bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and characterization of a novel surface antigen gene induced in mast cells activated through the high affinity IgE receptor.
Authors: Pirozzi G , Terry R W , Epstein D , Labow M A
Published: December 2022

In an effort to isolate novel genes involved in inflammation and/or mast cell activation, we have used a combination of differential screening and subtractive hybridization to isolate genes whose expression are induced upon activation of a transformed rat mast cell line. One of the isolated clones, pMCA-32, contained an open reading frame of 278 amino acids that included a putative hydrophobic transmembrane domain, a cysteine rich Ig-like extracellular domain, and a cytoplasmic domain containing three consensus SH2-domain phosphotyrosine binding sites. The MCA-32 gene is also highly conserved between rat and mouse, with the two coding regions being 73% identical. Although the MCA-32 coding region did not contain an obvious signal peptide, MCA-32 protein was detected on the surface of rat mast cells, and the cloned cDNA produced a cell surface protein when expressed in COS-7 cells. MCA-32 RNA from both mouse and rat undergoes alternative splicing, producing an mRNA containing an in-frame deletion of the TM domain, suggesting that a form of MCA-32 protein may be secreted. MCA-32 mRNA expression was up-regulated upon activation of RBL-2H3 cells and was highly abundant in primary peritoneal mast cells. Expression of MCA-32 RNA was only observed in primary and transformed mast cells from rat, while in the mouse MCA-32, RNA was also produced in significant amounts by a number of transformed monocyte cell lines. Thus, MCA-32 is a novel surface protein whose structure and expression suggest roles in the development and/or activation of mast cells and monocytes.
In vivo clonotypic regulation of human myelin basic protein-reactive T cells by T cell vaccination.
Authors: Zhang J , Vandevyver C , Stinissen P , Raus J
Published: December 2022

Autoreactive T cells specific for myelin basic protein (MBP) are implicated in the pathogenesis of multiple sclerosis. The mechanism by which MBP-reactive T cells are regulated in vivo remains unknown, but is thought to involve the clonotypic regulatory network that can be induced by immunization with attenuated T cells and TCR peptides. We reported previously that immunization of multiple sclerosis patients with irradiated MBP-reactive T cells (T cell vaccination) induced T cell responses to the immunizing clones, resulting in a clonal depletion of circulating MBP-reactive T cells. In this study, we demonstrated that in the majority of the recipients, MBP-reactive T cells remained undetectable in circulation over a period of 1 to 3 yr after vaccination, while they reappeared in some individuals (three of nine), coinciding with clinical exacerbation. The reappearing MBP-reactive T cells were found to originate from clonal origins different from those of T cells persisting before immunization, suggesting a shift of the T cell repertoire to other determinants of MBP. The immunization induces predominantly CD8+ regulatory T cells capable of lysing the immunizing clones in a clonotype-specific manner. The T cell responses induced by immunization were restricted to the immunizing clones and did not affect MBP-reactive clones not used for immunization. Our data further suggest that different hypervariable regions of the TCR may be involved in the observed clonotypic interaction. This study provides useful information for designing future clinical trials using T cell vaccination and other TCR-based therapeutic strategies.
Cytotoxic T cells directed to tumor antigens not expressed on normal melanocytes dominate HLA-A2.1-restricted immune repertoire to melanoma.
Authors: Anichini A , Mortarini R , Maccalli C , Squarcina P , Fleischhauer K , Mascheroni L , Parmiani G
Published: December 2022

To determine whether HLA-A21 restricted melanoma Ags exist that are not expressed on normal melanocytes, a panel of 478 T cell clones from six HLA-A21+ patients was selected for HLA-A2 restricted lysis of autologous tumor and then tested for differential recognition of HLA-A2.1+ melanomas and normal melanocytes. Four subsets of clones were identified in the panel of 107 HLA-A2-restricted CTL clones. CTL clones from three of the four subsets did not lyse melanocytes, but recognized fresh HLA-A2.1+ melanomas and defined three classes of epitopes, including unique Ags, common melanoma Ags, and Ags shared with neoplastic cells of different histologic origin. These CTL clones did not recognize any of the 10 peptides selected for specific association to HLA-A2.1 and derived from Melan-A/Mart-1, tyrosinase, gp100, or MAGE-3 proteins. By contrast, the fourth subset of HLA-A2.1-restricted CTl clones recognized both melanoma and melanocytes. These CTL clones were directed to a peptide from either Melan-A/Mart-1, tyronise, or gp100. By a limiting dilution assay, designed to evaluate the frequency of HLA-A2-restricted CTL precursors (CTLp) directed to melanoma but not to melanocytes, such precursors were found in the peripheral blood or tumor site of five of six HLA-A2.1+ melanoma patients, and their frequency was much higher than the frequency of CTLp recognizing both tumor cells and the melanocytes. These results suggest that in melanoma patients most of the HLA-A2.1-restricted immune repertoire to melanoma is directly to epitopes expressed in the neoplastic but not in the normal cells of the melanocyte lineage.
Catecholamines modulate human NK cell circulation and function via spleen-independent beta 2-adrenergic mechanisms.
Authors: Schedlowski M , Hosch W , Oberbeck R , Benschop R J , Jacobs R , Raab H R , Schmidt R E
Published: December 2022

