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PubMed2006

Development of hematopoietic stem cell gene therapy for Wiskott-Aldrich syndrome.

Wiskott-Aldrich syndrome (WAS) is a complex primary immunodeficiency disorder associated with microthrombocytopenia, autoinnmunity and susceptibility to malignant lymphoma. At the molecular level, this rare disorder is caused by mutations in the gene encoding the Wiskott-Aldrich syndrome protein (WASP). WASP is a cytosolic adaptor protein mediating the rearrangement of the actin cytoskeleton upon surface receptor signaling. Allogenic hematopoietic stem cell (HSC) transplantation represents a curative approach but remains problematic in light of severe risks and side effects. Recently, HSC gene therapy has emerged as an alternative treatment option. Cumulative preclinical data obtained from WASP-deficient murine models and human cells indicate a marked improvement of the impaired cellular and immunological phenotypes associated with WASP deficiency. The first clinical trial is currently being conducted to assess the feasibility, toxicity, and potential therapeutic benefit of transplanting autologous WASP-reconstituted hematopoietic stem cells.

Animals↗

PubMed1994

[Past and present of JSCP international exchange].

The Japan Society of Clinical Pathology was established in 1951 among laboratory physicians working in central clinical laboratories who discussed at the first several years on what clinical pathology and clinical pathologists mean. Many foreign guest lecturers were invited for its annual congresses. In 1962 returned from USA the three Japanese physicians who were certified in both anatomic and clinical pathology by the American Board of Pathology. Then, they also reported on various aspects in relation to clinical pathology, mainly in USA. The First International Symposium on Quality Control-Tokyo was held in 1974, inviting several laboratory scientists in Asia. In 1975, the Japanese-Korean Joint Conference in Clinical Pathology started, and in 1990 it was reorganized to the Asian Conference of Clinical Pathology, currently consisting of 4 national clinical pathology societies; Japan, Korea, China-Taipei and Indonesia. The official exchange program between the China-Beijing and JSCP started in 1992. JSCP joined to WASP (World Association of Societies of Pathology, Anatomic and Clinical) in 1957, and since then many members have been participating to various activities of the WASP. Dr. Tadashi Kawai, the Past President of JSCP, served the WASP President from 1991-1993. The permanent WASP Administrative Office was opened in Tokyo since 1992. The JSCP is expected to participate and contribute further for various WASP activities in future.

International Educational Exchange↗

PubMed1997

Tyrosine kinase signaling regulates Wiskott-Aldrich syndrome protein function, which is essential for megakaryocyte differentiation.

Platelets are produced from megakaryocytes differentiated from megakaryoblasts, but the differentiation mechanism still remains unknown. Here, we demonstrate that a tyrosine kinase signaling regulates Wiskott-Aldrich syndrome protein (WASP), which is essential for megakaryocyte differentiation. MEG-01 megakaryoblastic cells differentiate into large multinucleated megakaryocyte-like cells characterized by microvesicle formation with a protein kinase C (PKC) activator, 12-O-tetradecanoylphorbol 13-acetate. With parallel to the differentiation, WASP begins to accumulate at microvesicles with actin filaments and associates with tyrosine-phosphorylated Shc, the phosphorylation of which is induced. Moreover, a tyrosine kinase inhibitor, herbimycin A, blocked not only phosphorylation of Shc but also microvesicle formation without affecting cell enlargement and multinucleation, whereas a PKC inhibitor, H-7, completely inhibited all these processes. Because WASP also binds to Ash/Grb2 SH3 domains and the association of Ash/Grb2 and Shc is induced by 12-O-tetradecanoylphorbol 13-acetate treatment, a signaling pathway, PKC-tyrosine kinase-Shc-Ash/Grb2-WASP, is suggested for regulating megakaryocyte differentiation. In addition, WASP antisense oligonucleotide treatment abolishes both microvesicle formation and gathering of actin filaments. These data clearly show that WASP controls the assembly of actin filaments required for microvesicle formation downstream of PKC-tyrosine kinase.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1998

The Wiskott-Aldrich syndrome.

