bluuebunny/crossref_metadata_2025_split huggingface dataset:
Engraftment of human tumor-infiltrating lymphocytes and the production of anti-tumor antibodies in SCID mice.
Authors: Williams S S , Chen F A , Kida H , Yokata S , Miya K , Kato M , Barcos M P , Wang H Q , Alosco T , Umemoto T , Croy B A , Repasky E A , Bankert R B 
Published: December 2022

Abstract
We report here the placement of nondisrupted 1-mm3 pieces of fresh human lung tumor biopsy tissue into the subcutis of severe combined immunodeficient (SCID) mice results in the engraftment of tumor-infiltrating leukocytes (TIL) in all but 5 of 148 mice inoculated with 39 different biopsy tissue specimens. In mice coengrafted with tumor and TIL the normal histologic architecture of the tumor and TIL interface was maintained for up to 22 wk. The TIL in the xenograft were shown to divide and were maintained exclusively at the site of tumor inoculation. It is established here that plasma cells in the TIL population produce Abs that react in western blots with tumor cell lysates. These Abs were shown to react with high and low m.w. proteins derived from both the membrane and cytosolic fractions of tumor cell lysates. The production of human Ig was found to be T cell dependent, and immunohistochemistry and in situ hybridization of DNA, using a human-specific cDNA probe, established the human identity of the tumor and TIL. High levels of human Ig in the sera of mice inoculated with tumor biopsy tissue are associated with the growth arrest of adenocarcinoma xenografts. Our results establish the co-engraftment of human tumors and TIL into SCID mice as new animal model with which to evaluate TIL function and novel therapeutic strategies that are designed to augment TIL anti-tumor activity.
The phosphatidylinositol 3-kinase inhibitor wortmannin blocks mast cell exocytosis but not IL-6 production.
Authors: Marquardt D L , Alongi J L , Walker L L 
Published: December 2022

Abstract
Phosphatidylinositol 3-kinase (PI3-kinase) activity has been shown to be important in cellular signaling via receptors associated with tyrosine kinases and receptors coupled to small or heterotrimeric G proteins. The importance of this activity in mast cell degranulation, leukotriene C4 generation, and IL-6 production was examined in mouse bone marrow-derived mast cells stimulated by high affinity IgE receptor cross-linking, direct influx of calcium, and/or adenosine receptor agonist exposure. Wortmannin, a fungal metabolite that at nanomolar concentrations inhibits PI3-kinase relatively specifically, blocked the release of granule-associated mast cell mediators independent of the secretory stimulus used. This inhibition was most prominent after a 2- to 5-min preincubation with wortmannin and was equally effective in cells additionally treated with exogenous N-ethylcarboxamidoadenosine to potentiate preformed mediator release. Mast cell production of leukotriene C4 20 min after activation or IL-6 16 h after activation was unaffected by up to 100 nM of wortmannin exposure. Mast cells preincubated with wortmannin failed to develop the classic electronmicroscopic evidence of granule swelling and fusion, increased membrane ruffling, or exocytosis upon Ag challenge. Activation of PI3-kinase appears to be critical for mast cell degranulation but is not required for arachidonic acid metabolism or cytokine production to occur. Furthermore, the inhibition of mast cell secretion by wortmannin is not stimulus specific but is evident for both IgE receptor cross-linking and direct calcium influx.
Role for macrophage inflammatory protein-2 in lipopolysaccharide-induced lung injury in rats.
Authors: Schmal H , Shanley T P , Jones M L , Friedl H P , Ward P A 
Published: December 2022