Increases in catecholamines have been shown to induce changes in migration of lymphocytes, in particular NK cells. To analyze the mechanisms of catecholamine-induced NK cell trafficking, normal healthy male human subjects and splenectomized individuals were infused with either adrenaline (0.10 microgram/kg/min), noradrenaline (0.15 microgram/kg/min), or NaCl i.v. for 20 min. Lymphocyte subsets (CD3+, CD4+, CD8+) transiently increased after administration of both catecholamines, with most pronounced increases (up to 600%) in NK cell numbers (CD16+ or CD56+) after infusion of adrenaline. These changes in NK cell numbers and function were accompanied neither by alterations in expression of adhesion molecules (CD11a), CD11b, CD31, CD43, CD44, CD62L) on NK cells nor by changes in plasma concentrations of soluble (s) adhesion molecules (sVCAM-1, sICAM-1, sE-selectin). Comparable increases in lymphocyte subsets were observed in splenectomized subjects, suggesting lymphocyte recruitment from other sources than the spleen. Furthermore, catecholamine-induced increases in lymphocyte subsets could be inhibited by pretreatment with the nonselective beta-adrenoceptor antagonist propranolol, but not by the beta1-selective antagonist bisoprolol. These data demonstrate that adrenaline and noradrenaline modulate the migratory capacity of human NK cells via spleen-independent beta 2-adrenoceptor mechanism.
Reversal of 1,3-bis(2-chloroethyl)-1-nitrosourea-induced severe immunodeficiency by transduction of murine long-lived hemopoietic progenitor cells using O6-methylguanine DNA methyltransferase complementary DNA.
Authors: Maze R , Kapur R , Kelley M R , Hansen W K , Oh S Y , Williams D A
Published: December 2022

Bone marrow toxicity is a dose-limiting side effect of chloroethylnitrosourea (CNU) chemotherapeutic alkylating agents. A major determinant of CNU cytotoxicity is the methylation of guanine at the O6-position and the subsequent formation of interstrand DNA cross-links. O6-Methylguanine DNA methyltransferase (MGMT) removes alkyl groups from the O6 position of guanine and has been shown to repair CNU-induced DNA damage. We have previously demonstrated that transplantation of murine bone marrow cells transduced with a recombinant retroviral vector expressing MGMT via the human phosphoglycerate kinase promoter (PGK-MGMT) protects animals in vivo from acute myelotoxicity associated with CNU treatment. In the present study, we examined the effects of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), a commonly used CNU, on long term recovery of the lymphoid compartment, including thymus reconstitution, peripheral T and B cell populations, and lymphocyte mitogen responses in mice reconstituted with PGK-MGMT-transduced hemopoietic cells. Mice transplanted with either mock-infected control or PGK-MGMT-transduced stem cells were treated with five weekly doses of BCNU. Analysis of the lymphoid compartment demonstrated significant damage 3 mo after the last BCNU dose in control animals. In contrast, the profound deficiency in CD4+CD8+ double-positive thymocytes and mature lymphocytes observed in control mice surviving BCNU treatment was completely reversed in mice transplanted with PGK-MGMT-transduced bone marrow and was associated with molecular evidence of in vivo selection of transduced cells in the lymphoid compartment. Thus, long term immunodeficiency following CNU therapy may be prevented by genetic modification of murine hemopoietic stem cells with MGMT, leading to significant improvement in post-transplant immune function.
Dual role of HIV Tat in regulation of apoptosis in T cells.
Authors: McCloskey T W , Ott M , Tribble E , Khan S A , Teichberg S , Paul M O , Pahwa S , Verdin E , Chirmule N
Published: December 2022

Apoptosis has been suggested to be one of the major mechanisms of depletion of CD4+ T cells in HIV-1-infected individuals. Remarkably, HIV-1-infected cells appear protected from apoptosis, whereas bystander cells show increased apoptosis in lymph nodes of infected individuals. In this work, we present evidence that the trans-activating protein of HIV-1, Tat, has a dual role in regulation of apoptosis in T cells. While addition of exogenous Tat protein induced apoptosis in uninfected T cells, T cell clones stably expressing the Tat protein were protected from activation-induced apoptosis. The addition of exogenous Tat potentiated anti-CD3 mAb, anti-Fas IgM mAb, and TNF-alpha-induced apoptosis of T cells. Pretreatment of Tat with anti-Tat Ab abrogated Tat-induced apoptosis, but did not affect anti-Fas IgM Ab-induced apoptosis. Endogenously expressed Tat was analyzed in Jurkat T cell clones transfected with either full-length tat gene (101 amino acids), or in control cells containing an empty vector. The Tat101-transfected clones were resistant to anti-CD3-induced apoptosis, when compared with cells transfected with vector alone. Furthermore, cross-linking of CD4 molecules on T cells with gp160 and anti-gp160 Ab showed markedly decreased apoptosis in Tat101 cells compared with that induced in cells transfected with vector alone. Taken together, our results indicate that HIV-1 Tat can regulate apoptosis that may contribute to the immunopathogenesis of AIDS.
Homing potentials of circulating lymphocytes in humans depend on the site of activation: oral, but not parenteral, typhoid vaccination induces circulating antibody-secreting cells that all bear homing receptors directing them to the gut.
Authors: Kantele A , Kantele J M , Savilahti E , Westerholm M , Arvilommi H , Lazarovits A , Butcher E C , Mäkelä P H
Published: December 2022

