Richard Galinier- PhD
- French National Centre for Scientific Research
Richard Galinier
- PhD
- French National Centre for Scientific Research
About
144
Publications
44,140
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2,610
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Introduction
Current institution
Additional affiliations
July 2009 - present
Description
- Immuno-biological interactions in invertebrate host / pathogen. Invertebrate immunity.
December 2001 - June 2009
Description
- Immuno-biological interactions in host/parasite interactions. Schistosoma mansoni / Biomphalaria glabrata
March 1998 - April 2004
Description
- PhD in biology "Ubiquitin-ligases recruitment by human adenovirus protein responsible of viral entry".
Publications
Publications (144)
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 polymorph...
The freshwater snail Biomphalaria glabrata is an intermediate host of Schistosoma mansoni, the agent of human intestinal schistosomiasis. However, much is to be discovered about its innate immune system that appears as a complex black box, in which the immune cells (called hemocytes) play a major role in both cellular and humoral response towards p...
The genetic diversity of viral populations is a key driver of the spatial and temporal diffusion of viruses; yet, studying the diversity of whole genomes from natural populations still remains a challenge. Phylodynamic approaches are commonly used for RNA viruses harboring small genomes, but have only rarely been applied to DNA viruses with larger...
Host-parasite interaction can result in a strong alteration of the host-associated microbiota. This dysbiosis can affect the fitness of the host; can modify pathogen interaction and the outcome of diseases. Biomphalaria glabrata is the snail intermediate host of the trematode Schistosoma mansoni, the agent of human schistosomiasis, causing hundreds...
The genetic diversity of viral populations is a key to understanding the spatial and temporal diffusion history of viruses, but studying the diversity of whole genomes from natural populations remains a challenge. Phylodynamic approaches are commonly used for RNA viruses harboring small genomes, but have only rarely been applied to DNA viruses with...
Aerolysins initially characterized as virulence factors in bacteria are increasingly found in massive genome and transcriptome sequencing data from metazoans. Horizontal gene transfer has been demonstrated as the main way of aerolysin-related toxins acquisition in metazoans. However, only few studies have focused on their potential biological funct...
Biomphalaria glabrata is one of the snail intermediate hosts of Schistosoma mansoni , the causative agent of intestinal schistosomiasis disease. Numerous molecular studies using comparative approaches between susceptible and resistant snails to S. mansoni infection have helped identify numerous snail key candidates supporting such susceptible/resis...
In a number of species, individuals exposed to pathogens can mount an immune response and transmit this immunological experience to their offspring, thereby protecting them against persistent threats. Such vertical transfer of immunity, named trans-generational immune priming (TGIP), has been described in both vertebrates and invertebrates. Althoug...
Juvenile Pacific oysters (Crassostrea gigas) are subjected to recurrent episodes of mass mortalities that constitute a threat for the oyster industry. This mortality syndrome named “Pacific Oyster Mortality Syndrome” (POMS) is a polymicrobial disease whose pathogenesis is initiated by a primary infection by a variant of an Ostreid herpes virus name...
Planorbidae snails are the intermediate host for the trematode parasite of the Schistosoma genus, which is responsible for schistosomiasis, a disease that affects both humans and cattle. The microbiota for Schistosoma has already been described as having an effect on host/parasite interactions, specifically through immunological interactions. Here,...
Biomphalaria glabrata is a freshwater Planorbidae snail. In its environment, this mollusk faces numerous microorganisms or pathogens, and has developed sophisticated innate immune mechanisms to survive. The mechanisms of recognition are quite well understood in Biomphalaria glabrata, but immune effectors have been seldom described. In this study, w...
Thioester-containing proteins (TEPs) superfamily is known to play important innate immune functions in a wide range of animal phyla. TEPs are involved in recognition, and in the direct or mediated killing of several invading organisms or pathogens. While several TEPs have been identified in many invertebrates, only one TEP (named BgTEP) has been pr...