Abstract
Macrophage inflammatory protein-2 (MIP-2) is a C-X-C chemokine that possesses chemotactic activity for neutrophils. Rat MIP-2 was cloned and expressed as a 7.9-kDa peptide that exhibited dose-dependent neutrophil chemotactic activity at concentrations from 10 to 250 nM. Rabbit polyclonal Ab to the 7.9-kDa peptide showed reactivity by western blot analysis and suppressed its in vitro chemotactic activity. Cross-desensitization chemotaxis experiments suggested that the chemotactic responses elicited by MIP-2 and the related chemokine, cytokine-induced neutrophil chemoattractant, may be mediated through a common receptor. Also, chemotactic responses to human GRO-alpha were blocked by exposure of human neutrophils to either GRO-alpha or rat MIP-2, suggesting conservation of this receptor-mediated response. After LPS instillation into rat lung, mRNA for MIP-2 was up-regulated in a time-dependent manner, peaking at 6 h. MIP-2 protein was detected in bronchoalveolar lavage fluids of these animals and a significant amount of chemotactic activity present in these fluids was attributed to MIP-2. On the basis of intrapulmonary instillation of Ab to MIP-2, neutrophil accumulation in lungs after airway instillation of LPS was found to be MIP-2-dependent. These data indicate that MIP-2 plays a significant role in LPS-induced inflammatory response in rat lungs and is required for the full recruitment of neutrophils.
Protein S-thiolation and dethiolation during the respiratory burst in human monocytes. A reversible post-translational modification with potential for buffering the effects of oxidant stress.
Authors: Seres T , Ravichandran V , Moriguchi T , Rokutan K , Thomas J A , Johnston R B 
Published: December 2022

Abstract
Stimulation of the respiratory burst in mouse macrophages or human neutrophils results in the formation of disulfide bonds between low m.w. thiols and sulfhydryl groups on specific cytosolic proteins (S-thiolation). S-thiolation is reversible in certain chemical systems. The aim of the present study was to analyze the dynamic nature of this process in human monocytes under physiologic conditions. We report here that the extent of S-thiolation and the rate of respiratory burst stimulated by opsonized zymosan or phorbol diester increased for 10 to 20 min and then declined (dethiolation) in close association. Individual proteins underwent S-thiolation and dethiolation at different rates. H2O--appeared particularly effective in mediating S-thiolation, based on inhibition of S-thiolation by added catalase and accentuation by azide, which inhibits cellular catalase. S-thiolation did not occur in stimulated monocytes from patients with chronic granulomatous disease. The addition of H2O2 to monocytes or lymphocytes induced rapid S-thiolation (1 to 3 min); a subsequent dethiolation returned most of the proteins to baseline by 15 to 30 min. At 0 degrees C and after addition of 1,3-bis-(2-chloroethyl)-1-nitrosourea, there was effective S-thiolation on exposure to H2O2, but dethiolation was inhibited, suggesting a possible role for glutathione (GSH)/thioredoxin reductase systems in this process. GSH was determined to be the most abundant low m.w. thiol bound to S-thiolated proteins, but gamma-glutamylcysteine and cysteine were also bound. The time of maximal reduction in cytosolic GSH during the respiratory burst (10 min) coincided with the time at which protein-bound GSH was highest. S-thiolation-dethiolation represents a reversible post-translational modification that could protect cellular proteins from irreversible oxidative damage.
Up-regulation of VLA-5 expression during monocytic differentiation and its role in negative control of the survival of peripheral blood monocytes.
Authors: Terui Y , Furukawa Y , Sakai T , Kikuchi J , Sugahara H , Kanakura Y , Kitagawa S , Miura Y 
Published: December 2022

Abstract
Interaction between fibronectin (FN) and very late activation Ag-5 (VLA-5) integrin was recently reported to be involved in apoptosis of hematopoietic cells. In an effort to clarify the physiologic role of FN in the regulation of biologic behavior of terminally differentiated hematopoietic cells, we have examined the change of VLA-5 expression during myeloid cell differentiation and its effects on monocytes and granulocytes. VLA-5 alpha mRNA was up-regulated during monocytic differentiation, but not during granulocytic differentiation of HL-60 cells. Flow cytometric and immunocytochemical analysis revealed that surface expression of VLA-5 was selectively increased upon monocytic differentiation and that it was strongly positive on peripheral blood monocytes. Susceptibility to FN-induced apoptosis was greatly increased upon monocytic differentiation, and it was almost completely abrogated by anti-VLA-5 Ab or RGD peptide. Similarly, FN could significantly enhance apoptosis of normal monocytes but not of granulocytes. Finally, we have shown that anti-FN Ab could suppress spontaneous apoptosis of normal monocytes in culture and prolong their survival. These results suggest that FN might play an important role in negative regulation of the survival of monocytes through its interaction with VLA-5, which is selectively up-regulated during monocytic differentiation.
T lymphocyte indifference to extrathymic islet allografts.
Authors: Coulombe M , Gill R G 
Published: December 2022

