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N E Beckage

Publications and source records attributed to N E Beckage.

17 recordsLinked to original sources
PubMed2003

Persistence and expression of Cotesia congregata polydnavirus in host larvae of the tobacco hornworm, Manduca sexta.

The gregarious braconid wasp Cotesia congregata parasitizes host larvae of Manduca sexta, and several other sphingid species. Parasitism induces host immunosuppression due to the disruptive action of the wasp's polydnavirus (PDV) on host blood cells. During the initial stages of parasitism, these cells undergo apoptosis followed by cell clumping, which clears the hemolymph of a large number of cells. In this study, the persistence and expression of Cotesia congregata PDV (CcPDV) were examined using Southern and Northern blots, respectively. Digoxygenin-labelled total polydnaviral DNA was used to probe genomic DNA isolated from fat body and brains of hosts with emerged wasps taken 6 days following egress of the parasitoids, and significant cross-hybridization between the host fat body genomic DNA with viral DNA was seen. Thus, the virus persists in the host for the duration of parasitism, even during the post-emergence period, and may even be integrated in the host caterpillar DNA. Viral gene expression was examined using Northern blots and probes to the Cotesia rubecula CrV1 homolog, and the CrV1-like mRNAs were expressed as early as 4 h post-parasitization for at least 72 h and faint hybrization is even seen at the time the wasps eclose. In contrast, in Pieris rapae larvae the CrV1 transcript is expressed only for a brief time, during which time hemocyte function is disrupted. The effect is transitory, and hemocytes regain their normal functions after the parasites emerge as first instars.The genome of CcPDV contains one copy of the CrV1-like homolog as shown on Southern blots of viral genomic DNA. In conjunction with our earlier studies of the PDV-encoded early protein 1, the current work suggests multiple viral transcripts are produced following parasitization of the host, and likely target host hemocytes to induce their apoptosis, thereby preventing encapsulation of the parasitoid's eggs. Whether viral DNAs are integrated in the host's genomic DNA remains to be proven, but our results provide preliminary evidence that viral DNAs are detected in the host's fat body cells examined at the time of wasp emergence and several days later.

Animals↗
PubMed2002

Parasitoid-host endocrine relations: self-reliance or co-optation?

High titers of juvenile hormone (JH) maintain developmental arrest in Manduca sexta larvae parasitized by Cotesia congregata. Parasitized hosts exhibit up to 9.5 times greater amounts of total hemolymph JH (from 0.6+/-0.09 to 2.51+/-0.43ng/ml) compared to non-parasitized controls. Elevated titers are observed throughout the fifth instar, even beyond egression of the parasitoids on day 5. GC-MS analysis revealed that in hemolymph of unparasitized control larvae, JH I is the major homolog and levels of JH III are negligible; in parasitized individuals the amounts of JH I, II, and III rise, and JH III predominates. Neck ligation ensured separation of M. sexta's corpora allata from the posterior section, which contained most of the parasitoids in the infected insects. When the posterior region was sampled, JHs were not detected in the non-parasitzed larvae, but in those parasitized, JH III was found (1.98+/-0.29ng/ml, 24 h post-ligation). JH III was the only homolog produced and secreted by the parasitoid in in vitro culture. This is the first report stating that a parasitoid secretes JH III and may contribute, at least in part, to the circulating titer in the host hemocoel, concurrently promoting host production of JH I and II.

Animals↗
PubMed1999

The excision of polydnavirus sequences from the genome of the wasp Cotesia congregata (Braconidae, microgastrinae) is developmentally regulated but not strictly restricted to the ovaries in the adult.