Specific Ab-secreting cells (ASC) appear in the human blood as a response to oral and parenteral vaccination. The actual contribution of these cells to the defense of the body depends on their final effector site. The homing potentials of mucosally and parenterally induced ASC were compared by examining the homing receptor (HR) expression of circulating specific ASC in the blood of volunteers vaccinated orally or parenterally with the same Ag, Salmonella typhi Ty21a. Circulating lymphocytes were separated into receptor-positive and -negative populations, and the numbers of specific ASC were assayed. The alpha4 beta7 integrin, which acts as a gut HR, was expressed on all (99%) of the mucosally activated ASC, but on only 58% of the parenterally induced ASC or 58% of all Ig-secreting cells of the unvaccinated controls. L-selectin, the peripheral lymph node HR, showed an inverse distribution; it was found on 42% of mucosally activated ASC and on 86% of parenterally induced ASC. These results reveal that all of the circulating ASC after oral vaccination are committed to migrate to the mucosal compartment of the immune system, strongly arguing for a recirculation of activated mucosal cells in humans. By contrast, ASC induced by parenteral vaccination with the same Ag are mostly directed to the systemic compartment, yet a part of them has mucosal homing attitudes as well. These differences indicate that the site of Ag encounter determines the homing potential of the cell.
Staphylococcal enterotoxin B induces an early and transient state of immunosuppression characterized by V beta-unrestricted T cell unresponsiveness and defective antigen-presenting cell functions.
Authors: Muraille E , De Smedt T , Andris F , Pajak B , Armant M , Urbain J , Moser M , Leo O
Published: December 2022

Staphylococcal enterotoxin B (SEB) is a bacterial enterotoxin able to simultaneously bind to class II molecules on APCs and to selected V beta regions (including V beta 8) of the TCR complex. Administration of SEB to adult BALB/c mice results in clonal activation of T cells bearing V beta 8 receptors, leading to an excessive release of proinflammatory cytokines. This initial immune response is followed by a long-lasting state of V beta 8-specific unresponsiveness, thought to benefit both the host (as it contributes to the down-regulation of the inflammatory response) and the bacterium (through ligand-specific T cell anergy). However, it is not clear how this type of restricted unresponsiveness can effectively impair the generation of an antibacterial response. To gain insight into the mechanism by which Gram-positive bacteria subvert the host immune response, we have investigated the immune competence of SEB-treated mice 48 h following SEB administration. We demonstrate in this report that in vivo, SEB induces a transient but profound state of unresponsiveness affecting both T and Ag-presenting cell functions. Although in vivo activation by SEB appears to be V beta-restricted under our experimental conditions, SEB-treated mice displayed an early (lasting 48 to 72 h postinjection) and V beta-unrestricted unresponsive state characterized by the inability to produce IL-2 in response to polyclonal TCR mitogens including third party bacterial superantigens (staphylococcal enterotoxin A and toxic shock syndrome toxin 1, SEA and TSST-1, respectively), Abs to non-SEB reactive V beta regions (V beta 6), anti-CD3 epsilon Abs, and a lectin (Con A). Spleen cell populations from SEB-treated mice also displayed defective APC functions, possibly related to a selective decrease in splenic dendritic cells numbers. Taken together, these observations indicate that SEB induces an early and transient state of immunodeficiency in vivo, representing a potential mechanism for escaping host immune surveillance.
HAX-1, a novel intracellular protein, localized on mitochondria, directly associates with HS1, a substrate of Src family tyrosine kinases.
Authors: Suzuki Y , Demoliere C , Kitamura D , Takeshita H , Deuschle U , Watanabe T
Published: December 2022

Cross-linking of the Ag receptors on lymphocytes initiates activation of the receptor-coupled tyrosine kinases. HS1 is one of the substrates of these kinases and has been shown to transduce the signals for both clonal expansion and deletion in lymphoid cells. To gain further insight into the mechanism of action of HS1, we have tried to identify a protein that interacts with HS1 by yeast two-hybrid screening. The isolated cDNA, designated HAX-1, encodes a novel 35-kDa protein. The HAX-1 gene is expressed ubiquitously among tissues, and its protein is localized mainly in mitochondria, but also in endoplasmic reticulum and nuclear envelope in the cell. HS1/HAX-1 association is confirmed by coimmunoprecipitation of these proteins in the lysates of B lymphoma cells and COS-7 cells transfected with the corresponding cDNA expression vectors. Colocalization of these proteins in the cell is evident under confocal laser scanning microscope. Deletion mutant analysis of these proteins reveals that the association is mediated by the amino terminal region of HS1 and the carboxyl-terminal half of HAX-1. The potential role of the HAX-1/HS1 complex is discussed.
Human autoreactive CD4+ T cell clones use perforin- or Fas/Fas ligand-mediated pathways for target cell lysis.
Authors: Vergelli M , Hemmer B , Muraro P A , Tranquill L , Biddison W E , Sarin A , McFarland H F , Martin R
Published: December 2022

It is well established that target cell lysis by MHC class I-restricted CD8+ T cells is an important defense mechanism during infections with intracellular pathogens or against tumor targets. On the other hand, little is known about the physiologic role and the mechanisms of cytotoxicity of CD4+ MHC class II-restricted T cells. We have recently demonstrated that human autoreactive CD4+ T cells specific for one candidate autoantigen of multiple sclerosis, myelin basic protein, can mediate cytotoxicity. In the present report, we analyze the cytolytic mechanisms employed by these cells. We show that individual T cell clones, regardless of their cytokine phenotype, can be noncytotoxic or lyse target cells via either perforin- or Fas/Fas ligand-mediated cytotoxicity.
Roles of calnexin and Ig-alpha beta interactions with membrane Igs in the surface expression of the B cell antigen receptor of the IgM and IgD classes.
Authors: Wu Y , Pun C , Hozumi N
Published: December 2022

