bluuebunny/crossref_metadata_2025 huggingface dataset:
Induction of Cytotoxic T‐Cell Activity by the Protective Antigen of Schistosoma mansoni Sm28GST or its Derived C‐Terminal Lipopeptide
Authors: PANCRÉ V. , GRAS‐MASSE H. , DELANOYE A. , HERNO J. , CAPRON A. , AURIAULT C.
Published: November 2003

In a previous work the authors demonstrated that immunization with Schistosoma mansoni 28‐kDa glutathion‐S‐transferase (Sm28GST) was able to reduce hepatic damage in infected mice and that the adoptive transfer of Sm28GST‐specific T cells reproduced the protective effect obtained with the recombinant molecule. In the present paper, the authors show that Sm28GST is also able to stimulate an antigen‐specific, cytotoxic T‐cell response against Sm28GST‐pulsed P815 target cells in normal mice and that effector cells induced in vivo were classical Class I MHC‐restricted CD8+ lymphocytes. The authors found no spontaneous CTL activity against Sm28GST‐pulsed target cells during the course of the infection by S. mansoni although Sm28GST is expressed at different developmental stages of the parasite. It was observed, however, that immunization with Sm28GST is sufficient to elicit a significant level of CTL response for 6 weeks in infected mice. The role of these Class I MHC‐restricted CD8+ lymphocytes in the protection observed precisely at the same period in immunized mice remains to be elucidated. The authors also observe that immunization with the lipopeptidic form of the C‐terminal peptide of the molecule (190–211 peptide) led to a CTL activation comparable to that observed after immunization with the whole molecule demonstrating the feasibility of using a synthetic lipopeptide as immunogen for a CTL response against Sm28GST epitopes. Moreover, like Sm28GST‐specific CTLs, 190–211 lipopeptide‐specific cells were also Class I MHC‐restricted lymphocytes.
Human Brain Endothelial Cells and Astrocytes Produce IL‐1β but not IL‐10
Authors: CORSINI E. , DUFOUR A. , CIUSANI E. , GELATI M. , FRIGERIO S. , GRITTI A. , CAJOLA L. , MANCARDI G. L. , MASSA G. , SALMAGGI A.
Published: November 2003

The ability of human brain endothelial cells to produce mRNA for interleukin‐10, and release IL‐10 in culture supernatants after in vitro stimulation with LPS, TNF‐α and γ‐IFN was assessed and compared to that of astrocytes, peripheral blood mononuclear cells and human umbilical vein endothelial cells. IL‐1β and β2‐microglobulin release were also analysed. IL‐10 and TNF‐α mRNA presence was investigated in normal brain as well as in three plaques from two multiple sclerosis patients. While increased IL‐1β and β2‐microglobulin release in the supernatants of stimulated cells could be detected in all the studied cell lineages, IL‐10 mRNA and protein release was only seen in LPS‐stimulated PBMNCs. Similarly, mRNA for IL‐10 was not detected in CNS tissues, while TNF‐α was present in all plaques. The lack of production of significant amounts of IL‐10 by astrocytes and human brain endothelial cells suggests that these cells may not be the primary source of in vivo IL‐10‐mediated down‐regulation of immune reactions within the central nervous system.
Immunomodulation of LPS Ability to Induce the Local Shwartzman Reaction
Authors: AGUILLÓN J. C. , FERREIRA V. , NÚÑEZ E. , PAREDES L. , MOLINA M. C. , COLOMBO A. , HERMOSILLA T. , FERREIRA A.
Published: November 2003

Immunologically, the septic shock is a natural model of immunomediated vascular pathology where the interaction between cytokines and the endothelium mediates the syndrome and lethality. Tumour necrosis factor (TNF), a non‐species‐specific cytokine, has outstanding pleiotropic activities as an important mediator of the septic shock syndrome. In rabbits, passive immunization with anti‐lipopolysaccharide (LPS) polyclonal antibodies prior to the intravenous (i.v.) injection of LPS inhibits the haemorrhagic necrotic lesion characteristic of the local Shwartzman reaction (an excellent localized in vivo correlate of the septic shock). Paradoxically, tested in an ex vivo assay (short‐term whole human blood culture, stimulated with LPS), these antibodies mediated an increase in TNF production by mononuclear phagocytes and, in the rabbit model, they induced an increase in body temperature, as compared with the pre‐immune reagent. Although anchoring of immune complexes containing LPS to receptors (Fc or C4b‐C3b) on circulating monocytes may facilitate the access of LPS to these cells, access to localized, LPS‐sensitized macrophages may be impaired. Consequently inhibition of the local Shwartzman reaction and increased TNF production in the ex vivo system were observed. Concordantly, the higher temperature in the passively immunized animals may be a consequence of a higher, immune complex‐induced, systemic TNF production. These experimental results suggest that the use of anti‐LPS immunoglobulins, as a potential immunotherapy for septic shock syndrome in vertebrates, may lead to increased TNF production, with adverse effects such as the pyrogenic.
Auditory Stress Induces Changes in Membrane Functions of Mouse Peritoneal Macrophages
Authors: SPEHNER V. , DE WAZIERES B. , NICOD L. , HARRAGA S. , ROBERT J. F. , SEILLÈS E.
Published: November 2003

Stressful events induce responses in the endocrine and immune systems. The authors analysed the influence of repetitive noise stress on peritoneal macrophage oxidative and phagocytic responses. Plasma corticosterone levels were also measured. Different groups of 6‐ to 8‐week‐old C57BL/6 male mice were exposed for 1 night (n=14) and 3 nights (n=21) to a sound stress of 110 dB in an audiogenic stress chamber. Control animals were submitted to a sham stress for 1 night (n=13) and 3 nights (n=17). A marked decrease was observed in the phagocytic response to yeast (P=3×10−4) while a mild increase in the oxidative response stimulated by opsonized zymosan was noted only after the 3 night stress (P=0.02). Corticosterone levels of control and stressed mice did not differ. These results indicate that the stress resulting from repetitive noise causes modifications in peritoneal macrophage activity, and that these changes are dependant on the duration of stress. These functional alterations seem more complex than a simple general suppression or activation.
Contribution of Stabilizing Agents Present in Intravenous Immunoglobulin Preparations to Modulation of Mononuclear Cell Proliferation In Vitro
Authors: Alder L. B. A. , Morgan L. A. , Spickett G. P.
Published: November 2003

