment or before necropsy. Primary cultures were set up and shortly thereafter only viable glass-adhering tumor cells were utilized. The cells were first incubated with either media or various sera and then varying numbers of sensitized lymphocytes were added. The mixtures were incubated for 90 min in rotating test tubes and then plated into culture dishes or plates. Two and/or 5 days later, the cultures were fixed, stained, and viable target cells were counted. In vitro tumor cell destruction by high numbers of lymphocytes from dogs bearing tumors and the blocking of such reactivity by autologous serum were demonstrated. Serum from dogs with the same histologic type of neoplasm inhibited allogenic lymphocyte cytotoxicity, whereas serum from dogs with a different tumor, normal dogs, and a dog who had been clinically free of osteosarcoma after limb amputation 17 months earlier, did not significantly block the cellular-mediated reactivity. Stimulation of tumor growth in vitro by low ratios of sensitized lymphocytes to tumor cells was demonstrable. Autologous serum from dogs with progressively growing neoplasms appeared to potentiate the stimulation of tumor growth above a simple blocking of lymphocytic-mediated cytotoxicity. These results confirm our earlier reports of immune stimulation to tumor growth in vitro and support the hypothesis and work by Prehn that the early immune response to neoplasia might directly stimulate rather than inhibit tumor growth.
Mechanisms of Passive Sensitization
Authors: Ishizaka Teruko , Ishizaka Kimishige
Published: January 2023

The mononuclear cell fraction of human leukocytes was exposed to different pH at 0°C and the presence of cell-bound IgE molecules on basophil granulocytes was examined by autoradiography. The results showed that both cell-bound normal IgE and E-myeloma protein bound to the basophils by passive sensitization dissociated from the cells upon exposure to pH 4 or lower. The receptor molecules on basophils was not degraded by the treatment. Basophils exposed to acid pH combined more IgE than untreated cells upon incubation with E myeloma protein at neutral pH. The results indicated that the binding between IgE and receptor molecules was reversible and did not involve a covalent bond.
Purification of HL-A Antigens from Normal Serum
Authors: Billing Ronald J. , Terasaki Paul I.
Published: January 2023

A relatively simple procedure using non-denaturing techniques has been established to isolate HL-A antigens from serum. The procedure involves QAE Sephadex ion-exchange chromatography, Concanavalin A Sepharose 4B chromatography, and preparative polyacrylamide gel electrophoresis. There is approximately 17% recovery of the antigenic material in the final preparation with a 120-fold increase in specific activity over the original serum. The serum HL-A antigen is a glycoprotein containing 10 to 12% carbohydrate and having an approximate molecular weight of 130,000. The final antigen preparation migrates on polyacrylamide gels as a single band in the region of serum B1 globulins which can be entirely removed by pretreatment with specific HL-A antibodies.
In Vitro Induction of Specific Unresponsiveness Against the 2,4,6-Trinitrophenyl Determinant
Authors: Kettman J. , Davenport M.
Published: January 2023

A state of unresponsiveness against the hapten 2,4,6-trinitrophenyl (TNP) can be induced in a mouse spleen cell suspension by incubation in vitro with TMP human serum albumin (HSA), a polyvalent but apparently non-immunogenic (in vitro) molecule. The depression is specific for the hapten since cells treated with TNP-HSA and cultured with TNP sheep red blood cells (SRBC) show normal anti-SRBC responses. The precursors of cells making anti-TNP antibody of higher avidity appear to be suppressed selectively. The mechanism of the induction of this specific unresponsiveness remains obscure.
Effects of Staphylococcal Protein a on Heat Labile Opsonins
Authors: Forsgren Arne , Quie Paul G.
Published: January 2023

Staphylococcal protein A inhibited phagocytosis of Staphylococcus aureus and Escherichia coli in in vitro phagocytosis systems with normal human serum. Protein A had no effect on phagocytic cells per se, and produced inhibition of the heat labile opsonins, i.e., of the complement system in normal sera at low concentration. At higher concentration of protein A there was little complement fixation and little inhibition of serum opsonic activity.
Studies on the Control of Antibody Synthesis
Authors: Mond James , Kim Young Tai , Siskind Gregory W.
Published: January 2023

A progressive increase in antibody affinity occurred after immunization with dinitrophenylated keyhole limpet hemocyanin i.v. in the absence of adjuvants, provided a relatively low dose of antigen was used. When a higher dose of antigen was given i.v., very low affinity antibody was formed and no increase in affinity was observed. It is suggested that a high dose of antigen may stimulate lower affinity antibody-forming cells and induce tolerance in high affinity B lymphocytes. Adjuvants (complete Freund's adjuvant and endotoxin) increased the magnitude of the response and slightly increased the rate of increase in affinity. Depression of antibody synthesis with 6-MP had no effect on affinity unless treatment was begun before immunization. Under these conditions a marked depression in the magnitude of the immune response and a slight decrease in affinity was observed.

