bluuebunny/crossref_metadata_2025 huggingface dataset:
Migration of human helper/inducer T cells in response to supernatants from Con A-stimulated suppressor/cytotoxic T cells.
Authors: Van Epps D E , Durant D A , Potter J W
Published: December 2022

Previous studies have shown that supernatants from Con A-stimulated human mononuclear cells are chemotactic for T cells, and that the source of the activity is the Leu-2 (suppressor/cytotoxic) T cell. The specificity of this chemoattractant activity for Leu-2 and/or Leu-3 (helper/inducer) T cells was analyzed with isolated human T cell subsets in an in vitro chemotaxis assay system. These studies demonstrate that both human Leu-2 and Leu-3 cells migrate in response to the lymphocyte locomotor stimulus casein, whereas only Leu-3 cells migrate in response to supernatants from Con A-stimulated mononuclear cells. This migration of Leu-3 cells to the Con A supernatant was reflected in both the distance migrated and the number of migrating cells. When Leu-2 cell locomotion in response to Con A supernatants was analyzed, no significant differences from control were observed in either the distance migrated or the number of migrating cells. Supernatants from Con A-stimulated Leu-2 cells similarly attracted Leu-3 cells but not Leu-2 cells in a concentration-dependent manner. Supernatants from Con A-stimulated Leu-3 cells did not stimulate either Leu-2 or Leu-3 cell migration. These studies demonstrate that Leu-3 T cells migrate in response to supernatants from Con A-stimulated Leu-2 T cells. This specific response may promote cell-to-cell interaction by providing a means for Leu-2 T cells to recruit Leu-3 T cells.
Induction of proliferation and NK activity in human lymphocytes by mature myelomonocytic cells: evidence for an HLA-DR-independent MLR stimulatory ability of terminally differentiated nonlymphoid leukemic cell lines and of normal peripheral blood granulocytes.
Authors: Santoli D , Francis M K , Matera L , Ferrero D
Published: December 2022

Three human myeloid leukemic cell lines (HL60, KG1, and ML3) and one histiocytic lymphoma line (U937) were induced to differentiate terminally to mature myelomonocytic cells with either 12-O-tetradecanoylphorbol-13-acetate (TPA) or lymphocyte-conditioned medium (LCM), which is known to contain differentiation-inducing factors. HL60 cells were also forced to differentiate along the myeloid series with retinoic acid (RA) or dimethyl sulfoxide (DMSO). The striking morphologic changes and the expression of differentiated markers on the induced cells (whether macrophage- or granulocyte-like) were always associated with an acquired or dramatically increased ability to stimulate proliferation and natural killer cell (NK) activity in human lymphocytes. Like HL60 cells after granulocytic differentiation, granulocytes freshly separated from the peripheral blood of healthy donors were also strong inducers of mixed lymphocyte reaction (MLR) responses. Analysis of the expression of HLA-DR antigens on the surface of undifferentiated and mature cells with two monoclonal antibodies directed against HLA-DR monomorphic determinants, indicated that 1) upon differentiation induced with RA, DMSO, and TPA the cells never acquired surface DR antigens, and 2) normal peripheral blood granulocytes lacked these antigens. In contrast, treatment with LCM always resulted in the expression of high levels of DR antigens on the differentiated macrophage-like cells. Taken together, these findings indicate that all mature myelomonocytic cells, either freshly separated from peripheral blood or obtained after forced in vitro differentiation of leukemic cells, express MLR stimulatory antigens that appear to be unrelated to DR determinants. The possibilities discussed are that such antigens are associated with other molecules encoded by the D region or with new surface structures unrelated to Ia but dependent on the stage of differentiation.
Effect of thymectomy on peripheral lymphocyte subsets in myasthenia gravis: selective effect on T-cells in patients with thymic atrophy.
Authors: Haynes B F , Harden E A , Olanow C W , Eisenbarth G S , Wechsler A S , Hensley L L , Roses A D
Published: December 2022

Myasthenia gravis (MG) is an autoimmune disease affecting nicotinic acetylcholine receptors. The clinical improvement that follows thymectomy in some myasthenic patients implicates thymic factors as well in the pathogenesis of MG. We have studied circulating immunoregulatory T cell subsets before and after thymectomy in 11 adult patients with MG. Six patients had thymic hyperplasia and five patients had atrophic thymus at the time of thymectomy. Before thymectomy, patients who subsequently were shown to have an atrophic thymus, had lower lymphocyte counts than either patients later shown to have a hyperplastic thymus (1315 +/- 143 lymphocytes/mm3 vs. 2434 +/- 350 lymphocytes/mm3, p less than 0.01), or age-matched controls (1315 +/- 143 lymphocytes/mm3 vs. 2636 +/- 589 lymphocytes/mm3, p less than 0.02). Moreover, after thymectomy, in MG patients with an atrophic thymus, there was a significant rise in lymphocyte count (from 1315 +/- 143 lymphocytes/mm3 to 2279 +/- 292 lymphocytes/mm3, p less than 0.02) beginning 3 days postthymectomy and persisting for at least 6 weeks thereafter. In comparison, patients with a hyperplastic thymus showed no change in circulating lymphocyte counts (p greater than 0.1). Enumeration of lymphocyte subsets in MG patients with an atrophic thymus demonstrated normal B cell numbers before and after thymectomy (p greater than 0.1), whereas, T cells were significantly decreased before thymectomy compared with age-matched normal subjects (859 +/- 82 T cells/mm3 vs. 2215 +/- 545 T cells/mm3, p less than 0.05), and rose to near normal levels after thymectomy (1796 +/- 294 T cells/mm3, p less than 0.02 compared with prethymectomy levels). Using monoclonal anti-T cell antibodies 3A1, OKT4, and OKT8, we found that, before thymectomy in the atrophic thymus group, 3A 1+ T cells were significantly depressed compared with postthymectomy levels (620 +/- 173 cells/mm3 vs. 1627 +/- 331 cells/mm3, p less than 0.02) as were OKT4+ cells 436 +/- 88 cells/mm3 vs. 1112 +/- 63 cells/mm3, p less than 0.001), In contrast, no significant change was seen after thymectomy in the OKT8+ cell subset (p greater than 0.1). MG patients with an atrophic thymus had decreased plasma cortisol levels postthymectomy compared with prethymectomy levels, whereas thymectomy in MG patients with a hyperplastic thymus effected no change in plasma cortisol levels. These data demonstrate in MG patients with an atrophic thymus that thymectomy has an effect on the number of circulating T cells, and in particular, on those T cells expressing antigens 3A1 and OKT4. This effect may in part be mediated by changes in plasma adrenal corticosteroid levels after thymectomy or may be due to a factor produced by atrophic thymuses in MG.
Expansion of cyclophosphamide-resistant cytotoxic precursors in vitro and in vivo by purified human interleukin 2.
Authors: Merluzzi V J , Welte K , Savage D M , Last-Barney K , Mertelsmann R
Published: December 2022

