Compatibility Polymorphism Research Articles

Combining a transcriptomic approach and a targeted metabolomics approach for deciphering the molecular bases of compatibility phenotype in the snail Biomphalaria glabrata toward Schistosoma mansoni

Biomphalaria glabrata is a freshwater snail and the obligatory intermediate host of Schistosoma mansoni parasite, the etiologic agent of intestinal Schistosomiasis, in South America and Caribbean. Interestingly in such host-parasite interactions, compatibility varies between populations, strains or individuals. This observed compatibility polymorphism is based on a complex molecular-matching-phenotype, the molecular bases of which have been investigated in numerous studies, notably by comparing between different strains or geographical isolates or clonal selected snail lines. Herein we propose to decipher the constitutive molecular support of this interaction in selected non-clonal resistant and susceptible snail strain originating from the same natural population from Brazil and thus having the same genetic background. Thanks to a global RNAseq transcriptomic approach on whole snail, we identified a total of 328 differentially expressed genes between resistant and susceptible phenotypes among which 129 were up-regulated and 199 down-regulated. Metabolomic studies were used to corroborate the RNAseq results. The activation of immune genes and specific metabolic pathways in resistant snails might provide them with the capacity to better respond to parasite infection.

Multi-strain compatibility polymorphism between a parasite and its snail host, a neglected vector of schistosomiasis in Africa

Interactions between Schistosoma mansoni and its snail host are understood primarily through experimental work with one South American vector species, Biomphalaria glabrata. However, 90% of schistosomiasis transmission occurs in Africa, where a diversity of Biomphalaria species may serve as vectors. With the long-term goal of determining the genetic and ecological determinants of infection in African snail hosts, we developed genetic models of Biomphalaria sudanica, a principal vector in the African Great Lakes. We determined laboratory infection dynamics of two S. mansoni lines in four B. sudanica lines. We measured the effects of the following variables on infection success and the number of cercariae produced (infection intensity): (i) the combination of parasite and snail line; (ii) the dose of parasites; and (iii) the size of snail at time of exposure. We found one snail line to be almost completely incompatible with both parasite lines, while other snail lines showed a polymorphism in compatibility: compatible with one parasite line while incompatible with another. Interestingly, these patterns were opposite in some of the snail lines. The parasite-snail combination had no significant effect on the number of cercariae produced in a successful infection. Miracidia dose had a strong effect on infection status, in that higher doses led to a greater proportion of infected snails, but had no effect on infection intensity. In one of the snail-schistosome combinations, snail size at the time of exposure affected both infection status and cercarial production in that the smallest size class of snails (1.5–2.9 mm) had the highest infection rates, and produced the greatest number of cercariae, suggesting that immunity increases with age and development. The strongest predictor of the infection intensity was the size of snail at the time of shedding: 1 ​mm of snail growth equated to a 19% increase in cercarial production. These results strongly suggest that infection status is determined in part by the interaction between snail and schistosome genetic lines, consistent with a gene-for-gene or matching allele model. This foundational work provides rationale for determining the genetic interactions between African snails and schistosomes, which may be applied to control strategies.

Compatibility Polymorphism Based on Long-Term Host-Parasite Relationships: Cross Talking Between Biomphalaria glabrata and the Trematode Schistosoma mansoni From Endemic Areas in Brazil.

