Key Laboratory of Animal Epidemiology and Zoonosis

Beijing, China

Key Laboratory of Animal Epidemiology and Zoonosis

Beijing, China
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Liu Y.,China Agricultural University | Chen R.,China Agricultural University | Tariq M.,China Agricultural University | Xia C.,China Agricultural University | Xia C.,Key Laboratory of Animal Epidemiology and Zoonosis
Acta Crystallographica Section:F Structural Biology Communications | Year: 2014

In the process of antigen presentation, the MHCI-CD8 complex is important for immune signal transduction by the activation of cytotoxic T cells. Here, the expression, purification, crystallization and X-ray analysis of the complex of the chicken MHC class I molecule BF2 0401 and CD8αα (CD8αα-BF2 0401) are reported. This complex was verified by SDS-PAGE analysis of a CD8αα-BF2 0401 crystal, which showed three bands corresponding to the molecular weights of BF2 0401, β 2-microglobulin and CD8α, respectively. The crystal of CD8αα-BF2 0401 diffracted to 2.8Å resolution and belonged to space group P21, with unit-cell parameters a = 90.6, b = 90.8, c = 94.9 14;Å, β = 98°. The Matthews coefficient and solvent content were calculated to be 2.88 14;Å3 14;Da-1 and ∼57.3%, respectively. © 2014 International Union of Crystallography.


Jiang B.,China Agricultural University | Liu Y.,China Agricultural University | Chen R.,China Agricultural University | Wang Z.,China Agricultural University | And 3 more authors.
Acta Crystallographica Section F:Structural Biology Communications | Year: 2014

Amphioxus is regarded as an essential animal model for the study of immune evolution. Discovery of new molecules with the immunoglobulin superfamily (IgSF) variable (V) domain in amphioxus would help in studying the evolution of IgSF V molecules in the immune system. A protein was found which just contains only one IgSF V domain in amphioxus, termed Amphi-IgSF-V; it has over 30% sequence identity to the V domains of human immunoglobulins and mammalian T-cell receptors. In order to clarify the three-dimensional structure of this new molecule in amphioxus, Amphi-IgSF-V was expressed, purified and crystallized, and diffraction data were collected to a resolution of 1.95 Å. The crystal belonged to space group P3221, with unit-cell parameters a = b = 53.9, c = 135.5 Å. The Matthews coefficient and solvent content were calculated to be 2.58 Å3 Da-1 and 52.38%, respectively. The results will provide structural information to study the evolution of IgSF V molecules in the immune system. © 2014 CrossMark.


Yuan H.,China Agricultural University | Chen R.,China Agricultural University | Liu Y.,China Agricultural University | Tariq M.,China Agricultural University | And 3 more authors.
Acta Crystallographica Section F:Structural Biology Communications | Year: 2014

Complement 1q (C1q) is the first component of the complement system which can initiate the classical complement pathway. In human, C1q is composed of 18 polypeptide chains: six C1qA chains, six C1qB chains and six C1qC chains. Each chain has a signal peptide and is comprised of a collagen-like region and a C-terminal C1q globular domain (C1qgD), which is organized as a heterotrimer. C1qgD can recognize antigen-antibody complexes containing IgG and IgM or can bind directly to the C-reactive protein. Although the classical complement pathway is found from fish to mammals, only the human C1qgD structure has been determined. Compared with that of mammals, fish C1q exhibits similar immune functions and genome arrangement. In order to illustrate the structure of C1qgD in fish, zebrafish (Danio rerio) C1qA globular domain (Dare-C1qAgD) was expressed, purified and crystallized. X-ray diffraction data were collected from a crystal to a resolution of 2.05 Å; the crystal belonged to the orthorhombic space group P212121, with unit-cell parameters a = 50.347, b = 85.059, c = 95.560 Å. It contained three molecules in the asymmetric unit. The Matthews coefficient value V M was 2.31 Å3 Da -1, with a calculated solvent content of 46.7%. The data will help to give insight into the structural basis of C1qA in fish species. © 2014 International Union of Crystallography All rights reserved.


Wang Z.,China Agricultural University | Chen R.,China Agricultural University | Tariq M.,China Agricultural University | Jiang B.,China Agricultural University | And 3 more authors.
Acta Crystallographica Section F:Structural Biology Communications | Year: 2014

In order to clarify the structural characteristics of the bovine MHC class I molecule (BoLA-I) complexed with CD8αα (CD8αα-BoLA-I), bovine CD8αα, BoLA-I (BoLA-202201) and β2m were expressed and purified, and were then assembled with a peptide derived from Foot-and-mouth disease virus (FMDV-VP1YY9) and crystallized. The crystal diffracted to 1.7 Å resolution and belonged to space group P21, with unit-cell parameters a = 53.9, b = 103.8, c = 61.8 Å, α = γ = 90, β = 96°. The asymmetric unit contained one complex, with a Matthews coefficient of 2.41 Å3 Da-1 and a solvent content of 48.9%. The rotation-function Z-score and translation-function Z-score for molecular replacement were 3.4 and 8.9, respectively. In addition, SDS-PAGE analysis of CD8αα-BoLA-I crystals showed three bands corresponding to the molecular weights of BoLA-I heavy chain, β2m and CD8α. The structure of the CD8αα-BoLA-I complex should be helpful in obtaining insight into the interaction between bovine CD8αα and MHC class I molecules. Structure determination of BoLA-202201-FMDV-VP1YY9 will be useful in the design of vaccines for foot-and-mouth disease. © 2014 International Union of Crystallography All rights reserved.


