bluuebunny/crossref_metadata_2025 huggingface dataset:
The use of embryo genotyping in the propagation of genes involved in the immune response
Authors: SABERIVAND ADEL , OUTTERIDGE PM
Published: June 2007

Multiple ovulation and embryo transfer (MOET) now enables researchers to produce identical twin animals, to obtain progeny from pre‐pubertal females and to obtain more offspring from valuable animals. MOET and sexed semen have produced genetic progress of up to 60% of milk production. The oestrous cycles of animals are synchronized with progestagens before superovulation with gonadal hormones, pregnant mare serum gonadotrophin and follicle stimulating hormone. Surgical, non‐surgical and laparoscopic methods are applied to recover and transfer embryos. Sexing and genotyping of the pre‐implantation embryos is a key step in improving the management and breeding programmes for livestock, as well as in the human for the prenatal diagnosis of genetic disorders. Several serological and physiological methods have been used to determine the sex of the pre‐implantation embryos; none has had satisfactory results in terms of time and accuracy, Sexing by polymerase chain reaction (PCR) using male‐specific chromosome sequences alone or with female‐specific chromosomal DNA probes simultaneously has been sufficient to identify the sex of the embryos with 100% accuracy. However, caution should be taken against sources of the contamination. The MHC class I, class II and background genes have been implicated in resistance to internal parasites in animals, Biotechnological methods such as screening of embryos prior to transfer using PCR and primer extension pre‐amplification have already made it possible to detect transgenic or genetically disordered embryos and could be applied to select those embryos bearing immunological genotypes of interest, such as resistance to internal parasites. Ultimately, cloning and nuclear transplantation would provide the possibility of isolating these resistance genes and to transfer them to livestock pre‐implantation embryos to propagate these desirable traits.
ISCOMs (immunostimulating complexes): The first decade
Authors: BARR IAN G , MITCHELL GRAHAM F
Published: June 2007

A little over a decade ago, novel immunostimulating complexes (ISCOMs) were described. This review examines the position and progress that ISCOM technology has achieved in the fields of vaccine research and medicine over this period. Much of the work on ISCOMs has remained in the area of vaccine research where there is still an urgent need for improved adjuvants to help combat important diseases such as AIDS, malaria and influenza. Currently the only widely licensed adjuvants for human use are the aluminium salts, but with the trend towards highly purified subunit vaccines, which are inherently less immunogenic than some of the older vaccines, potent adjuvants capable of promoting specific immune responses are required. ISCOMs are one such technology that offers many of these requirements and as their use in vaccines enters its second decade clinical trials are commencing that will establish whether these submicron, non‐living particles composed of saponin, cholesterol, phospholipid and in many cases protein, are useful components for a range of human vaccines.
Populations of thymocytes and peripheral blood leucocytes in leukaemia‐bearing mice treated with G‐CSF
Authors: KAWIAK J , HOSER G , MIKS B , POJDA Z , SOBICZEWSKA A , MACHAJ E , WREMBEL A
Published: June 2007

The influence of granulocyle‐colony stimulating factor (G‐CSF) on thymocyte subsets and peripheral blood leucocytes in leukaemia L1210‐bearing mice was evaluated. Leukaemia‐bearing mice have a markedly reduced L3T4+Lyt2+ thymocyte subpopulation. We observed ‘correction’ of relative values of thymocyte subpopulations induced by treatment of mice with a G‐CSF preparation. The suggestion is presented that this could result from liberation of lymphocyte T precursor cells from the bone marrow and increased homing of them into the thymus. The direct or indirect influence of G‐CSF on the L3T4+Lyt2+subpopulation of thymocytes is discussed.
Immunologic responses following respiratory sensitization to house dust mite allergens in mice
Authors: O'BRIEN RICHARD M , OOI MARCUS A Y , CLARKE AMANDA H , THOMAS WAYNE R
Published: June 2007

Allergens from the house dust mite Dermatophagoides pteronyssinus are a major cause of human respiratory diseases, including asthma. In order to help in understanding the early events in allergen sensitization, a murine model of allergic respiratory disease was developed. Mice were immunized by intranasal inoculation of Der p 1 or Der p 2 on days 0, 3, 7, 10, 14, 17, 21 and 29. T cell reactivity was determined using in vitro assays of allergen‐specific cytokine production by cells from the spleen, the draining superficial cervical lymph nodes (SCLN) and the non‐draining brachial and inguinal nodes. The cytokines assayed in supernatants were IL‐4. as a measure of Th2 activation, IL‐2 as a measure of Th1 activation, and IL‐3/GM‐CSF as an overall marker of T cell stimulation. There was evidence of local and systemic T cell activation by day 7, with the release of IL‐2 and of IL‐3/GM‐CSF by SCLN and spleen cells, respectively. IL‐4 production by SCLN and spleen cells was not evident until day 21. T cell sensitization in non‐draining node groups was not detected. Intradermal skin tests were performed at the above specified times and positive wheal responses indicated that specific IgE was present from day 3. The above data suggest that respiratory immunization to allergen is rapid and is associated with early systemic sensitization. In this model both Th1 and Th2 responses were evident, with the Th1 response occurring early and the Th2 following after repeated immunizations.
Anti‐IgE in allergic sensitization
Authors: STADLER BEDA M , RUDOLF MICHAEL P , ZÜRCHER ADRIAN W , MIESCHER SYLVIA , VOGEL MONIQUE
Published: June 2007