Intravenous immunoglobulin (IVIG) preparations have been shown to suppress lymphocyte proliferation in vitro. This study aimed to investigate the effects of an IVIG produced by fractionation/DEAE‐Sephadex adsorption on lymphocyte proliferation in vitro, with particular reference to contributions of the stabilizing agents present in the IVIG to the modulation of mononuclear cell proliferation. It was found that glycine significantly inhibited stimulation by the mitogenic lectin phytohaemagglutinin (PHA; 58% inhibition, P<0.01). Glucose and human albumin also reduced the response to PHA but to a lesser extent (20% and 30%, respectively). In further experiments the effects of sucrose and maltose, two disaccharides used as stabilizing agents in IVIG preparations, were studied. Three doses were used (2.5mm, 25mm and 250mm), representing levels likely to be found in vivo after infusion of IVIG at immunotherapeutic doses, on four different proliferative stimuli. Maltose was found to inhibit proliferation to PHA, anti‐CD3 and PMA in a dose responsive manner. Sucrose also inhibited proliferation to these stimuli, but a dose response was not observed. For both sugars, only the highest dose (250mm) inhibited the proliferative response of mononuclear cells to PMA and a calcium ionophore (ionomycin). The repurified IgG component of the IVIG preparation did not inhibit PBMC responses to PHA in this system. Kinetic analyses, in which the sugars were added 24h after proliferative stimuli, indicated that both sugars still inhibited responses to PHA, and, to a lesser extent, PMA, but only maltose inhibited the anti‐CD3 response. These findings show that stabilizing agents currently found in commercial IVIG preparations make a significant contribution to modulating mononuclear cell proliferation and need to be considered when assessing the immunomodulatory role of IVIG in vitro.
Glutamine, Lymphocyte Proliferation and Cytokine Production
Authors: ROHDE T. , MACLEAN D. A. , KLARLUND PEDERSEN B.
Published: November 2003

The present in vitro study was conducted to examine how glutamine influences the lymphocyte function. Glutamine had no effect on the production of interleukin‐1β, interleukin‐6 or tumour necrosis factor‐α, but influenced the production of interleukin‐2 and interferon‐γ. Glutamate, leucine, isoleucine and valine (substrates for glutamine production), or the combination of glutamate and leucine, did not influence the lymphocyte proliferative response or the cytokine production. In conclusion, glutamine influenced the production of some T‐cell‐derived cytokines, and is thereby important for optimal lymphocyte proliferation. Furthermore, the results show that lymphocytes are not capable of producing glutamine.
Susceptibility of DA Rats to Arthritis Induced With Adjuvant Oil or Rat Collagen is Determined by Genes Both Within and Outside the Major Histocompatibility Complex
Authors: Lorentzen J. C. , Klareskog L.
Published: November 2003

Inbred DA rats are remarkably susceptible to arthritis induced both by non‐immunogenic mineral oil only (OIA) and by rat collagen II together with mineral oil (rCIA). This fact enables interesting studies concerning which DA genes are associated with the arthritogenicity of adjuvant oil and collagen, respectively. In this paper the authors have investigated the role of genes within and outside the major histocompatibility complex (MHC), in this respect by comparative susceptibility studies in inbred rat strains (DA, LEW) and MHC‐congenic strains (DA.1I, DA.1O, DA.1M, DA.1H, LEW.1AV1, PVG.1AV1). The authors confirm earlier reports on the arthritis‐promoting nature of the DA MHC haplotype (RT1av1), but demonstrate that neither OIA nor rCIA is restricted to this haplotype or its class II alleles (Ba,Da). A decisive role of non‐MHC genes in determining susceptibility is also clear since the PVG.1AV1 strain, with the permissive RT1av1 haplotype, is resistant to both arthritis models. In conclusion, the authors demonstrate that the susceptibility of DA rats to OIA and rCIA is determined similarly, by permissive genes both within and outside the MHC. Second, and more interesting from a general perspective, the investigation demonstrates a notable influence of genes within and outside the MHC on the pathogenic capacity of a non‐immunogenic adjuvant oil. This adjuvant is in turn a critical component in provocations leading not only to rCIA but also to many other experimental autoimmune diseases where the influence of ‘adjuvant response genes’ is rarely considered.
Anti‐Idiotypic T Cells in Early Stages of Myasthenia Gravis: Increase in the Number and Prevalence Correlated to Clinical Improvement in Patients
Authors: YI Q. , PIRSKANEN R. , LEFVERT A. K.
Published: November 2003

An idiotypic network involving T and B cells bearing complementary structures has been suggested to be important for the regulation of immune response in healthy and disease situations. A previous study by the authors has demonstrated the presence of a relatively higher concentration of anti‐idiotypic antibodies than of idiotypic antibodies in early myasthenia gravis (MG), suggesting that the development of an anti‐idiotypic immunity is important in early MG. The present study was conducted to examine the cellular components of the idiotypic network in the same situation. T and B cells reactive to acetylcholine receptor (AChR) or to a disease‐related idiotype and to an anti‐idiotype were analysed in seven patients with early MG at various times after the start of the disease. The results show that a significant increase in the number of idiotype‐reactive interferon‐γ‐secreting T cells and a shift from AChR‐reactive to idiotype‐ and/or anti‐idiotype‐reactive T cells in the patients at 6 month follow‐up were noted. Such changes seem to correlate to a clinical improvement in the patients. The enhanced anti‐idiotypic T‐cell response and the clinical improvement in the patients may speak in favour of a role for the anti‐idiotypic immunity in controlling the autoimmune response in MG, i.e., down‐regulating autoantibody‐producing B cells and idiotypic (AChR‐specific) T cells. Thus, an immune intervention towards the enhancement of the anti‐idiotypic immunity in patients might be a rewarding approach. Further studies with regard to cell interactions and immune regulations in the network are warranted.
IFN‐γ Restores HIV‐ and Non‐HIV‐Specific Cell Mediated Immune Response In Vitro and Its Activity is Neutralized by Antibodies from Patients with AIDS
Authors: DE FRANCESCO M. A. , CARUSO A. , DIMA F. , CANTALAMESSA A. , CANARIS A. D. , FOLGHERA S. , FIORENTINI S. , FLAMMINIO G. , LICENZIATI S. , PERONI L. , GAO JING , GAROTTA G. , TURANO A.
Published: November 2003

