bluuebunny/crossref_metadata_2025_split huggingface dataset:
The effect of dihydrofolate reductase-mediated gene amplification on the expression of transfected immunoglobulin genes.
Authors: Dorai H , Moore G P 
Published: December 2022

Abstract
Murine/human chimeric gamma 1 and K Ig genes were cloned adjacent to the gene coding for methotrexate-resistant dihydrofolate reductase. These constructs were introduced into myeloma cells, and lines containing stably integrated genes were selected. The integrated Ig genes were then amplified by selection of the cells in increasing concentrations of methotrexate. The extent of gene amplification, mRNA accumulation, and production of Ig was studied in transfectomas containing introduced light chain genes, heavy chain genes, or both. When the light chain gene was introduced alone, it was expressed at low levels, but after selection with methotrexate, light chain expression was increased as much as 63-fold. In contrast, the transfected heavy chain genes were highly expressed, but production of the corresponding protein was increased a maximum of only fourfold by methotrexate treatment. Cellular toxicity of unassembled heavy chain monomer was not observed, even at amounts equivalent to 2% of total cellular protein. Cointroduction of the heavy and light chain constructs with subsequent amplification resulted in as much as 25-fold increase in secretion of intact antibody relative to unamplified cells. The results demonstrate that amplification of Ig genes can induce transfectomas to secrete antibody at nearly the rate of hybridomas.
The immune response and the eye. I. The effects of monoclonal antibodies to T suppressor factors in anterior chamber-associated immune deviation (ACAID).
Authors: Ferguson T A , Kaplan H J 
Published: December 2022

Abstract
We report the effects of two monoclonal antibodies (mab) specific for murine T suppressor (Ts) factors (TsF) in anterior chamber (AC)-associated immune deviation (ACAID), as induced by AC inoculation of TNP-coupled syngeneic spleen cells (TNP-Spl). One mab (14-12) is specific for Ts effector factor and can block the induction of Ts cells in ACAID if given before or after AC injection of TNP-Spl. The other mab (14-30) is specific for Ts inducer factors and blocks suppression only after given after TNP-Spl. We also studied the surface phenotype of the Ts cells induced by AC injection of TNP-Spl. We show that at least two cells are required for the adoptive transfer of suppression in TNP-ACAID. One is Lyt-2+ and 14-12+, the other is I-J+. These Ts cells have the surface phenotype of Ts effector cells as seen in other systems. These results indicate that mab which bind TsF in other systems affect Ts cells in TNP-ACAID, and that the Ts cells induced in TNP-ACAID are only of the Ts effector type.
Dysfunction of irradiated thymus for the development of helper T cells.
Authors: Amagai T , Kina T , Hirokawa K , Nishikawa S , Imanishi J , Katsura Y 
Published: December 2022

Abstract
The development of cytotoxic T cells and helper T cells in an intact or irradiated thymus was investigated. C57BL/6 (H-2b, Thy-1.2) mice were whole body-irradiated, or were irradiated with shielding over either the thymus or right leg and tail, and were transferred with 1.5 X 10(7) bone marrow cells from B10.Thy-1.1 mice (H-2b, Thy-1.1). At various days after reconstitution, thymus cells from the recipient mice were harvested and a peanut agglutinin low-binding population was isolated. This population was further treated with anti-Thy-1.2 plus complement to remove host-derived cells and was assayed for the frequency of cytotoxic T cell precursors (CTLp) and for the activity of helper T cells (Th). In the thymus of thymus-shielded and irradiated mice, Th activity reached normal control level by day 25, whereas CTLp frequency remained at a very low level during these days. In the thymus of whole body-irradiated mice, generation of CTLp was highly accelerated while that of Th was retarded, the period required for reconstitution being 25 days and more than 42 days for CTLp and Th, respectively. Preferential development of CTLp was also seen in right leg- and tail-shielded (L-T-shielded) and irradiated recipients. Histological observation indicated that Ia+ nonlymphoid cells were well preserved in the thymus of thymus-shielded and irradiated recipients, whereas in L-T-shielded and irradiated recipients, such cells in the medulla were markedly reduced in number. These results suggest strongly that the generation of Th but not CTLp is dependent on radiosensitive thymic component(s), and that such components may represent Ia+ cells themselves in the medulla or some microenvironment related to Ia+ cells.
Dissociation between the expression of IL 2 receptor and IL 2 receptor mRNA in the antigen-specific T cell clone stimulated by the specific antigen with B cell APC.
Authors: Matsui M , Ariga H , Kakiuchi T , Nariuchi H 
Published: December 2022