Anti‐IgE autoantibodies exist predominantly in the sera of patients with atopic disease. For some time such anti‐IgE autoantibodies have been considered a phenomenon that may not be of clinical importance. The cloning of such anti‐IgE autoantibodies has eliminated doubts of whether these antibodies exist, but it is still unclear whether such autoantibodies play a pathophysiological role. However, there are ongoing clinical trials that use humanized anti‐IgE antibodies for passive immunization of atopic individuals. While this approach may not definitely clarify the role of anti‐IgE autoantibodies, it will nevertheless clarify the role of IgE.
The role of Th2 type CD4+ T cells and Th2 type CD8+ T cells in asthma
Authors: ERB KLAUS J , GROS GRAHAM LE
Published: June 2007

The cellular inflammatory responses which are seen in allergic and asthmatic diseases are viewed as being quite strongly dependent on the activities of T cells and their products. The major T cell subset involved appears to be the so‐called CD4+ Th2 subset which produces interleukin‐4 (IL‐4) and interleukin‐5 (IL‐5). In vitro and in vivo experiments have indicated that IL‐4 is a key regulator in these kinds of immune responses, not only switching B cells to IgE production, but acting on CD4+ T cells to drive their development towards a Th2 phenotype. Recent results have shown that the functional phenotype of CD8+ T cells can be switched from interferon γ production to IL‐4 and IL‐5 production by the presence of IL‐4. This could prove an especially important phenomenon since it is the production of interferon γ by CD8 T cells which is seen as necessary for protection against virus infection. This short review updates our current knowledge of how IL‐4 can act on CD4+ and CD8+ T cell subsets in in vivo models of asthma and allergic disease.
Interactions of tunicate immunomodulatory proteins with mammalian cells
Authors: RAFTOS DAVID
Published: June 2007

The ability of a tunicate immunomodulatory protein, tunIL1, to interact with mammalian cells was investigated. Tun IL1 is known to regulate inflammatory defence reactions in tunicates and to stimulate the proliferation of mammalian thymocytes. In the current study, tunIL1 was shown to enchance L929 fibroblast proliferation, induce IL‐2 secretion from human mononuclear cells and enhance IL‐2 receptor expression by EL‐4 murine lymphoma cells. These biological activities are comparable with those of the mammalian inflammatory cytokine, IL‐1. However, tunIL1 does not appear to stimulate its effects on mammalian cells by interacting with cell surface receptors in a manner analogous to mammalian IL‐1. TunIL1 cannot block the binding of anti‐IL‐1 receptor antibodies to EL‐4 cells, nor can anti‐IL‐1 receptor antibodies inhibit the capacity of tunILl to stimulate thymocyte proliferation. This indicates that tunIL1 does not induce its IL‐1‐like activities via structural homology to mammalian IL‐1.
The relevance of murine animal models to study the development of allergic bronchial asthma
Authors: HERZ UDO , LUMPP USCHI , DA PALMA JOAO CASIMIRO , ENSSLE KARLHEINZ , TAKATSU KIYOSHI , SCHNOY NORBERT , DASER ANGELIKA , KÖTTGEN ECKART , WAHN ULRICH , RENZ HARALD
Published: June 2007

Bronchial asthma (BA) develops on the basis of a genetic predisposition and involves a characteristic sequence of changes in immune functions. In the immunopathogenesis, several phases can be distinguished: the initial stage is defined as the development of allergic sensitization. This step is dependent on: (i) T cell activation; (ii) IL‐4 production; (iii) IgE synthesis; and (iv) mediator release by effector cells. The second phase of allergic inflammation as a consequence of the T cell dependent sensitization is characterized by IL‐5 production and eosinophil activation and recruitment. Airway mucosa remodelling is the consequence of chronic inflammatory processes and represents the final stage of BA. In this article animal models will be discussed with regard to their relevance for these different phases in development of chronic allergic BA.
T cell responses toMycobacterium bovis, in red deer, a large animal model for tuberculosis
Authors: CROSS MARTIN L , CHINN NGAIRE D , GRIFFIN J FRANK T , BUCHAN GLENN S
Published: June 2007

Red deer (Cervus elaphus) represent an appropriate large animal model to study the immunology of tuberculosis, being naturally susceptible toMycobacterium bovisinfection. Cell‐mediated immune responses were investigated in deer displaying protective‐ or disease‐type reactions, following immunization withM. bovisbacille Calmette‐Guerin (BCG) or infection with virulentM. bovis, respectively. T cell responses were measured as antigen‐dependent cell proliferation and production of T cell growth factor (TCGF) followingin vitrostimulation withM. bovisantigens (live or heat‐killed BCG, or PPD). T cells from immunized deer proliferated less in response to soluble denatured culture antigen (purified protein derivative, PPD) than to particulate BCG, although there were no differences in the magnitude of these responses between the two groups of animals. Cells derived from immunized deer produced less TCGF than cells from infected deer when stimulated with PPDin vitro, although responses to BCG antigens were similar between the two groups. The majority of TCGF activity was neutralized by anti‐IL‐2 antibodies, regardless of the animal group or source of antigen used forin vitrostimulation. After 7 daysin vitroculture with antigen, blast cells staining positively for αβ (CD4. CD8) and γδ T cell receptors were recorded. The majority of blasts were CD4+, although in immunized deer fewer CD4+blasts were produced followingin vitrostimulation with PPD than with BCG antigens. These results, together with previous reports from our laboratory, represent the only detailed examinations of T cell responses toM. bovisin this naturally‐susceptible ruminant species.
Mast cell‐derived histamine and tumour necrosis factor: Differences between SJL/J and BALB/c inbred strains of mice
Authors: JR BRUCE F BEBO , LEE CAROL H , ORR EDWARD L , LINTHICUM D SCOTT
Published: June 2007