Thus, nonspecific modifications of the magnitude of the immune response with adjuvants or cytotoxic drugs can affect, but only slightly, the affinity of the antibody produced. This is consistent with the concept that the process of selection of cells making high affinity antibody proceeds somewhat more efficiently when there is a vigorous immune response. However, the major factors determining antibody affinity appear to be the antigen dose and the “selective pressure” of decreasing antigen concentration.
Kinetics and Clonal Restriction of the Anti-Dinitrophenyl Antibody Response to Dinitrophenyl-6-Amino-Caproyl-L-Tyrosine-Azobenzene-p-Arsonate in Guinea Pigs
Authors: Roelants Georges E. , Goodman Joel W.
Published: January 2023

The anti-DNP responses in guinea pigs to the small bifunctional antigen DNP-SAC-RAT and to DNP-protein were compared. The response to DNP-SAC-RAT is more restricted both in terms of the anti-DNP titers reached in serum and of the number of DNP specific clones expressed. Priming with DNP-SAC-RAT led to an accelerated secondary response reaching similar levels of anti-DNP antibody as in the primary response. In sera showing restricted clonal patterns by isoelectric focusing, it was found that once a clone dominated the anti-DNP response it remained stable during the primary response and was the only one to reappear after secondary stimulation.
Antibody-Dependent Cell-Mediated Cytotoxicity in Prospective Kidney Transplant Recipients
Authors: Lightbody James J. , Rosenberg Jerry C.
Published: January 2023

Four types of cytotoxicity were observed in prospective kidney transplant recipients: a) antibody-mediated complement-dependent, b) cell-mediated, c) antibody-dependent cell-mediated (allogeneic effector cells), and d) antibody-dependent cell-mediated (recipient effector cells). The two types of cell-mediated assays which were antibody-dependent could detect the presence of preformed antibodies in recipients when the antibody-mediated complement-dependent cross-match was negative. The use recipient lymphocytes rather than allogeneic lymphocytes as effector cells generally resulted in higher 51Cr release. This appeared to be the result of antibody present in the recipient plasma directed toward determinants on the allogeneic effector cell. In some instances, however, the higher amount of cell lysis with recipient effector cells did not appear to be due to antibody inhibition of allogeneic effector cells.
The First Component of Human Complement (C1): Kinetics of Reaction with its Natural Substrates
Authors: Strunk Robert , Colten Harvey R.
Published: January 2023

Classical enzyme kinetic analyses were applied to define the mechanism of the effect of the fourth component (C4) on cleavage of the second component (C2) by the first component (C1) of human complement. The data indicated that the increased rate of cleavage of C2 by C1 in the presence of C4 was due to availability of a site for product deposition; the effect of C4 was reversed by blocking the site on C4 for C2 deposition. C2 cleavage by C1 followed first order kinetics.

In addition, our findings support the hypothesis that there are separate enzymatic sites on the C1 molecule for its natural substrates.
Interaction of the First (C1̄), the Second (C2) and the Fourth (C4) Component of Complement with Different Preparations of Bacterial Lipopolysaccharides and with Lipid A
Authors: Loos Michael , Bitter-Suermann Dieter , Dierich Manfred
Published: January 2023

Lipopolysaccharides of Gram-negative bacteria (LPS) known to activate the complement system via the alternate pathway bypassing C1̄, C4, and C2 had a direct effect on purified C1̄, but no effect on purified C4 and C2. These studies were performed with different preparations of LPS and of Lipid-A isolated from smooth (S-) and rought (R-) forms of Escherichia coli and Salmonella strains. It was shown that the effect of LPS on C1̄ depends upon the concentration and type of LPS applied: 0.77 µg of LPS from E. coli 0 75, 2.5 µg of LPS from S. minnesota R 595, 24 µg of Lipid-A and 55 µg of Lipid-A/BSA were necessary to get 50% inhibition of free C1̄. In contrast, 100 µg of LPS from E. coli 0 111, from S minnesota S-form, or from S. minnesota R 345 showed no significant inhibition. Binding of C1̄ to EA or EAC4 markedly reduced the effect of LPS and of Lipid-A on C1̄. The inhibition of C1̄ was shown to be the result of a direct interaction of LPS and of Lipid-A with the C1 subunit C1q: LPS precipitated C1q. LPS and Lipid-A had no effect on the C1 esterase activity against C2.
Heterogeneity of human monocyte subsets in the promotion of B cell colonies and the role of interleukin 1.
Authors: Whisler R L , Newhouse Y G , Lachman L B
Published: January 2023

The abilities of human peripheral blood mononuclear-phagocyte (M phi) subpopulations and of interleukin 1 (IL 1) to support human B cell colony formation in semisolid cultures stimulated by staph protein A were analyzed. Human M phi subsets enriched for complement receptors (CR) effectively functioned as accessory cells supporting colony growth, whereas the responses obtained with CR-depleted M phi were 4.6-fold less. In experiments analyzing IL 1 production, CR-enriched M phi secreted four to 12 fold greater amounts of basal and stimulated IL 1 than CR-depleted M phi. Also, the addition of IL 1 to CR-depleted M phi resulted in a fourfold increase of colony numbers. The responses of cultures containing CR-depleted M phi plus IL 1, however, remained 30% less than those observed for cultures supplemented with CR-enriched M phi. Other studies showed that IL 1 was unable to substitute for M phi; the responses of cultures containing IL 1 and B cells were reduced 10-fold compared to cultures supplemented with autologous M phi. These findings indicate that human M phi subsets exist that differ in their ability to function as accessory cells. Although IL 1 can collaborate with certain M phi subsets to restore their accessory cell function, it cannot replace intact M phi. Thus, it is possible that other monokines or lymphokines play a role in M phi accessory cell function.
Separable helper factors support B cell proliferation and maturation to Ig secretion.
Authors: Parker D C
Published: January 2023

