bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nonopsonic and Opsonic Association ofMycobacterium tuberculosiswith Resident Alveolar Macrophages Is Inefficient
Authors: Stokes Richard W. , Thorson Lisa M. , Speert David P. 
Published: January 2023

AbstractThe association of Mycobacterium tuberculosis with alveolar macrophages (Mφ) in a serum-free environment is a crucial first step in the pathogenesis of this facultative intracellular pathogen. We present data demonstrating that freshly explanted alveolar Mφ do not efficiently bind M. tuberculosis in a serum-free system, although a small subpopulation of these Mφ (10–15%) can bind mycobacteria. In contrast, almost 100% of a peritoneal Mφ population bind mycobacteria under the same conditions. The poor binding of mycobacteria by alveolar Mφ does not reflect a general inability to associate with particles; binding and ingestion of latex beads and zymosan particles were comparable with that seen with peritoneal Mφ. Resident alveolar Mφ did not efficiently bind mycobacteria in the presence of serum and expressed poorly several Mφ surface receptors, including CR3. Furthermore, we demonstrate that bovine surfactant protein A does not enhance the association of M. tuberculosis with alveolar Mφ. Differentiation of alveolar Mφ in vitro resulted in increased expression of Mφ surface receptors and an increased capacity to bind mycobacteria in the presence and absence of serum. Evidence is presented that opsonic binding of M. tuberculosis by differentiated alveolar Mφ is mediated by complement and CR3, and that the poor binding by resident alveolar Mφ is due to their poor expression of CR3. The receptor mediating nonopsonic binding of M. tuberculosis to differentiated alveolar Mφ was not unequivocally identified in this study, but could also be CR3.
The Fas Counterattack In Vivo: Apoptotic Depletion of Tumor-Infiltrating Lymphocytes Associated with Fas Ligand Expression by Human Esophageal Carcinoma
Authors: Bennett Michael W. , O’Connell Joe , O’Sullivan Gerald C. , Brady Ciaran , Roche Desmond , Collins J. Kevin , Shanahan Fergus 
Published: January 2023

AbstractVarious cancer cell lines express Fas ligand (FasL) and can kill lymphoid cells by Fas-mediated apoptosis in vitro. FasL expression has been demonstrated in several human malignancies in vivo. We sought to determine whether human esophageal carcinomas express FasL, and whether FasL expression is associated with increased apoptosis of tumor-infiltrating lymphocytes (TIL) in vivo, thereby contributing to the immune privilege of the tumor. Using in situ hybridization and immunohistochemistry, respectively, FasL mRNA and protein were colocalized to neoplastic esophageal epithelial cells in all esophageal carcinomas (squamous, n = 6; adenocarcinoma, n = 2). The Extent of FasL expression was variable, with both FasL-positive and FasL-negative neoplastic regions occurring within tumors. TIL were detected by immunohistochemical staining for the leukocyte common Ag, CD45. FasL expression was associated with a mean fourfold depletion of TIL when compared with FasL-negative areas within the same tumors (range 1.6- to 12-fold, n = 6, p &lt; 0.05). Cell death of TIL was detected by dual staining of CD45 (immunohistochemistry) and DNA strand breaks (TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling). There was a mean twofold increase in detectable cell death among TIL in FasL-positive areas compared with FasL-negative areas (range 1.6- to 2.4-fold, n = 6, p &lt; 0.05). In conclusion, we demonstrate a statistically significant, quantitative reduction of TIL concomitant with significantly increased TIL apoptosis within FasL-expressing areas of esophageal tumors. Our findings suggest Fas-mediated apoptotic depletion of TIL in response to FasL expression by esophageal cancers, and provide the first direct, quantitative evidence to support the Fas counterattack as a mechanism of immune privilege in vivo in human cancer.
IL-3 Enhances Both Presentation of Exogenous Particulate Antigen in Association with Class I Major Histocompatibility Antigen and Generation of Primary Tumor-Specific Cytolytic T Lymphocytes
Authors: Yeh Kun-Yun , McAdam Alexander J. , Pulaski Beth A. , Shastri Nilabh , Frelinger John G. , Lord Edith M. 
Published: January 2023