Membrane Igs (mIgs) of all five classes are associated with Ig-alpha beta dimers on the B cell surface. While mIgM requires the presence of these two associated molecules for its surface expression, mIgD does not. To study the structural basis for this differential Ig-alpha beta dependence, we created mutant mIgM and mIgD molecules (chimeras and those with reciprocal point mutations in their transmembrane sequences) and identified two amino acid residues in the transmembrane region of the mIg heavy chains responsible for this transport difference. Without Ig-alpha beta, mIgM and mutant mIgD molecules remained endoglycosidase H sensitive, consistent with endoplasmic reticulum (ER) localization. The molecular chaperone calnexin has previously been implicated in retaining unassembled mIgs in the ER. However, we found that inhibition of the association between calnexin and newly synthesized mIgs by castanospermine treatment did not lead to surface expression of normally retained mIgs. Therefore, calnexin cannot be the only retention molecule. Furthermore, for both wildtype and mutant mIgDs, association with calnexin was rather transient, thereby ruling out calnexin being a major ER retention molecule for mIgDs. Our study with castanospermine also showed that calnexin is required for wildtype mIgD surface expression only if Ig-alpha beta is absent, while the latter alone can function to promote mIg folding, assembly, and transport. Further study using our system will help to identify novel molecules and characterize their involvement in the control of mIg transport.
CD28 ligation prevents bacterial toxin-induced septic shock in mice by inducing IL-10 expression.
Authors: Wang R , Fang Q , Zhang L , Radvany L , Sharma A , Noben-Trauth N , Mills G B , Shi Y
Published: December 2022

The pathogenesis of septic shock is due mainly to bacterial toxin stimulation of the immune system, resulting in an excessive production of proinflammatory cytokines. TNF-alpha has been implicated as a major mediator in septic shock. Coinjection of D-galactosamine and LPS or staphylococcal enterotoxin B induced a rapid-onset, low-dose form of septic shock syndrome and ultimately led to death. We found that both the septic shock syndrome and death could be prevented by administration of anti-CD28 Ab. The protection induced by anti-CD28 Ab was associated with a decrease in TNF-alpha levels in the circulation. In addition, serum from anti-CD28 Ab-treated mice was capable of inhibiting the production of TNF-alpha by bone marrow-derived macrophages following treatment with LPS, indicating that anti-CD28 Ab induced production of soluble factors that subsequently inhibited the production of TNF-alpha. We confirmed that one of the factors present in serum was IL-10, because anti-CD28 Ab treatment stimulated the expression of IL-10, both in splenocytes and in T cell lines. Furthermore, injection of anti-IL-10 Abs could abolish the protective effect of anti-CD28 Ab on septic shock. Anti-IL-10 Ab could also suppress the anti-CD28 Ab-induced inhibition of TNF-alpha production, either in vivo or in vitro. Thus, we conclude that ligation of CD28 induces expression of IL-10, which in turn suppresses TNF-alpha production and prevents septic shock.
Granulocyte-macrophage colony-stimulating factor and IFN-gamma restore the systemic TNF-alpha response to endotoxin in lipopolysaccharide-desensitized mice.
Authors: Bundschuh D S , Barsig J , Hartung T , Randow F , Döcke W D , Volk H D , Wendel A
Published: December 2022

The influence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and IFN-gamma on the restoration of impaired TNF-alpha release in LPS-desensitized mice or their refractory macrophages was investigated. Mice pretreated with GM-CSF or IFN-gamma (50 microg/kg i.v.) and injected with 3 mg/kg LPS i.p. displayed increased plasma TNF-alpha levels compared with LPS controls. IL-10 was marginally up-regulated by GM-CSF but abrogated by IFN-gamma pretreatment. LPS-tolerant mice (30 microg/kg LPS i.p., -24 h) showed an attenuated plasma TNF-alpha and IL-10 response to LPS and survived LPS shock. Pretreatment of such mice with GM-CSF or IFN-gamma restored the previously impaired TNF-alpha response. In cultures of murine monocyte/macrophage-containing cell populations, i.e., alveolar, peritoneal, spleen, bone marrow cells, or blood, the presence of GM-CSF or IFN-gamma (10 ng/ml) resulted in an enhanced release of TNF-alpha initiated by 1 microg/ml LPS. Cells from LPS-tolerant mice showed a diminished responsiveness to LPS. However, when exposed to GM-CSF or IFN-gamma ex vivo, their TNF-alpha response to LPS was partially restored. These findings characterize GM-CSF and IFN-gamma as potent enhancers of LPS-induced TNF-alpha production in normal as well as in experimentally immunocompromised mice and provide the rationale for further experiments to explore the pharmacologic use of these cytokines for restoration of immunocompetence in sepsis-associated immunosuppression.
Expression of chemokine receptors by subsets of neurons in the central nervous system.
Authors: Horuk R , Martin A W , Wang Z , Schweitzer L , Gerassimides A , Guo H , Lu Z , Hesselgesser J , Perez H D , Kim J , Parker J , Hadley T J , Peiper S C
Published: December 2022

IL-8 is expressed by activated and neoplastic astrocytes and enhances the survival of hippocampal neurons in vitro. Since mRNA encoding chemokine receptors have been demonstrated in brain, the expression of chemokine receptors by specific cell types in anatomic regions of the central nervous system (CNS) was investigated. Archival tissues from various regions of the CNS were stained with specific mAbs to the Duffy Ag/receptor for chemokines, a promiscuous receptor that binds selected chemokines; the specific receptor for IL-8 (CXCR1); and the receptor (CXCR2) shared by IL-8 and melanoma growth stimulatory activity. The Duffy Ag/receptor for chemokines was expressed exclusively by Purkinje cells in the cerebellum. Chemokine binding and radioligand cross-linking confirmed the presence of a high affinity, promiscuous chemokine receptor in the cerebellum. Although CXCR1 was not expressed in the CNS, CXCR2 was expressed at high levels by subsets of projection neurons in diverse regions of the brain and spinal cord, including the hippocampus, dentate nucleus, pontine nuclei, locus coeruleus, and paraventricular nucleus, and in the anterior horn, interomediolateral cell column, and Clarke's column of the spinal cord. Fibers that express CXCR2 included those in the superior cerebellar peduncle and the substantia gelatinosa. Immunohistochemical analysis of the involved brain tissues from patients with Alzheimer's disease revealed expression of CXCR2 in the neuritic portion of plaques surrounding deposits of amyloid. These data suggest that chemokines may play a role in reactive processes in normal neuronal function and neurodegenerative disorders.
Distribution of Antibodies in the Serum and Organs of Rabbits
Authors: Freund Jules , Whitney Caroline E.
Published: December 2022