Chen Z.,China Agricultural University | Zhang N.,China Agricultural University | Lu S.,China Agricultural University | Tariq M.,China Agricultural University | And 3 more authors.
Acta Crystallographica Section F:Structural Biology Communications | Year: 2015

β2-Microglobulin (β2m) noncovalently associates with the heavy chain of major histocompatibility complex class I (MHC I) molecules, which bind foreign antigen peptides to control the cytotoxic T lymphocyte (CTL) immune response. In contrast to mammals, there are distinct types of β2ms derived from two loci in a number of teleost species. In order to clarify the structures of the β2ms, the zebrafish (Danio rerio) β2ms Dare-β2m-I and Dare-β2m-II were expressed in Escherichia coli, purified and crystallized, and diffraction data were collected to 1.6 and 1.9 Å resolution, respectively. Both crystals belonged to space group P212121. The unit-cell parameters were determined to be a = 38.2, b = 50.4, c = 50.9 Å for Dare-β2m-I and a = 38.9, b = 52.7, c = 65.8 Å for Dare-β2m-II. Each asymmetric unit was constituted of one molecule, with Matthews coefficients of 2.22 and 3.01 Å3 Da-1 and solvent contents of 45 and 59% for Dare-β2m-I and Dare-β2m-II, respectively. These two β2m structures will provide relevant information for further studies of the structures of the MHC I complex. © 2015 International Union of Crystallography.


Chen K.,State Key Laboratory of Agrobiotechnology | Chen K.,Key Laboratory of Animal Epidemiology and Zoonosis | Chen K.,China Agricultural University | Luo Z.,State Key Laboratory of Agrobiotechnology | And 5 more authors.
Virology Journal | Year: 2011

Background: Chicken anemia virus (CAV) infection of newly hatched chickens induces generalized lymphoid atrophy and causes immunosuppressive. VP3, also known as Apoptin, is non-structural protein of CAV. Apoptin specifically induces apoptosis in transformed or tumor cells but not in normal cells. In particular, there are no known cellular homologues of Apoptin hindering genetic approaches to elucidate its cellular function. Although a number of Apoptin-interacting molecules have been identified, the molecular mechanism underlying Apoptin's action is still poorly understood. To learn more about the molecular mechanism of Apoptin's action, we searched for Apoptin associated proteins. Results: Using yeast two-hybrid and colony-life filter approaches we got five positive yeast clones. Through sequencing and BLASTed against NCBI, one of the clones was confirmed containing Gallus heat shock cognate protein 70 (Hsc70). Hsc70 gene was clone into pRK5-Flag plasmid, coimmunoprecipitation assay show both exogenous Hsc70 and endogenous Hsc70 can interact with Apoptin. Truncated Apoptin expression plasmids were made and coimmunoprecipitation were performed, the results show the binding domain of Apoptin with Hsc70 is located between amino acids 30-60. Truncated expression plasmids of Hsc70 were also constructed and coimmunoprecipitation were performed, the results show the peptide-binding and variable domains of Hsc70 are responsible for the binding to Apoptin. Confocal assays were performed and results show that under physiological condition Hsc70 is predominantly distributed in cytoplasm, whereas Hsc70 is translocated into the nuclei and colocalized with Apoptin in the presence of Apoptin in DF-1 cell. Functional studies show that Apoptin markedly down-regulate the mRNA level of RelA/p65 in DF-1 cell. To explore the effect of Hsc70 on Apoptin-mediated RelA/p65 gene expression, we have searched two Hsc70 RNAi sequences, and found that all of them dramatically inhibited the expression of endogenous Hsc70, with the #2 Hsc70 RNAi sequence being the most effective. Knockdown of Hsc70 show Apoptin-inhibited RelA/p65 expression was abolished. Our data demonstrate that Hsc70 is responsible for the down-regulation of Apoptin induced RelA/p65 gene expression. Conclusion: We identified Gallus Hsc70 as an Apoptin binding protein and showed the translocation of Hsc70 into the nuclei of DF-1 cells treated with Apoptin. Hsc70 regulates RelA/p65 gene expression induced by Apoptin. © 2011 Chen et al; licensee BioMed Central Ltd.