Mast cells cultured from bone marrow of BALB/c and SJL/J inbred strains of mice using IL‐3 showed distinct patterns of growth and marked differences in their content of TNF‐α and histamine. Mast cells derived from SJL/J mice grew and matured at a faster rate than those from BALB/c bone marrow. SJL/J mast cells were found to contain more than twice the amount of histamine and TNF‐α in their granules than BALB/c‐derived cells. In addition, when triggered by anti‐DNP IgE antibody and specific antigen (DNPalbumin), mast cells derived from SJL/J mice released more histamine and TNF‐α than mast cells derived from BALB/c mice. These results confirm previous observations regarding a genetic basis for mouse strain differences in mast cell growth rates, and extend previous observations to document differences in mast cell mediator contents. These results are consistent with the concept that genetically controlled differences in the numbers of central nervous system (CNS)‐associated mast cells and their vasogenic mediators may play an important role in modulating oedema and inflammation in CNS trauma and diseases in mice.
The phosphotyrosine phosphatase inhibitor‐phenylarsine oxide restores defective phosphoinositide hydrolysis response in anergic C3H‐gld/gld lymphocytes
Authors: LING WEIJUN , TIBBETTS DIANE J , CRAIN RICHARD C , LYNES MICHAEL A
Published: June 2007

Mice homozygous for the gld (generalized lymphoproliferative disease) mutation develop both lymphadenopathy and autoimmune disease. CD4− CD8− (double negative. DN) T cells comprise the major population of T cells in mature C3H‐gld/gld peripheral lymphoid tissues. These DN T cells are unresponsive to many forms of stimuli and have previously been shown to exhibit abnormally elevated levels of membrane phosphotyrosine phosphalase (PTPase) activity. In the present study, we demonstrate that IP, production in response to mitogenic stimulation with Con A or anti‐CD3 mAb (145‐2C11) is significantly diminished in C3H‐gld/gld lymphocytes when compared to that in congenic C3H‐ +/+ cells. The capacity to produce this second‐messenger can be restored by pretreating C3H‐gld/gld cells with the PTPase inhibitor, phenylarsine oxide (PAO). Although the inhibition of PTPase activity by treatment with PAO did restore C3H‐gld/gld cell ability to produce IP3 the signal did not lead to lymphocyte proliferation, but instead to cell death. Our results suggest that the altered phosphoinositide hydrolysis observed in the mutant cells is related to their elevated membrane PTPase activity and that the anergy in these cells is at least in part related to the abnormally high activity of endogenous PTPases.
The effects of four neuropeptides on the degranulation of mucosal mast cells from sheep
Authors: STEWART MELINDA J , EMERY DAVID L , McCLURE SUSAN J , BENDIXSEN TUAN
Published: June 2007

Isolated mucosal mast cells (MMC) were used to examine the ability of four neuropeptides. substance P, vasoactive intestinal peptide. β‐endorphin and somatostatin, to release mediators in the presence or absence of parasite antigen. None of the neuropeptides induced the release of sheep mast cell protease (SMCP) or histamine from MMC of helminth‐immune sheep in the absence of parasite antigen. Incubation of immune MMC with 100 and 1.0μg/mL parasite antigen induced 32.1 and 15.5% specific SMCP release, respectively. While the neuropeptides did not augment SMCP release at 100 μg/mL parasite antigen, significant enhancement (40‐98%) of SMCP release at 1 μg/mL antigen was obtained by each neuropeptide at concentrations from 10−8 to 10−12 mol/L. The results provide additional support for modulation of MMC degranulation by neural activity in sheep and, to our knowledge, this is the first demonstration that the threshold antigen concentration for allergic responses may also be lowered by neuropeptides to render the reaction more sensitive to antigen.
Anti‐receptor antibodies inhibit Pseudomonas aeruginosa binding to the cornea and prevent corneal perforation
Authors: HOBDEN JEFFERY A , GUPTA SURESH K , MASINICK SHARON A , WU XUAN , KERNACKI KAREN A , BERK RICHARD S , HAZLETT LINDA D
Published: June 2007

A polyclonal antibody (pAb) against gangliotetraosylceramide (asialo GM1), a glycolipid to which bacterial pili and LPS bind, and a mAb against a 66 kDa pilus‐binding protein purified from adult mouse corneal epithelium were used to determine if antibodies against host receptors for bacterial adhesins could inhibit bacterial binding to wounded corneal epithelium and protect ocularly challenged mice from corneal perforation when topically applied. Bacteria were mixed with anti‐66 kDa mAb, a mixture of anti asialo GM1 pAb and anti‐66 kDa mAb, an irrelevant control mAb (anti‐human histocompatibility Ag HLA‐DR5) or PBS prior to application to scarified corneas in organ culture. The combination of the two antibodies or the anti‐66 kDa mAb alone was effective in reducing bacterial adherence compared with either PBS or the antibody control. To determine if these antibodies were protective in vivo, corneas of C57BL/6J mice were scarified and inoculated with Pseudomonas aeruginosa. Eyes were treated topically with anti‐asialo GM1 pAb, anti‐66 kDa mAb, a mixture of the two or control mouse serum. More serum‐treated corneas perforated compared to corneas from any other group (p≤= 0.005) by 30 days post infection. Treatment with a combination of the two antibodies resulted in significantly less corneal pathology 30 days p.i. when compared to any other treatment (p≤= 0.005). These data provide evidence that antibodies against host corneal receptors significantly inhibit bacterial binding in vitro and when applied topically in vivo, lessen the severity of ocular disease characteristic of P. aeruginosa keratitis.
Distribution of antigen specific memory T cells in lymph nodes after immunization at peripheral or mucosal sites
Authors: PREMIER ROBERT R , JACOBS HELEN J , BRANDON MALCOLM R , MEEUSEN ELS NT
Published: June 2007