The Fasciola hepatica/Pseudosuccinea columella interaction in Cuba involves a unique pattern of phenotypes; while most snails are susceptible, some field populations are naturally resistant to infection and parasites are encapsulated by snail hemocytes. Thus, we investigated the hemocytes of resistant (R) and susceptible (S) P. columella, in partic...
Selective pressures between hosts and their parasites can result in reciprocal evolution or adaptation of specific life history traits. Local adaptation of resident hosts and parasites should lead to increase parasite infectivity/virulence (higher compatibility) when infecting hosts from the same location (in sympatry) than from a foreign location...
Mortality of B. glabrata snails after infections.
The survival rates of B. glabrata subjected to infection by different S. mansoni strains were observed over 14 weeks. Kaplan Meier graphs were generated using xlstat, and the log-rank test (p < 0.05) was used to test for significant between-group differences. Colored curves indicate the mortality ra...
List of differentially represented transcripts in RNAseq clusters.
Quality reads (Phred score >29) were aligned on the transcriptome assembly using the C++ script Bowtie2 (v2.0.2) (255 score) running thanks local engine using Galaxy Project server (Giardine, Riemer et al. 2005). The DESeq2 software (Love, Huber et al. 2014) (v2.12;http://www.biocon...
List of transcripts express by Schistosoma within Biomphalaria glabrata tissues highlight in RNAseq clusters.
The Biomphalaria (v1) and Schistosoma (v5.2) genome have been concatenate to mapped the RNAseq reads of each experimental condition. Only quality reads (Phred score >29) were aligned to the concatenate genomes using Bowtie2 (v2.0.2), which...
Blast-like cells are non-proliferative cells.
In vitro EdU labeling of hemocytes collected for in vitro analysis. Confocal microscopy of EdU-labeled hemocytes from snails subjected to the allopatric interaction BgBRE/SmVEN at 24 h post-infection (BV24). Pictures corresponded to the merge of DAPI labelling (blue); EdU labelling (green) and phase con...
Biomphalaria and Schistosoma compatibility between African and South-American strains.
The prevalence (P %: percentage of snail infected) and intensity (I: number of parasites per infected host) of infection are presented for each experimental infection. The indicated values correspond to 10 miracidia. Each pairwise combination of Biomphalaria glab...
Clustering of all differential represented transcripts.
Clustering of differentially represented transcripts. Heatmap representing the profiles of the 1,895 differentially represented immune-related transcripts in the BB, BV, or BR interactions along the time course of infection (at 24 and 96 h). Each transcript is represented once and each line re...
miRNAs precursor identified in Biomphalaria glabrata RNAseq data.
(XLSX)
Invertebrate immune response may be primed by a current infection in a sustained manner, leading to the failure of a secondary infection with the same pathogen. The present study focuses on the Schistosomiasis vector snail Biomphalaria glabrata, in which a specific genotype-dependent immunological memory was demonstrated as a shift from a cellular...
Infectious diseases are mostly explored using reductionist approaches despite repeated evidence showing them to be strongly influenced by numerous interacting host and environmental factors. Many diseases with a complex aetiology therefore remain misunderstood. By developing a holistic approach to tackle the complexity of interactions, we decipher...
The importance of the microbiota in host-parasite interactions has taken a deeper interest recently. Microbiota has to be considered as a third partner playing a central role in these interactions. Biomphalaria glabrata is the vector snail of the trematode Schistosoma mansoni, the agent of human schistosomiasis causing hundreds of thousands of deat...
Background: Selective pressures between hosts and their parasites can results in reciprocal evolution or adaptation of specific life history traits. Local adaptation of resident hosts and parasites should lead to host-parasite systems performing better in sympatry when compared to allopatry. Between-population variations in parasite infectivity/vir...
Insect thioester-containing protein (iTEP) is the most recently defined group among the thioester-containing protein (TEP) superfamily. TEPs are key components of the immune system, and iTEPs from flies and mosquitoes were shown to be major immune weapons. Initially characterized from insects, TEP genes homologous to iTEP were further described fro...
