bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Vitro and In Vivo Dependency of Chemokine Generation on C5a and TNF-α
Authors: Czermak Boris J. , Sarma Vidya , Bless Nicolas M. , Schmal Hagen , Friedl Hans Peter , Ward Peter A. 
Published: January 2023

AbstractUnder a variety of conditions, alveolar macrophages can generate early response cytokines (TNF-α, IL-1), complement components, and chemotactic cytokines (chemokines). In the current studies, we determined the requirements for TNF-α and the complement activation product C5a in chemokine production in vitro and in vivo. Two rat CXC chemokines (macrophage inflammatory protein (MIP)-2 and cytokine-induced neutrophil chemoattractant (CINC)) as well as three rat CC chemokines (MIP-1α, MIP-1β, and monocyte chemoattractant protein (MCP)-1) were investigated. Chemokine generation in vitro was studied in rat alveolar macrophages stimulated with IgG immune complexes in the absence or presence of Abs to TNF-α or C5a. The rat lung injury model induced by IgG immune complex deposition was employed for in vivo studies. Abs to TNF-α or C5a were administered intratracheally or i.v., and effects on chemokine levels in bronchoalveolar lavage fluids were quantitated by ELISA. Both in vitro and in vivo studies demonstrated the requirements for TNF-α and C5a for full generation of CXC and CC chemokines. In vitro and in vivo blockade of TNF-α or C5a resulted in significantly reduced production of chemokines. Supernatant fluids from in vitro-stimulated macrophages revealed by Western blot analysis the presence of C5a/C5adesArg, indicating intrinsic generation of C5a/C5adesArg by alveolar macrophages and explaining the higher efficiency of intratracheal vs i.v. blockade of C5a in reducing chemokine production. These results underscore the central role of both TNF-α and C5a, which appear to function as autocrine activators to promote CXC and CC chemokine generation by alveolar macrophages.
CD95 (Fas/Apo-1) as a Receptor Governing Astrocyte Apoptotic or Inflammatory Responses: A Key Role in Brain Inflammation?
Authors: Saas Philippe , Boucraut José , Quiquerez Anne-Lise , Schnuriger Valérie , Perrin Gaelle , Desplat-Jego Sophie , Bernard Dominique , Walker Paul R. , Dietrich Pierre-Yves 
Published: January 2023

AbstractAstrocytes are a major cellular component of the brain that are capable of intense proliferation and metabolic activity during diverse inflammatory brain diseases (such as multiple sclerosis, Alzheimer’s dementia, tumor, HIV encephalitis, or prion disease). In this biological process, called reactive gliosis, astrocyte apoptosis is frequently observed and could be an important mechanism of regulation. However, the factors responsible for apoptosis in human astrocytes are poorly defined. Here, we report that short term cultured astrocytes derived from different brain regions express significant levels of CD95 at their surface. Only late passage astrocytes are sensitive to CD95 ligation using either CD95 mAb or recombinant CD95 ligand. Blocking experiments using caspase inhibitors with different specificities (DEVD-CHO, z-VAD-fmk, and YVAD-cmk), an enzymatic activity assay, and immunoblotting show that CPP32/caspase-3 play a prominent role in CD95-induced astrocyte death. In contrast, early passage astrocytes are totally resistant to death, but a significant increase in astrocytic IL-8 secretion (p &lt; 0.001, by Wilcoxon’s test for paired samples) is observed after CD95 triggering. Production of IL-8 contributes to the resistance of astrocytes to CD95 ligation. Furthermore, in the presence of IFN-γ, resistant astrocytes became sensitive to CD95-mediated death. These data suggest that microenvironmental factors can influence the consequences of CD95 ligation on astrocytes. Therefore, we propose that CD95 expressed by human astrocytes plays a pivotal role in the regulation of astrocyte life and death and may be a key factor in inflammatory processes in the brain, such as reactive gliosis.
Phosphorylation of Cytosolic Phospholipase A2 and the Release of Arachidonic Acid in Human Neutrophils
Authors: Syrbu Sergei I. , Waterman Waltraut H. , Molski Thaddeus F. P. , Nagarkatti Deepa , Hajjar Jean-Jacques , Sha’afi Ramadan I. 
Published: January 2023