Abstract
An antigen-presenting capacity of B cells was analyzed in comparison with that of whole spleen cells in terms of IL 2R expression of a human IgG-specific T cell clone, 24-2C.3. When the clone was stimulated with the antigen and irradiated spleen cells, the clone expressed an adequate amount of IL 2R on the surface in flow cytometric analysis. On the other hand, the clone expressed the receptor only poorly, being stimulated with B cell APC. The findings were confirmed by Western blot analysis on the clone solubilized by Nonidet P-40 and also by fluorescent antibody technique on the fixed clone. 24-2C.3 clone stimulated with B cell APC was also suggested not to secrete IL 2R in culture medium. In dot blot hybridization analysis performed with cDNA probe, however, 24-2C.3 clone stimulated with B cell APC was shown to express a comparable amount of IL 2R mRNA to the clone stimulated with whole spleen cells as APC. The mRNA expressed in the clones stimulated with B cells and with spleen cells could not be distinguished from each other in Northern blot analysis. The culture supernatant of 24-2C.3 clone incubated with the antigen and irradiated spleen cells was shown to enhance the expression of IL 2R on the clone stimulated with B cell APC. These results suggest that there is some defect in the antigen-presentation capacity of B cells in terms of the elicitation of the intracellular signal for IL 2R expression in T cells and that the humoral factor(s) produced in the presence of splenic adherent cells could supply the defect.
rIL 2-induced proliferation of human circulating NK cells and T lymphocytes: synergistic effects of IL 1 and IL 2.
Authors: Ben Aribia M H , Leroy E , Lantz O , Métivier D , Autran B , Charpentier B , Hercend T , Senik A 
Published: December 2022

Abstract
Cells participating in the rIL 2-induced proliferation of resting PBMC were identified by using different methods of cell purification. NK cells recovered in the light density fraction of Percoll gradients responded, as already known, directly to rIL 2 by strong proliferation. In contrast, large T lymphocytes co-purifying with NK cells, and small T cells sedimenting in the high density area of the Percoll gradients, were virtually unresponsive when cultivated in the sole presence of rIL 2. However, the addition of either irradiated autologous monocytes or highly purified IL 1 allowed both kinds of T cells to undergo cell division. Stringent elimination of possibly contaminating NK cells (NKH-1+) and/or activated T cells (TNKTAR, Tac+, HLA-DR+) from the high density T cells by complement lysis did not impair rIL 2-induced cell proliferation, indicating entire responsiveness of these cells to the synergistic action of IL 1 plus IL 2. Both high density CD4+ and CD8+ participated in this phenomenon, with an apparent advantage for CD4+ cells. All Tac+ cells emerging in a 6-day culture of these cells expressed the WT31 antigen, which indicates that T cells involved in rIL 2-induced proliferation are conventional mature T cells. The relative precursor frequencies of NK cells, large T lymphocytes, and small T lymphocytes that proliferated in response to rIL 2 were analyzed by limiting dilution analysis. The frequencies of clonal growth of NK cells and low density T lymphocytes were approximately the same (1/103 vs 1/185), whereas that of high density T cells was four times lower (1/458). Thus, we clearly demonstrate that resting T cells, defined as such by morphological, density, and phenotypic criteria, are able to proliferate in response to IL 2 in the presence of IL 1 without antigenic or mitogenic triggering.
Interleukin 1 stimulates human endothelial cells to produce granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor.
Authors: Broudy V C , Kaushansky K , Harlan J M , Adamson J W 
Published: December 2022