Cotesia congregata polydnavirus (CcPDV) is essential for the successful parasitism of Manduca sexta larvae by the braconid wasp Cotesia congregata. In the absence of PDV, parasitoid eggs are encapsulated. Molecular analysis has demonstrated that polydnavirus sequences are integrated in the wasp chromosomes, and an ultrastructural analysis has shown that PDV replication occurs in the calyx region in the ovaries of the wasp. The bracovirus sequences appear to be excised from the wasp genome in the calyx cells where the virus replicates. Following excision of the virus sequences, the flanking sequences are rejoined. We analysed the production of two polydnavirus circles during wasp development and in different body parts of the adults of both sexes. Our study indicates that the excision of viral sequences is developmentally regulated, beginning in the pupal stage. In the adult wasp, excision occurs ubiquitously. However, regulation in the adult seems to occur only in diploid individuals, as no excision is detected in haploid males produced from virgin females.

Animals↗
PubMed1998

Production of early expressed parasitism-specific proteins in alternate sphingid hosts of the braconid wasp Cotesia congregata.

Parasitism of Manduca sexta larvae by the braconid wasp Cotesia congregata or injection of C. congregata polydnavirus (CcPDV) causes numerous alterations in host physiology, including developmental arrest, abrogation of host immunity, and the production of three abundant early expressed proteins (EP1, EP2, and EP3) that are secreted in large amounts into the host's hemolymph. Here we compare the levels of these proteins present in the hemolymph of three other sphingid species that vary in their compatibility for C. congregata. Hyles lineata was found to be permissive for C. congregata and EP1, EP2, and EP3 were present in larval hemolymph at levels comparable to those found in hemolymph from parasitized M. sexta larvae. By contrast, the lowest levels of EP proteins were found in hemolymph from parasitized Pachysphinx occidentalis larvae and this species was found to be completely refractory, since C. congregata eggs were invariably encapsulated. Parasitism of Sphinx vashti by C. congregata resulted in moderate levels of EP production. While the observed immune response was incomplete and some encapsulation of C. congregata eggs and/or larvae was observed, low numbers of S. vashti nevertheless were able to complete their development and emerge as adults. Thus, a correlation was established between host compatibility and induction of synthesis of the three parasitism-specific proteins, although the linkage between quantitative levels of EP production and the extent of encapsulation was variable.

Animals↗
PubMed1998

Modulation of immune responses to parasitoids by polydnaviruses.

Parasitoids are parasites that invariably kill their host. Polydnaviruses are injected by parasitoid wasps into the body cavity of their insect host and cause immunosuppression, allowing the parasitoid to develop in the absence of encapsulation. One of the targets of the polydnaviruses are the haemocytes of the host, which undergo significant changes in response to entry of the virus. In some systems, haemocyte apoptosis is induced, or haemocyte clumping may be seen; in others, the cells round up and fail to adhere to a substrate. Effects on haemocytes may be transitory or permanent (cell death). Various polydnavirus gene products have been identified that interfere with normal haemocyte function. Phenoloxidase activity also is inhibited during parasitism, and the effect is inducible by polydnavirus. In some systems, venom components may act synergistically with polydnavirus in mediating the virally-induced effects on the host immune system. Polydnaviruses are powerful influences on the host immune system, which serve to permit successful development of the parasitoid without triggering the host immune response.

Animals↗
PubMed1997

Identification of neuropeptides in the midgut of parasitized insects: FLRFamides as candidate paracrines.

Parasitism of Manduca sexta (Lepidoptera: Sphingidae) larvae by the braconid wasp Cotesia congregata (Hymenoptera: Braconidae) leads to accumulation of peptides in host neurons and neurosecretory cells of the central nervous system (CNS) and neurons and endocrine/paracrine cells of the midgut. This accumulation has now facilitated the characterization of two new members of the FLRFamide family from midguts of parasitized larvae. The peptides, given the names F24 and F39, are 24 and 39 amino acids in length with the sequences VRDYPQLLDSGMKRQDVVHSFLRFamide and YAEAAGEQVPEYQALVRDYPQLLDSGMKRQDVVHSFLRFamide. The sequence of F24 is identical to the C-terminal 24 amino acids of F39. The C-terminal 10-mer of each is identical to a previously characterized decapeptide neurohormone (F10). This sequence is preceded by a potential processing site. In nonparasitized insects F39 was present at several-fold the amount of F24. In parasitized insects F24 and F39 accumulate in the middle and posterior regions of the midgut, which are enriched in endocrine/paracrine cells reacting with FLRFamide antisera. In the combined brain and subesophageal ganglion F39 was not detected and the amount of F24 never exceeded 2 fmol per Br/SEG. Of the three peptides, only F10 was found in the hemolymph. Thus, F24 and F39 may be intermediates in the biosynthesis of F10 and may themselves be released locally from endocrine/paracrine cells in the midgut epithelium.