In 1937, Wiskott described three brothers with congenital thrombocytopenia, bloody diarrhea, eczema, and recurrent ear infections. Seventeen years later, Aldrich showed X-linked (a gene carried on the X chromosome) inheritance. Subsequently, the characteristic immune defects of Wiskott-Aldrich syndrome (WAS) were reported, including lymphopenia, lymphocyte depletion in the thymus, T-dependent pericortical areas of lymph nodes, defective delayed type hypersensitivity, and failure to produce antibodies to polysaccharides and to a variety of bacterial, protein, and viral antigens. The consistent platelet abnormalities were explained by ineffective thrombocytopoiesis. The increased risk of autoimmune diseases and malignancies was recognized. In addition to the classic WAS phenotype, a milder form designated as hereditary X-linked thrombocytopenia (XLT) has been described. The genes for both WAS and XLT have been mapped to Xp11.22 and sequence analysis has identified mutations of the same gene in both phenotypes. The gene coding for the WAS protein (WASP) is composed of 12 exons containing 1,823 base pairs and encodes a 502-amino acid protein. WASP is expressed in the cytoplasm of all hematopoietic stem cell-derived lineages. Although the precise function of WASP is unknown, several unique binding domains have been identified, and WASP appears to play a critical role in signal transduction by interacting with SH3-containing molecules and in the regulation of the cytoskeletal reorganization. The identification of the WASP gene allows the diagnosis of WAS on a molecular basis, carrier detection, and prenatal diagnosis. Treatment is largely symptomatic and includes antibiotics, prophylactic intravenous immunoglobulin (i.v.IG) and splenectomy in selected cases to reduce hemorrhages. Stem cell transplantation corrects the defect and should be considered in younger patients.

Animals↗

PubMed2000

Parasitism of Lacanobia oleracea (Lepidoptera) by the ectoparasitoid, Eulophus pennicornis, is associated with a reduction in host haemolymph phenoloxidase activity.

When haemolymph from fifth instar Lacanobia oleracea was incubated in vitro, rapid melanization occurred. Similar levels of melanization occurred in haemolymph from larvae that had been experimentally injected with venom from the ectoparasitic wasp, Eulophus pennicornis. In contrast, haemolymph from larvae parasitized by this wasp melanized more slowly and less extensively. Phenoloxidase assays indicated that enzyme activity was present in haemocyte lysate supernatants, serum and plasma from L. oleracea and that on day 5 post-parasitization, fractions prepared from parasitized larvae had significantly less phenoloxidase activity than similar fractions from untreated or experimentally envenomated larvae. In addition, no PO activity was detectable in wasp venom, and the venom had no effect on L. oleracea plasma phenoloxidase activity in vitro. These results indicate that parasitism of L. oleracea by E. pennicornis suppresses host haemolymph phenoloxidase activity and that this suppression is not induced by adult wasp venom. The results are discussed with reference to the survival advantages of suppressing the activity of this host enzyme, and to the possible source(s) of putative suppressive factors.

Animals↗

PubMedDate not supplied

Allergy to stinging and biting insects in Queensland.

Over an eight and a half year period 742 patients were assessed for allergy to stinging and biting insects in Queensland; 452 (61%) had allergic reactions to honey bees, 244 (33%) to wasps, 30 (4%) to various ants, 11 (1.5%) to march flies (Tabanus sp.) and five to tick infestation. One hundred and fifty one patients (20%) presented with large local swelling only (RXN1), 98 (13%) with urticaria and/or facial angioedema distant from the sting site (RXN2) and 492 (66%) with subjective or objective evidence of dyspnoea or hypotension (RXN3). Allergy testing was performed with honey bee and wasp venoms by skin testing or by Radioallergosorbent testing. Fifty nine patients (30%) with RXN3 responses to wasps failed to react to either test, while this applied to only 19 (6%) of the patients with RXN3 responses to bee stings. Thus, a large number of wasp-allergic patients with RXN3 responses could not be offered immunotherapy. A similar problem exists in the lack of availability of specific reagents for anti- and tick-induced dyspnoea or hypotension. A whole-body insect extract of march fly, however, appears useful.

Adolescent↗

PubMed1998

Prolonged oviposition decreases the ability of the parasitoid Leptopilina boulardi to suppress the cellular immune response of its host Drosophila melanogaster.