Precursors of cytotoxic lymphoid cells obtained from mice treated with cyclophosphamide (CY) can be expanded in culture by alloantigens in the presence of purified human interleukin 2 (IL 2). Similarly, IL 2 delivered in vivo in a rate-controlled manner enhances cytotoxic activity in mice that are immunosuppressed by high doses of CY. The effector cells are Thy-1.2+ and are not elicited in the absence of antigen.
Tryptic peptide disparity between serologically indistinguishable guinea pig Ia.3,5 antigens.
Authors: Fuge Abruzzini L N , Zacheis M P , Schwartz B D
Published: December 2022

Serological studies have suggested that the DHCBA strain guinea pig expresses an I region which is identical to that of strain 13. However, chemical characterization of Ia.3,5 molecules isolated from these two strains has indicated that these serologically indistinguishable Ia molecules are actually chemically distinct. Ia.3,5 molecules biosynthetically labeled with either [3H]leucine, [3H]arginine, or [3H]lysine were purified by ricin affinity chromatography and isolated by indirect immunoprecipitation with specific alloantisera. Initial examination of the two Ia.3,5 molecules by SDS-PAGE, isoelectric focusing, and two-dimensional gel analyses revealed no strain-specific differences. Furthermore, comparative peptide mapping of the DHCBA and strain 13 radiolabeled Ia.3,5 alpha-chains demonstrated complete peptide homology. In contrast, tryptic peptide maps of amino acid radiolabeled beta-chains revealed two peptides unique to the strain 13 beta-chain and one peptide unique to the DHCBA beta-chain. Analysis of [3H] mannose-labeled beta-chain tryptic peptides verified that the peptide differences observed using 3H-amino acids were not due to variation in N-linked glycosylation. However, strain-specific variability was also noted in the profiles of [3H]mannose-labeled beta-chains. These data indicate that the strain 13 and DHCBA alpha-chains are probably structurally identical, while the beta-chains show strain specific alterations in their chemical structure.
IgE-dependent and ionophore-induced generation of leukotrienes by dog mastocytoma cells.
Authors: Phillips M J , Gold W M , Goetzl E J
Published: December 2022

Isolated dog mastocytoma cells sensitized with dog anti-ragweed IgE and challenged with ragweed antigen generate the C-6 peptide leukotriene (LT) constituents of the slow-reacting substance of anaphylaxis (SRS-A), LTC4 and LTD4, and the potent leukocyte chemotactic factor, LTB4, as well as 15-hydroxyeicosatetraenoic acid (15-HETE), 12-HETE, and 5-HETE. Antigen challenge evoked a mean maximum production of LTC4, LTD4, and LTB4, respectively, of 16.3 ng, 4.6 ng, and 6.7 ng per 10(6) mastocytoma cells within 5 min at 37 degrees C, which was maintained for up to 45 min. The maximum production of leukotrienes by mastocytoma cells activated with optimum concentrations of 0.2 to 1 microM calcium ionophore A23187 was 35% to 50% greater than that achieved by antigen challenge, without alterations in the relative quantities of leukotrienes, but was realized only after 30 min at 37 degrees C. The leukotriene products of mastocytoma cells were identified by high-performance liquid chromatography, spectral properties, and immunoreactivity with mono-specific antisera, and exhibited the same concentration-dependence of biologic activity as authentic synthetic standards. The ratio of quantities of C-6 peptide leukotrienes generated was attributable in part to the rate of peptidolytic conversion of LTC4 to LTD4 in the mastocytoma cells, which was 33% per 30 min at 37 degrees C. The rapid maximum response to IgE-dependent stimulation and the unique spectrum of products distinguishes the secretion of leukotrienes by dog mastocytoma cells from that by basophils and some other types of mast cells and suggests diverse contributions of mast cell leukotrienes to immediate and late hypersensitivity reactions.
Frequency of mast cell precursors in normal tissues determined by an in vitro assay: antigen induces parallel increases in the frequency of P cell precursors and mast cells.
Authors: Crapper R M , Schrader J W
Published: December 2022