The 24-hr culture supernatant of Con A-activated spleen cells (SN) contains helper factors that enable maturation to high-rate polyclonal Ig secretion and enhance proliferation in cultures of mouse B cells activated with the F(ab')2 fragment of class-specific rabbit antimouse IgM antibody (anti-Ig). When interleukin 2 (IL 2), also called T cell growth factor, is removed from SN by absorption with an IL 2-dependent cell line at either 4 degrees C or 37 degrees C, all the helper activity for anti-Ig-activated B cells is also removed. Partial removal of IL 2 results in partial removal of helper activity for B cells. However, the IL 2-depleted SN appears to contain another helper factor, TRF, that enables anti-Ig-activated B cell cultures to mature to high-rate Ig secretion. This TRF activity is revealed by adding purified human IL 2 or an IL 2-containing supernatant of a cloned, lectin-activated T cell hybridoma line (FS6-14.13) to Il 2-depleted SN, which restores the polyclonal antibody response to anti-Ig. The hybridoma supernatant by itself supports proliferation of anti-Ig-activated B cell cultures, as measured by an increase in cell number, but not maturation to Ig secretion. This proliferative response is likewise IL 2 dependent, although purified IL 2 with anti-Ig is not sufficient. These experiments define separable combinations of factors acting on anti-Ig-activated B cell cultures, one of which (SN) results in both proliferation and maturation to high-rate Ig secretion, whereas the other (hybridoma supernatant) results in proliferation only. IL 2 appears to be an essential component of both combinations, although the target cell for IL 2 action in this system remains to be determined.
Selective effects of interferon on distinct sites of the T lymphocyte triggering process.
Authors: Leanderson T , Hillörn V , Holmberg D , Larsson E L , Lundgren E
Published: January 2023

Lectin- and antigen-induced proliferation of murine T cells consists of two major events, namely, a rapid induction of susceptibility to growth factors and a later-occurring, accessory cell-dependent production of T cell growth factors (TCGF). The mechanism by which interferon (IFN) inhibits T cell responses was studied accordingly. A decrease of Con A-induced proliferation was observed in the presence of increasing amounts of IFN. The reduced proliferative response in such cultures was found to be due to an accumulation of cells in the G0/G1 phase of the cell cycle. Furthermore, the results show that IFN did not inhibit the early events in T cell triggering, because the acquisition of responsiveness of resting T cells to TCGF was unaltered in the presence of IFN, nor did it interfere with production of TCGF. In contrast, IFN was found to interfere with the TCGF-dependent T cell blast growth. Cytofluorometric analysis of the proliferative phase revealed that IFN exerts its effect on T cells, which have entered the proliferative cycle, by a postmitotic accumulation in G0/G1, thus reducing the proliferating population. The results demonstrate that IFN primarily affects the later phase of proliferative activity after T cell triggering, leaving the helper cell functions untouched.
Formation of rat IgE-binding factors by rat-mouse T cell hybridomas.
Authors: Huff T F , Uede T , Ishizaka K
Published: January 2023

Mesenteric lymph node (MLN) cells of rats infected with Nippostrongylus brasiliensis (Nb) were obtained 8 days after infection and were incubated overnight with rat IgE for the formation of IgE-binding factors. These cells were then fused with BW 5147 mouse T lymphoma cells. Two hybrid clones (A4 and B6) obtained by the fusion formed IgE-binding factors upon incubation with IgE. It was found that 1-hr incubation with 3 micrograms/ml of rat IgE was sufficient to induce hybridoma cells to form IgE-binding factors. Both hybrid clones express rat T cell markers; the A4 clone bears Fc epsilon R, whereas the B6 clone bears Fc gamma R on their surface. The IgE-binding factors formed by both clones bound to rat IgE-coated Sepharose and could be eluted from the beads at pH 3.0. The factors also have affinity for mouse IgE but not human IgE nor rat IgG. IgE-binding factors formed by the A4 clone had a m.w. between 26,000 and 30,000; the B6 clone formed IgE-binding factors of 26,000 and 13,000 daltons. The factors of both high and low m.w. failed to bind to lentil lectin and Con A, but had affinity for peanut agglutinin. Purified IgE-binding factors of 13,000 daltons selectively suppressed the IgE-forming cell response of DNP-ovalbumin-primed MLN cells to homologous antigen, whereas the factor of 26,000 to 30,000 daltons neither suppressed nor enhanced the IgE response.
Immunosuppressive ATS. V. Analysis of the effect of anti-thymocyte serum on T lymphocyte subsets.
Authors: Zimmerman B , Swain S L , Dutton R W
Published: January 2023