Abstract
Kinases mediating phosphorylation and activation of cytosolic phospholipase A2 (cPLA2) in intact cells remain to be fully characterized. Platelet-activating factor stimulation of human neutrophils increases cPLA2 phosphorylation. This increase is inhibited by PD 98059, a mitogen-activated protein (MAP)/extracellular signal-regulating kinase (erk) 1 inhibitor, but not by SB 203580, a p38 MAP kinase inhibitor, indicating that this action is mediated through activation of the p42 MAP kinase (erk2). However, platelet-activating factor-induced arachidonic acid release is inhibited by both PD 98059 and SB 203580. Stimulation by TNF-α increases cPLA2 phosphorylation, which is inhibited by SB 203580, but not PD 98059, suggesting a role for p38 MAP kinase. LPS increases cPLA2 phosphorylation and arachidonic acid release. However, neither of these actions is inhibited by either PD 98059 or SB 203580. PMA increases cPLA2 phosphorylation. This action is inhibited by PD 98059 but not SB 203580. Finally, FMLP increases cPLA2 phosphorylation and arachidonic acid release. Interestingly, while the FMLP-induced phosphorylation of cPLA2 is not affected by the inhibitors of the p38 MAP kinase or erk cascades, both inhibitors significantly decrease arachidonic acid release stimulated by FMLP. SB 203580 or PD 98059 has no inhibitory effects on the activity of coenzyme A-independent transacylase.
Modulation of the Humoral Immune Response by Antibody-Mediated Antigen Targeting to Complement Receptors and Fc Receptors
Authors: Baiu Dana C. , Prechl Jozsef , Tchorbanov Andrey , Molina Hector D. , Erdei Anna , Sulica Andrei , Capel Peter J. A. , Hazenbos Wouter L. W. 
Published: January 2023

AbstractDuring an ongoing immune response, immune complexes, composed of Ag, complement factors, and Igs, are formed that can interact with complement receptors (CRs) and IgG Fc receptors (FcγR). The role of CR1/2 and FcγR in the regulation of the immune response was investigated using OVA that was chemically conjugated to whole IgG of the rat anti-mouse CR1/2 mAb 7G6. FACS analysis using the murine B cell lymphoma IIA1.6 confirmed that the 7G6-OVA conjugate recognized CR1/2. Incubating IIA1.6 cells with 7G6-OVA triggered tyrosine phosphorylation and Ag presentation to OVA-specific T cells in vitro. Immunizing mice with 7G6-OVA at a minimal dose of 1 μg i.p. per mouse markedly enhanced the anti-OVA Ig response, which was primarily of the IgG1 isotype subclass. The 7G6-OVA did not enhance the anti-OVA response in CR1/2-deficient mice. OVA coupled to an isotype control Ab induced a considerably lower anti-OVA response compared with that induced by OVA alone, suggesting inhibition by interaction between the Fc part of the Ab and the inhibitory FcγRIIb on B cells. This finding was supported by the observation that IIA1.6 cells which were incubated with 7G6-OVA lost the ability to present Ag upon transfection with FcγRIIb. In sum, 7G6-conjugated OVA, resembling a natural immune complex, induces an enhanced anti-OVA immune response that involves at least CR1/2-mediated stimulation and that may be partially suppressed by FcγRIIb.
Differential Responses to CD40 Ligation Among Burkitt Lymphoma Lines That Are Uniformly Responsive to Epstein-Barr Virus Latent Membrane Protein 1
Authors: Henriquez Nico V. , Floettmann Eike , Salmon Mike , Rowe Martin , Rickinson Alan B. 
Published: January 2023

AbstractLigation of CD40 on the surface of B cells induces multiple phenotypic effects, many of which are mimicked by the EBV latent membrane protein 1 (LMP1) through its interaction with downstream components of the CD40 signaling pathway. Because the effects of LMP1 have been most closely studied in human Burkitt Lymphoma (BL) cell lines retaining a tumor biopsy-like phenotype in vitro, we have examined the response of a panel of such lines to CD40 ligation. Two distinct patterns of response were observed that were unrelated to the surface level of CD40 or to the EBV genome status of the lines. Following exposure to either CD40-specific mAbs or the soluble trimeric ligand (sCD40L), high responder (HR) lines showed rapid aggregation, activation of NF-κB, up-regulation of cell surface markers ICAM-1/CD54 and Fas/CD95, and growth inhibition. Aggregation was seen at lower doses than those required to elicit the other effects. By contrast, low responder (LR) lines showed no detectable response to CD40 mAbs, while their responses to sCD40L were limited to activation of NF-κB and up-regulation of CD95 only. However, in transfection experiments, LMP1 uniformly induced the full spectrum of phenotypic effects in both HR and LR lines. We conclude that some BL cell lines show a highly restricted response to CD40 ligation but remain fully susceptible to LMP1.
Postthymic Development of CD28−CD8+ T Cell Subset: Age-Associated Expansion and Shift from Memory to Naive Phenotype
Authors: Nociari Marcelo M. , Telford William , Russo Carlo 
Published: January 2023