The addition of IFN‐γ to cultures of peripheral blood mononuclear cells (PBMCs) obtained from asymptomatic HIV‐infected patients increased cell proliferation in response to HIV envelope synthetic peptides (Env), influenza A virus (VIRUS), andallogeneic lymphocytes (ALLO) but not to phytohaemagglutinin (PHA) stimulation. F(Ab)2 fragments of IgG purified from the sera of HIV‐seropositive patients specifically interfered with IFN‐γ‐induced cell proliferation in response to recallantigens. Neutralization of the lymphokine activity was found to be sustained by specific IFN‐γ antibodies. Data obtained demonstrate that IFN‐γ can restore the cell‐mediated immunity of a number of asymptomatic HIV+ individuals invitro, while IFN‐γ antibodies present in sera of patients with AIDS interfere with the activity of the lymphokine.
Induction of Human Cytotoxic T Lymphocytes by Oxidized Low Density Lipoproteins
Authors: WU R. , GISCOMBE R. , HOLM G. , LEFVERT A. K.
Published: November 2003

Oxidized low density lipoproteins (oxLDL) stimulated human blood T cells to a specific cytotoxic response which autologous PHA activated blood mononuclear cells which had been pre‐incubated with oxLDL. The cytotoxic reaction was exerted by CD8+ enriched cells but not by CD4+ enriched cells. Allogeneic target cells from HLA mismatched donors were not damaged and the reaction was consistently inhibited by monoclonal antibody against HLA class I. Antigen‐specific cytotoxic T cells might contribute to lysis of cells in the atherosclerotic plaque that have taken up oxLDL and thus participate in the inflammatory process associated with atherosclerosis.
A Non‐Competitive P55 TNF Receptor Antibody Enhances the Specific Activity of Lymphotoxin‐α
Authors: Medvedev A. E. , Laegreid A. , Sundan A. , Espevik T.
Published: November 2003

In the present study the authors elucidated the involvement of the two TNF receptors (TNFR) in discriminating TNF and lymphotoxin‐α (LT‐α) effects in human SW480‐βGal and KYM‐1 cell lines. A non‐competitive p55 TNFR monoclonal antibody (MoAb) 44E strongly enhanced LT‐α‐mediated gene regulation and cytotoxicity up to the level of the responses caused by TNF. TNF‐induced biological responses were only weakly influenced by 44E. 44E did not affect both binding and the rate of dissociation of the cytokines. The combination of the two p55 TNFR MoAb 44E and Htr5 elicited strong TNF responses, while none of them were agonistic alone. When the p75 TNFR was blocked with Utr1, LT‐α was still less potent than TNF in mediating CMV promoter activation and cytotoxicity. However, the addition of 44E in the presence of Utr1 merged the LT‐α dose‐response curves with those obtained with TNF plus Utr1. Using antagonistic TNFR  MoAb, the authors further showed that TNF functions through both TNFR types while LT‐α mediates its effects largely via the p55 TNFR. These data suggest that LT‐α is less potent than TNF due to its lower ability to properly trigger the p55 TNFR and because of its lack of signalling through the p75 TNFR.
Antibody‐Dependent Cellular Cytotoxic Activity of Cells in the Rat Metrial Gland in Pregnancy
Authors: PEEL S. , STEWART I. J.
Published: November 2003

Antibody‐dependent cellular cytotoxic (ADCC) activity of cells in rat metrial glands at days 13 and 20 of pregnancy has been examined. A 51chromium‐release cytotoxicity assay was carried out using Yac‐1 cells and P815 cells as targets in the presence of the corresponding, heat inactivated rat antibody or non‐immune rat serum. No killing was observed when effector metrial gland cells and target cells were cultured in the presence of non‐immune rat serum. In the presence of rat antibody toYac‐1 cells, or rat antibody to P815 cells, cells from the metrial glands of rats killed at day 13 of pregnancy demonstrated ADCC activity, but cells from the metrial glands of rats killed at day 20 of pregnancy showed no ADCC activity. Spleen andperitoneal exudate cells displayed ADCC activity. Immunohistological and immunocytological studies showed OX‐6, OX‐34, OX‐35 and OX‐42 positive cells were present at days 13 and 20 of pregnancy, in sections of metrial glands and in the cell suspensionsprepared for the cytotoxicity assays. It is suggested that macrophages are the cells most likely to be responsible for the ADCC activity in the rat metrial gland.
Adjuvant Properties of Montanide CSA 720 with a Recombinant HIV P17 gag Protein and Synthetic Peptide Antigens
Authors: KIRKLEY J. E. , GOLDSTEIN A. L. , NAYLOR P. H.
Published: November 2003