The cellular immune response of Drosophila against metazoan parasites is characterized by the production of melanotic capsules comprised mostly of host blood cells (hemocytes). During the latter part of the ovipositional period of the cynipid wasp parasitoid Leptopilina boulardi, eggs are deposited into host larvae of Drosophila melanogaster that are more susceptible to destruction by melanotic encapsulation than are eggs laid earlier. The increase in parasitoid mortality is attributed to a decline in the wasp's ability to suppress the host immune response. The decrease in active immune suppression is dependent on the reproductive physiology of the wasp, and this correlates with the extent of her prior ovipositional experience and not on her chronological age nor on the number of eggs remaining in the ovarioles. Such females with prior ovipositional experience which lack the ability to immune suppress infect far fewer hosts than females with no prior ovipositional experience. The reluctance of experienced wasps to infect hosts is not due to egg depletion, but instead is attributed to a depletion in immune suppressive substances. Perhaps by ovipositional restraints, retaining eggs that would otherwise become encapsulated reduces selection pressure in host populations for specific immune reactivity.

Analysis of Variance↗

PubMed1994

An abundantly expressed hemolymph glycoprotein isolated from newly parasitized Manduca sexta larvae is a polydnavirus gene product.

Cotesia congregata polydnavirus (CcPDV) is essential for successful parasitism of Manduca sexta larvae by the braconid wasp C. congregata. CcPDV virions are present in large numbers in the oviducts of C. congregata and injected with eggs into the hemocoel of M. sexta larvae during parasitization. Injection of sucrose density purified virions into nonparasitized larvae causes several of the parasitism-induced alterations in the physiology of host larvae that occur as a result of natural parasitism, including the synthesis of novel hemolymph proteins and abrogation of the host's immune response against the developing parasites. One of these proteins, early-expressed protein 1 (EP1), is a 190-kDa molecule which constitutes up to 5% of the total hemolymph protein by 24 hr following oviposition by the wasp. Using N-terminal sequence data for EP1 to construct primers for use in the polymerase chain reaction, we amplified and cloned a cDNA corresponding to the gene encoding EP1. This cDNA hybridized to DNA of the CcPDV genome, but not to DNA isolated from M. sexta larvae, suggesting that EP1 is a CcPDV gene product. A cDNA clone was isolated from an expression library generated from RNA extracted from newly parasitized M. sexta larvae. Sequence analysis of the cDNA clone revealed the presence of an open reading frame of 819 bp encoding a protein of 30.7 kDa. In vitro transcription/translation of the cDNA clone produced a protein of approximately 31 kDa, which was immunoprecipitated by EP1-specific polyclonal antiserum generated against purified deglycosylated EP1. EP1-like sequences also were amplified from male wasp genomic DNA, suggestive of integration of EP1-like sequences in the genome. This report constitutes the first evidence that a specific protein isolated from a parasitized host insect is a wasp polydnavirus gene product.

Amino Acid Sequence↗

PubMed2000

A gene encoding a polydnavirus structural polypeptide is not encapsidated.

Polydnaviruses are symbiotic viruses associated with some parasitic Hymenoptera that are vertically transmitted as proviruses within wasp genomes. To study this symbiotic association a gene encoding an abundant Campoletis sonorensis polydnavirus virion protein was characterized. This gene is not encapsidated but resides in the wasp genome where it is expressed only during virus replication. Immunolocalization studies detected the encoded 44-kDa protein only in oviduct tissue with ultrastructural studies detecting epitopes between or on virion envelopes. Expression and localization of the 44-kDa protein are consistent with its being a viral structural protein but localization of the gene only within the wasp genome is atypical, raising the possibility that this protein is adventitiously packaged during virion assembly. To address this possibility, quantitative dot blot and genomic Southern blot hybridizations were performed to determine whether the copy number of the p44 gene increased disproportionately during replication, as would be expected for a gene encoding a virion protein. The copy number of the p44 gene increases in tissues supporting virus replication but is unchanged in other tissues, suggesting that this gene is amplified in replicative cells. The data indicate that genes encoding polydnavirus virion proteins may be distributed between wasp and encapsidated viral genomes.

Amino Acid Sequence↗

PubMed1989

Role of virus-like particles in parasitoid-host interaction of insects.