Abstract
Although the deletion of self-reactive T cells maturing in the thymus is well documented, the nature of tolerance to Ags expressed exclusively on peripheral (extrathymic) tissues is less clear. Numerous studies have examined T cell reactivity to transgene-encoded Ags expressed in the periphery through tissue-specific promoters. These studies have yielded varied results concerning the degree and nature of tolerance induced to these Ags, ranging from T cell clonal deletion or inactivation to clonal "ignorance" of the peripheral transgene-encoded Ag. Furthermore, the relationship of these findings to the generation and maintenance of transplantation tolerance to extrathymic tissue allografts remains unclear. Whereas transgenic studies represent a developmental form of tolerance, grafted tissues are introduced into adult, nontolerant recipients. To relate these two model systems, we determined whether extrathymic tissue allografts could induce a developmental form of tolerance. As a model of extrathymic alloantigen expression, pancreatic islet allografts, depleted of donor APCs, were established in the periphery of severe combined immune-deficient (SCID) mice. We then examined the potential donor reactivity of adoptively transferred bone marrow-derived T cell precursors maturing in the presence of the established peripheral allograft. Such T cells were neither activated nor tolerized either in vitro or in vivo, suggesting that T cells are indifferent to Ags expressed by extrathymic islet allografts when presented in the absence of a second costimulatory signal.
Quantitative analysis of T cell activation: role of TCR/ligand density and TCR affinity.
Authors: Kim D T , Rothbard J B , Bloom D D , Fathman C G 
Published: December 2022

Abstract
(B6 X A)F1 mice were immunized with sperm whale myoglobin, and T cell clones and hybridomas were generated. Hybridoma 74a.e9 was specific for the sperm whale myoglobin 67-79 peptide and could be partially activated by a peptide analogue, equine myoglobin with a natural 74G substitution. Using this hybridoma in T cell activation assays, we studied the effects of varying the avidity of the TCR for its ligand, the concentration of MHC:peptide complex on the APC, and the density of TCR on the surface. Varying ligand concentration on the surface of the APC, the TCR avidity, or the density of TCR on the T cell were equally important parameters in driving T cell activation. The mouse myoglobin (74T) analogue, however, acted as an antagonist to the T cell response. Its effectiveness was also partially determined by its ability to bind to MHC. By independently altering each of these variables and following T cell activation, we describe the interrelationships among these three components (MHC:peptide:TCR) that control the activation of the T cell.
Plasma from patients with severe invasive group A streptococcal infections treated with normal polyspecific IgG inhibits streptococcal superantigen-induced T cell proliferation and cytokine production.
Authors: Norrby-Teglund A , Kaul R , Low D E , McGeer A , Newton D W , Andersson J , Andersson U , Kotb M 
Published: December 2022

Abstract
Previous studies have suggested a central role for superantigen-induced immune responses in the pathogenesis of streptococcal toxic shock syndrome. The production of streptococcal superantigens by clinical group A streptococcal (GAS) isolates was studied, and the ability of plasma collected from patients with severe invasive GAS infections to neutralize the proliferative- and cytokine-inducing activities of these superantigens was investigated. Overnight culture supernatants from all GAS isolates obtained from patients with invasive disease were found to contain superantigenic activity, as evident from their ability to drive potent T cell proliferation, induce high production of cytokines, and stimulate T cells in a V beta-specific manner. Twelve patients with severe invasive GAS infections, including 11 streptococcal toxic shock syndrome cases and one necrotizing fasciitis without shock, were treated with i.v. infusions of normal polyspecific Ig (IVIG). Plasma samples collected from each patient before and after IVIG administration were analyzed for their ability to neutralize the activity of streptococcal superantigens produced by the GAS isolate that caused their disease. In all IVIG-treated patients, the capacity to neutralize the superantigenic activity, produced by their respective GAS isolate or by purified streptococcal pyrogenic exotoxins, increased in plasma following IVIG administration. Of particular clinical relevance, post-IVIG plasma from each patient completely blocked cytokine production elicited by their respective GAS culture supernatants or by purified streptococcal pyrogenic exotoxins. This study shows that IVIG treatment confers in vivo inhibitory activity against a large array of streptococcal superantigens and suggests that IVIG may be useful in the treatment of severe invasive streptococcal infections.
IL-12 synergizes with IL-2 and other stimuli in inducing IL-10 production by human T cells.
Authors: Jeannin P , Delneste Y , Seveso M , Life P , Bonnefoy J Y 
Published: December 2022