AbstractDuring human aging, one of the major changes in the T cell repertoire is a dramatic expansion of T cells with the atypical CD28−CD8+ phenotype. In this study, we show that this increase is a consequence not only of an expansion in the CD28−CD8+ population but also of a decrease in the number of CD28+CD8+ T cells. The decrease in circulating CD28+CD8+ T cells is dramatically accelerated after the age of 50 and is not accompanied by an equivalent reduction in the CD28+CD4+ subset. Our findings confirm that aging leads to an accumulation of CD45RO+ T cells within the CD28+CD8+ subset as previously observed. Surprisingly, we found an increase in CD45RA+ expression with age in the CD28−CD8+ subset. Immune-phenotyping for activation markers, measurement of telomere DNA content, and cytokine production analysis indicate that the large majority of CD28−CD8+ T cells are Ag-experienced, despite their CD45RA+ phenotype. Our study further demonstrates that the poor proliferative response displayed by CD28−CD8+ T cells is not a consequence of telomere shortening. Also, analysis of cytokine production at the single cell level revealed that the proportions of IFN-γ+, IL-4+, and IL-10+ T cells are considerably higher among the CD28−CD8+ than the CD28+CD8+ subset. In summary, these data explain the presence of CD45RA+ T cells in the elderly, shed light on the phylogenetic origin of CD28−CD8+ T cells, and suggest a role for these cells in the immune senescence process.
Presentation of Antigens Internalized Through the B Cell Receptor Requires Newly Synthesized MHC Class II Molecules
Authors: Forquet Frédérique , Barois Nicolas , Machy Patrick , Trucy Jeannine , Zimmermann Valérie S. , Leserman Lee , Davoust Jean 
Published: January 2023

AbstractExogenous Ags taken up from the fluid phase can be presented by both newly synthesized and recycling MHC class II molecules. However, the presentation of Ags internalized through the B cell receptor (BCR) has not been characterized with respect to whether the class II molecules with which they become associated are newly synthesized or recycling. We show that the presentation of Ag taken up by the BCR requires protein synthesis in splenic B cells and in B lymphoma cells. Using B cells transfected with full-length I-Ak molecules or molecules truncated in cytoplasmic domains of their α- or β-chains, we further show that when an Ag is internalized by the BCR, the cytoplasmic tails of class II molecules differentially control the presentation of antigenic peptides to specific T cells depending upon the importance of proteolytic processing in the production of that peptide. Integrity of the cytoplasmic tail of the I-Ak β-chain is required for the presentation of the hen egg lysozyme determinant (46–61) following BCR internalization, but that dependence is not seen for the (34–45) determinant derived from the same protein. The tail of the β-chain is also of importance for the dissociation of invariant chain fragments from class II molecules. Our results demonstrate that Ags internalized through the BCR are targeted to compartments containing newly synthesized class II molecules and that the tails of class II β-chains control the loading of determinants produced after extensive Ag processing.
Identification of a Second Major Tumor-Specific Antigen Recognized by CTLs on Mouse Mastocytoma P815
Authors: Bilsborough Janine , Van Pel Aline , Uyttenhove Catherine , Boon Thierry , Van den Eynde Benoı̂t J. 
Published: January 2023

Abstract
Murine mastocytoma P815 induces CTL responses against at least four distinct Ags (AB, C, D, and E). Recent studies have shown that the main component of the CTL response against the P815 tumor is targeted against Ags P815AB and P815E. The gene P1A has been well characterized. It encodes the P815AB Ag in the form of a nonameric peptide containing two epitopes, P815A and P815B, which are recognized by different CTLs. Here, we report the identification of the P815E Ag. Using a cDNA library derived from tumor P815, we identified the gene coding for P815E. We also characterized the antigenic peptide that anti-P815E CTLs recognize on the MHC class I molecule H-2Kd. The P815E Ag results from a mutation within an ubiquitously expressed gene encoding methionine sulfoxide reductase, an enzyme that is believed to be important in the protection of proteins against the by-products of aerobic metabolism. Surprisingly, immunizing mice i.p. with syngeneic tumor cells (L1210) that were constructed to express B7-1 and P815E did not induce resistance against live P815, even though a strong anti-P815E CTL response was observed with splenocytes from immunized animals.
Paradoxical Preservation of a Lipopolysaccharide Response in C3H/HeJ Macrophages: Induction of Matrix Metalloproteinase-9
Authors: Jin Fen-yu , Nathan Carl F. , Ding Aihao 
Published: January 2023