Insect endoparasitoids are capable of suppressing the immune reaction of their habitual hosts in a specific way. Salt (1968) characterized some of the implications: This seeming contradiction--that defence reactions against all kinds of foreign bodies are available to insects and that endophagous parasitoids are nevertheless able to develop in insect hosts--is resolved by recourse to one of the principles of host specificity. Although insects as a group react to every foreign body in the sense that any organism or substance evokes a reaction in most insects, each species of insect fails to make a reaction (or makes an ineffective reaction) to a small group of organisms, its habitual parasites. It is the common paradox of parasitology that defence reactions are least effective against the most noxious parasites, involving the tautology that the most noxious parasites are those against which defence reactions are least effective. Recently, VLP of hymenopteran wasps have been shown to play a crucial part in suppressing the cellular encapsulation reaction (Stoltz and Vinson, 1979a). In some parasitoid wasps, polydnavirus particles are involved in the phenotypic transformation of hemocytes, reducing the capability of the host to mount an immune reaction towards the parasitoid egg (Stoltz and Guzo, 1986; Davies et al., 1987). However, at least in Venturia, the eggs are effectively protected by VLP that lack significant amounts of nucleic acids, precluding any virus expression in the host. The question was raised whether VLP could have acquired properties of the host immune system, which allows specific suppression of the immune response. The finding of structural similarities between VLP proteins and a host component indicated that a host function is expressed in VLP (Feddersen et al., 1986) and this observation has subsequently permitted the identification and characterization of a protein in caterpillars, which appears to inhibit cellular defense reactions (Berg et al., 1987). On the basis of these results we continue to approach this parasitoid-host interaction, assuming that VLP have evolved in the host organism and eventually acquired the coding sequences of a host protein with properties of an inhibitor of encapsulation. Although there are several ways to explain the emergence of VLP in endophagous parasitoid wasps, a simple proposal would be that such hypothetical viruses, which were able to suppress immune reaction in lepidopteran hosts, were incorporated into a parasitoid wasp to become part of the life cycle of the parasitoid.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1998

Development of polyembryonic insects: a major departure from typical insect embryogenesis.

The parasitic wasp Copidosoma floridanum represents the most extreme form of polyembryonic development known, forming up to 2000 embryos from a single egg. To understand the mechanisms of embryonic patterning in polyembryonic wasps and the evolutionary changes that led to this form of development we have analyzed embryonic development at the cellular level using confocal and scanning electron microscopy. C. floridanum embryogenesis can be divided into three phases: (1) early cleavage that leads to formation of a primary morula, (2) a proliferative phase that involves partitioning of embryonic cells into thousands of morulae, and (3) morphogenesis whereby individual embryos develop into larvae. This developmental program represents a major departure from typical insect embryogenesis, and we describe several features of morphogenesis unusual for insects. The early development of polyembryonic wasps, which likely evolved in association with a shift in life history to endoparasitism, shows several analogies with mammalian embryogenesis, including early separation of extraembryonic and embryonic cell lineages, formation of a morula and embryonic compaction. However, the late morphogenesis of polyembryonic wasps proceeds in a fashion conserved in all insects. Collectively, this suggests a lack of developmental constraints in early development, but a strong conservation of the phylotypic stage.

Animals↗

PubMed1980

The influence of X-rays on reproduction in Pseudeucoila bochei (Hymenoptera Cynipidae), a parasite of Drosophila melanogaster.

The parasitic wasp Pseudeucoila bochei was the subject in experiments designed to detect a reliable method of mutation induction. Radiation doses of 360, 720, 1080, 1440, 1800 and 2160 R were given to pupae 15 or 18 days old or to freshly eclosed male and female wasps. The irradiation did not influence the survival of pupae. Dose-action responses were established and appeared to be dependent on the stage of development of the wasps at the moment of irradiation. The production of sons by virgin females showed that the germ cells of the adults were the most sensitive to irradiation. In contrast, the sperms of the male wasps irradiated as adults were the most resistant. Irradiation of males as pupae with 1880 or 2160 R on the 15th day or as adults led to a high frequency of amle progeny. The elimination of sperm cells from fertilization is discussed. Comparisons with Habrobracon, Nasonia and Drosophila showed that the germ cells of Pseudeucoila are more sensitive to irradiation. In the progeny from irradiated females and males 2 types of mutant were found, with yellow eye colour and with aberrations in the antennae.

Animals↗

PubMed1989

Comparative studies on the protein composition of hymenopteran venom reservoirs.