In a previous study of the distribution of agglutinins in the serum and organs of rabbits (1), it has been shown that the liver, spleen, kidney, lung, uterus and skin of actively or passively immunized rabbits accumulate approximately the same amount of antibodies per gram of organ. A decided difference was observed, however, in the rate of accumulation in these organs. In passively immunized animals the accumulation of agglutinins is rapid in the liver, spleen, kidney and lung. In these organs the accumulation was completed within from ten to fifteen minutes after the immune serum had been injected into the ear vein. On the other hand, in the uterus and particularly in the skin, the accumulation of antibodies continued for several hours.

In the first paper of this series it was suggested that the relatively early appearance of antibodies in the liver, spleen, kidney and lung may be connected with the relatively greater rate of lymph flow from these organs.
On the Racial Distribution of Some Agglutinable Structures of Human Blood
Authors: Landsteiner K. , Levine Philip
Published: December 2022

There are recorded in the literature some attempts to make a serological distinction of races with the methods that had proved successful in the differentiation of species. One may state that these efforts have not led to quite definite results. The claim of Bruck (1) that the sera of Caucasians, Chinese and Malays can be distinguished by means of complement fixation tests could not be confirmed by later workers.1 Marshall and Teague (6) observed some differences in similar tests but in their opinion the data are not so clear cut as to permit of a practical application. Negative results were obtained by Fitzgerald (7) who feels justified in concluding that with the method employed “the existence of specific racial differences … has not been proven.” Taking into account the difficulty of distinguishing the sera of closely related species, e.g., man and chimpanzee (Nuttal (8)), one would scarcely expect a more favorable outcome without resorting to very sensitive methods.
The Reticulo-Endothelial System and Antibody Production
Authors: Roberts Edward F.
Published: December 2022

An ever-increasing number of investigators are accepting the viewpoint that antibodies are accompaniments rather than causes of increased resistance. The validity of this opinion, however, will, undoubtedly, not be established until the site and mechanism of antibody elaboration have been determined. The few facts available indicate that the process of manufacture must be a function of cells or systems of cells widely distributed throughout the animal organism and of fixed-tissue origin. Such a system of cells is provided by the reticulo-endothelial system of Aschoff (1).

Many efforts have been made to link up the reticulo-endothelial cells with resistance to infection as well as to antibody production by methods of splenectomy or blockade, or a combination of the two. While these avenues of attack are admittedly weak, results fairly consistent, but not uniform, indicating a participation of the reticulo-endothelial system in resistance to infection have been obtained, although it must not be overlooked that the vast majority of experiments directed toward this problem have not included a sufficient number of cases to warrant the formulation of general conclusions.
Mathematico-Statistical Considerations on the Inheritance of the Human Blood Groups
Authors: Ichida Kenkichi
Published: December 2022

There are two principal hypotheses concerning the inheritance of human blood groups; one assumes the existence of two independent pairs of factors (the old hypothesis; v. Dungern-Hirszfeld (5), Ottenberg (34) (35) (36), Kirihara (28) (29), etc.) and the other is based on the assumption of triple allelomorphs (the new hypothesis; Bernstein (2) (3), Furuhata-Ichida-Kishi (8) (9) (10) (11) (12) (13), Furuhata (14) (15) (16) (17) (18) (19), Snyder (38). There is some essential difference between Bernstein's hypothesis and Furuhata's second hypothesis. Furuhata (12) (13) (16) (17) (19) maintains that the biochemical characters of the blood cells and sera in human blood are inherited always in a certain relationship, but he does not recognize the group-specific antigenicity of blood cells of the group O, as Bernstein supposes. They agree, however, in a point that they assume the three standard blood groups (ab/ab, Ab/Ab, aB/aB (Furuhata) RR, AA, BB (Bernstein)), and only a genotype in group AB.
Cutting Edge: The Chicken TCR ζ-Chain Restores the Function of a Mouse T Cell Hybridoma
Authors: Göbel Thomas W. F. , Bolliger Luca
Published: December 2022

The TCR/CD3 complex has been intensively studied in mammals, but it has been difficult to isolate homologues in other vertebrates. Here, we characterize the chicken ζ-chain, the first nonmammalian homologue identified. The comparison of mammalian and chicken ζ proteins revealed high identity of the transmembrane and the C-terminal cytoplasmic domains. Transfection of a mouse ζ-deficient cell line, with the chicken ζ gene, restored surface expression of the murine TCR/CD3 complex. The chicken ζ-chain was stably associated with the mouse TCR/CD3 components and fully restored its signaling capacity upon stimulation with Ab, superantigen, and peptide Ag. This is the first report of a nonmammalian TCR component that is capable of fully restoring a mammalian TCR in every aspect analyzed, thus demonstrating the enormous selective pressure to maintain the ζ-chain as a structural and signaling component over a period of 300 million years.
NK Cell Natural Cytotoxicity and IFN-γ Production Are Not Always Coordinately Regulated: Engagement of DX9 KIR+ NK Cells by HLA-B7 Variants and Target Cells
Authors: Kurago Zoya B. , Lutz Charles T. , Smith Kelly D. , Colonna Marco
Published: December 2022