The effect of anti-thymocyte serum (ATS) on functional T cell subsets within the immunoregulatory network was examined. We found that low doses of ATS depleted the IL 2-producing subset of T helper cells and higher doses of antiserum also eliminated cytotoxic precursors. It was estimated that a standard low dose of ATS, which reduced T helper cells 80-fold, depleted cytotoxic precursors only about four-fold. The data indicate that ATS acts on different T cell subsets in a dose-dependent fashion and suggest that monoclonal antibodies might be used to preferentially deplete specific T cell subpopulations. Two monoclonal rat anti-mouse Lyt sera, however, proved ineffective in inducing functional changes in vivo, although an alloantiserum to Lyt1.1 was able to reduce cell-mediated cytotoxicity in a dose-dependent fashion. The results support the feasibility of specific depletion of T cell subsets with anti-lymphoid antibodies.
T cell control of the antibody response to the T-independent antigen, DAGG-Ficoll.
Authors: Nordin A A , Schreier M H
Published: January 2023

C57BL/6J nu/nu mice respond to the type 2 TI antigen DAGG-Ficoll, but not to the TD antigen SRC. A comparable difference can also be seen in vitro, but only at high spleen cell density and in the presence of selected batches of FBS. At low spleen cell density and in the absence of FBS, the DAGG-Ficoll-induced B cell response is strictly dependent on soluble helper factors or cloned specific helper T cells. The B cell response so induced requires that the T cell-depleted spleen cells be compatible in the I-A subregion of the H-2 complex. These helper factors, induced by antigen in an I-A-restricted T cell-macrophage interaction, provide helper for T cell-depleted spleen cells irrespective of their H-2 haplotype. Under conventional culture conditions, the stringent requirement for helper factors in the in vitro response to DAGG-Ficoll is obscured by FBS. In vitro culture of low numbers of spleen cells, in serum-free medium instead of FBS, provides a sensitive assay for helper factors. We have compared the helper activity for a B cell response to SRC or DAGG-Ficoll as provided by antigen-induced supernatants of various individual EA-specific T cell clones. There was a remarkable and consistent heterogeneity among individual T cell clones: their helper activity in the response to TI and TD antigens did not correlate, nor was there any correlation between helper activity and antigen-induced TCGF (interleukin 2) activity.
Neutral glycolipid antigens as developmental markers of mouse teratocarcinoma and early embryos: an immunologic and chemical analysis.
Authors: Willison K R , Karol R A , Suzuki A , Kundu S K , Marcus D M
Published: January 2023

Purified rabbit antibodies to neutral glycolipids were analyzed for their binding to mouse embryonal carcinoma cells (ECC) and preimplantation mouse embryos. Antibodies to globotetraosylceramide first bind to 2 to 4-cell embryos and reach a peak of staining intensity with morulae, whereas anti-Forssman antibodies first bind to late morulae and then, most intensely, to early blastocysts. We compared the binding of a monoclonal anti-Forssman antibody with that of rabbit anti-Forssman antibodies and show that although they react similarly with ECC, they do not do so with morulae: The monoclonal antibody stained weakly and unevenly, whereas the rabbit antiserum produced a uniformly bright immunofluorescent staining. Chemical analyses revealed that globotetraosylceramide is the most abundant glycolipid of F9 ECC and that there is poor correlation between the concentration of individual glycolipids in these cells and their reactivity with antibodies to glycolipid molecules. Interestingly, the 2 Forssman antibody-reactive glycolipids of F9 ECC differ in their mobility on TLC plates from the classical Forssman antigen extracted from sheep red blood cells. This illustrates the potential problems in extrapolating from the coincident binding properties of an anti-glycolipid antibody to the chemical structure or abundance of an antigen in different cell types.
Metabolism and clearance of antibody-excess immune complexes in lactating mice.
Authors: Kim Y W , Halsey J F
Published: January 2023

The metabolism of antibody-excess immune complexes was compared in lactating and normal female BALB/c mice. The initial rate of clearance of the antigen, dinitrophenyl- (DNP) ovalbumin, when complexed with mouse anti-DNP antibody was considerably faster in lactating mice. The half-life was 4 min in lactating mice compared with 8 min in normal female mice. Tissue distribution studies revealed that by 2 hr the mammary gland had sequestered immune complexes in threefold to fourfold excess over the levels in the liver. These lactating mice were shown to transfer immune complexes to their milk, and small quantities of the complexes could be demonstrated in the circulation of the nursing neonates.
Characterization of antigen recognized by the monoclonal antibody (4F2): different molecular forms on human T and B lymphoblastoid cell lines.
Authors: Hemler M E , Strominger J L
Published: January 2023