Persisting (P) cells, homogeneous populations of cells that grow in vitro for prolonged periods provided a specific growth factor is present, resemble mast cells in many respects. An in vitro assay based on limit dilution was used to determine the frequency of precursors capable of giving rise to P cells. The incidences of P cell precursors per 10(6) cells in tissues of CBA mice in representative experiments were as follows: bone marrow, 291; spleen, 30; mononuclear blood cells, 11; popliteal lymph node, 0.5; and mesenteric lymph node, 18. P cell precursors appeared to be relatively undifferentiated, non-granulated cells; no cells with metachromatically staining granules were detected in the bone marrow or peripheral blood. Furthermore, mice of the Wf/Wf genotype that were grossly deficient in mast cells had the same frequencies of P cell precursors in bone marrow and spleen as their normal +/+ littermates. In many tissues in which we found P cell precursors, pluripotential hemopoietic stem cells are present. Among nonepithelial cells from the gut mucosa, however, in which there was a 10-fold higher frequency of P cell precursors than in bone marrow cells, pluripotential hemopoietic stem cells were undetectable, indicating the existence of committed P cell precursors distinct from pluripotential hemopoietic stem cells. The frequency of P cell precursors in mesenteric lymph nodes was more than 30-fold higher than in the popliteal lymph nodes, suggesting that antigenic stimulation influences their numbers. This latter notion is supported by the observation that after immunization in the footpad, the number of P cell precursors in ipsilateral popliteal lymph nodes rose about 35-fold. Immunization was also accompanied by a rise in mast cell numbers in draining popliteal nodes. This correlation between P cell precursors and the local production of mast cells was strengthened by the observation that the frequency of P cell precursors in cells from the gut mucosa of mice of Wf/Wf genotype, which are unable to mount an intestinal mastocytosis, was more than 1000-fold lower than in wild type mice. Thus, the precursors of P cells and probably of at least the T cell-dependent subset of mast cells appear to be generated in the bone marrow and seed as non-granulated cells via the blood to peripheral tissues such as spleen, lymph node, and mucosal surfaces. P cells appear to be in vitro counterparts of the mucosal subset of mast cells.
Macrophages as effector cells of protective immunity in murine schistosomiasis. V. Variation in macrophage schistosomulacidal and tumoricidal activities among mouse strains and correlation with resistance to reinfection.
Authors: James S L , Skamene E , Meltzer M S
Published: December 2022

Mice of several inbred strains previously characterized as defective in various aspects of macrophage activation for tumor cell killing were utilized to compare the genetic control of induction of macrophage tumoricidal and schistosomulacidal activity and to examine the role of activated macrophages in concomitant immunity to Schistosoma mansoni infection. In all mouse strains tested, macrophage larvicidal activity developed concurrently with tumoricidal activity after in vivo bacillus Calmette Guérin or Corynebacterium parvum treatment. Lpsd strains, A/J, and P/N mice failed to develop macrophages capable of killing either tumor cells or helminth larvae in vitro, whereas C57BL/6J, C3H/HeN, and BALB/c mice all demonstrated strongly cytotoxic cells. In fact, analysis of the reactivity of peritoneal cells from C. parvum-treated F1 hybrids between H-2 congeneic high responder (B10.A) and low responder (A) strains of mice, as well as from F1 hybrid X parent backcross animals, suggested that macrophage activation for killing of these two unrelated extracellular targets is controlled by the same autosomal dominant gene. Strains of mice that failed to develop activated macrophages after in vivo treatment with bacterial stimuli also failed to develop tumoricidal and larvicidal macrophages as a result of S. mansoni infection. The addition of antischistosome sera to the cultures did not reverse the unresponsiveness of macrophages from S. mansoni-infected C3H/HeJ, A/J, or P/N mice in in vitro larvicidal assays. Furthermore, the pattern of concomitant immunity at 6 wk after primary S. mansoni infection correlated closely with the pattern of induction of activated macrophages in these strains of mice. Whereas animals with normal macrophage function developed highly significant levels of concomitant immunity, strains with the Lps defect demonstrated lower levels of acquired resistance, and A/J and P/N mice were not significantly resistant to S. mansoni challenge infection. These results indicate that the genes controlling macrophage activation for the killing of extracellular targets also influence the development of concomitant immunity in schistosomiasis, and suggest that macrophages activated as a consequence of primary S. mansoni infection may be involved in the in vivo effector mechanism of resistance to challenge infection.
Trypanosoma cruzi: description of a highly purified surface antigen defined by human antibodies.
Authors: Scharfstein J , Rodrigues M M , Alves C A , de Souza W , Previato J O , Mendonça-Previato L
Published: December 2022

A glycoprotein of 25,000 daltons (G25) purified from T. cruzi extracts is recognized by serum antibodies of Chagas' disease patients. These human antibodies were isolated by affinity chromatography and were used to demonstrate that G25 antigenic determinants are i) represented at the parasite surface, and ii) are expressed in all developmental stages of the parasite's life cycle, as well as in several T. cruzi strains. This antigen-antibody system may be useful for the diagnosis of Chagas' disease because antibodies to radiolabeled G25 are found in the serum of 96.5% of 173 chagasic patients from different endemic areas, but are not found in the serum from other individuals. Taken collectively, the data suggest that antibodies to G25 define highly conserved determinants of the species T. cruzi. Moreover, its remarkable immunogenicity to infected humans offers an opportunity to investigate the role of specific immunologic responses in the pathogenicity of Chagas' disease.
Epinephrine-induced changes in the distribution of lymphocyte subsets in peripheral blood of humans.
Authors: Crary B , Hauser S L , Borysenko M , Kutz I , Hoban C , Ault K A , Weiner H L , Benson H
Published: December 2022