Sympatric snail populations have been kept in the laboratory since the isolation of the parasite from the field. To evaluate the influence of the intermediate host in the infectivity of S. mansoni, this allopatric strain was compared to two sympatric strains, from different geographical origins, and with different time of maintenance in the laboratory. Snail–trematode compatibility was accessed for a total of nine possible combinations (three snail populations, three schistosome strains), using different charges of parasite: 1, 5, 10, and 15 miracidia/snail. Each S. mansoni strain was characterized according to its infectivity phenotype that reflects the efficiency of their infection mechanism and all B. glabrata populations were characterized according to its (in)compatible phenotype that reflects the level of (un)susceptibility they display. For all host-parasite combinations tested the dose-response relation indicated a trend for an increase in the infectivity of S. mansoni when higher miracidial doses were used. SmRES-2 presented the highest overall infectivity rate, especially in the SmRES-2/BgRES interaction with 15 miracidia/snail. However, SmRES was more infective to BgBAR than SmRES-2, indicating that SmRES strain was more infective at the first contact with this new host than after 2 years of interaction (SmRES-2). BgBAR presented the highest susceptibility to infection. SmRES and SmRES-2 are the same parasite strains. It seems that during these 2 years of interaction, BgBAR acted like a filter and shifted the compatibility polymorphism of the strain SmRES. SmRES-2 became more infective to BgRES (sympatric) than to BgBAR (allopatric), and conversely, SmRES was more infective to BgBAR (allopatric) than to BgRES (sympatric). This interplay suggests that epigenetic mechanisms are prompting these changes. This study concerns with infection of B. glabrata snails from different Brazilian localities with S. mansoni in allopatric and sympatric associations that will partially help in understanding the natural epidemiology of schistosomiasis within natural snail populations in watercourses. This work demonstrates that there is a shift on the compatibility polymorphism profile resulting from sympatric and allopatric interactions of B. glabrata and S. mansoni that constantly change during the time of interaction.

Integrated multi-omic analyses in Biomphalaria-Schistosoma dialogue reveal the immunobiological significance of FREP-SmPoMuc interaction

The fresh water snail Biomphalaria glabrata is one of the vectors of the trematode pathogen Schistosoma mansoni, which is one of the agents responsible of human schistosomiasis. In this host–parasite interaction, co-evolutionary dynamic results into an infectivity mosaic known as compatibility polymorphism. Integrative approaches including large scale molecular approaches have been conducted in recent years to improve our understanding of the mechanisms underlying compatibility.This review presents the combination of integrated Multi-Omic approaches leading to the discovery of two repertoires of polymorphic and/or diversified interacting molecules: the parasite antigens S. mansoni polymorphic mucins (SmPoMucs) and the B. glabrata immune receptors fibrinogen-related proteins (FREPs). We argue that their interactions may be major components for defining the compatible/incompatible status of a specific snail/schistosome combination.

The Compatibility Between Biomphalaria glabrata Snails and Schistosoma mansoni: An Increasingly Complex Puzzle.

This review reexamines the results obtained in recent decades regarding the compatibility polymorphism between the snail, Biomphalaria glabrata, and the pathogen, Schistosoma mansoni, which is one of the agents responsible for human schistosomiasis. Some results point to the snail's resistance as explaining the incompatibility, while others support a "matching hypothesis" between the snail's immune receptors and the schistosome's antigens. We propose here that the two hypotheses are not exclusive, and that the compatible/incompatible status of a particular host/parasite couple probably reflects the balance of multiple molecular determinants that support one hypothesis or the other. Because these genes are involved in a coevolutionary arms race, we also propose that the underlying mechanisms can vary. Finally, some recent results show that environmental factors could influence compatibility. Together, these results make the compatibility between B. glabrata and S. mansoni an increasingly complex puzzle. We need to develop more integrative approaches in order to find targets that could potentially be manipulated to control the transmission of schistosomiasis.

Epigenetic origin of adaptive phenotypic variants in the human blood fluke Schistosoma mansoni.

BackgroundAdaptive evolution is not possible without the generation of phenotypic variants. The origin of these variations has been a central topic in evolutionary biology. Up to now, it was commonly accepted that standing genetic variation is the only cause of phenotypic variants. However, epigenetic information is emerging as a complementary source of heritable phenotypic variation that contributes to evolution. The relative importance of genetics and epigenetics in generating heritable phenotypic variation is nevertheless a matter of debate.ResultsWe used a host–parasite system to address this question. The human blood fluke Schistosoma mansoni can adapt rapidly to new intermediate snail hosts. The interaction between parasite and mollusk is characterized by a compatibility polymorphism illustrating the evolutionary dynamics in this system. The principal molecular marker for compatibility (infection success) is the expression pattern of a group of polymorphic mucins (SmPoMuc) in the parasite. We show here that chromatin structure changes as the SmPoMuc promoters are the cause for SmPoMuc transcription polymorphism leading to phenotypic novelty and increase in infection success, i.e., fitness.ConclusionWe establish that epigenetic changes can be the major if not only cause of adaptive phenotypic variants in Schistosoma mansoni, suggesting that epimutations can provide material for adaptive evolution in the absence of genetic variation in other systems. In addition, our results indicate that epidrugs can be used to control parasite development but also parasite evolution.Electronic supplementary materialThe online version of this article (doi:10.1186/s13072-016-0076-2) contains supplementary material, which is available to authorized users.