The monoclonal antibody 4F2 recognizes a disulfide-linked ricin-binding glycoprotein complex (Mr congruent to 125,000) composed of a sialylated heavy subunit (Mr congruent to 85,000 on T cell lines) and an unsialylated light subunit (Mr congruent to 41,000). The antigen (T85,41) recognized by 4F2 on T cell lines is structurally distinct from the antigen (B93, 41) on B cell lines. The heavy subunits, but not the light subunits, from all T cell lines examined were uniformly smaller in size than the heavy subunits from several B cell lines. This reflects differences in carbohydrate rather than protein represent in B93,41 compared with T85,41, because both heavy subunits have a common unglycosylated form (p65) and a common partially glycosylated precursor form (p68). Among non-T, non-B hematopoietic cell lines, the monocytoid line U-937 expressed an antigen that resembles B93,41, whereas the erythroleukemic line K-562 expressed an antigen more similar to T85,41. 4F2 recognizes a protein determinant on the heavy subunit (with or without N-linked glycosylation) and also the unglycosylated heavy subunit retains the ability to associate with light subunit. The light subunit itself contains no detectable N-linked carbohydrate. Unlike the transferrin receptor, synthesis of the antigen recognized by 4F2 on the promyelocytic cell line HL-60 did not diminish upon dimethylsulfoxide-induced differentiation, and thus is not tightly correlated with cell proliferation.
Noncovalent association of heavy and light chains of human immunoglobulins. III. Specific interactions between VH and VL.
Authors: Horne C , Klein M , Polidoulis I , Dorrington K J
Published: January 2023

Mildly reduced monoclonal human IgM proteins have been cleaved at cysteinyl residues to give VH fragments after S-cyanylation with 2-nitro-5-thiocyanobenzoic acid. The noncovalent interaction between the VH fragments and autologous kappa-chains was studied by ultraviolet difference spectroscopy and circular dichroism. A bimolecular complex was formed with an association constant in excess of 10(7) M-1 at 23 degrees C. Complex formation was accompanied by burial of tryptophan and tyrosine side-chains. In contrast to the studies with autologous species, the VH fragments did not associate with heterologous kappa-chains as judged both by difference spectroscopy and gel filtration using radiolabeled VH fragments. This specificity in the association between VH and VL has been attributed to interactions contributed to by residues in the third hypervariable region of VH encoded by the DH and JH genes.
I-J restriction of T cells that suppress in vivo antibody responses to Thy-1.
Authors: Brunswick M , Sainis K B , Bromberg J S , Lake P
Published: January 2023

The contact-sensitizing haptens dinitrophenyl (DNP) and oxazalone (Ox) act as helper determinants for antibody responses to Thy-1 when conjugated to donor thymus cells. The helper effect is transferrable from primed to naive mice with spleen cells, producing specific augmentation of in vivo PFC responses to Thy-1. The helper cells are hapten-specific and require associative recognition of hapten and Thy-1, excluding a role for nonspecific B cell activation. The phenotype of the helper cells is Thy-1+ and Lyt-1+2-. Antigen-specific suppression could be readily generated by using an inoculum of DNP-modified syngeneic RBC. T cells from these suppressed donors (Ts) were shown to abolish the helper effects of TH in adoptive transfer experiments in vivo. These Ts were characterized as Thy-1+ and Lyt-1-2+. A requirement for MHC compatibility at the I-J subregion was necessary between the Ts and the recipient to obtain a transfer of suppression.
Conditioned immune response associated with allogeneic skin grafts in mice.
Authors: Gorczynski R M , Macrae S , Kennedy M
Published: January 2023

Groups of CBA mice were given sham skin grafts (conditioned stimulus, CS) or alloantigen only (unconditioned stimulus, US; C57BL/6 lymphoid cells inoculated i.p.). A further group of animals received simultaneous US and CS (alloantigen (US), in this case actually given in the context of a C57BL/6 skin graft). The peripheral blood leukocyte (PBL) pool of all animals was analyzed in tissue culture for the frequency of cytotoxic T lymphocyte precursors (CTLp) specific for alloantigens on the grafted tissue. After three "conditioning" trials, spaced at intervals to allow recovery in all groups of the CTLp frequency to that of naive control mice, individuals in all groups were sham-grafted (given CS alone). PBL of these mice were tested 2 days after removal of the casts. Only group CS + US, and only some 50 to 60% of these, were able to show a "conditioned" increase in the CTLp frequency specific for alloantigens of the grafted tissue after sham-grafting. When these "responder" mice were subdivided into two groups that were subsequently given two trials of CS only or CS + US, followed by retesting with CS alone, either extinction or reinforcement of the previously conditioned response was observed.
Transplantation in miniature swine. XII. N-terminal sequences of class I histocompatibility antigens (SLA) and beta 2-microglobulin.
Authors: Metzger J J , Lunney J K , Sachs D H , Rudikoff S
Published: January 2023