Abstract
Recent studies have reported that APC can present particulate exogenous Ag in the context of class I MHC to CD8+ CTL, and our laboratory demonstrated that IL-3 could enhance CTL generation to exogenous Ag. In this paper, we wished to determine whether presentation of particulate Ag could be enhanced by IL-3. A T cell hybridoma, B3Z86/90.14 (B3Z) restricted to Ova/Kb, was used as an indicator for presentation of particulate Ag with class I MHC. When activated, this hybridoma expresses lacZ, allowing a simple colorimetric measurement of Ag-specific T cell stimulation. We demonstrated that bone marrow cells stimulated by IL-3 in vivo and in vitro exhibited significantly increased presentation of exogenous OVA linked to beads. Lysate from OVA-transfected line 1 murine lung adenocarcinoma cells (line 1/OVA) was also presented by IL-3-stimulated bone marrow cells, suggesting that these APC can process tumor fragments or debris. Studies using TAP1/2-deficient mice and Ag presentation inhibitors indicate that this exogenous Ag presentation is mediated via the conventional class I MHC pathway. Adoptive transfer of IL-3-stimulated bone marrow cells pulsed with lysate from line 1/OVA tumor cells into naive recipient mice led to the generation of a potent CTL response. These observations indicate that use of such cells may provide a new avenue for development of tumor vaccines.
Relationships Among TCR Ligand Potency, Thresholds for Effector Function Elicitation, and the Quality of Early Signaling Events in Human T Cells
Authors: Hemmer Bernhard , Stefanova Irena , Vergelli Marco , Germain Ronald N. , Martin Roland 
Published: January 2023

AbstractDetermining how receptor ligand quality and quantity together control the biologic responses of T cells is central to understanding normal and pathologic T cell immunity. Here we have carefully examined how variations in antigenic peptide structure and dose affect multiple functional responses of human T cell clones and have correlated these observations with proximal TCR signaling events induced by the same set of related ligands. As the Ag concentration increases, effector functions are elicited according to a clone-specific hierarchy. The absolute amount of each peptide required to stimulate the entire set of effector functions (potency) differs markedly among ligands for a single TCR, correlating with the efficiency of TCR down-modulation and the extent of ZAP-70 activation. However, distinct patterns of TCR ζ-chain phosphorylation were observed, with the ratios of TCRζ isoforms relating to ligand agonist potency. The appearance of partially phosphorylated TCRζ isoforms was paralleled by relative changes in certain response thresholds within the hierarchy. Thus, a combination of density, potency, and quality of signaling all contribute to the distinct effects of agonist ligands on T cell immunity.
Expression and Characterization of Recombinant Soluble Peptide: I-A Complexes Associated with Murine Experimental Autoimmune Diseases
Authors: Radu Caius G. , Ober Bertram T. , Colantonio Lucia , Qadri Ayub , Ward E. Sally 
Published: January 2023

Abstract
Structural and functional studies of murine MHC class II I-A molecules have been limited by the low yield and instability of soluble, recombinant heterodimers. In the murine autoimmune diseases experimental autoimmune encephalomyelitis and collagen-induced arthritis, MHC class II molecules I-Au and I-Aq present peptides derived from myelin basic protein and type II collagen, respectively, to autoreactive T cells. To date, systems for the expression of these two I-A molecules in soluble form for use in structure-function relationship studies have not been reported. In the present study, we have expressed functional I-Au and I-Aq molecules using a baculovirus insect cell system. The chain pairing and stability of the molecules were increased by covalently linking the antigenic peptides to β-chains and adding carboxyl-terminal leucine zippers. Peptide:I-Aq complex quantitatively formed an SDS-stable dimer, whereas peptide:I-Au formed undetectable amounts. However, the two complexes did not show any significant difference in their response to thermal denaturation as assessed by circular dichroism analyses. The autoantigen peptide:I-A complexes were highly active in stimulating cognate T cells to secrete IL-2 and inducing Ag-specific apoptosis of the T cells. Interestingly, the T cells were stimulated by these soluble molecules in the apparent absence of experimentally induced cross-linking of TCRs, indicating that they may have therapeutic potential in autoimmune disease models.
Structural and Functional Analysis of J Chain-Deficient IgM
Authors: Wiersma Erik J. , Collins Cathy , Fazel Shafie , Shulman Marc J. 
Published: January 2023