We have previously demonstrated that mitogen responsiveness of mononuclear cells (MNC) from peripheral blood is reduced after a single injection of epinephrine to human subjects. The purpose of the present study was to characterize the relative distributions of MNC subsets after epinephrine administration using monoclonal antibodies and conventional cell markers. The absolute number of circulating MNC increased 64% within 30 min after injection of epinephrine, and returned to baseline by 2 hr. Analysis of MNC subsets revealed that there were no changes in the relative percentages of total T lymphocytes [T3+ cells, or neuraminidase-treated sheep red blood cell rosettes (EN-rosettes)], B lymphocytes (B1+, or cells with surface-bound immunoglobulin), or monocytes (by morphologic criteria) after epinephrine administration. The percentage of inducer T cells (T4+) declined at 30 and 60 min postinjection. Overall, the percentage of suppressor/cytotoxic T cells (T8+) did not change after injection of epinephrine; however, analysis of individual subjects revealed opposing responses of this subset. The T4:T8 ratio was 2.19 before injection, declined to 1.56 at 60 min, then increased to 3.10 2 hr postinjection. The percentage of natural killer/killer cells (HNK-1+) increased from a baseline of 15.5% before epinephrine injection to 29.6% at 30 min postinjection, then declined to 11.4% at 2 hr. Therefore, the administration of physiologic doses of epinephrine results in changes in the relative proportions of lymphocyte subsets in peripheral blood, in addition to reduced mitogen responsiveness as reported previously.
Two phenotypically distinct suppressor T cell subpopulations inhibit the induction of B cell differentiation by phytohemagglutinin.
Authors: Clement L T , Dagg M K , Lehmeyer J E , Kiyotaki M
Published: December 2022

Human B lymphocytes can be induced to differentiate into antibody-secreting plasma cells by Leu-3+ T lymphocytes stimulated with pokeweed mitogen (PWM), a polyclonal T cell activator. In contrast, other polyclonal T cell mitogens, such as phytohemagglutinin (PHA), also activate Leu-3+ T cells but are relatively ineffective inducers of B cell differentiation. We have performed a series of experiments to investigate the mechanism underlying this apparent paradox. When human B cells were cultured with unfractionated T cells and PWM or PHA, only PWM was able to induce plasma cell formation and immunoglobulin (Ig) secretion. However, when the T cells were treated with mitomycin C (MMC) before culture, both PWM and PHA were able to induce significant B cell differentiation. These data indicated that both mitogens were able to activate the helper T cells required for B lymphocyte differentiation and suggested that MMC-sensitive suppressor T cells were responsible for inhibiting the induction of antibody-secreting cells by MMC-untreated T cells stimulated with PHA. Phenotypic analysis of the T cells capable of suppressing PHA-induced B cell differentiation revealed that small numbers of either Leu-2+ or Leu-3+ T cells could profoundly suppress the B cell differentiation induced by PHA. In contrast, significant suppression of PWM-stimulated B cell differentiation was observed only with relatively large numbers of Leu-2+ T cells. These data confirm previous reports that OKT4+/Leu-3+ T cells can suppress human B cell differentiation and indicate that the difference in B cell differentiation induced by PWM and PHA with MMC-untreated T cells is largely a reflection of the relative potency of these mitogens to activate these phenotypically distinct suppressor T cell subpopulations.
Defective B cell function associated with inherited interstitial deletion of the short arm of the X chromosome.
Authors: Eskola J , Nurmi T , Ruuskanen O
Published: December 2022

The B cell function of a patient with low serum immunoglobulin (Ig) levels and with an interstitial deletion in the short arm of one of her X chromosomes (del(Xp] and the B cell function of her relatives were analyzed by indirect protein A plaque-forming cell (PFC) assay and by measuring immunoglobulin secretion in vitro by ELISA. B cells were activated by pokeweed mitogen (PWM) with or without hydrocortisone (HC) to inhibit HC-sensitive suppressor cells. The patient, her mother, and one sister, all of them having del(Xp), generated very few PFC induced by PWM, even in the presence of HC, whereas normal PFC responses were found in the patient's cytogenetically normal sisters. The B cells of the subjects with del(Xp) secreted very low amounts, if any, of IgA, IgG, and IgM. Co-culture experiments with B cells from del(Xp) subjects and normal OKT 4+ cells revealed no or very low Ig production. The function of the del(Xp) subjects' OKT 4+ cells was slightly reduced, whereas the activity of their OKT 8+ cells was normal. The cell subset analysis in the peripheral blood revealed decreased OKT 4+:8+ ratios in all del(Xp) subjects. The results indicate an intrinsic B cell defect with a possible concomitant immunoregulatory defect associated with del(Xp). Moreover, the results support the hypothesis that antibody production is at least partially controlled by genes located in the short arm of the X chromosome.
Monocyte receptors for the Fc portion of IgG increase in number in autoimmune hemolytic anemia and other hemolytic states and are decreased by glucocorticoid therapy.
Authors: Fries L F , Brickman C M , Frank M M
Published: December 2022

Peripheral blood monocyte receptors for the Fc portion of IgG were quantitatively studied in 43 normal subjects, in 14 patients with warm antibody autoimmune hemolysis (AIHA), and in nine individuals with nonantibody-mediated hemolysis. Monocytes of normal females expressed significantly greater numbers of Fc gamma receptors than did similar cells from male subjects, with no difference in affinity for the IgG1 probe. Monocyte Fc gamma receptor number was increased in patients of both sexes with AIHA; a similar, but smaller, increase in monocyte Fc gamma receptor number was noted in patients with nonantibody-mediated hemolysis. Glucocorticoid administration was associated with a dose-dependent decrease in monocyte Fc gamma receptor number in normal volunteers and patients. Possible etiologic mechanisms and pathogenetic consequences of enhanced monocyte Fc gamma binding in AIHA are discussed.
Regulatory function of Thy-1- cells. IV. Biologic and Biochemical characterization of B cell clone-derived lymphokine (BEF).
Authors: del Guercio P , Poirier M F
Published: December 2022