The distribution of antigen‐specific memory T cells in different lymph nodes of sheep was determined using an antigen‐specific in vitro proliferation assay. Lymph nodes were collected from sheep immunized simultaneously with avidin or ovalbumin in a peripheral tissue site (hind leg muscle) and keyhole limpet haemocyanin (KLH) in an intestinal tissue site (gut wall or colonic mucosa). The results showed a consistently high proliferative response in typical peripheral lymph nodes (popliteal and prescapular) and a low or negative response in gastrointestinal lymph nodes (abomasal and jejunal) while the response in other nodes was variable. The low proliferative response in the gastrointestinal lymph nodes was not due to the presence of supressor CD8+ lymphocytes and the proliferative response could not be raised to peripheral lymph nodes levels with the addition to cultures of IL‐2 or mitomycin‐C treated peripheral lymph node cells. The high proliferative response in the peripheral lymph nodes was not suppressed by the addition of mitomycin‐C‐treated gastric lymph node cells but was dramatically reduced by the addition of mAb against the IL‐2‐receptor or by depletion of CD4+ T cells. The results suggest that antigen‐specific proliferative memory T cells, which may be Th1‐like memory cells, preferentially migrate to peripheral lymph nodes independent of their site of induction.
Dissociation of production of interleukin‐4 and interleukin‐5
Authors: SEWELL WILLIAM A , MU HONG‐HUA
Published: June 2007

Interleukin‐4 (IL‐4) and IL‐5 are cytokines with important roles in IgE production and eosinophilia. Interleukin‐4 is essential for IgE production, and IL‐5 is the major factor involved in the production and activation of eosinophils. These two phenomena commonly occur together in parasitic infestation and allergic disease. Both cytokines are produced by T helper 2 (Th2) and Th0 cells but not by Th1 cells, and in a number of experimental systems IL‐4 is required for the production of IL‐5. This article presents evidence that IL‐4 and IL‐5 are not always co‐ordinately produced. There is evidence for selective production of either IL‐4 or IL‐5 in response to immune stimulation by different adjuvants. Dissociation of production of these two cytokines has also been reported in several pathological situations. An example is intrinsic or non‐atopic asthma, with eosinophilic bronchitis but without elevated IgE production, where there is evidence for excessive production of IL‐5 but not IL‐4. Different microenvironmental factors may favour production of either IL‐4 or IL‐5. For example, IL‐2 stimulates the production of IL‐5 but not IL‐4. Therefore the Th2 model does not account for all immune responses involving IL‐4 or IL‐5. Responses characterized by IL‐4 without IL‐5, and IL‐5 without IL‐4, can also occur.
Signal minus 1: A key factor in immunological tolerance to tissue‐specific self antigens?
Authors: PARISH CHRISTOPHER R
Published: June 2007

Recent data suggest that many autoreactive T cells, particularly to tissue‐specific self antigens, can escape thymic deletion. The current dogma is that these autoreactive T cells are silenced by the failure of most tissues to provide co‐stimulation (signal 2), antigen alone (signal 1) inducing T cell unresponsiveness. However, I propose that activation of autoreactive T cells frequently occurs but autodestruction by effector T cells is tightly regulated. This phenomenon is most evident with lymph node metastasizing tumour cells where the regional lymph node can mount a vigorous response to the invading tumour cells but tumour growth is unimpaired. I suggest that autodestruction is prevented by inhibitory receptors on T cells which recognize class I MHC structures on target cells. These receptors, which I propose deliver 'signal minus T to T cells, were recently described on NK cells and a subpopulation of peripheral T cells. They are also strikingly similar to a family of anti‐self receptors that my laboratory described on murine T and B cells 15 years ago. In the ‘signal minus 1’model, antigen‐activated T cells acquire the inhibitory receptors when they become co‐stimulation independent and gain the ability to exit lymphoid organs and enter non‐lymphoid tissues. Thus, if autoreactive effector T cells encounter autoantigen in tissues they are functionally silenced by inhibitory receptor engagement and signal minus 1 delivery. In contrast, I propose that in response to intracellular infections, cells down‐regulate expression of their ligands for inhibitory receptors. Such a model allows infected cells to be selectively eliminated by effector T cells. If correct, the model predicts that effector T cells, whether foreign antigen‐ or autoantigen‐specific, can selectively respond to infected cells. This apparent‘usefulness’of autoreactive T cells may explain their observed persistence even after an encounter with autoantigen. It is also suggested that signal minus 1 may silence autoreactive B cells specific for tissue‐specific cell surface antigens and lack of signal minus 1 may partially explain the vigorous T cell response to allogeneic MHC. Finally, it is hypothesized that, in evolutionary’terms, inhibition of autodestruction by the recognition of a 'self marker’and delivery of signal minus 1 is an ancient process which probably emerged in early metazoans.
Antigen‐specific IgA‐secreting cells induced by peripheral vaccination
Authors: ROTHEL JS , CORNER LA , SEOW H‐F , WOOD PR , LIGHTOWLERS MW
Published: June 2007

Antigen‐specific IgA‐secreting cells were shown to be generated in peripheral lymph nodes following s.c. vaccination. The efferent duct of prefemoral lymph nodes of sheep were cannulated prior to vaccination at a site draining to the cannulated node. Vaccines were contained a recombinant protein (Taenia ovis45W‐GST) and either incomplete Freund's, Quil A or A1(OH)3 as adjuvant. Lymph fluid was examined for the presence of 45W‐GST‐specific antibody by ELISA and antibody‐secreting cells by enzyme‐linked immuno‐spot (ELISPOT) assay. Large numbers of anti‐45W‐GST IgA‐secreting cells were detected at various times after vaccination as were IgM, IgG1, and IgG2‐secreting cells. For sheep vaccinated using incomplete Freund's adjuvant, up to 11% (> 1 million/day) of 45W‐GST‐specific ELISPOT were IgA‐secreting cells.
Malaria vaccine trials: The missing qualitative data
Authors: FERREIRA MU
Published: June 2007