AbstractPrevious studies have discerned two forms of polymeric mouse IgM: moderately cytolytic (complement-activating) pentamer, which contains J chain, and highly cytolytic hexamer, which lacks J chain. To investigate the relationships among polymeric structure, J chain content, and cytolytic activity, we produced IgM in J chain-deficient and J chain-proficient mouse hybridoma cell lines. Both hexamer and pentamer were produced in the absence as well as the presence of J chain. Hexameric IgM activated (guinea pig) complement approximately 100-fold more efficiently than did J chain-deficient pentamer, which, in turn, was more active than J chain-containing pentamer. These results are consistent with the hypothesis that J chain-containing pentamer cannot activate complement. We also analyzed the structure of IgM-S337, in which the μ-chain bears the C337S substitution. Like normal IgM, IgM-S337 was formed as a hexamer and as both J chain deficient- and J chain-containing pentamers. Unlike normal IgM, IgM-S337 dissociated in SDS into various subunits. For IgM-S337 pentamer, the predominant subunits migrated as μ2κ2 and μ4κ4, and the subunit distribution was unaltered by J chain. However, J chain was found only in the μ2κ2 species, suggesting that some arrangement of inter-μ bonds directs incorporation of J chain. IgM-S337 hexamer also dissociated to μ2κ2 and μ4κ4, but also yielded several species migrating much more slowly in SDS-PAGE than wild-type μ12κ12. To account for these forms, we propose that each μ-chain can interact with three other μ-chains and that some hexameric molecules contain two catenated μ6κ6 circles.
T Lymphocytes Are Required for Protection of the Vaginal Mucosae and Sensory Ganglia of Immune Mice Against Reinfection with Herpes Simplex Virus Type 2
Authors: Milligan Gregg N. , Bernstein David I. , Bourne Nigel 
Published: January 2023

Abstract
Intravaginal inoculation of mice with an attenuated strain of herpes simplex virus type 2 (HSV-2) resulted in vigorous HSV-specific immune responses that protected against subsequent challenge with fully virulent HSV-2 strains. Even in the presence of high titers of HSV-specific Ab, T cell-dependent mechanisms were required for protection of the vaginal mucosae of HSV-immune mice and could be detected by 24 h after intravaginal reinoculation. Depletion of specific T cell subsets from HSV-immune mice before HSV-2 reinoculation demonstrated that CD4+ T cells were primarily responsible for this protection. Similarly, optimal protection of the sensory ganglia against reinfection with HSV-2 was dependent on the presence of T cells. Infectious HSV-2 was not detected in the sensory ganglia or spinal cord of HSV-immune mice depleted of only CD4+ or CD8+ T cells, suggesting that the T cell-mediated protection could be provided by either subset. Similarly, neutralization of IFN-γ during challenge of HSV-immune mice resulted in diminished protection of the vaginal mucosa, but not of the sensory ganglia. These results suggest that the ability to induce vigorous HSV-specific T cell responses is an important consideration in the design of vaccines to protect both the vaginal mucosa and sensory ganglia against HSV-2.
Immunization with Purified Natural and Recombinant Allergens Induces Mouse IgG1 Antibodies That Recognize Similar Epitopes as Human IgE and Inhibit the Human IgE-Allergen Interaction and Allergen-Induced Basophil Degranulation
Authors: Vrtala Susanne , Ball Tanja , Spitzauer Susanne , Pandjaitan Budhi , Suphioglu Cenk , Knox Bruce , Sperr Wolfgang R. , Valent Peter , Kraft Dietrich , Valenta Rudolf 
Published: January 2023

AbstractMolecular characterization of allergens by recombinant DNA technology has made rapid progress in the recent few years. In the present study we immunized mice with aluminum hydroxide-adsorbed purified recombinant major timothy grass pollen allergens (rPhl p 1, rPhl p 2, rPhl p 5), dog albumin, a major animal dander allergen, and proteins with low (β-lactoglobulin) or no (ribulose diphosphate carboxylase) allergenic potential in humans. Allergens that bind high levels of IgE in humans (Phl p 1, Phl p 5, dog albumin) induced high IgE and IgG1 levels in mice, whereas proteins with little or no allergenic activity in humans failed to induce significant IgE and IgG1 levels in mice. Continuous immunization for a period of 27 wk resulted in the production of mouse IgG1 Abs that recognized recombinant allergen fragments/epitopes defined by IgE Abs of allergic patients. As a consequence, allergen-specific mouse Abs strongly inhibited human IgE binding to the allergens and suppressed the allergen-induced histamine release from human basophils. In summary, our data indicate that 1) the allergenic potency of a protein may be related to its overall immunogenicity and 2) prolonged immunization with single purified recombinant allergens induces protective IgG Abs. The presented experimental in vivo/in vitro system allows the evaluation of Ag preparations (e.g., recombinant allergens) to be used for immunotherapy in humans.
Physiologic Regulation of Postovulatory Neutrophil Migration into Vagina in Mice by a C-X-C Chemokine(s)
Authors: Sonoda Yasuo , Mukaida Naofumi , Wang Jian-bin , Shimada-Hiratsuka Motoko , Naito Makoto , Kasahara Tadashi , Harada Akihisa , Inoue Masaki , Matsushima Kouji 
Published: January 2023

