Embryonic entorhinal transplants project selectively to the deafferented entorhinal zone of adult mouse hippocampi, as demonstrated by the use of Thy-1 allelic immunohistochemistry. Effect of timing of transplantation in relation to deafferentation

The mouse Thy-1.1/Thy-1.2 allelic marking system is used to show that transplanted embryonic entorhinal cortex can reinnervate adult host hippocampi. The projection is limited to the appropriate terminal zones--viz. the outer two-thirds of the stratum moleculare of the dentate gyrus, and the stratum lacunosum-moleculare of the hippocampus--and extends for up to about 2 mm into the denervated host terminal field. The reconstruction of the entorhinal projections to the host requires direct contact between the embryonic donor tissue and the denervated adult host terminal field, and is dependent upon removal of the ipsilateral host entorhinal area. In the absence of an overall deafferenting host entorhinal lesion the transplanted entorhinal area forms only small local projections which are confined to areas which would have been locally deafferented as a result of direct damage to the host entorhinal afferents (i.e. during their intrahippocampal course) by the hippocampal lesion caused at the time when the transplant was inserted. The correct relative timing of deafferentation and transplantation is vital for the formation of the transplant-to-host projection. The host dendrites can be made receptive to entorhinal transplant projections by removal of the host entorhinal area at the time of transplantation. When deafferentation is performed first and transplantation is delayed, it is found that the deafferented host dendrites retain this receptivity even when deafferentation has been performed as much as two months before transplantation. Reversing the order of transplantation and deafferentation, however, shows that the transplants have only a transient ability to project to the deafferented host territory. Thus, transplants inserted and allowed to become established for one week before host deafferentation make very much reduced projections to the host, and from two weeks onwards are incapable of any detectable response to subsequent removal of the host entorhinal area. Coextensive with the formation of transplant-to-host entorhinodentate projections, the host entorhinal lesion also induces an intensification of the acetylcholinesterase staining of the host septodentate afferents in the denervated outer dentate stratum moleculare. The findings demonstrate the accurate reconstruction of a lost projection in adult brain by transplanting the appropriate type of embryonic tissue, but the results of altering the relative timing of deafferentation and transplantation raise currently unsolved questions about the nature of the competitive interactions between transplant and host axons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Attachment of Toxoplasma gondii to a specific membrane fraction of CHO cells

We have observed previously that attachment of Toxoplasma gondii to synchronized host cells is considerably increased at the mid-S phase (4 h postrelease). Synchronized CHO host cells at the mid-S phase were fractionated by molecular weight, and the antigens were used to produce a panel of polyclonal mouse antisera. The polyclonal antisera raised against fraction 4 with molecular mass ranging approximately from 18 to 40 kDa significantly reduced attachment to mid-S-phase host cells. Immunofluorescence assays demonstrated strong reactivity to mid-S-phase host cells and identified a number of potential receptors on Western blots. These data indicate that there is a specific host membrane receptor for parasite attachment that is upregulated during the mid-S phase of the host cell cycle.     

Interactions of bacterial effector proteins with host proteins

Pathogenic bacteria have evolved several clever survival strategies for manipulating host cell signaling pathways to establish beneficial replicative niches within the host. Recent literature has revealed novel mechanisms adopted by bacteria to manipulate host responses. For instance, host signaling pathways that were traditionally thought to be regulated by phosphorylation events have now been shown to be irreversibly blocked by bacterially-mediated acetylation, beta-elimination, and lytic modifications. This review highlights some of the common host proteins and signaling cascades targeted by such pathogens.     

Presence of host immunoglobulin in the gut of Sarcoptes scabiei (Acari: Sarcoptidae)

Sarcoptes scabiei (De Geer) mites burrow in the nonliving stratum corneum of the epidermis of their mammalian hosts. These mites ingest extracellular fluid (serum) that seeps into the burrow from the lower vascular dermis. A strong host antibody response occurs when mites die in the skin. This suggests internal immunogenic proteins are released into the host at this time. Vaccination with internal antigens may be an approach to protect against this mite if host antibody to internal antigens that regulate key physiological processes is ingested along with serum. Our study clearly showed that scabies mites ingest host immunoglobulin as evidenced by the localization of fluorescent-labeled antibody to host immunoglobulin in the anterior midgut and esophagus of fresh mites removed from the host. This is the first study that demonstrates that this nonblood-feeding ectoparasitic mite ingests host antibody while feeding on tissue fluid that seeps into the stratum corneum.     