Abstract
Endothelial cells are a potent source of hematopoietic growth factors when stimulated by soluble products of monocytes. Interleukin 1 (IL 1) is released by activated monocytes and is a mediator of the inflammatory response. We determined whether purified recombinant human IL 1 could stimulate cultured human umbilical vein endothelial cells to release hematopoietic growth factors. As little as 1 U/ml of IL 1 stimulated growth factor production by the endothelial cells, and increasing amounts of IL 1 enhanced growth factor production in a dose-dependent manner. Growth factor production increased within 2 to 4 hr and remained elevated for more than 48 hr. To investigate the molecular basis for these findings, oligonucleotide probes for granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), macrophage colony-stimulating factor (M-CSF), and multi-CSF were hybridized to poly(A)-containing RNA prepared from unstimulated and IL 1-stimulated endothelial cells. Significant levels of GM-CSF and G-CSF, but not M-CSF or multi-CSF, mRNA were detected in the IL 1-stimulated endothelial cells. Biological assays performed on the IL 1-stimulated endothelial cell-conditioned medium confirmed the presence of both GM- and G-CSF. These results demonstrate that human recombinant IL 1 can stimulate endothelial cells to release GM-CSF and G-CSF, and provide a mechanism by which IL 1 could modulate both granulocyte production and function during the course of an inflammatory response.
Androgen receptors in human thymocytes.
Authors: Kovacs W J , Olsen N J 
Published: December 2022

Abstract
On the basis of a variety of evidence androgenic hormones are thought to exert modulatory effects on immune function. Because specific actions of these hormones are exerted through receptor-mediated mechanisms, we have examined the human thymus gland for the presence of specific high-affinity androgen receptors. Cytosolic extracts of whole human thymus were found to contain 6.5 (+/- 2.6) fmol of specific [3H]methyltrienolone binding per mg protein. The receptor sedimented at 8s on sucrose density gradient centrifugation analysis. Thymic fibroblast lines derived from these glands did not express sufficient receptor content to account for the findings in whole thymus cytosolic extracts. Purified thymocyte preparations (greater than 99.3% pure as assessed by flow cytometry with monoclonal anti-thymocyte marker OKT11) contained 3.34 (+/- 1.2) fmol receptor per mg cytosolic protein. Mature peripheral blood lymphocytes, human T cells obtained from thoracic duct drainage, and the human T cell line Jurkat were not found to contain these receptors. These studies demonstrate for the first time that a high affinity androgen receptor is present in maturing human thymocytes. A functional role for these receptors is suggested by dihydrotestosterone-mediated decreases in interleukin 2-like activity produced by cultured human thymocytes.
Molecular mapping class II polymorphisms in the human major histocompatibility complex. II. DQ beta.
Authors: MacMurray A J , Bell J I , Denney D , Watling D , Foster L S , McDevitt H O 
Published: December 2022

Abstract
The restriction fragment length polymorphisms have been determined for six restriction enzymes (Bam HI, Bg1 II, Eco RI, Hinc II, Hind III, and Pvu II) and a DQ beta probe on 25 cell lines that are homozygous by consanguinuity at the MHC. These patterns reflect both DR haplotypes and DQ types of the cells tested. At least one non-polymorphic band is present in all the cell lines with every restriction enzyme except Hinc II. This band most probably represents DX beta hybridization. The polymorphic bands indicate that more polymorphism exists in the DQ subregion than is predicted serologically. Each DR haplotype is associated with a unique set of restriction fragments except for DR2 and DR6. The patterns are largely consistent within each DR haplotype. In addition, some bands reflect the established DQ specificities DQw1 and DQw2. Individual bands can be identified that are unique to the haplotypes DR1, DR4, DR5, and DR6 and the DQw1- and DQw2-associated haplotypes. Subdivisions of haplotypes can be identified with this probe. In particular, MVL (DR1), Akiba (DR2), QBL (DR3), FPF (DR5), and APD (DR6) have polymorphisms that distinguish them from other members of their DR haplotype.
Molecular analysis of heavy and light chains used by primary and secondary anti-(T,G)-A--L antibodies produced by normal and xid mice.
Authors: Busto P , Gerstein R , Dupre L , Giorgetti C A , Selsing E , Press J L 
Published: December 2022