Abstract
Leukocytes, particularly neutrophils, infiltrate into female genital organs after ovulation in both humans and mice. In mice, a female sexual cycle consists of 5 phases: proestrus, estrus, metestrus-1, metestrus-2, and diestrus. Ovulation occurs at estrus; at metestrus-2, a large number of neutrophils infiltrate into the vaginal epithelium accompanied by an increased neutrophil number in vaginal lavage fluid. Concomitantly, concentrations of a functional IL-8 homologue, murine macrophage inflammatory protein (MIP)-2, were increased significantly in vaginal lavage fluid at metestrus-2 as compared with other phases. On the contrary, MIP-2 was not detected in plasma during the whole course of a sexual cycle. Moreover, immunohistochemical analyses demonstrated that MIP-2 protein expression was prominent at the upper layer of the vaginal epithelium at metestrus-2, in contrast to a marginal staining in the vaginal epithelium at proestrus and estrus. These results suggest that a C-X-C chemokine, MIP-2, was produced physiologically in the vaginal epithelium in a sexual cycle-dependent manner. Furthermore, the administration of neutralizing anti-IL-8R homologue Abs at proestrus abrogated leukocyte infiltration into the vagina at metestrus. However, anti-MIP-2 Abs reduced leukocyte influx at metestrus by ∼50%. Thus, a murine IL-8 homologue, MIP-2, and its related molecules physiologically regulate neutrophil migration into the vagina in a sexual cycle-dependent manner.
Antibody-IL-12 Fusion Proteins Are Effective in SCID Mouse Models of Prostate and Colon Carcinoma Metastases
Authors: Gillies Stephen D. , Lan Yan , Wesolowski John S. , Qian Xiuqi , Reisfeld Ralph A. , Holden Sylvia , Super Michael , Lo Kin-Ming 
Published: January 2023

AbstractIL-12 is a complex cytokine in both its structure and its range of biologic activities. Fusions of this heterodimeric molecule with an intact antitumor Ab were made to test the feasibility and efficacy of targeting IL-12 to tumors to elicit a local immune response. Fusion proteins composed of the human p35 and p40 subunits had IL-12 bioactivities that were nearly as potent on human immune cells as the rIL-12 standard, but were inactive on mouse cells. Hybrid IL-12 fusion proteins composed of mouse p35 and human p40, fused to Ab, were capable of inducing IFN-γ, but were much less active on mouse spleen cells than a mouse IL-12 standard. Despite this relatively low activity, the hybrid fusion protein was as effective in a SCID mouse model as a fully active Ab-IL-2 fusion protein in eliminating established pulmonary metastases of CT26 colon carcinoma. Specific targeting of a human IL-12 fusion protein to metastatic prostate carcinoma xenografts was also shown to be effective in SCID mice transplanted with human lymphocyte-activated killer cells. These results demonstrate the importance of directing this potent cytokine to the tumor microenvironment and suggest an important alternative to systemic IL-12 administration or gene therapy for increasing its therapeutic index.
Reconstitution of EBV-Specific T Cell Immunity in Solid Organ Transplant Recipients
Authors: Haque Tanzina , Amlot Peter L. , Helling Nabeela , Thomas J. Alero , Sweny Paul , Rolles Keith , Burroughs Andrew K. , Prentice H. Grant , Crawford Dorothy H. 
Published: January 2023