DX9 mAb-binding killer cell-inhibitory receptors (KIR) recognize HLA-B molecules that express the Bw4 public serologic epitope. We assessed DX9+ NK cell fine specificity recognition of HLA-B7 variants and HLA-B27 alleles by 51Cr release natural cytotoxicity assays and by flow cytometry and enzyme-linked immunospot (ELISPOT) IFN-γ synthesis and release assays. 721.221 target cell expression of Bw4+ HLA-B27 alleles specifically inhibited DX9+ NK cell natural cytotoxicity and IFN-γ synthesis and release. A triple substitution of HLA-B7 at residues 80, 82, and 83 known to induce expression of the Bw4 serologic epitope also specifically inhibited DX9+ NK cell natural cytotoxicity and IFN-γ responses. Single HLA-B7 amino acid substitution variants were recognized in the same decreasing rank order by DX9+ NK cells and Bw4-reactive mAbs: G83R > R82L > N80T = HLA-B7. Natural cytotoxicity inhibition was reversed by the presence of blocking DX9 mAb. Natural cytotoxicity and IFN-γ production were coordinately regulated by a panel of HLA-B7 variants expressed on 721.221 cells, suggesting that these two effector functions are inhibited by the same KIR-mediated signaling mechanisms. In contrast, some NK cell clones killed 721.221 and K562 target cells equally well but released much more IFN-γ in response to K562 target cells. Differential regulation of natural cytotoxicity and IFN-γ release shows that NK cell effector functions respond to distinct signals.
TGF-β Promotes Immune Deviation by Altering Accessory Signals of Antigen-Presenting Cells
Authors: Takeuchi Masaru , Alard Pascale , Streilein J. Wayne
Published: December 2022

Macrophages incubated with OVA in the presence of TGF-β2 induce immune deviation in vivo (impaired delayed hypersensitivity and IgG2a Ab production) when injected into naive, syngeneic mice. OVA-specific TCR transgenic naive T cells (DO11.10 T cells) produce Th1-type cytokines when stimulated in vitro with OVA-pulsed peritoneal exudate cells (PEC), but if PEC are first treated with TGF-β2 and then pulsed with OVA, the T cells secrete Th2-type cytokines instead. In this study, we investigated the mechanisms that are involved in the modified Ag-presenting functions of macrophages by TGF-β2 pretreatment. We have found that: 1) TGF-β2 impaired the capacity of PEC to produce IL-12 and to express CD40; 2) reduced CD40 expression on TGF-β2-treated PEC impaired IL-12 production when the cells were cocultured with DO11.10 T cells; 3) the failure of TGF-β2-treated PEC to stimulate DO11.10 T cells to secrete IFN-γ was due to their impaired IL-12 production. From these results, we conclude that TGF-β2 treatment impairs the ability of macrophages to produce IL-12 and to express CD40. As a consequence, TGF-β2-treated PEC fail to promote development of pT cells toward the Th1 phenotype.
Non-MHC-Restricted Cell-Mediated Lysis of Human Oligodendrocytes In Vitro: Relation with CD56 Expression
Authors: Antel Jack P. , McCrea Ellie , Ladiwala Uma , Qin Yu-fen , Becher Burkhard
Published: December 2022

Oligodendrocytes and their myelin membranes are the apparent target of the autoimmune response that characterizes the human adult central nervous system-demyelinating disease multiple sclerosis. Human oligodendrocytes do not express MHC class II molecules, a requirement for MHC-restricted injury mediated by myelin-reactive CD4+ T cells, the cell type implicated in initiating the disease process. In this study we observed that human adult central nervous system-derived oligodendrocytes can be susceptible to non-MHC-restricted lysis mediated by myelin basic protein-reactive CD4+ T cell lines. Cytotoxicity was significantly greater (37 ± 4 vs 7 ± 3%) with cell lines in which a high proportion of cells (mean, 28 ± 6%) expressed CD56 compared with cytotoxicity mediated by low CD56 cell lines (8 ± 2%). High CD56 cell lines, when rested in IL-2, lost cytotoxic activity and had reduced expression of CD56 (mean, 5 ± 2%). CD4+ T cells isolated from short term (3-day) anti-CD3/IL-2-activated mononuclear cell cultures did not express CD56 and were not cytotoxic to oligodendrocytes unless lectin was added. In contrast, an enriched population of non-T cells extracted from the same activated MNC cultures expressed CD56 as well as other NK cell-associated surface molecules and was cytotoxic. These results indicate the potential susceptibility of human oligodendrocytes to non-MHC-restricted injury mediated by both Ag-reactive and nonspecific cellular immune effector cells, with CD56 expression being a common feature of the effector cells.
IL-12 Receptor (IL-12R) Expression and Accumulation of IL-12Rβ1 and IL-12Rβ2 mRNAs in CD4+ T Cells by Costimulation with B7-2 Molecules
Authors: Igarashi Osamu , Yamane Hidehiro , Imajoh-Ohmi Shinobu , Nariuchi Hideo
Published: December 2022