Abstract
C3H/HeJ mice carry a mutant allele (Lpsd) of a recently identified gene whose normal allele (Lpsn) confers responsiveness to bacterial LPS in C3H/HeN and most other mouse strains. Recently we reported a differential display analysis of matched macrophage-derived cell lines from C3H/HeJ and C3H/HeN mice under LPS-free conditions. Of the ∼12,000 transcripts evaluated, 4 were differentially expressed. One transcript represented secretory leukocyte protease inhibitor. In this study, we report another differentially expressed transcript, mouse matrix metalloprotease-9 (MMP-9). Like secretory leukocyte protease inhibitor, MMP-9 was expressed constitutively in the Lpsd macrophage cell line and not in the Lpsn cell line. Similarly, two additional macrophage cell lines that respond readily to LPS by producing nitric oxide and TNF expressed no MMP-9 under LPS-free conditions. However, in all four cell lines, LPS induced MMP-9 or augmented its expression. In primary macrophages, concentrations of LPS in the ng/ml range augmented the expression of MMP-9 mRNA. Paradoxically, macrophages from Lpsd mice expressed more MMP-9 transcripts than macrophages from Lpsn mice. In contrast, the induction of TNF in response to LPS was much more pronounced in Lpsn macrophages. The present findings with MMP-9 suggest that homozygosity at Lpsd does not so much prevent a response to LPS as dysregulate it, resulting in the suppression of some LPS signaling pathways and the preservation of others.
Clonal Dominance Patterns of CD8 T Cells in Relation to Disease Progression in HIV-Infected Children
Authors: Than Soe , Kharbanda Monica , Chitnis Vivek , Bakshi Saroj , Gregersen Peter K. , Pahwa Savita 
Published: January 2023

Abstract
CD8 T cells are important mediators of cellular immune responses as evidenced by clonal expansions in the CD8 TCR Vβ repertoire during primary HIV infection in adults. This study investigated the CD8 TCR Vβ repertoire by complementarity-determining region 3 length analysis using multiplex PCR in purified peripheral blood CD8 T cells of 22 HIV-infected children (age range was 0.75–15 yr, mean was 8.2 ± 4.1 yr). Evidence of clonal dominance in one or more Vβ families was obtained in 15 of 22 children. The patterns of clonal dominance were designated as major, minor, single, and none to indicate the involvement of three or more, two, one, or no Vβ families, respectively. A pattern of major or minor clonal dominance was observed in 12 children (group 1), whereas 10 children had single or no clonal dominance (group 2). In comparison with group 2, the children in group 1 had a higher percentage of CD4 cells (28.3 ± 11.6 vs 8.6 ± 4.8, p &lt; 0.001); a higher stimulation index in lymphoproliferative responses to Candida (92.0 ± 59.5 vs 12.3 ± 14.4, p = 0.002), tetanus (76.3 ± 51.2 vs 11.2 ± 12.7, p = 0.002), and alloantigens (178.3 ± 298.9 vs 32.9 ± 35.2, p &lt; 0.001); and a lower percentage of CD8+HLA-DR+CD38+ cells (37.4 ± 13.1 vs 54.6 ± 14.2, p &lt; 0.01). The number of dominant CD8 T cell clones was significantly correlated with the percentage of CD4 T cells (r = 0.669, p &lt; 0.001) but not with plasma HIV RNA. Compared with group 1, patients in group 2 had a 4.8 times greater probability of having &lt;15% CD4 cells. These findings indicate that CD8 clonal dominance in HIV-infected children reflects robustness of immune responses, regardless of time since infection and virus load.
Cutting Edge: LFA-1 Is Required for Liver NK1.1+TCRαβ+ Cell Development: Evidence That Liver NK1.1+TCRαβ+ Cells Originate from Multiple Pathways
Authors: Ohteki Toshiaki , Maki Chikako , Koyasu Shigeo , Mak Tak W. , Ohashi Pamela S. 
Published: January 2023

AbstractUsing mice deficient for LFA-1, CD44, and ICAM-1, we examined the role of these adhesion molecules in NK1.1+TCRαβ+ (NKT) cell development. Although no defect in NKT cell development was observed in CD44−/− and ICAM-1−/− mice, a dramatic reduction of liver NKT cells was observed in LFA-1−/− mice. Normal numbers of NKT cells were present in other lymphoid organs in LFA-1−/− mice. When LFA-1−/− splenocytes were injected i.v. into wild-type mice, the frequency of NKT cells among donor-derived cells in the recipient liver was normal. In contrast, when LFA-1−/− bone marrow (BM) cells were injected i.v. into irradiated wild-type mice, the frequency of liver NKT cells was significantly lower than that of mice injected with wild-type BM cells. Collectively, these data indicate that LFA-1 is required for the development of liver NKT cells, rather than the migration to and/or subsequent establishment of mature NKT cells in the liver.
Activation of CD4 T Cells by Somatic Transgenesis Induces Generalized Immunity of Uncommitted T Cells and Immunologic Memory
Authors: Gerloni Mara , Miner Kent T. , Xiong Sidong , Croft Michael , Zanetti Maurizio 
Published: January 2023