The proteins of venom reservoirs from 25 hymenopteran species from 21 genera were investigated with regard to their protein composition and immunological similarities. It was found that low mol. wt proteins and polypeptides are typical for the venoms of ants, social wasps and bees. Six species of ichneumonoid parasitic wasps lack these low mol. wt proteins. Except for bee venoms none of the low mol. wt proteins contain mannose. The venoms of ichneumonoid parasites and ants contained primarily acidic proteins, whereas those of social wasps and bees contained abundant neutral and basic proteins. Immunological analysis showed that a number of venom proteins from parasitic braconid (Chelonus) wasps shared conserved immunoreactive determinants with the venom proteins of higher Hymenoptera, including some not of parasitic habit. Implications of phylogenetic relationships among hymenopteran species are discussed.

Animals↗

PubMed2003

Characterization of Campoletis sonorensis ichnovirus unique segment B and excision locus structure.

Polydnaviruses (PDVs) are segmented, symbiotic, double-stranded DNA viruses that are vertically transmitted as proviruses within the genomes of some parasitoid Hymenoptera. The PDV associated with the ichneumonid wasp Campoletis sonorensis (CsIV) consists of 24 non-redundant DNA segments varying in size from approximately 6 to 20 kbp. CsIV segment B, one of the smallest genome segments, was sequenced and the excision sites of the proviral segment were characterized. The segment B sequence was 83.2% non-coding with only two open reading frames (ORFs). Some non-coding sequences have similarities to database sequences and were likely pseudogenic, but most were unrelated to known nucleic acid or predicted protein sequences. One ORF, BHv0.9, encodes a member of the rep gene family and was expressed only in parasitized insects while transcription of the other ORF could not be detected. Previously, a third region of the segment was shown to hybridize to 0.6 and 1.2 kb poly A+ RNAs from female wasps during virus replication (Theilmann and Summers, 1988) but this region did not have an identifiable ORF in the determined sequence. In contrast to CsIV segment W, segment B had little repetitive sequence. The segment B proviral integration locus contains a 59 bp direct imperfect repeat. Further analyses of this integration locus demonstrated that segment B was excised from wasp genomic DNA with flanking sequences at the integration site rejoined after segment excision. The segment B "excision locus" retained one of the two copies of the 59 bp repeat sequence with the other repeat present in the excised segment. The data indicate that Ichnovirus segments have distinctive characteristics possibly reflecting functional co-evolution between the wasp and individual types of polydnavirus segments.

Animals↗

PubMed1999

A 24 kDa parasitism-specific protein from the Caribbean fruit fly, Anastrepha suspensa: cDNA and deduced amino acid sequence.

A 24 kDa parasitism-specific protein (PSP24) was previously reported from the hemolymph of the Caribbean fruit fly, Anastrepha suspensa (Diptera: Tephritidae) after parasitization by the wasp Diachasmimorpha longicaudata (Hymenoptera: Braconidae). This study was designed to sequence the open reading frame of PSP24 and to determine whether it is encoded by the wasp, fruit fly host or by the entomopoxvirus D1EPV which is normally injected into the host with the wasp's egg. Utilizing an existing partial amino acid sequence of PSP24, we obtained two cDNAs by reverse transcription-polymerase chain reaction, from the host hemolymph 48 h post parasitization. The smaller cDNA has an open reading frame (ORF) that encodes 85 amino acids (aa) with a molecular mass of 9711.33 Da and the larger encodes 203 aa with a molecular mass of 23,076 Da. Both cDNAs share a common N-terminus with a signal peptide predictive of secreted proteins, a characteristic that agrees with the observed nature of PSP24. The mature proteins have 39 and 157 aa with deduced molecular masses of 4286.86 Da and 17,651 Da, respectively. Western blots of host hemolymph probed with the anti-PSP24 serum reveal proteins of 0.10 and 0.24 kD, respectively. The discrepancy between the deduced and the observed molecular masses may be explained by their predicted O-linked glycosylation. The amino acid sequences are not homologous with any protein in the available databases. Southern blot hybridization experiments revealed that the proteins are encoded by both the host and the parasite. Furthermore, injection of D1EPV into healthy fruit fly puparia induces the two proteins. Thus, in surprising contrast to an earlier hypothesis that D1EPV encodes PSP24, these results clearly demonstrate that the PSP24 proteins are encoded by wasp and fruit fly but not D1EPV genes. However, their expression is D1EPV induced.