Methylamine treatment effect on BgTEP binding to Sp1. 500 Sp1 were incubated 30 min at 26°C with either (1) 1 mL of Chernin’s balanced salt solution (CBSS) buffer, (2) 1 mL of cell-free plasma, or (3) 1 mL of cell-free plasma + methylamine (2 mM). Sp1 were further centrifuged and washed with CBSS buffer. Proteins from Sp1 pellet were subsequently e...
Web-logo representation of the consensus sequence of intron–exon junction in BgTEP gene.
Histological section (10 µm thickness) of Biomphalaria glabrata paraffin embedded tissue in which can be observed the encapsulation of a Schistosoma mansoni parasite by hemocytes 24 h post-infection. White arrows show the BgTEP-positive cells (red) around and in the capsule. The BgTEP protein is detected by immunolocalization using anti BgTEP-PEP a...
May–Grünwald–Giemsa staining of Biomphalaria glabrata hemocytes revealing the three major hemocyte subtypes: hyalinocytes, granulocytes, and blast-like cells.
Insect Thioester-containing protein (iTEP) is the most recently defined group among the TEP superfamily. TEPs are key components of the immune system, and iTEPs from flies and mosquitoes were shown to be major immune weapons. Initially characterised from insects, TEP genes homologous to iTEP were further described from several other invertebrates i...
Protein identification of the 109 spots revealed only in “pathogens + hemolymph” gels as compared to “pathogens only” gels. For each spot, the −10logP values of proteins and peptides are indicated, together with the top BLAST hit in NCBI nr database, the conserved domains of the sequence retrieved (performed with NCBI CD-search available at https:/...
2D-PAGE gels of “pathogens + hemolymph” and “pathogens only” for each of the five pathogens used. Arrows are indicating spots exclusively present in the “pathogens + hemolymph” profiles but not in the proteomic profiles of “pathogens only,” which represents proteins from Biomphalaria glabrata hemolymph that participated in the recognition of pathog...
Detection of pathogens by all living organisms is the primary step needed to implement a coherent and efficient immune response. This implies a mediation by different soluble and/or membrane-anchored proteins related to innate immune receptors called PRRs (pattern-recognition receptors) to trigger immune signaling pathways. In most invertebrates, t...
This corrects the article DOI: 10.1038/ncomms15451.
Freshwater ecosystems face very strong anthropogenic pressures, among which overexploitation, habitat degradation, flow modification, species invasion, and water pollution lead to growing threats on biodiversity. Urbanization through wastewater treatment, industry through the release of inorganic and organic chemicals, and agriculture through the u...
Biomphalaria snails are instrumental in transmission of the human blood fluke Schistosoma mansoni. With the World Health Organization's goal to eliminate schistosomiasis as a global health problem by 2025, there is now renewed emphasis on snail control. Here, we characterize the genome of Biomphalaria glabrata, a lophotrochozoan protostome, and pro...
Secreted_proteins SCW&ES_Topblast_results
HSP annotation
HSP expression in Bge cell-line
Pattern recognition receptors (PRRs) and cytokines
Supplementary Figures, Supplementary Notes and Supplementary References
Biomphalaria glabrata linkage groups
Proteome InterProscan annotation
fasta sequences of predicted secreted proteins (fasta)
secretome B. glabrata, Interproscan annotation
Pfam domain annotation of predicted secretome
Transcripts, tissue distribution
Predicted GPCR genes of Biomphalaria glabrata
Annotation unknown microarray features
Conserved precursor miRNAs, Structural, location and thermodynamic
Novel predicted miRNA and target genes
Neuropeptides precursors and prepropeptides
Sequence data sets
proteome KEGG/SNV mapping
Mycoplasma/Mollicute annotation
GREPs bind to S.mansoni
Expression patterns of previous unknowns.
neuropeptides, tissue distribution
Predicted bioactive cleavage products of neuropeptides
Biomphalaria glabrata actins
Biomineralization transcripts from B. glabrata mantle tissue.