Purified class I histocompatibility antigens (SLA) from three haplotypes were prepared by papain treatment of lymphoid cell membranes obtained from spleens and lymph nodes of miniature swine homozygous at their major histocompatibility complex. Antigens were purified by ion-exchange chromatography followed by gel filtration. Purity was analyzed by SDS-PAGE, and antigenic specificity by inhibition of complement-dependent, alloantiserum-mediated cytotoxicity. The SLA antigens were reduced and alkylated, and the component heavy and light chains were isolated by gel filtration under dissociating conditions. N-terminal amino acid sequences were obtained for SLAaa, SLAcc, and SLAdd heavy chains, as well as for the light chain, beta 2-microglobulin. The swine antigens showed high levels of homology with class I antigens from other animal species. Heterogeneity was observed among the swine haplotypes, and several of the positions at which substitutions were found are apparently invariant in other animal species. In contrast, only minimal sequence heterogeneity was detected within haplotypes, the basis of which may be of relevance to understanding the evolutionary development of these molecules.
Lack of specificity for viral and H-2 antigens by anomalous T killer cells generated in murine leukocyte cultures.
Authors: Shain B , Holt C A , Lilly F
Published: January 2023

Mouse spleen cells, cultivated in vitro for 6 days in the presence or absence of syngeneic irradiated virus-induced tumor cells, lysed a variety of virus-induced tumor cells and normal cells in a long (19-hr) 51Cr-release cytotoxicity assay. Little or no specificity for viral or H-2 antigens was detected. This anomalous killing (AK) activity contrasted with H-2-restricted cytotoxic T lymphocyte (CTL) activity generated in secondary mixed leukocyte-tumor cell cultures and detected in a short (4- to 6-hr) 51Cr-release assay. Studies of competitive inhibition by unlabeled target cells in the long assay suggested that AK effectors were a broadly heterogeneous mixture of cells representing many clones, each recognizing a different antigenic determinant. AK effector cells shared properties with CTL: they could be generated from spleen cells of natural killer cell-deficient mice bearing the beige mutation but not from those of T cell-deficient nude mice; they expressed Thy-1 and Lyt-2 antigens; and they were nonphagocytic. AK activity could be generated from normal spleen cells of mice of all inbred strains examined, but the levels of activity generated varied in a strain-specific manner and were not significantly influenced by H-2 genotype.
Delineation of immunologically and biochemically distinct HLA-A2 antigens.
Authors: Biddison W E , Kostyu D D , Strominger J L , Krangel M S
Published: January 2023

Cytotoxic T cell (CTL) recognition of influenza virus in conjunction with HLA-A2 was examined in a population study. Virus-infected target cells from three unrelated A2-positive donors were not lysed by virus-immune CTL from any donor matched only for A2. The A2 antigens of these three donors were indistinguishable from the A2 antigens of other A2-positive donors as assessed by extensive serologic analyses; however, isoelectric focusing (IEF) of A2 molecules from these three donors demonstrated that their A2 heavy polypeptide chains are structurally distinct from those of "normal" A2-positive donors. To date 11% of all A2-positive donors tested exhibited a "variant" A2-associated CTL restriction antigen, and IEF of A2 heavy chains from all "variant" A2-positive cells revealed structural differences in each of these polypeptides. These results suggest there may be considerably greater polymorphism of HLA-A gene products than has been revealed by current serologic techniques.
The alloantibody response in the allogeneically pregnant rat. I. The primary and secondary responses and detection of Ir gene control.
Authors: Smith R N , Sternlicht M , Butcher G W
Published: January 2023

We compared the primary pregnancy-induced alloantibody responses with secondary pregnancy-induced responses and with conventional immunologic responses and found there are profound differences. Conventional and secondary responses produce strong lytic sera and readily detectable immunologic memory. The primary pregnancy-induced responses lack memory and produce alloantisera or widely variable titer. Such sera were rarely directly lytic. Pregnancy-induced primary responses detect MHC Ir gene control, even when the allogeneic difference is a complete HMC haplotype. The kinetics of the primary and secondary pregnancy-induced alloantibody responses against DA were different. We conclude that the primary pregnancy-induced alloantibody response is very different immunologically from conventional and secondary pregnancy-induced alloimmunizations.
Immune response of the mouse to gram-negative bacterial outer membrane extracts as assessed with monoclonal antibodies.
Authors: Mackie E B , Longenecker B M , Rabin H R , Di Ninno V L , Bryan L E
Published: January 2023

Neisseria meningitidis outer membrane extracts of known serotype antigens as well as isolates from a linear carriage study were used to immunize mice for subsequent production of monoclonal antibodies. As well, membrane extracts from one strain of Pseudomonas aeruginosa, serotype M, were used. The degree of specificity of the resulting monoclonal antibodies to the two genera differed markedly when tested with known serotype antigens. That is, antibody from a given clone reacting with more than one serotype specificity was a more frequent occurrence among clones resulting from the Pseudomonas fusion. Preexisting "natural" antibody to several Pseudomonas species is considered at least partially responsible for the diversity of the response to this genus. Evidence is presented for the divergence of specificity with time after immunization with Neisseria.
Antibody response to stage-specific Trichinella spiralis surface antigens in strong and weak responder mouse strains.
Authors: Jungery M , Ogilvie B M
Published: January 2023