Abstract
EBV-specific autologous CTL were grown in vitro from three adults (two liver transplant recipients and one patient on hemodialysis awaiting kidney retransplant). All CTL lines were TCR αβ, CD8 positive cells, EBV specific, and MHC class I restricted. The CTL lines were expanded in vitro and infused in three escalating doses (5 × 107, 1 × 108, and 2 × 108) at monthly intervals. Weekly blood samples were collected following each infusion. EBV-specific CTL precursor cells in peripheral blood were quantitated by limiting dilution analysis, and their effect on EBV load in vivo was assessed by semiquantitative PCR. In all three patients, the numbers of CTL precursor cells increased during the weeks following the infusions and were highest after the third infusion. This level gradually declined but remained above the preinfusion levels for up to 3 mo. EBV genome copy number, on the other hand, dropped following the first infusion and became undetectable thereafter. The EBV DNA level remained lower than the pretransplant level in all patients for up to 3 mo after the last infusion. Our study shows that it is feasible to generate and expand EBV-specific CTL from pretransplant blood samples of solid organ transplant recipients, that these CTL can be stored and infused posttransplant, and that they remain cytotoxic and EBV specific in vivo. The aim of this study is to use these CTL for prevention and treatment of lymphoproliferative disease in solid organ transplant recipients.
Thymosin α1 Stimulates Endothelial Cell Migration, Angiogenesis, and Wound Healing
Authors: Malinda Katherine M. , Sidhu Gurmel S. , Banaudha Krishna K. , Gaddipati Jaya P. , Maheshwari Radha K. , Goldstein Allan L. , Kleinman Hynda K. 
Published: January 2023

Abstract
In wound healing, lymphoid cells release soluble factors that attract fibroblasts and macrophages, initiating repair, endothelial cell migration, angiogenesis, and matrix production. We analyzed the effect of thymosin α1 (Tα1) on endothelial cell migration, angiogenesis, and wound healing. Tα1, a 28 amino acid peptide initially isolated from the thymus, enhanced the morphologic differentiation of endothelial cells and was a potent chemoattractant for endothelial cells and monocytes in vitro. In vivo, Tα1 stimulated angiogenesis in a subcutaneous model. When given either topically or i.p., it accelerated wound healing in a punch model, demonstrating that Tα1 promotes angiogenesis and wound healing.
Rejection of Allogeneic and Syngeneic But Not MHC Class I-Deficient Tumor Grafts by MHC Class I-Deficient Mice
Authors: Freland Sofia , Chambers Benedict J. , Andersson Malena , Van Kaer Luc , Ljunggren Hans-Gustaf 
Published: January 2023

Abstract
The ability of TAP1−/−, β2m−/−, and TAP1/β2m−/− mice to mount rejection responses against allogeneic, syngeneic, and MHC class I-deficient tumor grafts was examined. The results demonstrate a potent ability of TAP1−/− and β2m−/− as well as TAP1/β2m−/− mice to reject allogeneic tumors. In contrast to published data, rejection of syngeneic MHC class I-expressing tumors was also observed. This response was specific for the MHC class I-deficient mice, since wild-type mice did not reject syngeneic MHC class I-positive tumors under identical experimental conditions. The rejection response of syngeneic tumors required preimmunization of the mice and was MHC class I specific at the level of priming as well as at the level of the tumor target. Finally, MHC class I-deficient tumor grafts were accepted in MHC class I-deficient mice while similar grafts were rejected in wild-type mice. In summary, while MHC class I-deficient mice have retained a capacity to reject allogeneic tumors, they have gained an ability to reject syngeneic MHC class I-positive tumors and lost the ability to reject MHC class I-negative tumors. The present results are discussed in relation to the role of MHC class I molecules in selecting functional CD8+ T and NK cell repertoires, and the development of cell-mediated immunity.
MHC Class II Expression in Double Mutant Mice Lacking Invariant Chain and DM Functions
Authors: Kenty George , Martin W. David , Van Kaer Luc , Bikoff Elizabeth K. 
Published: January 2023

Abstract
Invariant (Ii) chain and DM functions are required at distinct stages during class II maturation to promote occupancy by diverse peptide ligands. The class II molecules expressed by mutant mouse strains lacking Ii chain or DM activities display discrete structural and functional abnormalities. The present report describes the cellular and biochemical characteristics of Ii−DM− doubly deficient mice. As for Ii chain mutants, their mature AαbAβb dimers similarly exhibit reduced mobilities in SDS-PAGE, and in functional assays these molecules behave as if empty or occupied by an easily displaced peptide. Additionally, the present experiments demonstrate that the production of floppy AαbAβb dimers is TAP independent. In comparison with Ii chain mutants, Ii−DM− doubly deficient cell populations exhibit increased peptide binding activities and consistently greater presentation abilities in T cell stimulation assays. These functional differences appear to reflect higher class II surface expression associated with their increased representation of B lymphocytes. We also observe defective B cell maturation in mice lacking Ii chain or DM expression, and interestingly, B cell development appears more severely compromised in Ii−DM− double mutants. These mutant mice lacking both Ii chain and DM activities should prove useful for analyzing nonconventional class II Ag presentation under normal physiological conditions in the intact animal.