Low-copy transposable elements (TE) abundance
Prediction of secreted proteins
Cytochrome P 450 (CYP) genes of B. glabrata.
TLR pathway signaling
Epigenetic toolbox genes
miRNA and piRNA pathway factors
Conserved mature miRNAs B. glabrata and homologs
conserved precursor miRNAs tissue distribution in B. glabrata
Conserved miRNA target gene predictions
Novel miRNAs predictions
Steroidogenesis pathway genes
Complement-like factors.
Antimicrobial factors
Antimicrobial factors expression profiles
Cardiac regulatory and structural genes in B. glabrata
Identity of 34 B. pfeifferi and B. glabrata sequence homologs
Sequence Identity of B. pfeifferi homologs to 162 B. glabrata genes from
In recent decades, numerous studies have sought to better understand the mechanisms underlying the compatibility between Biomphalaria glabrata and Schistosoma mansoni. The developments of comparative transcriptomics, comparative genomics, interactomics and more targeted approaches have enabled researchers to identify a series of candidate genes. Ho...
Transcriptome statistics.
The characteristics of BgBAR, BgBRE1, BgBRE2, BgVEN and BgGUA transcriptomes were detailed in terms of reads, contig length, contig number and nucleotide numbers. BgBRE1 and BgBRE2 transcriptomes came from 2 independent biological replicates.
(XLSX)
Variants of cDNA obtained from 11 individual sporocysts of S. mansoni strains.
(XLS)
Annotation of the 24 classes of FREP molecules identified.
The longest transcript number of each class is mentioned, as are: its length in nucleotides and in amino acids (after virtual translation); its new FREP class and standard classification when possible; its IgSF domain number; its position on the B. glabrata genome draft (genome assembly ver...
Validation of FREP by alignment of sequences obtained from Sanger sequencing and from computational assembly of de novo transcripts.
To confirm that de novo assembly will not generate miss assemblies for highly diverse FREP molecules, assembled FREP transcripts were validated by traditional Sanger sequencing of PCR products. Sanger sequences and co...
B glabrata immunome.
List of molecule categories previously shown to play roles in snail immunity, and the corresponding loci in the BgBRE transcriptome. Underlined lanes correspond to transcripts that are differentially expressed across the four B. glabrata strains. Molecules were selected from: B. glabrata transcriptomes and the literature [11,17...
We report the genome sequence and organization of five viruses infecting snails of both Biomphalaria glabrata and Biomphalaria pfeifferi, which are vectors of the intestinal schistosomiasis. Four viruses presented a polyadenylated positive single strand RNA genome encoding one or two large open reading frames (ORFs) flanked by untranslated region....
Questions
Questions (2)
Hi everybody,
I have tested a kit for cell free protein expression (Next generation cell free protein expression kit, wheat germ CFPS 700) from Merck and I didn't get the expected yield for protein production.
In the procedure of this kit you have to prepare DNA template by a game of several PCR, then in vitro transcription is realized from PCR template, and finally cell free translation using wheat germ extract.
All is good until transcription (agarose gel checking)
But after that the protocol is a mRNA purification using amonium acetate salt and ethanol.
I think these step is the problem because I loose a lot of mRNA.
Can somebody tell me if this step is necessary or if I can try to translate without mRNA purification? Or else, is there another methode for mRNA purification, that preserve its quality for the following transcription (the kit exclude phenol, trizol or ammonium sulfate purification that rendered mRNA unsuitable for translation)?
I have produced a recombinant protein with a His6 tag at C-ter position. The produced protein is not soluble and needs denaturing conditions for extraction (Urea or guanidine thiocyanate). Purification is then performed with Ni-NTA resin (affinity for nickel ions). The protein binds to the column. But it is impossible for me to elute it. I have tried all the recommended protocols: elution with acidic conditions, with imidazole, with EDTA, a combination of these three protocols. I have also tried a renaturation of the protein on the column (hybrid conditions) followed by imidazole elution. However, the protein is still bound to the resin, as I have checked by SDS-PAGE.
Does anyone have any suggestions or solutions to this problem?