The adjuvant properties of Montanide CSA 720 were assessed in a comparison with alum. BALB/c mice were immunized with recombinant HIV‐1 gag protein p17 administered in either of the two adjuvants. The serum antibody response to p17 with Montanide CSA 720 appeared faster and reached a higher titre than with alum. The serum antibody response to p17 in Montanide CSA 720 was further characterized by a higher titre antibody directed against a 30 amino acid segment from the entire protein. The Montanide CSA 720 adjuvant was sufficiently strong to induce an antibody response against a weak synthetic peptide immunogen after two immunizations, while immunization with the peptide in alum generated no detectable serum antibody. The p17‐specific proliferative response of splenocytes from animals immunized with recombinant protein in either adjuvant was similar.
Heterogenous Expression of the Related MBP70 and MPB83 Proteins Distinguish Various Substrains of Mycobacterium bovis BCG and Mycobacterium tuberculosis H37Rv
Authors: WIKER H. G. , NAGAI S. , HEWINSON R. G. , RUSSELL W. P. , HARBOE M.
Published: November 2003

MPB70 and MPB83 are homologous cross‐reactive secreted mycobacterial proteins with very limited species distribution. The expression of these two proteins was compared between several substrains of Mycobacterium bovis BCG, virulent M. bovis and Mycobacterium tuberculosis H37Rv. A polyclonal antibody specific for MPB70 in Western blotting, and a monoclonal antibody, MBS43, found to be specific for MPB83 in ELISA and Western blotting, were used for the comparison. The previously established pattern of high‐ and low‐producing substrains of BCG for MPB70 is only partially applicable for MPB83. MPB70 low‐producing strains are also MPB83 low‐producing, but the expression of MPB83 is much more variable than the expression of MPB70 in the MPB70 high‐producing strains. Purified MPB83 (23 kDa) was found to be glycosylated. A band in SDS‐PAGE at 1–2 kDa lower than that of purified MPB83 may represent unglycosylated MPB83. Furthermore, it was confirmed that purified MPB70 (22 kDa) is unglycosylated. There is cross‐reactive antigen at 26 kDa. The MPB83 related antigen at 26 kDa was found to be the most abundant. These findings indicate greater heterogeneity between different substrains of BCG than previously realized. Virulent M. bovis produce and secrete large amounts of MPB70 and MPB83 while both these proteins occur in a far lower concentration in M. tuberculosis
The Interferon‐α/β Responses of Mice to Herpes Simplex Virus Studied at the Blood and Tissue Level In Vitro and In Vivo
Authors: ELORANTA M.‐L. , SANDBERG K. , ALM G. V.
Published: November 2003

Murine mononuclear leucocytes from bone marrow, spleen, lymph node and blood stimulated in vitro by UV‐irradiated Herpes simplex type I virus (HSV) produced about equal proportions of IFN‐α and ‐β determined by immunoassay. Thymocytes produced only IFN‐α. The frequency of IFN‐α/β mRNA containing cells detected by in situ hybridization was highest with bone marrow (15 per 104 cells), followed by spleen (4/104), lymph node (2/104), blood (1/104) and thymus (0.2/104). Such IFN‐α/β producing cells (IPCs) were heavily labelled in autoradiographs, each producing about 0.4 U of IFN. After one intravenous injection of UV‐irradiated HSV in mice, high levels of IFN‐α and ‐β were present in blood at 3–9 h and little or none at 24 h or later. Frequent cells strongly positive for IFN‐α mRNA at in situ hybridization and for IFN‐α/β at immunohistochemical staining were found almost exclusively in the marginal zones of spleens. Occasional IPCs were detected in lymph nodes but not in bone marrow, liver and kidneys. The marginal zone IPCs may be the major source of IFN in blood, and high splenic levels of IFN‐α/β should have efficient antiviral and immunoregulatory functions.
Decrease in cAMP Levels Promoted by CD48–CD2 Interaction Correlates with Inhibition of Apoptosis in B Cells
Authors: BAIXERAS E. , GARCÍA‐LOZANO E. , MARTINEZ‐A C.
Published: November 2003

The authors recently reported that CD2 ligation rescues B cells from antigen‐induced apoptosis by up‐regulation of intracellular Bcl‐2 levels. However, the characterization of the early signals involved in apoptosis rescue by CD2 ligation has not been well established. In this context, CD2 does not promote either phosphatidylinositol turnover or Ca2+ mobilization in B cells. In this paper the authors show that CD2 interaction with its ligand CD48 also reduces the apoptosis induced by forskolin and the phosphodiesterase inhibitor 3‐isobutyl‐1‐methylxanthine and, to a much lesser extent, the apoptosis induced by cholera toxin in murine B splenocytes. Using a cAMP detection system sensitive to the picomolar range, the authors demonstrate that CD2–CD48 interaction decreases the intracellular cAMP concentrations induced by forskolin but not by cholera toxin. In comparison with the CD2–CD48 interaction, CD40–CD40 ligand interaction completely inhibits the apoptosis induced by cAMP increases without affecting the intracellular cAMP levels promoted by forskolin or cholera toxin. These results indicate that CD2 can also control the apoptosis at the very early steps after receptor signalling, such as the adenylate cyclase activity. Given that heterotrimeric G‐proteins can mediate the adenylate cyclase activity the authors suggest that CD2 signalling could act through these small proteins, which would explain the inability of CD2 signalling to rescue from the apoptosis induced by cholera toxin, a Gs‐protein activator. Conversely, CD40 seems to control apoptosis further downstream of the cAMP‐PKA pathway where the survival and apoptotic signals are confluent, which might therefore render it a more efficient system to block apoptosis.
Secretion of Tumour Necrosis Factor α and Lymphotoxin α in Relation to Polymorphisms in the TNF Genes and HLA‐DR Alleles. Relevance for Inflammatory Bowel Disease
Authors: BOUMA G. , CRUSIUS J. B. A. , OUDKERK POOL M. , KOLKMAN J. J. , VON BLOMBERG B. M. E. , KOSTENSE P. J. , GIPHART M. J. , SCHREUDER G. M. TH. , MEUWISSEN S. G. M. , PEÑA A. S.
Published: November 2003