IL-12 is a crucial cytokine for the determination of a Th1/Th2 balance. It is important, therefore, to elucidate the mechanisms of IL-12R expression on Th cells. In this report, we present evidence to show that B7-2 costimulation plays a pivotal role in the expression of IL-12R on Th cells. A Th1 clone expressed a low density of IL-12R in a resting condition, the expression was enhanced by stimulation with specific Ag on splenic adherent cells and the enhancement was inhibited by anti-B7-2 or CTLA-4-Ig. When stimulated with anti-CD3 plus B7-2-transfected Chinese hamster ovary (CHO) cells, the clone strongly expressed IL-12R, although anti-CD3 by itself only weakly enhanced the expression. We obtained results that were similar to those in the Th1 clone in CD4+CD45RBlow memory T cells. In CD4+CD44low naive T cells, costimulation with B7-2-CHO was found to play a more important role in IL-12R expression. The accumulation of both IL-12Rβ1 and -β2 chain mRNAs was detected in naive T cells only when they were costimulated with anti-CD3 and B7-2-CHO, but β2 mRNA was not expressed upon anti-CD3 stimulation alone. On the other hand, both Th1 clones and memory T cells expressed low amounts of these mRNA without any stimulation, and the expression was weakly enhanced by anti-CD3 stimulation alone. For the maximum expression of these mRNAs, however, these cells also required costimulation with anti-CD3 and B7-2-CHO.
IL-1α and TNF-α Are Required for IL-12-Induced Development of Th1 Cells Producing High Levels of IFN-γ in BALB/c But Not C57BL/6 Mice
Authors: Shibuya Kazuko , Robinson Douglas , Zonin Francesca , Hartley Suzanne B. , Macatonia Steven E. , Somoza Chamorro , Hunter Christopher A. , Murphy Kenneth M. , O’Garra Anne
Published: December 2022

The development of Th1- or Th2-type responses determines the type of immune response that is elicited in response to Ag. Responsiveness to IL-12 is critical for the development of Th1-type CD4+ T cells required for cell-mediated immune responses. Addition of IL-12 to primary cultures of CD4+ T cells stimulated with OVA and splenocytes or dendritic cells resulted in the development of a Th1 phenotype with the capacity to secrete high levels of IFN-γ upon restimulation with splenic APC. The present study shows that using dendritic cells to present Ag upon restimulation reveals a requirement for additional cofactors, including IL-1α and TNF-α, which were provided by spleen cells but not dendritic cells. Furthermore, these cofactors are required for optimal IL-12-induced Th1 development in BALB/c but not C57BL/6 mice. This differential requirement for such cofactors in IL-12-driven Th1 development may play a role in genetic predisposition to Th1 or Th2 responses to infectious agents.
A Tyrosine-Based Signal Present in Ig α Mediates B Cell Receptor Constitutive Internalization
Authors: Cassard Sylvanie , Salamero Jean , Hanau Daniel , Spehner Daniele , Davoust Jean , Fridman Wolf Herman , Bonnerot Christian
Published: December 2022

B lymphocytes express Ag receptors (BCR) that are composed of ligand binding subunits, the membrane Igs, associated with Ig α/Ig β heterodimers. One main BCR function is to bind and to internalize Ags. Peptides generated from these internalized Ags may be presented to T lymphocytes. Here, we have analyzed the involvement of BCR Ig α/Ig β components in BCR constitutive endocytosis. The role of Ig α subunit in BCR constitutive endocytosis was first determined in the context of an IgM-based BCR. In contrast with BCR that contain wild-type Ig α, surface BCR lacking Ig α cytoplasmic domain were not constitutively internalized. The respective roles of Ig α and Ig β subunits were then analyzed by expressing chimeric molecules containing the cytoplasmic domains of either subunits in a B cell line. Only the Ig α cytoplasmic domain contained an internalization signal that allowed constitutive endocytosis of Ig α chimeras via coated pits and accumulation in sorting-recycling endosomes. This internalization signal is contained in its immunoreceptor tyrosine-based activation motif. These results indicate that Ig α, through its immunoreceptor tyrosine-based activation motif, may account for the ability of IgM/IgD BCR to constitutively internalize monovalent Ags.
Pentoxifylline Inhibits Ig κ Gene Transcription and Rearrangements in Pre-B Cells
Authors: Wang Weihong , Rath Satyajit , Durdik Jeannine M. , Sen Ranjan
Published: December 2022

Pentoxifylline (PF) has been used in a wide variety of clinical situations; however, the molecular consequences of this drug are not well characterized. In this paper we assayed the effects of PF in two models of pre-B differentiation. In 70Z pre-B cells, transcriptional induction of rearranged Ig κ-chain gene in response to LPS was suppressed by PF, without affecting the induction of Rel family proteins. In contrast, κ induction by IFN-γ was not suppressed by PF, indicating that the drug inhibited certain activation pathways. We also found that LPS-induced activation of germline κ transcription and Vκ to Jκ recombination were inhibited by PF in the pre-B cell line 38B9. These observations suggest that PF may adversely affect B lymphopoiesis during chronic administration.
Effects of Aminoguanidine on Latent Murine Tuberculosis
Authors: Flynn JoAnne L. , Scanga Charles A. , Tanaka Kathryn E. , Chan2 John
Published: December 2022

A unique feature of Mycobacterium tuberculosis is its ability to establish latent infection in the human host, which can reactivate to cause disease years later. In the present study, the mechanisms involved in the control of latent tuberculous infection were examined using two murine experimental tuberculosis models. Analysis of the model involving infection of mice with a relatively low inoculum of the virulent Erdman strain of M. tuberculosis indicated that in vivo inhibition of reactive nitrogen intermediate (RNI) production by the nitric oxide synthase inhibitor aminoguanidine resulted in reactivation. This reactivation was evidenced by hepatosplenomegaly, a robust tissue granulomatous reaction, and increased bacillary load. IFN-γ, TNF-α, and inducible nitric oxide synthase were all expressed throughout the latent phase of infection. Reactivation of latent tuberculous infection by aminoguanidine treatment was confirmed using a second murine tuberculosis model based on treatment with antimycobacterial drugs. Results obtained using this drug-based model also suggested the existence of an RNI-independent antimycobacterial mechanism(s) operative in the latent phase of infection. Together, these data suggest that both RNI-dependent and -independent mechanisms contribute to the prevention of tuberculous reactivation.
Identification of a T Cell Subset Capable of both IFN-γ and IL-10 Secretion in Patients with ChronicBorrelia burgdorferiInfection
Authors: Pohl-Koppe Annette , Balashov Konstantin E. , Steere Allen C. , Logigian Eric L. , Hafler David A.
Published: December 2022