Molecular mechanisms of hookworm disease: stealth, virulence, and vaccines

Hookworms produce a vast repertoire of structurally and functionally diverse molecules that mediate their long-term survival and pathogenesis within a human host. Many of these molecules are secreted by the parasite, after which they interact with critical components of host biology, including processes that are key to host survival. The most important of these interactions is the hookworm's interruption of nutrient acquisition by the host through its ingestion and digestion of host blood. This results in iron deficiency and eventually the microcytic hypochromic anemia or iron deficiency anemia that is the clinical hallmark of hookworm infection. Other molecular mechanisms of hookworm infection cause a systematic suppression of the host immune response to both the parasite and to bystander antigens (eg, vaccines or allergens). This is achieved by a series of molecules that assist the parasite in the stealthy evasion of the host immune response. This review will summarize the current knowledge of the molecular mechanisms used by hookworms to survive for extended periods in the human host (up to 7 years or longer) and examine the pivotal contributions of these molecular mechanisms to chronic hookworm parasitism and host clinical outcomes.     

Elimination of allogeneic multipotent stromal cells by host macrophages in different models of regeneration

Allogeneic multipotent stromal cells were previously thought to be poorly recognized by host immune system; the prolonged survival in host environments was explained by their immune privileged status. As long as the concept is currently reconsidered, the routes of elimination of allogeneic multipotent stromal cells by host immunity must be taken into account. This is necessary for correct comprehension of their therapeutic action. The study was focused upon survival of umbilical cord-derived allogeneic multipotent stromal cells in different rat models of tissue regeneration induced by partial hepatectomy or by critical limb ischemia. The observations were carried out by means of vital labeling of the cells with PKH26 prior to injection, in combination with differential immunostaining of host macrophages with anti-CD68 antibody. According to the results, allogeneic multipotent stromal cells are specifically eliminated by host immune system; the efficacy can reach 100%. Massive clearance of transplanted cells by host macrophages is accompanied by appropriation of the label by the latter, and this is a pronounced case of misleading presentation of exogenous label by host cells. The study emphasizes the role of macrophages in host response and also the need of additional criteria for correct data interpretation.     

泛素系统在结核分枝杆菌与宿主相互作用中的调控机制研究进展

结核分枝杆菌(Mycobacterium tuberculosis,Mtb)是一种极其成功的胞内病原菌,可通过多种策略实现免疫逃逸,从而在宿主巨噬细胞中长期存活。在对抗病原菌的防御过程中,泛素系统(Ubiquitin system)在激活宿主炎症免疫反应、细胞自噬、吞噬体成熟和细胞死亡等天然免疫功能及相关信号通路中发挥了重要的调控作用。而另一方面,近年的研究表明Mtb 等胞内病原菌可通过分泌效应蛋白(Effector proteins)挟持并利用宿主泛素系统进而抑制宿主的免疫功能,这些病原-宿主互作的界面有望成为抗结核药物研发的新靶点。     

Identification of the host determinant of two prolate-headed phages infecting Lactococcus lactis

A gene responsible for host determination was identified in two prolate-headed bacteriophages of the c2 species infecting strains of Lactococcus lactis. The identification of the host determinant gene was based on low DNA sequence homology in a specific open reading frame (ORF) between prolate-headed phages with different host ranges. When a host carrying this ORF from one phage on a plasmid was infected with another phage, we obtained phages with an altered host range at a frequency of 10(-6) to 10(-7). Sequencing of phage DNA originating from 10 independent single plaques confirmed that a genetic recombination had taken place at different positions between the ORF on the plasmid and the infecting phage. The adsorption of the recombinant phages to their bacterial hosts had also changed to match the phage origin of the ORF. Consequently, it is concluded that this ORF codes for the host range determinant.     

Ultrastructural study of entry of Chlamydia strain TWAR into HeLa cells.

Ultrastructural studies of the entry of Chlamydia strain TWAR into HeLa cells showed that the elementary bodies first attach to host cells by the pointed end, secure other binding sites on the host cells by forming cell wall protrusions, enter host cells by invaginating the host cell membrane, and form vacuolated endocytic vesicles. Differences were demonstrated between TWAR and other chlamydiae in the mode of attachment and endocytosis.     

Regulation of Salmonella-host cell interactions via the ubiquitin system

Salmonella infections cause acute intestinal inflammatory responses through the action of bacterial effector proteins secreted into the host cytosol. These proteins promote Salmonella survival, amongst others, by deregulating the host innate immune system and interfering with host cell ubiquitylation signaling. This review describes the recent findings of dynamic changes of the host ubiquitinome during pathogen infection, how bacterial effector proteins modulate the host ubiquitin system and how the host innate immune system counteracts Salmonella invasion by using these pathogens as signaling platforms to initiate immune responses.     