Fibrinogen-Related Proteins (FREPs) in Mollusks.

Anti-parasite responses of the snail Biomphalaria glabrata involve antigen-reactive plasma lectins termed fibrinogen-related proteins (FREPs) comprising a C-terminal fibrinogen (FBG) domain and one or two upstream immunoglobulin domains. FREPs are highly polymorphic; they derive from several gene families with multiple loci and alleles that are diversified by exon loss, alternative splicing, and random somatic mutation (gene conversion and point mutations). Individual B. glabrata snails have dynamically distinct FREP sequence repertoires. The immune relevance of B. glabrata FREPs is indicated by FREP binding to polymorphic antigens of (snail-specific) digenean parasites and altered resistance of B. glabrata to digeneans following RNAi knockdown of FREPs. The compatibility polymorphism hypothesis proposes that FREP mutation increases the range of germline-encoded immune recognition in B. glabrata to counter antigenically-varied parasites. Somatic mutation may result from sequence exchange among tandemly arranged FREP genes in the genome, and analysis of sequence variants also suggests involvement of cytidine deaminase-like activity or epigenetic regulation. Without current indications of selection or retention of effective sequence variants toward immunological memory, FREP diversification is thought to afford B. glabrata immunity that is anticipatory but not adaptive. More remains to be learned about this system; other mollusks elaborate diversified lectins consisting of single FBG domains, and bona fide FREPs were reported from additional gastropod species, but these may not be diversified. Future comparative immunological studies and gene discovery driven by next-generation sequencing will further clarify taxonomic distribution of FREP diversification and the underlying mutator mechanisms as a component of immune function in mollusks.

Digenean-gastropod host associations inform on aspects of specific immunity in snails

Gastropod immunology is informed importantly by the study of the frequent encounters snails endure with digeneans (digenetic trematodes). One of the hallmarks of gastropod-digenean associations is their specificity: any particular digenean parasite species is transmitted by a limited subset of snail taxa. We discuss the nature of this specificity, including its immunological basis. We then review studies of the model gastropod Biomphalaria glabrata indicating that the baseline responses of snails to digeneans can be elevated in a specific manner. Studies incorporating molecular and functional approaches are then highlighted, and are further suggestive of the capacity for specific gastropod immune responses. These studies have led to the compatibility polymorphism hypothesis: the interactions between diversified fibrinogen-related proteins (FREPs) and diverse carbohydrate-decorated polymorphic parasite antigens determine recognition and trigger specific immunity. Complex glycan structures are also likely to play a role in the host specificity typifying snail–digenean interactions. We conclude by noting the dynamic and consequential interactions between snails and digeneans can be considered as drivers of diversification of digenean parasites and in the development and maintenance of specific immunity in gastropods.

Multi-parasite host susceptibility and multi-host parasite infectivity: A new approach of the Biomphalaria glabrata/Schistosoma mansoni compatibility polymorphism

In this study, we analyze the degree of susceptibility/un-susceptibility of five strains of Biomphalaria glabrata from different geographical origins successively challenged with a panel of 4 Schistosoma mansoni strains. A total of 20 homopatric and heteropatric host–parasite combinations were tested with exposure doses of 1, 10, 20, 30 and 50 miracidia per individual host. By doing this, we characterized each B. glabrata strain by its “multi-parasite susceptibility phenotype” that reflects better the efficiency of their defense mechanism against not only one, but a diversity of schistosome stocks. In the same time, all the S. mansoni strains used were characterized, by their “multi-host infectivity phenotype” that reflects the level of infectivity they display when confronted to diverse snail populations. Based on these results it is possible to select different homogenous stocks of snails with different spectrum of susceptibility/un-susceptibility for several parasite strains. This will be a useful tool for future functional studies conducted to understand the genetics and molecular basis of the compatibility polymorphism in this host/parasite model.