The genes for tumour necrosis factor alpha (TNFα) and lymphotoxin alpha (LTα; TNFβ) are tandemly arranged in the central region of the MHC. They may, therefore, be of importance for the aetiology of MHC‐associated diseases. The authors have prospectively studied the secretion of TNFα and LTα in relation to polymorphisms at positions ‐308 and ‐238 in the TNFα gene (TNFA), and two polymorphisms in the first intron of the LTα gene (LTA), as well as HLA‐DR in 30 patients with chronic inflammatory bowel diseases (IBD) and 12 healthy controls. In the Dutch population, the alleles of these four polymorphisms are present in only five combinations, called TNF‐haplotypes: TNF‐C, ‐E, ‐H, ‐I, and ‐P. Significant associations between TNF haplotypes and TNFα and LTα secretion were found when PBMC were cultured with T‐cell activators, irrespective of disease. Mean TNFα secretion of individuals carrying the HLA‐DR3 associated TNF‐E haplotype was significantly higher, as compared to individuals without this haplotype (26 441 pg/ml versus 19 629 pg/ml; P = 0.014). Individuals carrying the TNF‐C haplotype produced the lowest amount of TNFα (17 408 pg/ml; P = 0.022). The TNF‐C and TNF‐E haplotypes differ only at position ‐308 in the promoter of TNFA. Individuals carrying the HLA‐DR1 associated TNF‐I haplotype produced significantly less LTα when compared to those who lack this haplotype (1979 pg/ml versus 3462 pg/ml; P = 0.006). As the TNF‐I haplotype is also associated with low TNFα secretion, this haplotype thus defines a ‘low secretor phenotype’. In conclusion, this is the first study to show associations between TNF haplotypes and TNFα and LTα secretion when T‐cell stimulators are used. These findings will contribute to define disease heterogeneity in IBD and may be of relevance for understanding the pathogenesis of autoimmune diseases.
Characterization of a Human Basophil‐Like Cell Line (LAMA‐84)
Authors: Blom T. , Nilsson G. , Sundström C. , Nilsson K. , Hellman L.
Published: November 2003

LAMA‐84, a human leucocytic cell line, which upon establishment was described as having megakaryocytic, erythroid and granulocytic characteristics, was analysed for expression of various differentiation markers. In addition to some of the previously described phenotypic characteristics, this cell line was found to express mRNA for several proteins characteristic for basophilic leucocytes and mast cells. The authors show that LAMA‐84 cells express mRNA for the mast cell tryptase, the proteoglycan core protein, carboxypeptidase A and the α and β chains of the high affinity IgE receptor (FcεRI). The authors examined the potential of LAMA‐84 to differentiate in serum‐free medium or after DMSO or PMA treatment. Depending on the inducing factor, surface expression of the FcεRI α‐chain was increased from 20% to 35–50% of the cells and mRNA levels for tryptase were increased in serum‐free medium and after DMSO treatment. LAMA‐84 was found to express CD13, CDw17, CD29, CD33, CD40, CD45 and CD117. Furthermore, mRNA for the eosinophil/basophil markers Charcot–Leyden crystal (CLC) protein and the major basic protein (MBP), as well as the erythrocyte differentiation marker α‐globin, was detected. However, the authors observed only trace amounts of mRNA for another erythroid differentiation marker (glycophorin), trace amounts of the megakaryocytic marker GPIIIa, and no detectable level of GPIbα. By comparing the expression pattern of a panel of differentiation markers in LAMA‐84, and a second human cell line (KU812) expressing a basophil phenotype, it is evident that these cell lines, which presently are the only two cell lines identified with basophilic characteristics, share a large number of phenotypic characteristics.
Interleukin‐2 (IL‐2) Receptor α, β and γ Subunit Expression as a Function of B‐Cell Lineage Ontogeny: the Use of IL‐2‐PE664Glu to Characterize Internalization via IL‐2 Receptor Subunits
Authors: STEINBERGER I. , BEN‐BASSAT H. , HOCHBERG E. , LORBERBOUM‐GALSKI H.
Published: November 2003

Interleukin‐2 (IL‐2) is a pluripotent cytokine which plays a crucial role in the immune system response. Although the IL‐2/IL‐2 receptor (IL‐2R) system has been well characterized in cells of the T lineage it is less known in B lymphocytes. The authors therefore studied the expression of the IL‐2Rα, β and γ subunits in human B‐cell lines at different stages of maturation, by the polymerase chain reaction technique. The authors found that the α and β subunits are expressed in the final stages of B‐cell lineage maturation, whereas the γ subunit is constitutively expressed during B‐lymphocyte differentiation. The results indicate that the IL‐2/IL‐2R system, most probably, does not have a role in the early stages of B‐cell differentiation, but may be involved only in the final stages of B‐cell lineage ontogeny. Moreover, the ability of the different forms of IL‐2R to internalize the IL‐2 ligand was investigated, using the chimeric protein IL‐2‐PE664Glu. Cell lines bearing the αγ, βγ and αβγ forms of IL‐2R were inhibited by the chimeric protein, while those bearing the γ subunit alone did not respond to the chimera. Thus, internalization of IL‐2 is most likely mediated via the αγ form of the IL‐2R, as shown here for the first time, as well as through the βγ and αβγ IL‐2R forms. However, IL‐2 cannot be internalized through the IL‐2R γ subunit alone.
Reactivity Patterns of Synovial T‐Cell Lines Derived from a Patient with Rheumatoid Arthritis. I. Reactions with Defined Antigens and Auto‐Antigens Suggest the Existence of Multireactive T‐Cell Clones
Authors: MELCHERS I. , JOOß‐RÜDIGER J. , PETER H. H.
Published: November 2003