Recent population‐based efficacy trials of the synthetic malaria vaccine SPf66 have shown restricted, if any, clinical protection against Plasmodium falciparum infection. Despite the well‐established role of antibodies in effector responses against asexual blood‐stage malaria parasites, the titres of anti‐SPf66 IgG antibodies do not correlate with the ability of sera from vaccine recipients to inhibit parasite growth in vitro nor with partial clinical protection which could be detected in some trials. Qualitative or functional parameters of SP66‐induced antibody responses, such as IgG subclass composition and affinity, may be more predictive of clinical protection against malaria than quantitative estimates of antibody concentration or titre. Since these parameters are readily estimated by laboratory techniques currently available, and may be modulated by changes in vaccination protocols and by the use of different adjuvants, a better understanding of qualitative antibody responses induced by SPf66 and other asexual blood‐stage malaria vaccine candidates, and of their relationship with clinical protection in vivo, is urgently needed for the improvement of currently used immunization schedules.
Quantifying phagocytosis of Mycobacterium avium complex by human monocytes in whole blood
Authors: HEWISH MJ , MEIKLE AM , HUNTER SD , CROWE SM
Published: June 2007

Studies of phagocytic efficiency in cells of the macrophage lineage have assumed additional importance since the discovery that HIV infection of these cells impairs their immune function. A rapid method has been developed for measuring phagocytosis of the opportunistic pathogen Mycobacterium avium complex by human monocytes. Fluoresceinated M. avium complex (F‐MAC) was incubated with whole blood at 37°C and the fluorescence of extra‐cellular F‐MAC was quenched using a vital blue stain. Monocytes were then stained with a monoclonal antibody (mAb) to human CD14 conjugated to phycoerythrin (PE) red cells were lysed, and the percentage of monocytes which had phagocytosed F‐MAC was measured by flow‐cytometry. The results were reproducible in samples of blood taken from individual donors over a period of 1or 2 weeks, and optimum F‐MAC concentrations and an optimum incubation time were determined by experiment. This method has the advantages of requiring only a small volume of blood, not necessitating manipulation of cells before testing, and using a phagocytic target relevant to the pathogenesis of HIV infection.
Exacerbation of invasive aspergillosis by the immunosuppressive fungal metabolite, gliotoxin
Authors: SUTTON PHILIP , WARING PAUL , MÜLLBACHER ARNO
Published: June 2007

Invasive aspergillosis is a significant cause of death in immunocompromised individuals. The majority of strains of the main causative agent, Aspergillus fumigatus, produce gliotoxin, a secondary metabolite with demonstrated in vitro immunosuppressive activity. Pretreatment of normally resistant mice with a single injection of a sublethal dose of gliotoxin was sufficient to make them susceptible to infection and subsequent death, after challenge with A. fumigatus spores. Animals infected with the non‐gliotoxin producing strain survived significantly longer than those infected with a gliotoxin producer. We propose that the release of gliotoxin by A. fumigatus hyphae during infection can exacerbate the pathogenesis of aspergillosis.
Modulation of immune responses by bovine β‐casein
Authors: WONG CHUN W , SEOW HENG F , LIU AI H , HUSBAND ALAN J , SMITHERS GEOFFREY W , WATSON DENNIS L
Published: June 2007

The present study reports the influence of bovine β‐casein on in vitro and in vivo immune responses. Bovine β‐casein showed an inhibitory effect on ovine neutrophil chemotaxis but had an enhancing effect on superoxide production by neutrophils. In response to mitogenic stimulation, the proliferative response of both T and B lymphocytes was significantly enhanced by β‐casein. While β‐casein had no significant effects on IFNγ production by ovine blood lymphocytes, and TNFα production and MHC Class II antigen expression by ovine bronchoalveolar macrophages, it enhanced IL‐1β production by the macrophages. β‐casein also had no influence on bovine NK cell activity against a virally‐infected cell line. Interestingly, β‐casein was found to reduce the adjuvant effect of matrix immune stimulating complexes (ISCOM) on anti‐ovalbumin antibody response in mice when given intramuscularly. Taken together, the results suggest that bovine β‐casein had selective modulating effects in vitro on both innate and adaptive immune responses in ruminants, whereas systemic administration of β‐casein, that might have a depressive effect on adjuvant activity, requires further study.
Endogenous presentation of a nascent antigenic epitope to CD8+ CTL is more efficient than exogenous presentation
Authors: HAHN YOUNG S , HAHN CHANG S , BRACIALE THOMAS J
Published: June 2007