Amino Acid Sequence↗

PubMed2006

Ultrarush venom desensitization after systemic reactions during conventional venom immunotherapy.

BACKGROUND: Rush and ultrarush venom immunotherapy (VIT) protocols are safe and effective in patients with Hymenoptera hypersensitivity. However, these protocols have typically been used instead of conventional VIT and not in patients who have experienced adverse reactions during conventional VIT. To date, there are no reports of using an ultrarush VIT protocol to desensitize patients with a history of severe systemic reactions during conventional VIT. OBJECTIVE: To determine whether ultrarush VIT can be safely administered to a high-risk patient with a history of severe systemic reactions to conventional VIT. METHODS: Premedication with 40 mg of prednisone, 180 mg of fexofenadine, and 150 mg of ranitidine orally twice daily was initiated. The patient received VIT to mixed vespid and wasp in a medical intensive care unit via a 13-step buildup on day 1 followed by a 2-step buildup on day 2. Immunotherapy was begun with a dose of 0.005 microg of mixed vespid and 0.002 microg of wasp venom and achieved a total dose of 300 microg of mixed vespid and 100 microg of wasp venom. RESULTS: The patient tolerated the procedure with minimal adverse effects. She subsequently received maintenance dosing in the outpatient clinic weekly for 4 weeks and bimonthly for 8 weeks, and she continues monthly maintenance VIT. CONCLUSIONS: We report the first successful use of ultrarush VIT in a high-risk patient with a history of severe systemic reactions during conventional VIT. This protocol should be considered in patients with a history of allergy to vespids or wasps who require VIT but cannot reach a maintenance dose with conventional VIT owing to systemic reactions.

Adult↗

PubMed2004

Behaviour and development of Elasmosoma sp. (Neoneurinae: Braconidae: Hymenoptera), an endoparasite of Formica ants (Formicidae: Hymenoptera).

The behaviour and development of Elasmosoma sp., near pergandei Ashm. (Neoneurinae: Braconidae: Hymenoptera), a parasite of the ant, Formica obscuriventris clivia Creighton, was studied in a coastal sand dune habitat in central Oregon over a 2-year period. Female wasps oviposited into the abdomen of major worker ants after alighting briefly on the host. Embryonic development of the wasp occurred inside the serosal membrane or trophamnion of the egg in the abdomen of the ant host. First instar wasp larvae possessed a large head capsule with falcate mandibles, 11 ventral body lobes, an elongate tail and a dorsally located anus. Second instar larvae were unspecialized and lacked the large head capsule, elongate tail and paired ventral body lobes. Third instar larvae were hymenopteriform with a distinct head capsule. Cocoon formation and adult emergence is described. Observations on the percentage parasitism and altered behaviour of parasitized ants are presented. The association between neoneurine wasps and ants is a well-balanced system that has been in existence for at least 40 million years.

Abdomen↗

PubMed2002

Learning of herbivore-induced and nonspecific plant volatiles by a parasitoid, Cotesia kariyai.

Learning of host-induced plant volatiles by Cotesia kariyai females was examined with synthetic chemicals in a wind tunnel. Wasps were preconditioned by exposure to volatiles and feces simultaneously. A blend of four chemicals, geranyl acetate, beta-caryophyllene, (E)-beta-farnesene, and indole, which are known to be specifically released from plants infested by host larvae Mythimna separata (host-induced blend), elicited a response in naive C. kariyai, but did not enhance the response after conditioning. A blend of five chemicals, (E)-2-hexenal, (Z)-3-hexen-1-ol, (Z)-3-hexen-1-yl acetate, beta-myrcene, and linalool, which are known to be released not only from plants infested by the host larvae, but also from artificially damaged plants or undamaged ones (unspecific blend), elicited little response in naive wasps, but significantly enhanced the wasps' response after conditioning. With a blend of the above nine chemicals, wasps could learn the blend at lower concentrations than they did in the nonspecific blend. Hence, both the host-induced and nonspecific volatile compounds appear to be important for C. kariyai females to learn the chemical cues in host location.

Animals↗