Schistosoma mansoni Mucin Gene (SmPoMuc) Expression: Epigenetic Control to Shape Adaptation to a New Host

The digenetic trematode Schistosoma mansoni is a human parasite that uses the mollusc Biomphalaria glabrata as intermediate host. Specific S. mansoni strains can infect efficiently only certain B. glabrata strains (compatible strain) while others are incompatible. Strain-specific differences in transcription of a conserved family of polymorphic mucins (SmPoMucs) in S. mansoni are the principle determinants for this compatibility. In the present study, we investigated the bases of the control of SmPoMuc expression that evolved to evade B. glabrata diversified antigen recognition molecules. We compared the DNA sequences and chromatin structure of SmPoMuc promoters of two S. mansoni strains that are either compatible (C) or incompatible (IC) with a reference snail host. We reveal that although sequence differences are observed between active promoter regions of SmPoMuc genes, the sequences of the promoters are not diverse and are conserved between IC and C strains, suggesting that genetics alone cannot explain the evolution of compatibility polymorphism. In contrast, promoters carry epigenetic marks that are significantly different between the C and IC strains. Moreover, we show that modifications of the structure of the chromatin of the parasite modify transcription of SmPoMuc in the IC strain compared to the C strain and correlate with the presence of additional combinations of SmPoMuc transcripts only observed in the IC phenotype. Our results indicate that transcription polymorphism of a gene family that is responsible for an important adaptive trait of the parasite is epigenetically encoded. These strain-specific epigenetic marks are heritable, but can change while the underlying genetic information remains stable. This suggests that epigenetic changes may be important for the early steps in the adaptation of pathogens to new hosts, and might be an initial step in adaptive evolution in general.

A Large Repertoire of Parasite Epitopes Matched by a Large Repertoire of Host Immune Receptors in an Invertebrate Host/Parasite Model

For many decades, invertebrate immunity was believed to be non-adaptive, poorly specific, relying exclusively on sometimes multiple but germ-line encoded innate receptors and effectors. But recent studies performed in different invertebrate species have shaken this paradigm by providing evidence for various types of somatic adaptations at the level of putative immune receptors leading to an enlarged repertoire of recognition molecules. Fibrinogen Related Proteins (FREPs) from the mollusc Biomphalaria glabrata are an example of these putative immune receptors. They are known to be involved in reactions against trematode parasites. Following not yet well understood somatic mechanisms, the FREP repertoire varies considerably from one snail to another, showing a trend towards an individualization of the putative immune repertoire almost comparable to that described from vertebrate adaptive immune system. Nevertheless, their antigenic targets remain unknown. In this study, we show that a specific set of these highly variable FREPs from B. glabrata forms complexes with similarly highly polymorphic and individually variable mucin molecules from its specific trematode parasite S. mansoni (Schistosoma mansoni Polymorphic Mucins: SmPoMucs). This is the first evidence of the interaction between diversified immune receptors and antigenic variant in an invertebrate host/pathogen model. The same order of magnitude in the diversity of the parasite epitopes and the one of the FREP suggests co-evolutionary dynamics between host and parasite regarding this set of determinants that could explain population features like the compatibility polymorphism observed in B. glabrata/S. mansoni interaction. In addition, we identified a third partner associated with the FREPs/SmPoMucs in the immune complex: a Thioester containing Protein (TEP) belonging to a molecular category that plays a role in phagocytosis or encapsulation following recognition. The presence of this last partner in this immune complex argues in favor of the involvement of the formed complex in parasite recognition and elimination from the host.