The functional T‐cell repertoire in the inflamed joint of a patient with rheumatoid arthritis was analysed at the clonal level. Using limiting dilution techniques and selecting for growth of in vivo activated and/or autoreactive T cells, 149 T‐cell lines were established. They were tested in a proliferation assay for reactivity against an autologous Epstein–Barr virus (EBV)‐transformed B‐cell line and a panel of auto‐antigens and foreign antigens. Seventy‐five lines (∼50%) could be stimulated. Thirty‐six lines (∼24%) were antigen‐reactive. They were stimulated by human collagens type I (15), II (10), IV (7) or V (4), cartilage proteoglycans (4), Mycobacterium tuberculosis (15), the 60 kDa heat‐shock protein of M. bovis (13) or tetanus toxoid (10). T‐cell lines were either monoreactive (19), bireactive (6), or multireactive (11), i.e. they were stimulated by either one, by two, or by more antigens in the panel. About half of the antigen‐reactive lines were at the same time autoreactive towards the autologous B‐cell line. These data suggest the existence of multispecific autoreactive T‐cell receptors comparable to multireactive or natural autoantibodies and prove the presence of autoantigen‐reactive T cells in the inflamed joints of patients with rheumatoid arthritis.
Changes in the Specificity of Antibodies in Mice Infected with Lactate Dehydrogenase‐Elevating Virus
Authors: Gómez K. A. , Coutelier J.‐P. , Retegui L. A.
Published: November 2003

Infection with lactate dehydrogenase‐elevating virus (LDV) modifies the isotypic distribution of antibodies (Ab) directed to several antigenic proteins with a preferential production of IgG2a. Because it was not known whether the virus could also affect the Ab specificity, the authors addressed this point using human growth hormone (hGH) as a model antigen. Anti‐hGH monoclonal antibodies (MoAb) were used as probes to study the occurrence of Ab to three native hGH epitopes (3C11, F11 and 10D6) in sera from LDV‐infected CBA/Ht and BALB/c mice immunized with hGH. Competition ELISA was used to determine the extent of Ab directed to cryptic hGH epitopes, i.e. antigenic determinants hidden in the native hormone. Results indicated that in LDV‐infected CBA/Ht mice the titres of anti‐hGH Ab were lower than in controls, although a consistent isotypic shift to IgG2a subclass was observed. Concurrently, the presence of Ab to epitopes 3C11, F11 and/or 10D6 were markedly reduced in infected animals and most anti‐hGH Ab were directed to hGH cryptic epitopes. By contrast, LDV infection increased the amount of anti‐KLH Ab elicited by CBA/Ht mice and did not affect Ab specificity, whilst control and LDV‐infected BALB/c mice showed similar concentrations of anti‐hGH Ab. Furthermore, the proportion of Ab to cryptic hGH epitopes did not change in infected animals even though an important shift to IgG2a was detected. Thus, data presented herein suggest that LDV infection modifies Ab specificity depending on the mice genetic background and on the antigenic characteristics of the immunogen.
Generation and Characterization of Rat Monoclonal Antibodies Against Human Serum Amyloid A
Authors: YAMADA T , HIRANO N , KURODA T , OKUDA Y , ITOH Y
Published: November 2003

Monoclonal antibodies against human serum amyloid A (SAA) were generated in rat (which seems not to have mature SAA proteins) by immunizing intact human SAA. Thirteen clones selected by initial screening were analysed based on reactivity with synthetic peptides of SAA and with carboxyl‐terminal truncated recombinant SAA. Antibodies were divided into four types, i.e. those recognizing the area around residue 18, 30, 90, and 100, respectively, of SAA. The antibody to the carboxyl terminus (around residue 100) of SAA, when subjected to immunohistochemistry for amyloid deposits in specimens from patients with reactive amyloidosis, always yielded negative reactivity, supporting the general concept that the carboxyl terminus of SAA is absent from human AA deposits.
Immune Responses in Mice After Gastric and Subcutaneous Immunization with BCG
Authors: IBSEN M. W. , BAKKEN V. , JONSSON R. , HORDNES K.
Published: November 2003

Mucosal immunization can induce immune responses different from those induced by systemic immunization. In this study, murine immune responses were analysed after immunization through gastric and subcutaneous routes with Mycobacterium bovis BCG. The number of circulating cells secreting antigen‐specific immunoglobulins (Ig), and the number of gamma interferon (IFN‐γ) secreting cells in spleen‐cell cultures after in vitro stimulation with mycobacterial antigens were analysed at the single cell level by the ELISPOT method. Levels of antigen‐specific antibodies in sera were determined by ELISA. In the subcutaneously immunized mice the authors found approximately 100 times more splenic cells secreting antigen‐specific IgG than in gastrically immunized mice or control mice. Their levels of antigen‐specific IgG in sera were 66–6700 times higher than in sera from gastrically immunized mice or control mice. In contrast, the numbers of IFN‐γ producing cells in spleen‐cell cultures after in vitro activation with BCG were equally high in the immunized groups of mice, and for both groups higher than in non‐immunized controls. Furthermore, IFN‐γ producing cells could be demonstrated in gastrically immunized animals even without in vitro activation. The results demonstrate that gastric immunization with the BCG vaccine can induce a systemic T cell‐mediated immune response against mycobacterial antigens.
Immaturity, Ageing and Oral Tolerance
Authors: VAZ N. , FARIA A. M. C. , VERDOLIN B. A. , CARVALHO C. R.
Published: November 2003

Founding studies of cellular immunology emphasized that tolerance to allografts could only be achieved early in the embryonic or neonatal period, suggesting that the establishment of self‐tolerance, a main event in the organization of the immune system, would necessarily take place in immature hosts. Contradicting these ideas, oral tolerance is a common, daily phenomenon, easily achieved by a physiological route in adult immunocompetent animals. Furthermore, there is solid evidence that, after the neonatal period, the susceptibility to oral tolerance induction also wanes and that it may be restored by adoptive transfer of cells from young hosts. These findings are briefly reviewed here to emphasize that immunological activity is a continuous and ongoing epigenesis extending throughout the entire life of the organism, far beyond the early phases of ontogenesis.
A Unique Response to Staphylococcal Enterotoxin B by Intrahepatic Lymphocytes and its Relevance to the Induction of Tolerance in the Liver
Authors: MATSUI S. , TERABE M. , MABUCHI A. , TAKAHASHI M. , SAIZAWA M. , TANAKA S. , YOKOMURO K.
Published: November 2003