A novel population of both IFN-γ- and IL-10-secreting human T cells that differentiate in the presence of exogenous IL-12 in vitro has recently been described. Whether this T cell population exists in vivo is unknown. Borrelia burgdorferi, the etiologic agent of Lyme disease, can induce a chronic infection in the presence of a vigorous humoral immune response. We established T cell lines specific for B. burgdorferi and tetanus toxoid from subjects with chronic B. burgdorferi infection and healthy controls in limiting dilution experiments and assessed proliferation and cytokine secretion. As expected, higher frequencies of B. burgdorferi-specific precursor T cells were observed in Lyme patients compared with controls. In both groups of subjects, T cell lines specific for B. burgdorferi secreted high amounts of IFN-γ. However, in patients with Lyme disease, 27% of T cell lines secreted not only IFN-γ but also IL-10, which was only observed in 0.6% of B. burgdorferi-reactive T cell lines generated from controls and in none of the tetanus toxoid-reactive T cell lines generated from either Lyme patients and controls. Single cell PHA cloning confirmed that both cytokines were secreted from one clonally expanded precursor cell. Whole mononuclear cells from B. burgdorferi-infected individuals, but not from controls, secreted IL-12. Moreover, neutralizing anti-IL-12 mAbs inhibited the generation of the IFN-γ/IL-10 population. These data demonstrate that this novel population of IL-12-induced IFN-γ/IL-10-secreting T cells is generated in response to chronic B. burgdorferi infection.
Th2 Cells Are Required for theSchistosoma mansoniEgg-Induced Granulomatous Response
Authors: Kaplan Mark H. , Whitfield Joel R. , Boros Dov L. , Grusby Michael J.
Published: December 2022

The development of Schistosoma mansoni ova-induced granulomas is regulated by cytokines secreted by distinct Th cell subsets. To determine the importance of Th1 and Th2 cells in granuloma formation, we have studied the immune response to S. mansoni ova in Stat4- and Stat6-deficient mice, which lack Th1 and Th2 cells, respectively. Lymphocytes from both naive and infected Stat6-deficient mice produced minimal levels of Th2 cell cytokines and Ag-specific IgG1 and IgE, but showed enhanced production of IFN-γ and Ag-specific IgG2a and IgG2b following schistosome egg injection. This shift away from a Th2 cell-mediated immune response was coupled with the development of pulmonary and hepatic granulomas that were greatly decreased in size compared with those in control littermates. Hepatic granulomas in Stat6-deficient mice were composed of predominantly mononuclear cells with very sparse appearance of eosinophils, and their diminished size was accompanied by decreased amounts of liver hydroxyproline content as a measure of collagen deposition. In contrast, lymphocytes from infected Stat4-deficient mice produced Th2 cell cytokines in amounts comparable to those produced by control littermates, but low levels of IFN-γ. While infected Stat4-deficient mice developed pulmonary granulomas following schistosome egg injection that were modestly impaired in size, the granuloma size and amount of collagen deposition in the liver were equivalent to those seen in control littermates. These studies demonstrate that Th2 cells are required for the full development of the granulomas and tissue-destructive fibrotic pathology associated with the immune response to S. mansoni ova.
LeishmaniaLipophosphoglycan Reduces Monocyte Transendothelial Migration: Modulation of Cell Adhesion Molecules, Intercellular Junctional Proteins, and Chemoattractants
Authors: Lo Siu K. , Bovis Lisa , Matura Rosemarie , Zhu Baixin , He Suhui , Lum Hazel , Turco Salvatore J. , Ho John L.
Published: December 2022

We previously identified the structural requirement for the inhibitory activity of Leishmania lipophosphoglycan (LPG) to block endothelial adhesion to monocytes. Here we showed that LPG reduces transendothelial migration of monocytes. LPG pretreatment of endothelial cells (2 μM, 1 h) reduced monocyte migration across endothelial cells activated by bacterial endotoxin (LPS) or IL-1β (60 and 46%, respectively). A fragment of LPG (i.e., repeating phosphodisaccharide (consisting of galactosyl-mannose)) and LPG coincubated with LPG-neutralizing mAb lacks inhibitory activity on monocyte migration. Pretreatment of monocytes with LPG (2 μM, 1 h) also did not affect monocyte migration through control or LPS-activated endothelial cells. FACS analysis reveals that LPG treatment blocked the LPS-mediated expression of E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 on endothelial cells and monocyte adhesion without altering the integrity of the endothelial monolayer. LPG (2 μM, 1 h) alone was capable of altering the expression and distribution of two junctional adhesion molecules, CD31 and vascular endothelium cadherin, as well as reversing the effects of LPS on these proteins. The induction of endothelial cells by LPS to transcribe and release monocyte chemoattractant protein-1 (MCP-1) was significantly reduced by LPG (40–65%). LPG treatment of nonactivated endothelial cells also suppressed by 55 to 75% the monocyte migration triggered by a MCP-1 chemoattractant gradient, and coincubation of LPG with neutralizing mAb abrogated the inhibitory activity. Together, these data point to a novel anti-inflammatory function of LPG in reducing monocyte migration across endothelial cells via a mechanism of inhibition of endothelial expression of cell adhesion molecules, modulation of intercellular junctional proteins, and synthesis of MCP-1.