Studies were made on the immunoregulatory activity of a lymphokine produced by a B cell clone that does not express immunoglobulin heavy or light chains. The B factor (BEF; B cell-derived enhancing factor), was effective in augmenting the primary and the anamnestic response to SRBC and/or in prolonging the IgG anamnestic response to SRBC and the response to DNP-LE, provided it was added at early stages of culture. The BEF seems to act mainly by preventing the activation of T suppressor cells rather than by counteracting the activity of already activated suppressor cells. Indeed, thymocytes and virgin T cells, but not Con A-activated thymocytes, Con A-activated T cells or B cells, express acceptors for the BEF. The biochemical properties of the BEF were also reported. The apparent m.w. of the factor was 450,000 to 500,000 by gel chromatography. The BEF was sensitive to digestion by papain, trypsin, and subtilisin, indicating that it was protein in nature. The regulatory activity was precipitated by 50% (NH4)2SO4 and was destroyed by heating at 70 degrees C for 30 min or by exposure to pH 2.2 for 6 hr.
Restoration of LPS responsiveness of C3H/HeJ mouse lymphocytes by microinjection of cytoplasmic factor(s) from LPS-stimulated normal lymphocytes.
Authors: Eda Y , Ohara J , Watanabe T
Published: December 2022

Cytosol fractions of lymphocytes from LPS-responder mice (C3H/HeN) were prepared and injected into B lymphocytes of LPS-nonresponder mice (C3H/HeJ) by a microinjection technique utilizing polyethyleneglycol-mediated cell fusion. The B lymphocytes of C3H/HeJ mice microinjected with cytosol prepared from LPS-stimulated C3H/HeN cells became normally responsive to LPS. Microinjection of cytosol itself did not stimulate C3H/HeJ cells to proliferate or differentiate into immunoglobulin-producing cells, and the cells injected with cytosol had to be restimulated with LPS in order to proliferate and differentiate. These data suggested that C3H/HeJ B cells acquired LPS responsiveness by microinjection of cytoplasmic factor(s) from LPS-stimulated C3H/HeN cells and that these factor(s) may be one of the components involved in normal signal transmission from cell surface to nucleus in the early stages of the LPS response. The apparent m.w. of the cytoplasmic factor(s) is 100,000 by gel filtration. Chromatofocusing analysis suggested that these factors may consist of two components with the same m.w.
Preferential association of kappa IIIb light chains with monoclonal human IgM kappa autoantibodies.
Authors: Ledford D K , Goñi F , Pizzolato M , Franklin E C , Solomon A , Frangione B
Published: December 2022

The predominance of the relatively uncommon V region subgroup isotype kappa III among the light chains of human monoclonal (IgM kappa) anti-IgG antibodies, (i.e., rheumatoid factors), was further documented through sequence analyses of ten such autoantibodies isolated from IgM-anti-IgG cold-insoluble immune complexes (mixed cryoglobulins). The amino-terminal sequence of all ten kappa-chains was characteristic for kappa III proteins and virtually identical to that of a prototype kappa III light chain. Similar sequence identity was found for kappa-chains isolated from three IgM kappa autoantibodies that formed cold-insoluble immune complexes with low-density lipoprotein (LDL). The thirteen light chains were found to be virtually identical in sequence for the first framework region (FR); ten of these proteins sequenced through the first complementarity-determining region (CDR) and into the second FR were markedly similar. The second CDR of five proteins was almost identical in sequence to that of the prototype kappa III-chain. Concordance was also demonstrated between the structural classification of the light chains as kappa III and their immunochemical classification as members of this V region subgroup. Serologic analyses of light chains isolated from seven IgM kappa autoantibodies (six anti-IgG, one anti-LDL) and of one intact IgM kappa anti-LDL antibody showed that each had antigenic determinants common to kappa II proteins. These light chains also expressed the antigenic determinant(s) of a V-region sub-subgroup of kappa III proteins designated kappa IIIb. Our studies confirm the preferential association of kappa III (and kappa IIIb) light chains with IgM kappa anti-IgG antibodies and demonstrate a similar association for IgM kappa anti-LDL antibodies. The finding that these and other types of IgM kappa autoantibodies, e.g., cold agglutinins, have remarkably similar light chains suggests an inherent restriction in the immune response to self-antigens.
Syngeneic anti-idiotypes against monoclonal anti-HLA-DR antibodies: characterization of recurrent idiotopes.
Authors: Piatier-Tonneau D , Boyer B , Debre P , Charron D J
Published: December 2022

Monoclonal anti-HLA-DR antibody (anti-DR mAb) reactivity was investigated at the idiotypic level; three syngeneic anti-idiotype (Id) sera were raised against three monomorphic anti-DR mAb (206, BM 50, and D 1.12). The syngeneic anti-Id responses exhibited some differences in their intensity and the idiotypic determinants recognized by the three anti-Id sera were localized, at least in part, on the antigen combining site of the respective anti-DR mAb. Idiotype analysis of 24 anti-DR mAb was performed with these anti-Id sera by Id-binding inhibition assays. Idiotypic cross-reactivity was demonstrated for 12 out of the 24 anti-DR mAb, indicating a large degree of idiotypic recurrence. Analysis of the cross-inhibition patterns led us to define at least five sets of recurrent idiotopes: one set cross-reacting with anti-206 Id, the second with both anti-BM 50 and anti-D 1.12 Id, the third with anti-BM 50 Id only, the fourth with anti-D 1.12 Id only, and a fifth set was shared by D 1.12 and NE 4. Eleven monomorphic anti-DR mAb and one polymorphic anti-DR 3 mAb (16.23) did not cross-react with the three anti-Id sera. These results suggest that the mouse antibody response to monomorphic determinants on human Ia antigens is based on a limited number of recurrent idiotopes. In two cases, recurrent idiotopes could be related to a similar anti-DR mAb fine specificity. These anti-Id provide useful reagents to classify and compare anti-DR mAb.
Variation in the glycosylation of the murine sex-limited protein: comparison with the fourth component of murine complement.
Authors: Karp D R
Published: December 2022