Cuticular surface lipids are responsible for disguise properties of an entomoparasite against host cellular responses

Entomopathogenic nematodes are widely used as alternatives to chemicals for the biological control of insects. These endoparasites are symbiotically associated with bacteria that are lethal for the host; however, parasites need to overcome the host immune defences to complete a successful life cycle. The processes parasites employ to escape or depress host immunity are targeted at deceiving non-self recognition as well as inactivating defence reactions.

The purpose of this paper is to investigate the interactions between the entomopathogenic nematode Steinernema feltiae and the lepidopteran Galleria mellonella, focusing on the role of the parasite's body-surface compounds in the immunoevasion of host cell-mediated responses.

To evaluate host self/non-self discrimination and encapsulation efficiency, we carried out a series of interaction assays between cultured host hemocytes and parasites or isolated cuticles. The data obtained suggest that the parasite cuticular lipids (PCLs) are able to bind a variety of host hemolymph molecules; PCLs attract host proteins from the hemolymph creating a coat around the parasite, thus, enabling Steinernema to disguise itself against hemocytes recognition. The role of parasite lipids in the disguise process was also investigated by simulating the nematode body surface with agarose microbeads covered with purified cuticular components; when the beads were coated with cuticular lipids, host hemocytes were not able to recognize and encapsulate.Results suggest that by means of attracting host hemolymph components onto its cuticular surface, S. feltiae prevents hemocytes attachment to its cuticle and inhibits melanization by depleting hemolymph components.     

Transmission and scanning electron microscopy of host cell entry by Toxoplasma gondii.

Entry by tachyzoites of Toxoplasma gondii, the causative agent of toxoplasmosis, into peritoneal cells was investigated with transmission and scanning electron microscopy. The process of entry is initiated by the parasite contacting the host cell with its anterior end, creating a small depression in the plasmalemma of the host cell. Occasionally, a small portion of the host cell cytoplasm protrudes and contaccts the anterior end of the parasite. A cylindrical structure (35 nm in diameter) extends from the pellicle of the parasite to the host cell. Such structures appear to assist host cell entry by T. gondii. As the entry process progresses, pseudopods of the host cell surround theparasite and finally T gondii becomes intracellular, being located in a vacuole separated from the host cell cytoplasm by a unit membrane. (Am J. Pathol 87:285-296, 1977).     

Molecular mechanisms of host cell egress by malaria parasites

Egress is a crucial step for malaria parasites to progress from one host cell to another. The rapid transition between host cells is mediated by the invasive merozoite stages. Merozoite egress from the enveloping cell includes the rupture of two membranes, the membrane of the parasitophorous vacuole and the host cell membrane. Egress from the host cell is also of importance for the intraerythrocytic gametocytes in order to undergo gametogenesis following their transmission to the mosquito during the blood meal. An increasing number of studies have aimed to identify the molecules involved in host cell egress by malaria parasites and decipher the sequence of membrane rupture. Recent work has acknowledged the crucial roles of plasmodial and host-derived proteases in membrane rupture and has indicated the involvement of secretory vesicles in priming the enveloping membranes for egress. This review highlights recent insight into the mechanisms of host cell egress by Plasmodium parasites. We will discuss the mode of egress of intrahepatic and intraerythrocytic parasites and their measures to evade the host immune system during this process.     

Characterization of rickettsial attachment to host cells by flow cytometry.

The mechanisms of rickettsial attachment have been studied by measuring quantitative changes in rickettsial binding to host cells by flow cytometry after different treatments of the rickettsiae and host cells. Time-dependent binding of Rickettsia conorii to host cells was demonstrated by the increasing intensity of host cell surface fluorescence of rickettsia-host cell combinations when examined with a rickettsia-specific monoclonal antibody. More than 70% of host cells had intensity of fluorescence above the threshold value after 10 min of incubation, owing to rickettsiae bound to the cell surface, and the greatest fluorescence intensity indicative of binding occurred at 20 min. The binding kinetics was rickettsial dose dependent. The binding of rickettsiae to host cells was greatly decreased when host cells or rickettsiae were treated with 1% paraformaldehyde for 30 min or 0.25% trypsin for 5 or 15 min, respectively. Rickettsiae that were heated at 56 degrees C for 15 min lost more than 80% of their ability to attach to host cells. R. rickettsii, an organism closely related to R. conorii, competitively inhibited the attachment of R. conorii (51% inhibition when mixed in equal numbers). These results indicate that the rickettsial binding structures are trypsin and heat sensitive and likely to be surface proteins.     