Cytotoxic T lymphocytes (CTL) recognize short antigenic peptides in association with class I MHC molecules at the cell surface. Newly synthesized viral polypeptides are processed in the cytoplasm and the fragments of antigen are transported into the endoplasmic reticulum (ER) via a peptide transporter where they complex with nascent class I molecules. The peptide‐MHC complex is transported to the cell surface and presented to CTL. Sequence analysis of endogenously expressed, MHC‐associated self or viral antigens indicates that the naturally processed peptides bound to class I MHC molecules are in general 9 ± 1 residues long. Peptides bound to specific class I MHC molecules have in common allele‐specific motifs of conserved residues. The motif for the class I Kd molecules has been shown to be nine or 10 residues with the sequence X‐Tyr‐(X)6‐I/L or X‐Tyr‐(X)7‐I/L. The Tyr residue at the second position and the I/L residue at the ninth position are allele‐specific anchor residues which appear to be required for binding of the peptide to Kd. To examine the stringency of the requirement for Tyr at the second position, we have performed saturation mutagenesis of a minigene encoding the class I Kd‐restricted influenza HA210‐219 site at the Tyr residue 211. A series of 10 mutants was tested for effects on target‐cell sensitization. Most amino acid substitutions for the Tyr residue resulted in a loss of endogenous peptide recognition by HA210‐219 reactive CTL, consistent with the critical role of the Tyr at the second position for interaction with Kd molecules. One mutant gene‐product encoding a His substitution for the Tyr residue was recognized by CTL. However, the corresponding synthetic peptide containing a His substitution at the dominant anchor position bound only weakly to Kd, and target cells treated with the peptide were poorly recognized by CTL. The endogenous His‐containing peptide was also less stably associated with class I MHC Kd molecules at the cell surface than the wild‐type Tyr peptide. These data indicate that endogenous antigenic peptides may bind newly‐synthesized class I MHC molecules in the ER more efficiently than fully formed class I molecules at the cell surface and that endogenous peptides may dissociate from class I MHC molecules at different rates. The implications of these findings for CTL recognition and epitope mapping are discussed.
Regulation of autoimmune diabetes: Characteristics of non‐islet‐antigen specific therapies
Authors: GAZDA LAWRENCE S , BAXTER ALAN G , LAFFERTY KEVIN J
Published: June 2007

Non‐islet‐antigen specific treatments have been shown to alter the natural history of insulin dependent diabetes in both the non‐obese diabetic (NOD) mouse and in recently diagnosed patients. However, concerns have been raised regarding the possibility that non‐islet‐antigen specific therapy may trade cell mediated autoimmunity for antibody dependent autoimmunity. Female NOD mice at approximately 70 days of age were treated with the non‐islet‐antigen specific agents complete Freund's adjuvant (CFA) and Bacillus Calmette‐Guérin (BCG) and assayed for the development of antibody mediated autoimmunity at 300 days of age. Autoantibodies to red cells were not detected in any of the BCG (n= 19) or CFA (n= 15) treated animals, while 2 of 13 age‐matched NOD animals had autoantibodies to red cells, shown by a positive direct Coomb's test. Anti‐nuclear autoantibodies and complement deposition in the renal glomeruli were not significantly increased in the treated animals as compared to age‐matched non‐diabetic mice. The relative effectiveness of CFA and BCG treatment was examined in terms of the ability of these agents to preserve insulin containing islets. Complete Freund's adjuvant treatment was found to be more effective in preserving insulin containing islets when compared to BCG treatment. This study demonstrates that it is possible to inhibit the development of autoimmune diabetes without increasing the probability that treated animals will develop antibody dependent autoimmunity.
Expression of the recombinase‐activating gene (RAG‐1) in murine early embryogenesis
Authors: HAYAKAWA SATOSHI , TOCHIGI MEIJIN , CHISHIMA FUMIHISA , SHIRAISHI HISAMI , TAKAHASHI NORIKO , WATANABE KOH , FUJII KIYOSHI T , SATOH KAZUO
Published: June 2007

The recombinase activation genes, RAG‐1 and RAG‐2, are expressed together in immature T or B lymphocytes and possess activity to induce V(D)J rearrangement in T cell receptor (TCR) and Ig genes. In vertebrates, only Ig and TCR molecules are reported to have recombination in their development using multiple V, D, J component gene segments. Thus, expression of RAG genes are localized only in lymphoid organs and sites of extrathymic T cell differentiation. In this study, we have used RAG‐1 and RAG‐2 genes as markers of possible genetic recombination in developing murine preimplanation embryos, using the highly sensitive reverse transcriptase polymerase chain reaction (RT‐PCR) technique and in situ hybridization. From 40 preimplantation embryos of various developmental stages we extracted RNA. reverse‐transcribed it into cDNA and used it in RT‐PCR studies. A PCR of 35 cycles disclosed expression of RAG‐1 but not RAG‐2 in morulae and blastocysts. Southern blot hybridization using a specific synthetic oligonucleotide probe for RAG‐1 and RT‐PCR with another primer pair identified RAG‐1 expression in developing embryos. In situ hybridization using a cooled CCD camera also revealed localization of RAG‐1 mRNA in blastocysts. We propose possible genetic recombination during late preimplantation murine embryogenesis which may contribute to the loss of totipotency and differentiation of inner cell mass and trophoectoderm.
In vivo studies of macrophages and intercellular adhesion molecule‐1 following lipopolysaccharide treatment in tumourbearing rats
Authors: NILSSON TM , WOODS AE , ROFE AM
Published: June 2007

Histological, haematological and immunocytochemical analyses were performed on a mammary adenocarcinoma implanted in Dark Agouti rats treated with LPS to further elucidate the role of TNF‐α in tumour biology. Tumours were examined 4–12 days after implantation with parallel studies on rats treated with LPS (200μg/100g bodyweight [b.w.J) 24 h before removal. Tumour tissue from rats with 12‐day‐old lesions was also examined over the 24 h period after LPS administration. Haemorrhagic necrosis was evident 6 h after endotoxin treatment and became profound by 24 hours, at which stage only a thin band of viable cells was seen at the rim of the tumour. Using immunocytochemical techniques and a monoclonal anti‐macrophage antibody, macrophage numbers were seen to increase throughout the tumour following LPS administration at each stage of tumour growth. Immunocytochemical staining for intercellular adhesion molecule‐1 (ICAM‐I), an adhesion molecule normally involved in monocyte recruitment, could not be detected in vascular channels within the tumours investigated. However, subcutaneous vessels at the tumour periphery produced a strong linear reaction. The results show that macrophage numbers in tumours increase after LPS administration, concurrently with the development of haemorrhagic necrosis. This cellular infiltration was associated with peripheral monocytopenia. ICAM‐I expression on the vascular endothelium within tumours was deficient.
Cloning of novel kinectin splice variants with alternative C‐termini: Structure, distribution and evolution of mouse kinectin
Authors: LEUNG EUPHEMIA , PRINT CRISTIN G , PARRY DAVID AD , CLOSHY DAVID N , LOCKHART PETER J , SKINNER STEPHEN JM , BATCHELOR DAVID C , KRISSANSEN GEOFFREY W
Published: June 2007