The murine sex-limited protein (Slp) is a hemolytically nonfunctional homologue of the fourth component of complement (C4). Two congenic mouse strains, B10.BUA1 (H-2w16) and B10.KPB128 (H-2w19), which have been previously shown to share a variant form of C4 (Karp et al., J. Biol. Chem., 257: 7330-7335), were examined and found to also produce a variant form of Slp. Slp molecules isolated from the plasma or peritoneal macrophage cultures from these strains have an alpha-chain approximately 2,000 daltons smaller than the alpha-chain of Slp from H-2d or H-2w7 mice as judged by sodium dodecyl sulfate (NaDodSO4)/polyacrylamide gel electrophoresis. Expression of this Slp was constitutive, i.e., not regulated by androgen, and is cis-dominant in F1 hybrid mice. Autolysis of the different relative molecular mass (Mr) alpha-chains at the internal thiolester produced similar Mr amino-terminal fragments and different Mr carboxy-terminal fragments. Deglycosylation of the alpha-chains with trifluoromethanesulfonic acid eliminated most, if not all, the Mr difference. The Mr difference was also manifested by the intracellular precursors of Slp and could be eliminated by endoglycosidase H (endo H) treatment. The number of oligosaccharides on the Slp alpha-chain was deduced by limited endo H treatment of Slp synthesized in the presence of swainsonine, a plant alkaloid that prevents maturation of complex-type oligosaccharides. This method is a simple way to enumerate the complex-type, N-linked oligosaccharides on glycoproteins. The genetic variation in the glycosylation of Slp was compared with the known variation in glycosylation of C4, and a scheme depicting some of the structural differences among these molecules was developed.
Inhibition of mitogen-induced lymphocyte proliferation by ouabain: interference with interleukin 2 production and interleukin 2 action.
Authors: Stoeck M , Northoff H , Resch K
Published: December 2022

Since the discovery of IL 2 it has been possible to dissect the mitogenic activation of T lymphocytes into two steps: first, the production of IL 2; and second, the response of IL 2-dependent T cell blasts to IL 2. We investigated the influence of ouabain, a known inhibitor of T cell activation, on the production of and the response to IL 2 in mouse T lymphocytes. Ouabain dose-dependently inhibited DNA synthesis in Con A activated C57BL/6 spleen cells; inhibition below background levels occurred at a concentration of 5 X 10(-4) mol/liter. The action of IL 2 was assayed on a mouse T cell line, named M 2, derived from C57BL/6 spleen cells and maintained in continuous IL 2-dependent culture for more than 21 mo. Ouabain dose-dependently inhibited the response of these cells to mouse IL 2 with optimal inhibition at 10(-3) mol/liter ouabain. The production of IL 2 by Con A-stimulated C57BL/6 spleen cells was also totally inhibited by 10(-3) mol/liter ouabain; 10(-4) mol/liter ouabain inhibited IL 2 production partially, and 10(-5) mol/liter and 10(-6) mol/liter ouabain had no inhibitory effect. The ouabain-mediated inhibition of IL 2 production and of IL 2 action was not due to a nonspecific toxicity, because the preincubation of spleen cells with 10(-3) mol/liter ouabain for 24 hr did not impair their ability to produce IL 2, nor was the response of M 2 cells to exogenously added IL 2 abrogated by the preincubation of the cells with 10(-3) mol/liter ouabain. The data suggest that ouabain acts on two steps during the induction of T cell activation.
Antigen and MHC restriction specificity of two types of cloned male-specific T cell lines.
Authors: Tomonari K
Published: December 2022

Two types of H-Y antigen-specific cloned T cell lines were established. Clone 2-1 is an IAb restricted proliferating T cell line and clone 10-2 is an H-2Db restricted proliferating and cytotoxic T cell line. Clone 2-1 proliferated in response to H-Y antigen with an IAb restriction in the absence of IL 2. IAb mutant B6.C-H-2bm12 (bm12) mice did not stimulate this clone. This defect may be the cause of the unresponsiveness to H-Y antigen with cytotoxic T lymphocytes (CTL) in bm 12 mice (i.e., the failure of activation of H-Y-specific helper T cells). Although (B10.A X bm12)F1 mice are known to show functional complementation, as demonstrated by the presentation of insulin to helper T cells and the H-Y-specific CTL response, the F1 male cells could not stimulate this clone. The IA determinant restricting H-Y antigen presentation to helper T cells and/or the H-Y antigenic determinant seems to differ between C57BL/6 (B6) and the F1 mice. Monoclonal anti-IAb antibodies but not anti-H-Y antibodies blocked the proliferation of this clone. The clone had no cytotoxic activity unless Con A was added to the cytotoxicity assay culture. Clone 10-2 proliferated in response to H-Y antigen with an H-2Db restriction. Proliferation was not observed unless specific stimulator cells and IL 2 were supplied to the culture; IL 2 alone could not support this clone. The clone did not respond to male cells from the H-2Db mutant B6.C-H-2bm14 (bm14). The clone had cytotoxic activity against male H-2Db+ cells but not against male bm14 cells. This evidence indicates that the mutation of bm14 occurred in the structural gene coding for a molecule on which a determinant restricting the recognition of H-Y antigen by B6 CTL exists. The phenotypes of clone 2-1 were Thy-1.2+, Ly-1.2-, Ly-2.2-, IAb-, and H-2Kb+, and the phenotypes of clone 10-2 were Thy-1.2+, Ly-1.2-, Ly-2.2+, IAb-, and H-2Kb+.
Modulation of B and T cell subsets in mice treated with fractionated total lymphoid irradiation (TLI). IV. Expression of TL4-bearing cells in spleen and bone marrow of TLI-treated mice.
Authors: Zan-Bar I
Published: December 2022