Elfeil W.K.,Jilin University | Elfeil W.K.,Suez Canal University | Abouelmaatti R.R.,Jilin University | Abouelmaatti R.R.,Key Laboratory of Animal Epidemiology and Zoonosis | And 12 more authors.
Journal of Animal and Veterinary Advances | Year: 2012

Toll like receptor mediate immune response through recognition of Pathogen-Associated Molecular Patterns (PAMPs) thus play important roles in host defense. Little is known about the Avian Immune System structure and the majority of it based on chicken as a Universal Avian Model while recent researches showed that a lot of factor not common between chicken and other birds. This research aimed to identify and clone the duck TLR4 as a model to Anseriformes species, researchers success to clone a novel functional dTLR4 mRNA and deposit into the NCBI GenBank database where the mRNA consist of 2529 nucleotide and it showed 81 % identity with chicken and 78% identity with zebra finch while showed 95% identity with goose. The translated amino acid showed that the gene consist of 843 amino acid and the transmembrane structure analysis reveal a novel unique LRR motifs in duck and absent in both chicken and zebra finch at the position 56-80 also showed that TIR domain consist of 146 amino acid not 145 like chicken and zebra finch. © 2012 Medwell Journals.


Hao P.,Key Laboratory of Animal Epidemiology and Zoonosis | Yang N.,Key Laboratory of Animal Epidemiology and Zoonosis | Cui X.,Key Laboratory of Animal Epidemiology and Zoonosis | Liu J.,Key Laboratory of Animal Epidemiology and Zoonosis | And 2 more authors.
Journal of Parasitology | Year: 2014

Neospora caninum is an important cause of abortion in cattle worldwide, but the isolation of a viable parasite from an abortus is difficult, and viable N. caninum has not been isolated from any host in China. In the present study, peripheral blood samples were collected from a jugular vein of an adult dairy cow that had aborted; the cow was seropositive to N. caninum antibodies by ELISA. White blood cells were separated and seeded onto Vero cell monolayer cultures for parasite isolation. Tachyzoites were first observed in cell culture on day 84 after initial inoculation. The parasite was confirmed to be N. caninum by gene sequencing and immunofluorescence, and by bioassays in BALB/c mice. The new N. caninum isolate (NC-Bj) has a unique pattern on microsatellite Cont-14. To our knowledge, this is the first successful isolation of N. caninum in China from any host. © 2014 American Society of Parasitologists.


Elfeil W.M.K.,Suez Canal University | Elfeil W.M.K.,King Faisal University | Algammal A.M.,Suez Canal University | Abouelmaatti R.R.,Jilin University | And 3 more authors.
Central European Journal of Immunology | Year: 2016

The rabbit has great commercial importance as a source of meat and fur, as well as its uses as a laboratory animal for the production of antibodies, used to detect the presence or absence of disease and for research in infectious diseases and immunology. One of the most critical problems in immunology is to understand how the immune system detects the presence of infectious agents and disposes the invader without destroying the self-tissues. Genetic characterization of Toll-like receptors has established that innate immunity is a skillful system that detects invasion of microbial pathogens. Our work aimed to identify, clone and express the Oryctolagus cuniculus (rabbit) TL R-1 mRNA and its encoding protein. We cloned the complete mRNA sequence of Oryctolagus cuniculus TLR-1 and deposit it in the GenBank under accession number (KC349941), which has 2388 base pair and it encodes encode an open reading frame (ORF) translated into 796 amino acids mRNA and consist of 20 types of amino acids. The analysis of amino acid sequence revealed that the rabbit TLR-1 has a typical protein components belonging to the TLR family. Rabbit TLR-1 was expressed in a wide variety of rabbit tissues, which indicate an important role in immune system in different organs.


Abouelmaatti R.R.,Jilin University | Abouelmaatti R.R.,Wenzhou University | Abouelmaatti R.R.,Key Laboratory of Animal Epidemiology and Zoonosis | Algammal A.M.,Suez Canal University | And 8 more authors.
Central-European Journal of Immunology | Year: 2013

Toll-like receptors (TLRs) are the best understood of the innate immune receptors that detect infections in vertebrates. However, the fish TLRs also exhibit very distinct features and a large diversity, which is likely derived from their diverse evolutionary history and the distinct environments that they occupy. Little is known about the fish immune system structure. Our work was aimed to identify and clone the Nile tilapia TLR-3 as a model of fresh water fish species; we cloned the full-length cDNA sequence of Nile tilapia (Oreochromis niloticus) TLR-3 and according to our knowledge, it is the first report illustrating tilapia TLR-3. The complete cDNA sequence of Nile tilapia TLR-3 was 2736 pair base and it encodes a polypeptide of 912 amino acids. Analysis of the deduced amino acid sequence indicated that Nile tilapia TLR-3 has typical structural features and main component of proteins belonging to the TLR family. Our results illustrate a complete and functional Nile tilapia TLR-3 and it is considered an ortholog of the other vertebrate's receptor.

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