Follow-up of the genetic diversity and snail infectivity of a Schistosoma mansoni strain from field to laboratory

Schistosoma mansoni is an endoparasite causing a serious human disease called schistosomiasis. The quantification of parasite genetic diversity is an essential component to understand schistosomiasis epidemiology and disease transmission patterns but some studies on parasite genetic diversity are performed using parasite laboratory strains. However, a potential discrepancy in level of genetic variation between field populations and laboratory strains may have various implications in our deductions. In this study, 246 adult worms were analysed on 15 microsatellite markers to investigate variation of genetic diversity between a founder field isolate and the nine successive laboratory generations during three years of laboratory maintenance. In parallel, we measured a parasite life trait (snail infectivity) at each generation in order to test a potential link between inbreeding and snail infectivity. Our genetic analyses demonstrate a significant genetic differentiation between all parasite generations and a significant isolation by time associated with a decrease in neutral genetic diversity that is likely to be the result of successive bottleneck events. However, while snail infectivity decreases sharply between field isolate and the first laboratory generation, this parasite life trait does not evolve between other laboratory generations and appeared disconnected from this continuous neutral genetic diversity loss. We hypothesize that a sufficient level of compatibility polymorphism at a genomic level is maintained independently of an increase of inbreeding, ensuring the stability in the parasite life trait.

The compatibility polymorphism in invertebrate host/trematodes interactions: research of molecular determinants

The co-evolutionary dynamics that exist in many host-parasite interactions sometimes leads to compatibility polymorphism. This phenomenon is well documented in mollusc/trematodes interactions but its molecular base is unknown. In order to identify key molecules involved in this phenomenon, we developed several molecular approaches comparing compatible or incompatible strains of mollusc or parasite. These comparisons led to the identification of numerous candidate genes listed and discussed (some of them) in the present review.

Expression analysis of highly polymorphic mucin proteins ( Sm PoMuc) from the parasite Schistosoma mansoni

The co-evolutionary dynamics that exist in many host-parasite interactions sometimes lead to a compatibility polymorphism, of which the molecular bases are unknown. To identify key molecules involved in this phenomenon in the S. mansoni/ B. glabrata model, we developed a comparative proteomics approach for the larval stages that interact with the invertebrate host. The comparison of the proteomes of compatible and incompatible parasite strains led to the identification of a new family of schistosome antigens that share molecular characteristics with the molecules of the mucin family. In particular, they possess a domain containing a variable number of tandem repeats (VNTR). The pronounced polymorphism of these proteins, that distinguishes compatible and incompatible parasite strains, led us to further investigate the role that this protein family plays in the compatibility polymorphism in our model. In the present study, we examine precursor structure, report analysis of mucin-like expression and describe their polymorphism. Our data show that these proteins share structural characteristics with highly glycosylated secreted mucins. The proteins are (i) only expressed in larval stages that interact with the mollusc, (ii) located in the apical gland of miracidia and sporocysts and (iii) secreted and released in excretion–secretion products. Finally, we show that these mucins display a high degree of polymorphism and that extensive differences are observed between S. mansoni strains. These different characteristics led us to name this novel gene family “ S. mansoni polymorphic mucins” ( Sm PoMuc).

Molecular determinants of compatibility polymorphism in the Biomphalaria glabrata/ Schistosoma mansoni model: New candidates identified by a global comparative proteomics approach

The co-evolutionary dynamics that exist in host–parasite interactions sometimes lead to compatibility polymorphisms, the molecular bases of which are rarely investigated. To identify key molecules that are involved in this phenomenon in the Schistosoma mansoni/ Biomphalaria glabrata model, we developed a comparative proteomics approach using the larval stages that interact with the invertebrate host. We used qualitative and quantitative analyses to compare the total proteomes of primary sporocysts from compatible and incompatible parasite strains. The differentially expressed proteins thus detected belong to three main functional groups: (i) scavengers of reactive oxygen species, (ii) components of primary metabolism, and (iii) mucin-like proteins. We discuss the putative roles played by these protein families as determinants of compatibility polymorphism. Since mucins are known to play key roles in the host–parasite interplay, we consider the newly discovered S. mansoni mucin-like proteins ( SmMucin-like) as the most promising candidates for influencing the fate of host–parasite interactions. An analysis of their expression is presented in a paper published in the same journal issue.