The pre-thymic cell phenotypes arising in spleen and bone marrow (BM) of fractionated total lymphoid irradiation (TLI)-treated mice are characterized in this paper. Different strains of mice received 200 rad lymphoid irradiation daily for 8 to 10 days. At different time intervals after treatment, BM or spleen cells of treated mice were stained with a battery of antisera directed specifically to TL and Thy-1 antigens. Analysis of the stained cells on a fluorescence-activated cell sorter (FACS) apparatus revealed that 10 to 30% of spleen or BM cells of TLI-treated mice were TL+ cells whereas no TL+ cells could be found in the same organs in the normal untreated mice. The appearance of TL+ cells in the spleen and BM of TLI-treated mice was found to be thymic independent, as thymectomy of mice neither prevented nor altered the kinetics of TL+ cell appearance. The sequence of antigenic marker appearance reveals that TL markers precede Thy-1 markers and that TLaa markers precede TL4 antigen. The relevance of the TL4+ cells to the development of leukemias is discussed.
Differential production of deoxyadenosine by human T and B lymphoblasts.
Authors: Iizasa T , Kubota M , Carson D A
Published: December 2022

The association of a genetic deficiency of adenosine deaminase (ADA) with immunodeficiency disease has emphasized the importance of deoxyadenosine and adenosine metabolism for human lymphocyte function. However, information concerning the endogenous production and metabolism of deoxyadenosine and adenosine in normally growing human T and B lymphoblasts is lacking. In the present experiments, we used a diverse series of cell lines deficient in individual enzymes of purine metabolism to quantitate the de novo formation of deoxyadenosine and adenosine in human T lymphoblasts (CEM), B lymphoblasts (WI-L2), and histiocytic lymphoma cells (DHL-9). The B lymphoblasts and histiocytic lymphoma cells generated deoxyadenosine at a rate of 60 to 80 pmol/hr/10(7) cells. This value was several fold greater than the rate of production of deoxyadenosine by T cells (6 to 7 pmol/hr/10(7) cells). Deoxyadenosine synthesis required ribonucleotide reductase activity, and was maximal during the S-phase of the cell cycle. The T and B lymphoblasts formed relatively similar amounts of adenosine (870 to 1620 pmol/hr/10(7) cells) throughout the cell cycle. In ADA-deficient cells, a major fraction of the deoxyadenosine synthesized de novo was excreted into the extracellular space. These results establish that the endogenous synthesis and metabolism of deoxyadenosine (but not adenosine) is distinctly different in T and B lymphoblasts.
Interleukin 2 receptor expression by activated HNK-1+ granular lymphocytes: a requirement for their proliferation.
Authors: Abo T , Miller C A , Balch C M , Cooper M D
Published: December 2022

A subpopulation of HNK-1+ cells was found to express interleukin 2 receptor (IL 2R) as monitored by the anti-Tac monoclonal antibody after lectin or allogeneic cell stimulation. When blood mononuclear cells were stimulated with phytohemagglutinin (PHA) or concanavalin A for 3 days, or with pokeweed mitogen or allogeneic mononuclear cells for 6 days, virtually all of the HNK-1+ cells remained as small resting lymphocytes. One-fourth of these HNK-1+ cells expressed IL 2R, however, usually within 18 hr after stimulation. Neither circulating HNK-1+ cells nor unstimulated HNK-1+ cells in short-term culture expressed IL 2R. Only the subset of HNK-1+ cells that expressed T cell surface markers, such as the sheep erythrocyte receptor and Leu-4 antigen, could be induced to express IL 2R. When long-term cultures of HNK-1+ cells were established with initial PHA stimulation and the continued presence of IL 2, a majority of the cells was found to express the IL 2R. When anti-Tac antibody was added to the long-term cultures, proliferation of the HNK-1+ cells was completely inhibited. These results suggest that entry into growth cycle by this subpopulation of HNK-1+ granular lymphocytes may require two signals: a relatively nonspecific ligand interaction with cell surface glycoprotein(s) to induce expression of IL 2R, and subsequent IL 2 interaction with these specific cell surface receptors.
Preparation of goat antibodies against interleukin 1: use of an immunoadsorbent to purify interleukin 1.
Authors: Mizel S B , Dukovich M , Rothstein J
Published: December 2022

Antibodies with specificity for interleukin 1 (IL 1) were produced in a goat immunized with purified IL 1 alpha obtained from the murine macrophage cell line, P388D1. The anti-IL 1 IgG were capable of completely inhibiting the biologic activity of IL 1 in the murine thymocyte assay but had no effect on IL 2-driven T cell responses. Although the anti-IL 1 IgG were produced by using mouse IL 1, these antibodies also recognized IL 1 prepared from a human monocyte leukemia cell line. An immunoadsorbent column was prepared and used for the large-scale purification of IL 1 in relatively high yield. Approximately 250 micrograms of purified IL 1 were obtained from 50 liters of culture fluid, the yield being 20% of the initial activity. Six major species of IL 1 were resolved by using tris-glycinate discontinuous polyacrylamide gels. The purified IL 1 exhibited 50% of its maximal activity in the thymocyte proliferation assay at a concentration of approximately 1 X 10(-11) M.
Biochemical and physicochemical characterization of mouse B cell growth factor: a lymphokine distinct from interleukin 2.
Authors: Farrar J J , Howard M , Fuller-Farrar J , Paul W E
Published: December 2022