Amino Acid Sequence↗
PubMed1995

Accumulation of neuropeptides in the cerebral neurosecretory system of Manduca sexta larvae parasitized by the braconid wasp Cotesia congregata.

Fifth instar larvae of Manduca sexta that were parasitized by the braconid wasp Cotesia congregata failed to develop after the parasitoid larvae emerged, and these host larvae lingered for 2-3 weeks in a quiescent, nonfeeding state without initiating a larval molt or metamorphosis. This study was focused on the neuroendocrine changes associated with the host's developmental arrest. Immunohistochemical studies suggested that the host brain neurosecretory cells as well as their axon terminals in the corpora cardiaca-corpora allata complex accumulated multiple neuropeptides. The extent of accumulation in cells and axons increased with time, so that hosts examined 7-14 days after the wasps emerged showed the most intense staining with antibodies against prothoracicotropic hormone, bombyxin, allatotropin, allatostatin, diuretic hormone, eclosion hormone, proctolin, and FMRFamide. Increased levels of prothoracicotropic hormone and FMRFamide-like peptides in the brains of parasitized larvae were confirmed using Western blots and enzyme-linked immunosorbent assay (ELISA), respectively. Starvation of the unparasitized larvae induced some accumulation of the neuropeptides; however, the intensity of staining and number of immunopositive cells and axons were in most cases clearly higher in the parasitized larvae. Our results suggest that accumulation of the neuropeptides is associated with developmental arrest of parasitized larvae. Because a similar developmental arrest occurs in a wide range of parasitized insects, our findings may have relevance for many other species. Moreover, these data illustrate the potential value of using parasitized M. sexta larvae as a model for studying the mechanisms governing the rates of neuropeptide expression, processing, packaging, and release, as well as providing a rich source of neuropeptides, thus facilitating their isolation and characterization.

Animals↗
PubMed1995

Polydnaviruses: potent mediators of host insect immune dysfunction.

Endoparasitic insects are used as biological control agents to kill many species of insect pest. One key to the success of parasitoids that develop in the hemocoel of their host is their ability to knock out the host's immune system, inducing a decline in the responsiveness of a variety of cellular and humoral components so that parasitoid eggs are not encapsulated. In many species parasitized by braconid and ichneumonid wasps, host immunosuppression appears to be mediated by polydnaviruses (PDVs) injected by the female parasitoid into the host hemocoel. The viruses exhibit a complex and intimate genetic relationship with the wasp, since viral sequences are integrated within the wasp's chromosomal DNA. Here Mark Lavine and Nancy Beckage summarize the current evidence for mechanisms of virally induced host immunosuppression in parasitized insects, as well as the roles of other factors including wasp ovarian proteins and venom components, in suppressing hemocyte-mediated and humoral immune responses. Interestingly, in some species, the PDV-induced host immunosuppression appears transitory, with older parasitoid larvae probably exploiting other mechanisms to protect themselves from the host's immune system during the final stages of parasitism. During the final stages of parasitism, the parasitoids likely exploit other mechanisms of immunoevasion via antigen masking, antigen mimicry, or production of active inhibitors of the hemocyte-mediated encapsulation response as well as inhibiting melanization.

Journal Article↗
PubMed1995

Parasitism-induced accumulation of FMRFamide-like peptides in the gut innervation and endocrine cells of Manduca sexta.