It has been reported that the intrahepatic lymphocyte (IHL) population is somewhat differently constituted from lymphocytes in other lymphoid tissues. Staphylococcal enterotoxin B (SEB) is a superantigen which can induce T‐cell tolerance in mice. The authors investigated the in vitro and in vivo responses of mouse IHL to SEB. An intravenous injection of SEB did not result in the augmentation of the proliferative response of IHL, while mesenteric and splenic lymphocytes (mLNC and SPC, respectively) had augmented responses. Interleukin‐2 (IL‐2) mRNA was clearly detected in mLNC and SPC by reverse transcriptase–polymerase chain reaction (RT‐PCR) shortly after the administration of SEB, but it was scarcely expressed in IHL. The expression of CD25 (IL‐2 receptor) was also augmented in mLNC and SPC in the early period, while it was not changed in IHL. These findings suggested that the time required for tolerance induction is different locally and that the loss of augmentation of IL‐2 and IL‐2 receptor production by IHL may be relevant to the rapid induction of T‐cell tolerance in the liver.
The Role of Natural Killer Cells in Viral Infections
Authors: SEE D. M. , KHEMKA P. , SAHL L. , BUI T. , TILLES J. G.
Published: November 2003

Natural killer (NK) cells are important effectors for the lysis of both neoplastic and virus‐infected cells. Lectin‐like receptors on human NK cells, such as NKR‐PIA and CD94, bind to target cell carbohydrate ligands and initiate the lytic process. In addition, P58 and P70 bind to major histocompatibility class I antigens on targets and mediate negative signals. Models using NK cell‐deficient mice have proven useful in elaborating the role of NK cells in the immune defence against multiple viral agents. In addition, studies in humans have suggested a vital role of NK cells in the host defence against human immunodeficiency virus, herpesviruses, hepatitis B and C and other viruses. Several genetic disorders, chronic illnesses and infections have been associated with decreased NK function.
Decreases in CD5+ B Lymphocytes in Children with the DiGeorge Anomaly
Authors: KOURTIS A. P. , IBEGBU C. C. , LEE F. K. , VOGLER L. , PHILIPS K. , NAHMIAS A. J.
Published: November 2003

The authors evaluated the lymphocyte subsets in eight children with the DiGeorge anomaly, compared with 48 age‐matched control infants. Of particular interest was the finding that the percentage and number of CD5+ B lymphocytes were decreased in seven of the eight cases. This observation may provide insight into thymic function and the interaction of the B and T cell systems in some forms of congenital and acquired immunodeficiencies.
Evaluation of the Role of Fcγ and Complement Receptors in the Decreased Phagocytosis of Hereditary Haemochromatosis Patients
Authors: MOURA E. , VERHEUL A. F. M. , MARX J. J. M.
Published: November 2003

Hereditary haemochromatosis (HH) monocytes have a decreased antibody mediated phagocytosis of rabbit erythrocytes and Staphylococcus aureus compared to control monocytes. In order to investigate whether this decrease could be attributed to a different level of expression of Fcγ receptors (FcγR) or complement receptors (CR), which cooperate even in the absence of complement, the surface expression of these receptors was determined on monocyte‐enriched suspensions. In contrast to what was expected, HH monocytes displayed a significantly higher level of FcγRI and FcγRIIa as compared to healthy donor monocytes, but these differences were very small. The expression of the other receptors studied were similar for both groups. The heat‐inactivated mouse serum used for opsonizing the erythrocytes mainly contained mouse IgG1. Two genetically different forms of FcγRIIa are known, each with a different affinity for mouse IgG1 antibodies. Therefore, the FcγRIIa polymorphism in monocytes (MN) of both groups was also investigated. A similar distribution was found for patients and healthy donors. In addition, the extent of erythrophagocytosis of both donors and patients was independent of FcγRIIa allotype. Our results indicate that the altered phagocytosis by HH monocytes cannot be attributed to a different level of expression of receptors involved in phagocytosis or to FcγRIIa polymorphism.
High Levels of Neutralizing Autoantibodies Against IL‐1α are Associated with a Better Prognosis in Chronic Polyarthritis: a Follow‐Up Study
Authors: JOUVENNE P. , FOSSIEZ F. , BANCHEREAU J. , MIOSSEC P.
Published: November 2003

Neutralizing autoantibodies to interleukin (IL)‐1α were detected in a subset of chronic polyarthritis patients characterized by an increased proportion of patients with primary Sjögren's syndrome or self‐limiting inflammatory arthritis, diseases with a much better prognosis than rheumatoid arthritis (RA). The evolution of anti‐IL‐1α antibody levels was followed over 3 years. Incidence and levels were higher in patients with a benign form of polyarthritis. In these patients levels remained stable or increased over the follow‐up period. In contrast, incidence and levels were lower and some RA patients became negative. Negative correlations were observed between the levels of anti‐IL‐1α antibodies and the clinical and biological indices of disease activity. The relative risk factor of developing RA was 12 in the absence of high anti‐IL‐1α antibody levels and 18.2 when associated with the presence of HLA‐DR4. In conclusion, the presence of anti‐IL‐1α autoantibodies appears to be protective and their detection could represent a marker of good prognosis for destruction.
Lymphoid Cell Accumulation in Salivary Glands of Autoimmune MRL Mice Can be Due to Impaired Apoptosis
Authors: SKARSTEIN K. , NERLAND A. H. , EIDSHEIM M. , MOUNTZ J. D. , JONSSON R.
Published: November 2003