A subline of the C57BL/6 mouse EL-4 thymoma produces upon stimulation with phorbol myristate acetate (PMA) a B cell growth factor (BCGF) that co-stimulates anti-IgM-stimulated splenic B cells. The BCGF present in the EL-4 thymoma culture supernatants is shown to be distinct from IL 2 by a variety of chromatographic criteria. Untreated BCGF exhibits a heterogeneity of isoelectric points (pH 6.4 to 6.6, 7.4 to 7.6, 8.5 to 8.7) all of which are more alkaline than that of IL 2 (pl 3.5 to 4.8). Neuraminidase-treated BCGF exhibits a single major pl of 9.3 that is also distinct from similarly treated IL 2. Moreover, IL 2 and BCGF exhibit distinct monomeric m.w. by SDS gel electrophoresis under nonreducing conditions (IL 2 is 21,000; BCGF is 11,000 and 15,000). The biologic activity of IL 2 is sensitive to trypsin inactivation as is BCGF activity. Thus, BCGF and IL 2 appear to be biologically and biochemically distinct glycoproteins.
Biosynthesis, glycosylation, and in vitro translation of the human T cell antigen Leu-4.
Authors: Bergman Y , Stewart S J , Levy S , Levy R
Published: December 2022

Two families of polypeptides were found in immunoprecipitates formed from human T cell lines by using the monoclonal antibody anti-Leu-4. When analyzed by one- and two-dimensional SDS-PAGE, the first family consisted of members with m.w. of 26,000 and isoelectric points that varied from very basic to acidic. The second family consisted of members with m.w. of 22,000 to 23,000 and isolectric points that varied from neutral to very acidic. The biosynthetic processing and intracellular glycosylation of several of these polypeptides were examined. Some, but not all, intracellular forms of the Leu-4 antigen contained N-linked oligosaccharides of the high mannose type. By using messenger RNA from human T cell lines, intracellular forms of Leu-4 were shown to be accurately translated and properly processed by Xenopus oocytes in vitro.
Regulation of azophenylarsonate-specific repertoire expression. II. Fine specificity and isotype heterogeneity of anti-arsonate antibodies.
Authors: Sigal N H
Published: December 2022

This report investigates the isotope and binding site heterogeneity of IgG anti-azophenylarsonate (ARS) monoclonal antibodies derived from splenic fragment cultures of nonimmune and immune A/J mice. By using five arsanilic acid derivatives in a hapten inhibition assay to determine fine specificity, the secondary ARS-specific repertoire was found to be extremely heterogeneous, with 52 unique patterns among the 81 antibodies analyzed. CRIA+ and CRIA+/- clonotypes were as heterogeneous as CRIA- antibodies, but a majority of CRIA+/- clones maintained a characteristic relative affinity for two of the haptens, p-arsanilic acid (p-ARS) and p-azobenene arsonic acid-N-tyrosine (ABA-tyr), which was identical to the prototype CRIA+ hybridoma protein R16.7. We also observed that the average relative affinity of CRIA- antibodies from immunized donors was higher than nonimmune mice with respect to p-ARS and ABA-tyr, and that there was a further increase in avidity to ABA-tyr over time after immunization. In contrast, no affinity maturation occurred among the CRIA+ and CRIA+/- clones. Rare antibodies were also identified within the nonimmune repertoire that had higher relative affinities than R16.7. Despite this, a greater proportion of CRIA+ and CRIA+/- clones produced IgG antibody in vitro. Thus, the predominance of CRIA+ antibodies in the ARS-specific immune repertoire and the preferential differentiation of CRIA+ precursor cells to IgG secretors cannot be entirely explained by the avidity of this clonotype family for p-ARS or ABA-tyr.
A circular antibody-antigen complex is responsible for increased affinity shown by mixtures of monoclonal antibodies to human chorionic gonadotropin.
Authors: Moyle W R , Anderson D M , Ehrlich P H
Published: December 2022

Mixtures of some pairs of monoclonal antibodies that have separate epitopes on the beta-subunit of hCG have increased affinity for the hormone relative to that of either antibody alone. A mathematical model developed to explain the phenomenon predicted that a circular tetrameric complex composed of each antibody and two molecules of hCG was responsible for the effect. This structure has now been identified experimentally by the following criteria: 1) the m.w. of the complex observed by electrophoresis (370,000 g/mol) and gel filtration (440,000 g/mol) was in agreement with the m.w. expected for a tetramer composed of two molecules of antibody and two molecules of hCG (i.e., 376,000 g/mol); 2) the ratio of individual antibodies to hCG measured with the use of 131I and 125I-labeled antibodies and/or hCG was 1:1:2; and 3) the complex failed to adhere to affinity columns containing either antibodies or hCG covalently coupled to Sepharose. These columns adsorbed B101, B102, hCG, and mixtures of B101 plus hCG or B102 plus hCG. The observations made with the affinity resins are compatible with a circular model for antigen-antibody complex in which the epitopes of the antigen and the binding site of the antibodies were mutually and completely obscured. Although not studied in detail, a similar complex was formed when the beta-subunit of hCG was substituted for the intact hormone. In addition, a mixture of antibodies that bound to the alpha- and beta-subunits of hCG (i.e., A102 and B102) and that had a higher affinity for the hormone than either antibody also gave rise to a similar species that could be detected after electrophoresis. A pair of antibodies that bind to separate epitopes on the beta-subunit (i.e., B101 and B103) and do not show enhanced affinity for hCG failed to form a stable complex that could be identified as a separate species after electrophoresis. Thus, the studies reported here confirm earlier theoretical predictions linking the increase in affinity observed on mixing monoclonal antibodies to the formation of a circular complex.