Abstract
IL-12, a potent inducer of IFN-gamma production by T cells and NK cells, has been recently reported to exacerbate an established Th2 response in vivo. However, the effect of IL-12 on Th2-lymphokine production remains unclear. Since IL-10 is a lymphokine associated with Th2 responses which decreases both IL-12-induced IFN-gamma production and IL-12 production by macrophages, we have analyzed here, in an APC-free system, the ability of IL-12 to modulate the production of human IL-10 by established Th0, Th1, and Th2 T cell clones (TCC), T cell lines, and purified peripheral blood T cells. IL-12 synergized with anti-CD3 mAb, Con A, or IL-2 in inducing IL-10 production by Th0, Th1, and Th2 TCC and by T cell lines. This effect was dose dependent (from 0.1 to 50 U/ml) and associated with an increase of IL-10 mRNA transcription. As previously reported, IL-12 also enhanced IFN-gamma production by stimulated Th1 and Th0 TCC and, to a lesser extent, IL-4 production by stimulated Th0 and Th2 TCC. These observations were extended to peripheral blood T cells stimulated in the presence of exogenous IL-2. Moreover, using neutralizing anti-IL-2 Ab, we report that endogenous IL-2 produced by stimulated Th0 TCC could in part contribute to the effect of IL-12 on IL-10 and IL-4 production. In conclusion, IL-12 synergizes with IL-2 and other stimuli in inducing IL-4 and IL-10 production by T cells. This property may help to explain why IL-12 does not efficiently down-regulate an established Th2 response.
Allele-specific differences in the interaction of MHC class I molecules with transporters associated with antigen processing.
Authors: Neisig A , Wubbolts R , Zang X , Melief C , Neefjes J 
Published: December 2022

Abstract
MHC class I molecules bind peptides that are translocated from the cytosol into the endoplasmic reticulum by the peptide transporter associated with antigen processing (TAP). Class I heterodimers have been shown to associate with TAP and are released when loaded with peptide. Here, we show the existence of two pools of class I heterodimers, one associated with TAP and one that is free. Whereas the free pool is recognized by the class I-specific Ab W6/32, the TAP-associated pool is not. Analysis of several class I alleles shows binding to TAP with different efficiencies, even at the earliest time points of MHC class I assembly. Most HLA-A and -C alleles tested interacted efficiently with TAP, whereas a considerable number of HLA-B alleles associated very inefficiently or not at all with TAP. This was also observed in cells with nonfunctional TAP. Sequence comparison of the different class I alleles allowed the definition of amino acids in the peptide binding groove that might be involved in TAP association. Binding of peptides to two different pools of class I heterodimers may ensure efficient peptide association in an environment where peptides have a short life span.
Enhanced development of Th2-like primary CD4 effectors in response to sustained exposure to limited rIL-4 in vivo.
Authors: Yoshimoto K , Swain S L , Bradley L M 
Published: December 2022

Abstract
Previous studies have established that cytokines regulate development of Th cell subsets from naive CD4 cells, suggesting promising therapeutic potential for cytokines that has not been fully realized, in part due to high dose toxicity. Here we examined effects of sustained delivery of low doses of rIL-4 on development of cytokine-secreting primary CD4 cells in vivo. Diffusion chambers injected with X63.Ag-653 plasmacytoma transfectants that constitutively produce rIL-4 were implanted s.c. in mice 24 to 48 h before immunization with keyhole limpet hemocyanin (KLH). Five days later at the peak of the response, KLH-specific CD4 cells from controls, which received chambers containing untransfected plasmacytoma cells, secreted primarily IL-2 and IL-4, with low levels of IFN-gamma and no IL-5 following Ag restimulation. Administration of rIL-4 enhanced IL-2 and IL-4 production by two- to fivefold, with a corresponding decrease in IFN-gamma. As frequencies of KLH-specific precursors that secreted IL-4 were unaltered, exogenous cytokine apparently affected the magnitude of IL-4 secretion by primed CD4 cells. Although IL-4 was undetectable in sera of mice carrying chambers, titers of KLH-specific IgG1 and IgG3 Ab were increased by rIL-4, whereas IgM, IgG2a, and IgG2b levels were unaltered. The results indicate that sustained delivery of low doses of cytokines at sites distant from Ag exposure can selectively potentiate development of CD4 subsets.
Immunogenicity of peptides bound to MHC class I molecules depends on the MHC-peptide complex stability.
Authors: van der Burg S H , Visseren M J , Brandt R M , Kast W M , Melief C J 
Published: December 2022