The analysis of cDNA clones encoding novel variant forms of mouse kinectin, an endoplasmic reticulum (ER)‐bound receptor for the motor protein kinesin, is reported. Kinesin and cytoplasmic dynein are involved in mediating the anterograde and retrograde movements of intracellular vesicles along the microtubule network. The amino acid sequence deduced from kinectin cDNA isolated from mouse spleen cell and testis libraries revealed a long signal peptide or transmembrane sequence, and a 328 amino acid residue globular N‐terminal domain adjacent to a much larger 858–999‐residue C‐terminal coiled‐coil rod domain. The C‐terminal domain was composed of 18 coiled‐coil regions formed from multiple contiguous heptad repeats which undergo alternative splicing as evidenced by the presence of at least five small (23‐33 amino acid residue) insertion sequences scattered throughout. The inserts are present in any one of a number of combinations, generating an array of novel kinectin variants. Insert 5 contains a termination codon, producing a C‐terminus that is highly homologous to that of human kinectin. Three out of five mouse kinectin clones lack insert 5, generating a novel eleven amino acid C‐terminus encoded by sequence that extends past the insertion site. The existence of alternative C‐termini may have functional relevance given that the C‐termini are exposed for interaction with kinesin, whereas the globular N‐terminus is embedded in the ER membrane. Alternative C‐termini represent candidate modifications that could determine specificity of binding to kinesin or cytoplasmic dynein, and the switching of directionality of movement. The cDNA hybridized to 4.5 kb transcripts expressed in ail mouse cell lines and tissues examined, which provides the first indication that the kinectins are very widely distributed. Mouse kinectin is 42% similar over a 203 amino acid region to the chicken extracellular cardiac morphogen ES/130, whose canine homologue containing an inserted sequence of 10 amino acids repeated 54 times in tandem, is a ribosome receptor expressed on the ER. Mouse kinectin shares 64 and 83% identity, respectively, with its M, 160 000 chicken and human kinectin homologues. There is a two‐fold molar excess of kinectin over kinesin in unextracted vesicles, suggesting that kinectin might be a dimer. The electrostatic properties of the coited‐coil region of mouse kinectin. together with the relative frequencies of residues in particular positions within the heptad repeats support this notion.
Breast cancer immunotherapy: Current status and future prospects
Authors: APOSTOLOPOULOS VASSO , McKENZIE IAN FC , PIETERSZ GEOFFREY A
Published: June 2007

The development of an immunotherapeutic approach to cancer is the concern for many immunologists, but despite the impressive progress over the past decade, such as the identification of tumour antigens and antigenic peptides as potential targets, there are still many obstacles in eliciting an effective immune response to eradicate cancer. Mucins have attracted interest as potential targets for immunotherapy in the development of vaccines for cancers expressing Mucin 1 (MUC1; e.g. breast, pancreas, ovary etc.). All of the identified targets for cancer, including MUC1, are normal proteins; however MUC1 expressed on tumours can be considered as tumour specific due to their overexpression, altered glycosylation and its ubiquitous distribution on the cell surface rather than at the secretory pole in adenocarcinomas. These observations have led to the development of several different approaches to immunize against breast cancer using synthetic carbohydrates or peptides conjugated to carriers and given together with a variety of adjuvants to elicit the appropriate immune response. Mannan, a polymannose carbohydrate isolated from the cell wall of yeast, is an appropriate and effective protein carrier for eliciting a cellular (T1‐type) or humoral (T2‐type) immune response depending on the mode of conjugation (oxidized or reduced). In addition, mannan holds promise and opens many avenues as a carrier for vaccine development for other antigens. Several clinical trials are in progress to evaluate the immunogenicity of MUC1 and its suitability as to use for immunotherapy/vaccine for breast cancer.
Tumour necrosis factor and lymphotoxin: Molecular aspects and role in tissue‐specific autoimmunity
Authors: KORNER HEINRICH , SEDGWICK JONATHON D
Published: June 2007

Tumour necrosis factor (TNF) is a highly potent, proinflammatory cytokine with broad‐ranging functions from the regulation of endothelial cell adhesion molecules to facilitate entry of leucocytes into tissues, to direct induction of cellular cytotoxicity. This diversity of function potentially attributable to TNF in the genesis of inflammatory disorders place TNF as a primary candidate for clinical targeting and considerable success in this regard has been achieved, particularly in rheumatoid arthritis (RA). In this article we provide a short overview of TNF and its homologue lymphotoxin (LT) α and β. Particular emphasis is placed on recent discoveries regarding the cell surface expression of these cytokines and the role of TNF/LT in experimental autoimmune encephalomyelitis (EAE), an animal model of the human demyelinating disease, multiple sclerosis (MS).
NK cell proliferation and inflammation
Authors: WARREN HILARY S
Published: June 2007