The antibody response to antigens on the cuticular surface of Trichinella spiralis was compared in two strains of mice, NIH mice, which control the parasite relatively strongly, and C3H, which reject this nematode more slowly. The evolution of the antibody response to the nematode surface was monitored by the appearance of antibodies that mediate the adherence of eosinophils to the worm and the appearance of antibodies that recognize molecules on the surface that can be labeled with 125I. Antibody responses were measured using three life cycle stages; muscle stage larvae, immature and mature intestinal stages, and newborn larvae. We found that NIH mice responded immunologically to the four molecules found in extracts of surface-labeled T. spiralis adult worms, but that C3H mice did not produce antibodies to one of these molecules until much later in the infection. NIH mice produced antibodies against both infective larvae antigens by day 4 after infection, whereas C3H recognized only one before day 30. Variations in antibody response also occurred against antigens on the surface of newborn larvae: antibody to one of these antigens was not present in C3H immune sera until day 30. Sera from the two strains likewise differed in their ability to mediate eosinophil adherence to each parasite stage, and in some of the life cycle stages, these differences corresponded to the recognition of specific antigens found on the parasites' surface. The links between recognition of defined parasitic antigens and the fate of the parasite in genetically different strains of mice are discussed.
CT hybridomas: tumor cells capable of lysing virally infected target cells.
Authors: Whitaker R B , Kauffman R S , Che M , Finberg R
Published: January 2023

By fusing primed murine lymphocytes with a syngeneic T cell lymphoma, we have been able to select for H-2-restricted, virus-specific cytotoxic T cell hybridomas (CTH). These T cell hybrids, which replicate in ordinary tissue culture medium or in ascites, are capable of lysing virally infected target cells, and their activity is facilitated by the presence of lectins in the assay medium. Unlike cells mediating lectin nonspecific lysis, these hybridomas are H-2 restricted and specific for single viral proteins. The ability to maintain these cells in culture for over 18 mo and to pass them in vivo without loss of activity or specificity indicates that they will provide sufficient material for the analysis of surface proteins and genetic information required for the recognition and lysis of virally infected cells by killer T cells.
Antigen-induced acceleration of shedding and replacement of B cell antigen receptors requires mature T cells.
Authors: Ashman R F , Noel E J
Published: January 2023

To determine which early and intermediate events in the response of antigen-binding B cells to a T-dependent antigen (sheep erythrocytes [SRC]) require T help, the antigen-induced changes in receptor turnover and surface IgD loss in BALB/c athymic nu/nu mice were compared with that of nu/+ littermates and +/+ BALB/c mice. Nonimmune SRC antigen-binding spleen B cells (ABC) from +/+, nu/+, and nu/nu BALB/c mice coexpressed IgM and IgD, and 85 to 95% retained receptors well when incubated for 2.5 hr in 100 micrograms/ml cycloheximide (which prevents receptor replacement). Also they were able to regain their ability to bind antigen by 18 hr after pronase treatment, but not by 2 hr. However, 5 days after in vivo immunization, 1) the proportion of ABC expressing surface IgD declined from around 90% to less than 50% in +/+ mice and nu/+ mice but not in nu/nu mice; 2) substantial recovery of antigen-binding occurred by 2 hr after pronase treatment in +/+ and nu/+ ABC but not in nu/nu ABC; and 3) when spleen cells were incubated in cycloheximide, uncompensated receptor shedding reduced +/+ and nu/+ ABC by around 80% but produced only about a 10% reduction in nu/nu ABC. Thus, although the ABC in nonimmune nu/nu mice appeared normal with respect to their surface Ig turnover and expression, they failed to undergo the normal antigen-induced loss of IgD or acceleration of surface Ig shedding and replacement, suggesting that these intermediate activation events require interaction with mature T cells. To determine whether this interaction had to occur during B cell development, during the development of the immune response, or during receptor shedding or replacement itself, cell transfer experiments were carried our wherein nu/+ T cells were transferred i.v. to nu/nu littermates 1 day before immunization with SRC. In the transfer recipients, pronase-treated day 5 ABC were then able to replace and shed their receptors at the accelerated rate, like ABC from +/+ and nu/+ mice. In contrast, the co-incubation of 5-day immune nu/+ T cells with nu/nu B cells did not alter the rate of shedding or replacement.
Anti-immunoglobulin in combination with cytochalasin stimulates proliferation of murine B lymphocytes.
Authors: Rothstein T L
Published: January 2023

The ability of cytochalasin to influence the stimulation of murine B lymphocytes through surface immunoglobulin was assessed during short term cultures. Modest doses of anti-immunoglobulin alone did not stimulate proliferation of mouse spleen cells at 2 days. Cytochalasin B alone also had no effect. However, anti-immunoglobulin in combination with cytochalasin B stimulated substantial proliferation as judged by [3H]thymidine incorporation. Cytochalasins A, E, and D, and dihydrocytochalasin B were all effective in promoting B cell proliferation. Spleen cells from xid-defective (CBA/N X DBA/2)F1 male mice failed to proliferate in response to anti-immunoglobulin plus cytochalasin, suggesting that this treatment affects the same subset of B cells as anti-immunoglobulin plus B cell growth factor. Moreover, proliferation that was stimulated by anti-immunoglobulin plus cytochalasin B was not affected by T cell depletion. Cytochalasin may circumvent the need for, or replace, a second signal for proliferation.
Phenotypic expression of B lymphocytes. III. Marginal zone B cells in the spleen are characterized by the expression of Tac and alkaline phosphatase.
Authors: Hsu S M
Published: January 2023