Fine structure of adrenal medullary grafts in the pain modulatory regions of the rat periaqueductal gray

Summary Recent findings in our laboratory indicate that adrenal medullary grafts produce significant alterations in pain sensitivity. Electron microscopic studies were undertaken to correlate these behavioral changes with the neural interactions of the host and graft tissue in the periaqueductal gray. A striking change found 8 weeks after transplantation is that pronounced myelination has taken place both in the graft and in the host tissue. The new myelin formation in the graft has the typical appearance of PNS myelination and, in the host the appearance of CNS myelination. The endothelial cells of the capillaries in the grafted tissue are attenuated and fenestrated in contrast to those of the surrounding parenchymal tissue of the host. By 8 weeks, the graft becomes heavily encapsulated with collagen, while the host CNS tissue develops layers of glial processes outlining the graft. However, collagen and glial layers apparently do not form an absolute barrier to either cellular or humoral interaction between the host and graft tissue. Chromaffin cells can be found protruding into the host CNS tissue and sometimes forming synapses with presumably the host neuronal processes. Grafted chromaffin cells may participate as both postsynaptic and, less often, as presynaptic components of synaptic junctions. The behavioral relevance of these synaptic contacts is unclear, since similar implants of adrenal medullary tissue into the dorsal spinal cord subarachnoid space, which also induce potent analgesia, do not contain synapses. Thus, it is more likely that behavioral changes are brought about by diffusion of neuroactive substances from grafted chromaffin cells to host receptors.     

Exploitation of host cell signaling machinery: activation of macrophage phosphotyrosine phosphatases as a novel mechanism of molecular microbial pathogenesis

Intracellular pathogens, particularly those that target host mononuclear phagocytes, have evolved strategies to either evade or inhibit cellular mechanisms of host defense. Mycobacterium tuberculosis and Leishmania donovani exemplify a diverse group of microorganisms that have developed the ability to invade and replicate within host macrophages, leading to disease expression. Recent studies have suggested that the pathogenesis of intracellular infection may involve interference with host cell signaling. Drawing upon examples from in vitro models that focused on M. tuberculosis and L. donovani, we review evidence that activation of host cell phosphotyrosine phosphatases may contribute to pathogenesis. A leading candidate appears to be the Src homology 2 domain containing phosphotyrosine phosphatase SHP-1, the activation of which may contribute to the development of infection and disease progression.     

Survival,replication, and antibody susceptibility of Ehrlichia chaffeensis outside of host cells

Ehrlichia chaffeensis, an obligate intracellular, tick-transmitted bacterium, is susceptible to antibody-mediated host defense, but the mechanism by which this occurs is not understood. One possible explanation is that antibodies directly access the bacteria in the extracellular environment of the host, perhaps during bacterial intercellular transfer. Accordingly, we investigated whether bacteria could be found outside of host cells during infection. Host cell-free plasma obtained from infected mice was found to contain ehrlichiae, and the host cell-free ehrlichiae readily transferred disease to susceptible SCID recipients. The host cell-free ehrlichiae were found during infection of both immunocompetent BALB/c and immunocompromised BALB/c-scid mice and reached levels as high as 10(8)/ml in plasma during persistent infection in SCID mice. Approximately 10% of the blood-borne bacteria were found outside of host cells. Although it is generally accepted that replication of ehrlichiae occurs only within host cells, the cell-free bacteria were shown to undergo DNA replication and cell division in vitro for 3 to 5 days when incubated at 37 degrees C in plasma. Paradoxically, both infectivity and virulence were lost after 24 h of ex vivo culture. The data indicate that E. chaffeensis is exposed to the extracellular milieu during infection, presumably during intercellular transfer, and reveal that these intracellular bacteria do not require the environment of the host cell for replication. Our findings reveal a possible mechanism by which antibodies can access the intracellular bacteria upon their release into the extracellular milieu and mediate host defense and also have implications for understanding the replication and transmission of this vector-borne pathogen.     

Dependence of MS2 and T4 phage growth upon host amino acid biosynthesis during infections of Escherichia coli

By manipulating the growth conditions of Escherichia coli both before and after phage MS2 or phage T4 infections, a dependence of phage growth upon postinfection host amino acid biosynthesis can be inferred. Cells grown under repressing conditions for amino acid biosynthesis shifted to amino acid-free medium postinfection (in the absence of further host gene expression) are poor hosts for phage growth, whereas cells grown under derepressing conditions for amino acid biosynthesis are good hosts regardless of postinfection conditions; thus postinfection biosynthesis of amino acids utilizes performed host enzymes which still function to carry out their biosynthetic pathways during phage infections. Phage MS2 also appears to permit the derepression of host amino acid biosynthetic operons during the infection. A functional dependence of MS2 growth upon postinfection host gene function is perhaps the strongest argument that the RNA phage do not shut off host messenger RNA and protein synthesis.