Abstract
The primary (1 degree) antibody response to (T,G)-A--L shows limited heterogeneity, consisting mostly of side chain-specific antibodies that bind GT and that express the TGB5 idiotype (Id). The secondary (2 degrees) response is very diverse: antibodies that bind the backbone A--L constitute a third of the response, and a high proportion of the side chain-specific antibodies do not bind GT and are TGB5 Id-. To provide a molecular basis for understanding this difference in repertoire expression, we analyzed the Ig genes used by heavy and light chains of 1 degree and 2 degrees side chain-specific anti-(T,G)-A--L hybridoma antibodies (HP). Southern blot restriction analysis and nucleotide sequence analysis of the expressed genes used by three TGB5 Id+ 2 degrees HP showed usage of three different VH genes in two VH gene families (36-60 and J558), different D segments, and two different Vk1 genes (the Vk1A and Vk1C subgroups). Thus, antibody heterogeneity in the 2 degrees response is contributed by combinatorial diversity of distinct germ-line genes. Nucleotide sequence analysis of the expressed genes used by TGB5 Id+ 1 degree HP showed use of highly homologous VH genes in the J558 VH gene family and highly homologous Vk1A genes. The majority of TGB5 Id+ 1 degree HP from different donors gave similar heavy and similar light chain gene rearrangements by Southern blot restriction analysis, after correction for known or potential J region differences. The combined nucleotide sequence and Southern blot restriction analysis data suggest that most 1 degree B cells use the same or very similar VH and Vk genes, i.e., the 1 degree response is paucigenic. Different D segments were used by the TGB5 Id+ 1 degree and 2 degrees HP that were sequenced, and there was no apparent correlation between TGB5 idiotypy and VH, D gene, or JH gene usage. However, all TGB5 Id+ HP sequenced used highly homologous genes from the Vk1 group. Expression of a Vk1 light chain correlates with, but is not sufficient for, TGB5 idiotypy, because one GT-binding, TGB5 Id- HP was found to use a Vk1C subgroup light chain. By Southern blot and nucleotide sequence analysis, the Vk genes used by two TGB5 Id+ 2 degrees HP from xid mice are highly homologous, if not identical to the Vk1A gene(s) used by 1 degree and 2 degrees Id+ HP from wild-type mice.
Induction of cytotoxicity in human peripheral blood mononuclear cells by monoclonal antibody OKT3.
Authors: Jung G , Martin D E , Müller-Eberhard H J 
Published: December 2022

Abstract
It has been previously reported from this laboratory that incubation of PBMC with OKT3 generates potent cytotoxic lymphocytes that can be targeted by using antibody heteroconjugates consisting of anti-target cell antibody and OKT3. In the present study these conjugates were used to explore the kinetics of induction of cytotoxicity in PBMC and the subpopulations of lymphocytes involved. It was found that in addition to conjugate-dependent cytotoxicity, a considerable amount of conjugate-independent cytotoxicity was generated during OKT3 stimulation. Although the conjugate-dependent activity resided in the CD8+ population, the conjugate-independent cytotoxicity was found to be a function of CD4-/CD8- natural killer-like cells. Being largely CD3-, those cells were most likely activated by lymphokines produced by OKT3-stimulated CD3+ cells. They were capable of killing not only tumor cells but also autologous lymphocytes. The CD4+ cells of some donors were found to exhibit low but clearly demonstrable cytotoxicity. Induction of cytotoxicity was characterized as an early event in T cell activation, correlating with the kinetics of RNA synthesis. Cytotoxicity, interleukin 2 receptor expression, and DNA synthesis declined after 3 days of activation with OKT3, indicating the existence of as yet undefined regulatory mechanisms.
Murine T lymphoma cells express a novel membrane-associated antigen with unique features.
Authors: Takei F 
Published: December 2022

Abstract
A novel membrane-associated antigen expressed on various murine T lymphoma cells has been detected by a rat monoclonal antibody. The antibody YE6/6 initially produced against Moloney leukemia virus-transformed T lymphoma line MBL-2, reacted with several other lymphoma lines including non-T lymphoma lines as well as thymocytes from leukemic AKR mice, but it did not show significant reactivities with resting or mitogen-activated normal lymphocytes by flow cytometric analysis. The antibody did not bind to some Abelson leukemia-transformed cells, which express Moloney virus antigens, suggesting that the antigen is unlikely to be encoded by Moloney virus genome. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the antigen molecules immunoprecipitated by the antibody revealed three major polypeptides. Two of the polypeptides, with approximate m.w. of 95,000 and 35,000, can be labeled by the cell surface iodination and, therefore, seem to be exposed on the cell surface. The third polypeptide, with approximate m.w. of 65,000, is not labeled by the surface iodination but it is readily detected by [35S]methionine labeling. The third polypeptide was labeled with [32P]orthophosphate indicating that it is a phosphoprotein. Western blot analysis showed that YE6/6 antibody primarily reacts with 35,000 m.w. polypeptide. Furthermore, the same 35,000 m.w. protein was also detected in concanavalin A-activated spleen cells at a low level by Western blot, but normal resting lymphocytes were negative. These results suggest that the antigen detected by YE6/6 antibody may be a cell proliferation-associated antigen and its expression is highly elevated on transformed lymphoma cells as compared to normal mitogen-activated lymphocytes.
Receptor-like role of HLA-class I antigens: regulation of T cell activation.
Authors: Dasgupta J D , Yunis E J 
Published: December 2022