Manduca sexta larvae that are parasitized by the braconid wasp Cotesia congregata enter a state of developmental arrest following emergence of the wasp larvae from the host. These fifth instar hosts linger for 2 to 3 weeks without resuming feeding, molting, or metamorphosis once the wasps emerge. Immunohistochemical staining with antiserum against FMRFamide revealed dramatic accumulation of FMRFamide-like peptide(s) in the gut nervous and endocrine systems of the developmentally arrested larvae when compared to that observed in unparasitized feeding or starved larvae. Specifically, the number of immunopositive cells and the intensity of staining was enhanced in the neurons of the frontal ganglion, the axons and axon terminals on the midgut surface, and in the gastric endocrine cells. These results were confirmed using ELISA to show that the relative amounts of FMRFamide-like peptides in midgut extracts were highly elevated in the parasitized larvae relative to the fed or starved unparasitized larvae. These data suggest that FMRFamide-like peptides in developmentally arrested larvae are produced in a significantly larger number of gastric endocrine cells, and that the rate of release of the peptides may be suppressed, or the rate of their synthesis may be elevated. Localization of FMRFamide-like peptides in the gastric endocrine cells of C. congregata is also described.

Animals↗
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↗
PubMed1994

Extraembryonic membranes of the endoparasitic wasp Cotesia congregata: presence of a separate amnion and serosa.

Little information is available describing the development of endoparasitic Hymenoptera prior to hatching of the first instar larva. The present study reveals that in Cotesia congregata, and likely other braconid wasp parasites, the serosa and amnion comprise separate layers encasing the developing first instar larva. Examination of newly hatched first instar parasites in the hemocoel of Manduca sexta confirmed that the larvae were ensheathed in the remnants of an "amnion" at hatching, with patches of nucleated cells seen adhering in thin sheets to the larva. The amnion is delaminated from a cellular serosal membrane that lies directly beneath the chorion, which subsequently gives rise to teratocytes. Scanning electron micrographs indicate that remnants of the amnion remained associated with the larvae for a longer period posthatch than did the teratocytes, which rapidly dispersed into the host hemolymph. The presence of an amnion-like layer in braconid endoparasitic wasps may prove to be significant with respect to our understanding of host-parasite immunological interactions occurring during the initiation of the relationship of the parasites with their host.

Amnion↗
PubMed1993

Effects of parasitism by the braconid wasp Cotesia congregata on host hemolymph proteins of the tobacco hornworm, Manduca sexta.

Parasitism by the braconid wasp Cotesia congregata causes major alterations in the hemolymph proteins of host tobacco hornworm larvae. Earlier studies showed that the total amount of hemolymph protein is reduced during parasitism, beginning almost immediately after the host is parasitized. Simultaneously, parasitism induces synthesis of large amounts of novel proteins that appear in the blood as early as 1-2 h post-parasitization. The present report confirms earlier studies describing the presence of novel proteins in last instar hosts, and also characterizes the effects of parasitism in altering the titers of several endogenous host hemolymph proteins normally produced by the fat body and other tissues. Analysis of hemolymph plasma using SDS-PAGE and densitometry, as well as immunodiffusion assays, showed that in terminal stage fifth instar host larvae, the titers of serpins and arylphorin were dramatically reduced relative to the levels of these proteins detected in nonparasitized gate II fifth instar larvae of the same age. The relative differences between parasitized and nonparasitized larvae increased with time following ecdysis to the fifth instar, so that the day 4 nonparasitized larvae had arylphorin titers of c. 30 mg/ml, whereas parasitized day 4 larvae with newly emerged wasps had only one sixth that amount of storage protein circulating in the hemolymph. Similarly, in nonparasitized larvae the hemolymph serpin concentration increased from c. 200 micrograms/ml (on day 0) to > 600 micrograms/ml (on day 4) in prewandering gate II larvae, but in parasitized larvae the hemolymph serpin concentration was maintained in the range of 100-200 micrograms per ml hemolymph until the pharate third instar parasites emerged from the host larva on day 4. In contrast, the level of hemolymph lipophorin was unaffected by parasitism, and lipophorin increased from c. 1.3 to > 3 mg/ml during the time interval between days 0 and 4 in both nonparasitized and parasitized larvae. Hemolymph titers of insecticyanin also were not significantly different in parasitized vs nonparasitized larvae, and in both types of larvae the concentration of this pigment decreased by c. 50% during the same time interval when lipophorin was increased significantly. Instead of causing a generalized inhibition of host hemolymph protein synthesis, parasitism causes a complex array of changes in the hemolymph protein profile of Manduca sexta, possibly via the mediation of hormonal modulators of host protein synthesis, or transcriptional or translational regulation of host gene expression by factors associated with the polydnavirus or molecules secreted by the parasites.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Endocrine and neuroendocrine host-parasite relationships.