MRL‐lpr mice and the congenic strain MRL +/+ exhibit pathological abnormalities in the salivary glands similar to Sjögren's syndrome in humans. The lpr genotype has been identified as a mutation in the gene encoding Fas which is a cell surface protein that mediates apoptosis. The mutation is leaky, allowing for low levels of the APO‐1/Fas (CD95) receptor and partial activity of Fas/Fas ligand‐mediated programmed cell death in this strain. To examine the expression of Fas in situ, the authors analysed thymus, lymph node and salivary gland tissue from BALB/c, MRL +/+ and MRL‐lpr mice by an immunohistochemical technique (ABC‐immunoperoxidase) using an anti‐Fas (Jo2) antibody. For detection of apoptotic cells the authors used the terminal deoxynucleotidyl‐transferase‐mediated dUTP‐digoxigenin nick end labelling (TUNEL) method. Thymus from MRL +/+ and normal BALB/c mice showed a higher frequency of Fas expression than was seen in the lpr mice, but the +/+ mice had similar expression of Fas in lymph nodes as lpr mice. The Fas protein was detected among infiltrating mononuclear cells in the salivary glands of both lpr and +/+ mice. Apoptotic cells were found in the thymus with similar frequency in all three strains, while in the lymph nodes only BALB/c mice showed apoptosis. There was no, or very low, frequency of apoptosis among infiltrating mononuclear cells in salivary glands of both MRL strains. In conclusion, despite mutation of the Fas gene in the MRL‐lpr strain, there was nevertheless an expression of the apoptosis‐related Fas protein in lymphoid tissue and salivary glands of these mice. Based on analysis of apoptotic activity, the impaired Fas in autoimmune MRL mice seems to affect primarily the peripheral organs.
A Novel Mycobacterial Antigen Relevant to Cellular Immunity Belongs to a Family of Secreted Lipoproteins
Authors: OFTUNG F. , WIKER H. G. , DEGGERDAL A. , MUSTAFA A. S.
Published: November 2003

The gene sequence of a novel 24.1 kDa Mycobacterium tuberculosis protein was identified within the Sanger Centre (UK) M. tuberculosis genome database (cosmid MTCY24G1) by searching with a 126 bp DNA sequence isolated from a genomic M. lepraeλgt11 library with M. leprae reactive human T cell clones as probes. The 24.1 kDa antigen is common to the vaccine strain Mycobacterium bovis BCG, as well as Mycobacterium leprae. The 699 bp open reading frame encodes a 233 amino acid long precursor protein with a signal peptide sequence for secretion and a consensus motif for lipid conjugation, which suggests that the mature protein is an exported lipoprotein antigen. The molecular mass of the mature protein antigen from M. leprae sonicate was shown to correspond to the deduced size of the M. tuberculosis protein by T cell Western analysis. Homology searches revealed two other similarly sized hypothetical secreted mycobacterial lipoproteins within the M. tuberculosis genome database.
How to Diagnose Chronic Rejection. A Study in Porcine Intestinal Allografts
Authors: KUUSANMÄKI P. , LAURONEN J. , PAAVONEN T. , PAKARINEN M. , YILMAZ S. , HÄYRY P. , HALTTUNEN J.
Published: November 2003

Porcine small bowel allografts were followed for 18 weeks during immunosuppression with cyclosporine‐A (CyA), azatioprine and prednisone. The mucosal alterations noted at the 12th week were epithelial vacuolation and loss of Goblet cells. Moderate infiltration of inflammatory cells, mainly lymphocytes, was found in the lamina propria. In addition, a few grafts exhibited oedema and fibrosis. Vessels already showed endothelial swelling and intimal proliferation at the 12th week. In the submucosa, the infiltration of inflammatory cells was not present till the 18th week. Further changes in the mucosa at the 18th week were the blunting of villi, cuboidal epithelium, crypt abscesses and epithelial atrophy. The histological alterations of mucosa and lamina propria existing in the full thickness biopsies were mostly also detectable in mucosal biopsies, provided that multiple biopsies were taken. Thus these parameters analyzed from mucosal biopsy material are suitable for the diagnosis and monitoring of chronic small bowel rejection. In autopsy, the most prominent features were in the mesenterial arteries: intimal proliferation, vasculitis, proliferation of media and endothelial alterations. The activity of the mucosal disaccharidases maltase and sucrase remained near the initial level till the 12th week and had decreased markedly by the 18th week.
Preferential Localization of Human CD5+ B Cells in the Peritoneal Cavity
Authors: NISITANI S. , MURAKAMI M. , AKAMIZU T. , OKINO T. , OHMORI K. , MORI T. , IMAMURA M. , HONJO T.
Published: November 2003

In a transgenic mouse model of autoimmune haemolytic anaemia, CD5+ B lymphocytes localized in the peritoneal cavity are shown to play an important role in the onset of autoimmune disease. The authors have examined whether CD5+ B cells are present in the peritoneal cavity of 12 human individuals with non‐invasive gastrointestinal tumours and found that in humans CD5+ B cells preferentially lodge in the peritoneal cavity as compared to the peripheral blood and spleen while the numbers of the peritoneal B lymphocytes in humans are much lower than in mice and vary widely between individuals.
Studies in Knockout Mice Reveal that Anti‐Chlamydial Protection Requires TH1 Cells Producing IFN‐γ: Is this True for Humans?
Authors: JOHANSSON M. , SCHÖN K. , WARD M. , LYCKE N.
Published: November 2003

Chlamydia trachomatis is one of the major causes of infertility and preventable blindness in the world. The organism is of particular interest from an immunological point of view because it is one of the few obligate intracellular bacterial pathogens. There is some evidence that repeated infections in humans stimulate protective immunity. However, until recently, it was unclear which components of the adaptive immune system give rise to protection. Studies in gene knockout mice reported here and elsewhere now give a coherent and cogent picture of the importance of the Th1 response, in particular IFN‐γ, for the localization and eradication of C. trachomatis genital tract infection. The key questions still to be addressed are the identity of the IFN‐γ responsive cells and whether the mouse is truly representative of host protection against chlamydiae in humans.