Abstract
The impact of the MHC class I peptide binding stability on the immunogenicity of particular peptide Ags in class I-restricted cytotoxic T lymphocyte responses is not clearly established. Therefore, we have determined the dissociation rate of each peptide from MHC class I at 37 degrees C and compared this to that of a consensus CTL epitope. Newly defined immunogenic peptides formed relatively stable MHC-peptide complexes as shown by their low dissociation rates, whereas nonimmunogenic peptides displayed high dissociation rates. In addition virtually all previously described HLA-A*0201-restricted T cell epitopes showed low dissociation rates. Furthermore, we show that the immunogenicity of HIV-1-derived peptides can be predicted more accurately by their dissociation rate than by the MHC class I binding affinity. Selection of peptides based on affinity and their dissociation rate leads to a more precise identification of candidate CTL epitopes than selection based on affinity alone. These results help to understand why some peptides are recognized by CTL and, along with detailed knowledge of protein processing rules, therefore have important implications for the selection of peptides in peptide-based vaccines.
Antibodies to an autostimulatory growth factor (IL-2) or its receptor induce death of leukemogenic cells.
Authors: Orban P C , Schrader J W 
Published: December 2022

Abstract
We show for the first time that Ab-mediated antagonism of growth factor activity can induce death of all cells in clonal populations surviving and growing by an autostimulatory mechanism. Models of autostimulatory leukemia were generated by transfecting a mouse IL-2-dependent cell line (FD.C/2) with vectors directing production of IL-2 by these cells. One series of clones grew in a density-dependent manner in the absence of exogenous IL-2 and produced tumors in syngeneic mice. Although these clones released relatively small amounts of IL-2, their growth and survival was only partially inhibited by Abs to IL-2 or the IL-2R. Another autostimulatory clone was derived, using a different vector, which produced significantly less IL-2. Treatment of cells of this clone with Abs to IL-2 or its receptor resulted in death of all cells. These data demonstrate that Abs that antagonize growth factor action can induce the death of cells transformed by autostimulatory mechanisms. They suggest that while autostimulatory tumors are relatively resistant to Abs that block growth factor action, this is a quantitative phenomenon, and competitive antagonists of growth factor action of sufficiently high affinity may provide effective and specific adjuvants to the treatment of such tumors.
Resistance of naive mice to murine hepatitis virus strain 3 requires development of a Th1, but not a Th2, response, whereas pre-existing antibody partially protects against primary infection.
Authors: Pope M , Chung S W , Mosmann T , Leibowitz J L , Gorczynski R M , Levy G A 
Published: December 2022

Abstract
Murine hepatitis virus strain 3 (MHV-3) produces a strain-dependent spectrum of disease. The development of liver necrosis has been shown to be related to production of a unique macrophage procoagulant activity (PCA), encoded by the gene fgl-2, in susceptible mice. These studies were designed to examine the influence of Th1/Th2 cells on resistance/susceptibility and production of macrophage PCA in resistant (A/J) and susceptible (BALB/cJ) strains of mice following infection with MHV-3. Immunization of A/J mice with MHV-3 induced a Th1 cellular immune response, and one Th1 cell line (3E9.1) protected susceptible mice and inhibited PCA production by macrophages both in vitro and in vivo. In contrast, immunization of BALB/cJ mice with an attenuated variant of MHV-3 derived from passaging MHV-3 in YAC-1 cells resulted in a Th2 response. Transfer of spleen cells and T cell lines from immunized BALB/cJ mice failed to protect naive susceptible syngeneic mice from infection with MHV-3 and augmented macrophage PCA production to MHV-3 in vitro. However, serum from immunized BALB/cJ mice contained high titrated neutralizing Ab that protected naive BALB/cJ animals from lethal primary MHV-3 infection. These results demonstrate that susceptible BALB/cJ mice generate a Th2 response following MHV-3 infection and that these Th2 cells neither inhibit MHV-3-induced macrophage PCA production nor protect naive mice from MHV-3 infection. The results suggest that Ab protects against primary infection but cannot eradicate ongoing infection. Thus, these data define the differential role of Th1/Th2 lymphocytes in primary and secondary MHV-3 infection and emphasize the importance of PCA in the pathogenesis of MHV-3 infection.