AbstractCellular immune responses were analyzed in vivo after a single intraspleen inoculation of DNA coding for a 12-residue Th cell determinant associated with a 12-residue B cell epitope, a process termed somatic transgene immunization. We show that CD4 T cells are readily activated and produce IL-2, IFN-γ and IL-4, characteristics of an uncommitted phenotype. Linked recognition of the two epitopes coded in the same transgene promoted IgM-IgG1 switch and enhanced the total Ab response but had no effect on IgG2a Abs. Although originating in the spleen, T cell responsiveness was found to spread immediately and with similar characteristics to all lymph nodes in the body. A single inoculation was also effective in establishing long term immunologic memory as determined by limiting dilution analysis, with memory T cells displaying a cytokine profile different from that of primary effector T cells. These studies provide evidence that by initiating immunity directly in secondary lymphoid organs, an immune response is generated with characteristics that differ from those using vaccines of conventional DNA or protein in adjuvant administered in peripheral sites. Somatic transgene immunization can therefore be used to probe T cell responsiveness in vivo and represents a tool to further understanding of the nature of the adaptive immune response.
Naive Human CD4+ T Cells Are a Major Source of Lymphotoxin α
Authors: Ohshima Y. , Yang L-P. , Avice M-N. , Kurimoto M. , Nakajima T. , Sergerie M. , Demeure C. E. , Sarfati M. , Delespesse G. 
Published: January 2023

AbstractIt is generally accepted that immunologically naive T cells display a very restricted cytokine production profile consisting mainly of IL-2, which is used as an autocrine growth factor. Here we report that activated naive CD4+ T cells, of neonatal or adult origin, express very high levels of soluble lymphotoxin (LT) α (LTα3), as determined by ELISA, RNase protection assay, and intracytoplasmic staining. Besides LTα3 and IL-2, these cells also produce high levels of TNF-α together with significant amounts of IFN-γ and IL-13. Naive cells also express LTβ mRNA and the membrane form of LTα (LTαβ). On average, naive CD4+ T cells secrete four times more LTα3 than Th1-like cells, twice more than naive CD8+ T cells, and ten times more than B cells. Thus, naive T cells express a large spectrum of cytokines, mainly of the Th1 type, and the very high levels of LTα3/TNF-α that they release may play an hitherto unsuspected role in the early stage of T cell-dependent immune responses.
p38 Mitogen-Activated Protein Kinase Mediates Signal Integration of TCR/CD28 Costimulation in Primary Murine T Cells
Authors: Zhang Jian , Salojin Konstantin V. , Gao Jian-Xin , Cameron Mark J. , Bergerot Isabelle , Delovitch Terry L. 
Published: January 2023

Abstract
Optimal T cell activation requires two signals, one generated by TCR and another by the CD28 costimulatory receptor. In this study, we investigated the regulation of costimulation-induced mitogen-activated protein kinase (MAPK) activation in primary mouse T cells. In contrast to that reported for human Jurkat T cells, we found that p38 MAPK, but not Jun NH2-terminal kinase (JNK), is weakly activated upon stimulation with either anti-CD3 or anti-CD28 in murine thymocytes and splenic T cells. However, p38 MAPK is activated strongly and synergistically by either CD3/CD28 coligation or PMA/Ca2+ ionophore stimulation, which mimics TCR-CD3/CD28-mediated signaling. Activation of p38 MAPK correlates closely with the stimulation of T cell proliferation. In contrast, PMA-induced JNK activation is inhibited by Ca2+ ionophore. T cell proliferation and production of IL-2, IL-4, and IFN-γ induced by both CD3 and CD3/CD28 ligation and the nuclear expression of the c-Jun and ATF-2 proteins are each blocked by the p38 MAPK inhibitor SB203580. Our findings demonstrate that p38 MAPK 1) plays an important role in signal integration during costimulation of primary mouse T cells, 2) may be involved in the induction of c-Jun activation and augmentation of AP-1 transcriptional activity, and 3) regulates whether T cells enter a state of functional unresponsiveness.