Considerable progress has been made in recent years in understanding the biology of NK cells. NK cells are no longer ‘null cells’, but express an array of functionally important molecules with which they mediate and regulate their cytolytic activity and the cytokines they secrete. Activation and proliferation of NK cells is influenced by cytokines produced by activated monocytes (IL‐15, IL‐12, 1L‐10) and activated T cells (IL–2). This paper reviews the phenotype and effector functions of NK cells, their tissue distribution, and evidence that NK cells proliferate in vivo as part of productive or pathologic consequences of immunity.
Characterization of IgG Fc receptors on CD34 antigen‐expressing cell lines (KG‐1 and KG‐la)
Authors: WU ZHANHE , MARKOVIC BOBAN , CHESTERMAN COLIN N , CHONG BENG H
Published: June 2007

Although Fcγ receptors (FcγR) on mature blood cells have been extensively studied, there are only limited data on FcγR expression in the early haematopoietic progenitor cells. In this study, we used the stem cell antigen (CD34)‐expressing cell line (KG‐1) and its less differentiated subline (KG‐1a) as a model for the study of FcγR in the early haematopoietic progenitors. Flow cytometry and immunoprecipitation studies on KG‐1 and KG‐1 a cells with anti‐FcγR mAb showed that FcγRII is the only FcγR expressed on the cell surface. Analysis of the steady‐state levels of FcγR mRNA in KG‐1 and KG‐la cells using a quantitative in situ hybridization assay revealed the presence of only FcγRII mRNA. On further analysis FcγRIIA mRNA but no FcγRIIB or FcγRIIC transcripts were found in these cells: FcγRIIA transcripts with and without the transmem‐brane exon were present in approximately equal amounts. These findings are surprisingly similar to those observed previously with FcγR in platelets and megakaryocytic cells but different from those found with FcγR in cells of other lineages. These data suggest that the FcγR transcript distribution pattern observed in the early haematopoietic progenitors (KG‐l cells) is retained in later stages of haematopoietic differentiation only in cells of megakaryocytic lineage.
Primary cytotoxicity of SEB‐activated human PBMC and separated CD8+ and CD4+ T lymphocytes elicited by two different stimulation protocols
Authors: SEPRÉNYI GYÖRGY , ABE JUN , KOHSAKA TAKAO
Published: June 2007

The primary cytotoxicity of Staphylococcal enterotoxin B (SEB)‐stimulated PBMC and separated CD8+ and CD4+ T cells against Burkitt's lymphoma target cells has been characterized by applying two stimulation protocols. In the bulk protocol, the PBMC were stimulated with 100 ng/mL SEB for 3 days before separation to CD4+ and CD8+ T subsets. In the direct protocol, the separated CD4+ and CD8+ T cells were stimulated with 100 ng SEB preabsorbed to mitomycin C (MMC)‐treated APC. Comparison of the results of the two different protocols revealed the following differences: (i) PBMC in the direct protocol provided greater cytolytic activity than in the bulk protocol; and (ii) the CD4+ T cells acquired cytotoxicity only in the direct protocol. Unexpectedly, the superantigen‐dependent cellular cytotoxicity (SDCC) of SEB‐stimulated cells was not dominant compared with the basal cytotoxicity. The classical NK target, K‐562 was also sensitive to SEB‐augmented cytotoxicity. The parallel stimulation with IL‐2 and SEB caused a similar extent of cytotoxicity enhancement against both types of target cells. However, the cyclosporin A (CSA) inhibited only the SEB‐induced cytotoxicity. The results suggest that SEB‐induced PBMC acquire mainly a LAK‐like cytotoxicity, as a consequence of newly produced lymphokines. This observation might propose a different approach in pathological studies.
Cloning of the mucosal addressin MAdCAM‐1 from human brain: Identification of novel alternatively spliced transcripts
Authors: LEUNG E , GREENE J , NI J , RAYMOND LG , LEHNERT K , LANGLEY R , KRISSANSEN GW
Published: June 2007

The mucosal addressin cell adhesion molecule‐1 (MAdCAM‐1). expressed selectively on high endothelial venules (HEV) and lamina propria venules, directs lymphocyte traffic by binding the lymphocyte Peyer's patch adhesion molecule‐1 (LPAM‐1, α4β7). Full‐length DNA encoding human MAdCAM‐1 was obtained by combining sequences from an expressed sequence tag (EST) identified in an early stage human brain cDNA library, a polymerase chain reaction‐derived clone, and a MAdCAM‐1 genomic clone. The deduced amino acid sequence revealed an 18 amino acid signal peptide, two N‐terminal immunoglobulin (Ig)‐like domains conserved (59–65%) in sequence with those of the mouse homologue, an 86 amino acid mucin‐like region rich in serine‐threonine residues, a 20 amino acid transmembrane domain and a 43 amino acid charged cytoplasmic domain. No counterpart to the third IgA‐like domain of mouse MAdCAM‐1 was present; however, the serine‐threonine‐rich mucin domain was extended as two distinguishable major and minor mucin regions unrelated to the mouse mucin domain. The major domain is formed from six tandem repeats of an eight amino acid sequence having the MUC‐2‐related consensus DTTSPEP/SP. Human MAdCAM‐1 mRNA transcripts were restricted to small intestine, colon, spleen, pancreas and brain. Alternatively spliced MAdCAM‐1 variants were identified that lack parts of the second Ig domain and all or part of the major mucin domain, indicating that the function of this vascular addressin is regulated by extensive modifications to its multi‐domain structure.
Masters of deception: A review of herpesvirus immune evasion strategies
Authors: DAVIS‐POYNTER NICHOLAS J , FARRELL HELEN E
Published: June 2007

Herpesviruses have acquired a variety of different mechanisms to avoid the damaging effects of host immunity. Frequently, these viruses subvert normal immune regulatory functions utilized by the host. The focus of this review is upon herpesvirus genes encoding known or potential immunomodulatory proteins. Areas covered include inhibition of complement and antibody function, herpesvirus‐encoded homologues of cytokines and chemokine receptors, and potential disruption of cellular recognition of virally infected targets.