The relationships between parasites and hosts are complex, with many of these interactions involving an amazing degree of biochemical coevolution and communication. Hormones, neurohormones, and growth factors figure prominently in these relationships. In vertebrate hosts, many parasites secrete hormones, neuropeptides, or cytokine-like molecules that influence the host's physiological and immunological responses. Alternatively, the parasites secrete factors that alter the host's hormone levels. Simultaneously, molecules emanating from the host strongly influence the parasites' success. In some cases the host's hormones directly influence the parasites; in others, effects are mediated indirectly via the host's immune system. In invertebrates, the presence of parasites likewise has a major influence on the host's endocrine status and the normal suite of processes governed by hormones, including host development, metamorphosis, and reproduction. In insects, interactions involving juvenile hormone and ecdysteroids are especially well-documented, and recent evidence suggests that neuropeptides may also be affected by parasitism. Moreover, recent data suggest that in some species, such as snails, the host's nervous, neuroendocrine, and immune systems are functionally linked, similar to the complex interactions seen in vertebrates.

Animals↗
PubMed1991

Baculovirus-mediated expression of human apolipoprotein E in Manduca sexta larvae generates particles that bind to the low density lipoprotein receptor.

Human apolipoprotein E (apoE) is a ligand for the low density lipoprotein (LDL) receptor and mediates the catabolism of several classes of lipoprotein particles. Binding of apoE to the LDL receptor requires association of apoE with lipid in a vesicle or a lipoprotein particle. Because of this requirement, purified apoE or apoE derived directly from bacterial expression systems does not bind to the LDL receptor. To overcome this problem and to facilitate analysis of apoE structure, recombinant baculoviruses containing the human apoE cDNA fused to the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus were constructed. The recombinant viruses were used to infect larvae of the tobacco hornworm Manduca sexta in vivo. High levels of lipoprotein particles containing human apoE were present in the hemolymph of infected larvae. In contrast to apoE produced by recombinant baculovirus-infected insect cells in vitro, these particles were excellent ligands for the LDL receptor.

Animals↗
PubMed1990

Hymenolepis diminuta: an investigation of juvenile hormone titre, degradation and supplementation in the intermediate host, Tenebrio molitor.

Metacestodes of Hymenolepis diminuta cause a perturbance of vitellogenesis in the intermediate host Tenebrio molitor. The reduction in host reproductive output associated with infection may be due to this pathophysiology. Many of these events are regulated by host juvenile hormone (JH). A comparison of the titre of JH and its rate of degradation in female control and parasitized 15-day-old insects has been made. Haemolymph from female beetles contained 1.27 pMol JH equivalents/100 microliters. No significant difference was associated with infection. Likewise, the activity of JH esterase in female haemolymph was not affected by infection. However, topical application of a JH analogue, methoprene, at the time of infection or 8 days post-infection reduced the significant accumulation of vitellogenin usually found in the haemolymph of females 12 days or more post-infection. These findings indicate that parasite-induced alteration of host vitellogenesis is not mediated via alteration in JH titres, although observations made after hormone supplementation suggest some form of interaction between the parasite and the host endocrine system.

Animals↗