The marginal zone of the spleen contains lymphocytes with an intermediate morphologic form between small lymphocytes and plasmablasts. The majority of cells can be stained by B cell monoclonal antibodies including B1, Leu-14, and Leu-12. The most significant finding is the expression of an interleukin 2 receptor (Tac) and an enzyme-alkaline phosphatase by the marginal zone B lymphocytes. The Tac antigen is not normally present in B cells, but can be found in a portion of in vitro-activated B lymphocytes. In conjunction with other evidence, the marginal zone B lymphocytes may represent the activated B cells in the early stages of B cells differentiation. Alternatively, the Tac-positive, marginal zone B lymphocytes may be a discrete subpopulation of activated B cells.
Influence of RU 41.740, a glycoprotein extract from Klebsiella pneumoniae, on the murine system. II. RU 41.740 facilitates the response to Con A in otherwise unresponsive T-enriched cells.
Authors: Wood C D , Möller G
Published: January 2023

RU 41.740, a glycoprotein extract from Klebsiella pneumoniae, is a polyclonal B cell activator in the thymus-independent category, and both RU 41.740 and its fraction F1 induce the production of interleukin 1 (IL 1). RU 41.740 alone appears to have no direct effect on T-enriched cells. However, we show here that when given in conjunction with concanavalin A (Con A), RU 41.740 enabled nylon wool-passed T cells, which had been accessory cell depleted and lost responsiveness to Con A, to proliferate in response to Con A. Analysis of this observation indicated that RU 41.740 probably acted via a residual accessory cell population and that contact of this cell with the T cell was necessary for Con A activation of the T cell. Because both lectin/antigen and a source of IL 1 are required to stimulate accessory cell-depleted T cells, the mechanism of action of RU 41.740 in this system may be by induction of IL 1 from residual accessory cells in the nylon wool-passed, T-enriched cell population. However, two other agents that stimulate IL 1 production, lipopolysaccharide (LPS) and F1, do not enable T-enriched cells to respond to Con A in this system.
Identification and purification of NK cells with lysosomotropic vital stains: correlation of lysosome content with NK activity.
Authors: Shau H , Dawson J R
Published: January 2023

The lysosome content of lymphocytes has been analyzed with lysosomotropic vital stains and the fluorescence-activated cell sorter (FACS). Large granular lymphocytes (LGL), which account for virtually all natural killing activity in peripheral blood, are quantitatively different from small lymphocytes (SL) in this respect. LGL obtained by Percoll gradient density centrifugation accumulate more of the lysosomotropic vital dyes than SL do, staining with either neutral red or mepacrine (quinacrine). Furthermore among the LGL-rich, low density lymphocyte population highly, granulated cells can be separated from less granulated ones by mepacrine staining and FACS. Thus, separated highly granulated LGL express very high natural killing, whereas the less granulated low density large lymphocytes do not kill.
Generation of functional I-Ed variants from an antigen-presenting macrophage cell line.
Authors: Schwarzbaum S , Diamond B
Published: January 2023

We report the ability of the BAC1.2 macrophage cell line to present antigen both to populations of antigen-primed lymph node cells and to T cell hybridomas. We also describe the selection of I-Ed variants derived from this cell line. These variants have lost the ability to present antigen to an I-Ed plus ovalbumin-specific T cell line, but retain the ability to stimulate an I-Ad plus ovalbumin-restricted T cell line, and a T cell line reactive with I-Ed alone. These variants should aid in the elucidation of structural elements of the I-E molecule involved in the interaction with helper T cells.
Studies on antigens associated with the activation of murine mononuclear phagocytes: kinetics of and requirements for induction of lymphocyte function-associated (LFA)-1 antigen in vitro.
Authors: Strassmann G , Springer T A , Adams D O
Published: January 2023

Macrophages activated and primed in vivo, although not resident or responsive macrophages, express the lymphocyte function associated (LFA)-1 antigen. By contrast, the biochemically related Mac-1 antigen is expressed on all populations of macrophages. In the present paper, we studied regulation of the LFA-1 antigen in vitro. LFA-1 could be induced in vitro on thioglycollate (TG)-elicited but not on proteose peptone (PP)-elicited or resident macrophages. Specifically, macrophage-activating factor (MAF), interferon-gamma (IFN-gamma), or picogram amounts of endotoxin (LPS) induced LFA-1 on TG-elicited macrophages following overnight incubation. Interferon, -alpha or -beta, fucoidin, and colony-stimulating factor were not effective. While some levels of LFA-1 could be detected as soon as 10 hr, peak expression was observed after 16 to 32 hr of incubation. The induction could be completely abrogated by cycloheximide, suggesting that protein synthesis was required. These results indicate that the induction of LFA-1 on mononuclear phagocytes is closely regulated and that the requirements for such induction are distinct from but share certain similarities with induction of cytotoxic functions and expression of Ia antigen.