Abstract
Class I major histocompatibility antigens are known to restrict the cytotoxic activity of T lymphocytes. However, experiments using monoclonal antibodies against class I antigens showed that these antigens also play some role in the regulation of T cell activation. Three monoclonal antibodies, namely W6/32 (anti-class I HLA-A, B, C, antigens), 4E (anti-class I HLA-B antigens), and BBM.1 (anti-beta 2-microglobulin) significantly suppressed the phytohemagglutinin-induced T cell proliferation. The inhibitory effect of anti-class I antibody was found to depend on the presence of monocyte/macrophage-type adherent cells. In the presence of antibody, adherent cells released a factor that suppressed T cell proliferation. These results suggest that HLA class I antigens on Mo1+ monocyte/macrophage cells function like ligand-receptor molecules, and regulate the secretion of suppressor factor(s).
Cell-associated IgM, but not IgD or I-A/E, is important in the activation of suppressor T cells by antigen-primed B cells.
Authors: Elkins K L , Stashak P W , Baker P J 
Published: December 2022

Abstract
The ability of transferred antigen-primed immune B cells to induce T cell-mediated suppression of the antibody response to Type III pneumococcal polysaccharide (SSS-III) could be blocked or eliminated by prior treatment of B cells with F(ab')2 anti-Ig or anti-IgM antibodies; however, F(ab')2 anti-IgD antibodies, or M5/114 (monoclonal anti-I-A/E antibody), had no effect on activation of suppression by SSS-III-primed B cells. Thus, cell-associated IgM antibody plays an important role in the activation of suppressor T cells during the antibody response to SSS-III.
A stathmokinetic study of B lymphocytopoiesis in rat bone marrow: proliferation of cells containing cytoplasmic mu-chains, terminal deoxynucleotidyl transferase and carrying HIS24 antigen.
Authors: Deenen G J , Hunt S V , Opstelten D 
Published: December 2022

Abstract
In rat bone marrow (BM), the B lineage surface antigen HIS24 is expressed by all surface mu chain-bearing (s mu+) B cells, by cytoplasmic mu chain-containing (c mu+s mu-) pre-B cells and TdT+ cells, and by lymphoid cells lacking both mu and TdT. Because TdT+ and HIS24+TdT-mu- cells may represent stages in B lymphocytopoiesis before mu chain expression, we investigated their kinetics. The metaphase arrest method was combined with immunofluorescence staining to detect proliferation and to quantitate cell production in the BM pre-B, TdT+, and HIS24+TdT-mu- compartments. Their apparent cell cycle times (tC(a)) were 38, 36, and 19 hr, and the number of cells produced per hour per femur were 58, 9, and 41 X 10(4), respectively. The HIS24+ compartments showed further phenotypic heterogeneity. Six percent of TdT+ cells expressed mu chains and were therefore pre-B cells. Twenty percent of HIS24+TdT-mu- cells expressed Ig other than mu chains, with size distribution and kinetics similar to HIS24+TdT-Ig- cells. Thus, the rate of production in the truly Ig-HIS24+ compartment was about 40 X 10(4)/hr/femur (8.5 by TdT+mu- and 33 by TdT-Ig-). In each phenotypic compartment, mitoses were confined to subsets of large (greater than 11 to 12 micron) cells with tC(a) of 13 to 15 hr. Surface mu+ B cells were essentially non-cycling. To quantitate whole body BM cell production, the recovery of marrow from bone and the distribution of BM were measured in 59Fe distribution experiments. The number of cells produced by whole body BM was estimated as follows: for pre-B cells, 4.5 X 10(8)/day; for TdT+mu-, 0.7 X 10(8)/day; and for HIS24+TdT-Ig- 2.6 X 10(8)/day. From the derived cell flux in these compartments we suggest that 1) many more pre-B cells are produced than needed by the peripheral B cell pool; 2) if TdT is an obligatory stage in B cell genesis, there must be at least two cell cycles in the pre-B cell compartment; 3) if it is not, the TdT+ stage may be bypassed, with HIS24+TdT-Ig- cells perhaps feeding directly into the pre-B cell compartment.