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Poultry Science May 2023Epidemiologic investigations in recent years have shown that the detection rate of avian hepatitis E virus (HEV) in chicken flocks is increasing in China. Nevertheless,...
Epidemiologic investigations in recent years have shown that the detection rate of avian hepatitis E virus (HEV) in chicken flocks is increasing in China. Nevertheless, effective prevention and control measures are still lacking. In this study, specific pathogen-free (SPF) chicken serum against HEV was prepared using recombinant HEV open reading frames (ORF2 and ORF3) proteins as immunogens. An SPF chicken infection model was established by intravenous inoculation of chick embryos. Swab samples were collected at 7, 14, 21, and 28 d of age and used to detect avian HEV load, along with other indicators, by fluorescence quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) assay. The therapeutic effects on blocking vertical HEV transmission were observed, by using the methods of antibody application alone, mixed, or combined application of each of the 2 antibodies with type I interferon. The results showed that type I interferon alone or in combination with antiserum reduced the positive rate of HEV from 100 to 62.5% and 25%, respectively. However, the avian HEV-positivity rate was reduced to 75, 50, and 37.5% after type I interferon was used alone or in combination with antisera against ORF2 and ORF3, respectively. The inhibitory effect of type I interferon alone or in combination with an antiserum, on HEV replication was more significant in cells than in vivo. In this study, the inhibitory effect of type I interferon alone or in combination with an antiserum on avian HEV replication was observed in vitro and in vivo, providing the necessary technical reserve for disease prevention and control.
Topics: Chick Embryo; Animals; Hepevirus; Chickens; Immunoglobulins; Immune Sera; Interferon Type I
PubMed: 36966643
DOI: 10.1016/j.psj.2023.102591 -
Toxins Apr 2022Sea snake venom is extremely toxic, and it can induce severe respiratory failure and cause high mortality. The most effective first aid treatment for sea snake bites is...
Sea snake venom is extremely toxic, and it can induce severe respiratory failure and cause high mortality. The most effective first aid treatment for sea snake bites is to inject antivenom as soon as possible. However, in China, there are only four types of terrestrial snake antivenoms, none of which are effective in the treatment of sea snake bites. In order to develop an antivenom for the dominant species of sea snakes in Chinese seas, venom (HcuV) was chosen as the antigen to immunize horses. From immune plasma, a high-titer antivenom (HcuAV) was prepared. In vitro assessment showed that HcuAV had a cross-neutralizing capacity against HcuV and venom (HcyV). In vivo assessment indicated that HcuAV injection could significantly improve the survival rates of the HcuV and HcyV envenomated mice (0% to 100% and 87.5%, respectively) when it was injected at a sufficient amount within the shortest possible time. In addition, HcuAV could also effectively alleviate multiple organ injuries caused by HcuV. These results provide experimental support for the future clinical application of HcuAV.
Topics: Animals; Antivenins; Elapid Venoms; Horses; Hydrophiidae; Immune Sera; Mice; Snake Bites
PubMed: 35448862
DOI: 10.3390/toxins14040253 -
PLoS Neglected Tropical Diseases Apr 2016Snakebite envenomation is a serious medical problem in many tropical developing countries and was considered by WHO as a neglected tropical disease. Antivenom (AV), the...
Snakebite envenomation is a serious medical problem in many tropical developing countries and was considered by WHO as a neglected tropical disease. Antivenom (AV), the rational and most effective treatment modality, is either unaffordable and/or unavailable in many affected countries. Moreover, each AV is specific to only one (monospecific) or a few (polyspecific) snake venoms. This demands that each country to prepare AV against its local snake venoms, which is often not feasible. Preparation of a 'pan-specific' AV against many snakes over a wide geographical area in some countries/regions has not been possible. If a 'pan-specific' AV effective against a variety of snakes from many countries could be prepared, it could be produced economically in large volume for use in many countries and save many lives. The aim of this study was to produce a pan-specific antiserum effective against major medically important elapids in Asia. The strategy was to use toxin fractions (TFs) of the venoms in place of crude venoms in order to reduce the number of antigens the horses were exposed to. This enabled inclusion of a greater variety of elapid venoms in the immunogen mix, thus exposing the horse immune system to a diverse repertoire of toxin epitopes, and gave rise to antiserum with wide paraspecificity against elapid venoms. Twelve venom samples from six medically important elapid snakes (4 Naja spp. and 2 Bungarus spp.) were collected from 12 regions/countries in Asia. Nine of these 12 venoms were ultra-filtered to remove high molecular weight, non-toxic and highly immunogenic proteins. The remaining 3 venoms were not ultra-filtered due to limited amounts available. The 9 toxin fractions (TFs) together with the 3 crude venoms were emulsified in complete Freund's adjuvant and used to immunize 3 horses using a low dose, low volume, multisite immunization protocol. The horse antisera were assayed by ELISA and by in vivo lethality neutralization in mice. The findings were: a) The 9 TFs were shown to contain all of the venom toxins but were devoid of high MW proteins. When these TFs, together with the 3 crude venoms, were used as the immunogen, satisfactory ELISA antibody titers against homologous/heterologous venoms were obtained. b) The horse antiserum immunologically reacted with and neutralized the lethal effects of both the homologous and the 16 heterologous Asian/African elapid venoms tested. Thus, the use of TFs in place of crude venoms and the inclusion of a variety of elapid venoms in the immunogen mix resulted in antiserum with wide paraspecificity against elapid venoms from distant geographic areas. The antivenom prepared from this antiserum would be expected to be pan-specific and effective in treating envenomations by most elapids in many Asian countries. Due to economies of scale, the antivenom could be produced inexpensively and save many lives. This simple strategy and procedure could be readily adapted for the production of pan-specific antisera against elapids of other continents.
Topics: Animals; Antivenins; Asia; Bungarus; Cross Reactions; Elapid Venoms; Elapidae; Enzyme-Linked Immunosorbent Assay; Epitopes; Freund's Adjuvant; Horses; Immune Sera; Immunization; Lethal Dose 50; Mice; Snake Bites
PubMed: 27058956
DOI: 10.1371/journal.pntd.0004565 -
Canadian Medical Association Journal Jun 1964A review of literature on snakebites was undertaken to evaluate the various recommended and practised methods of treatment used over the years, and to choose the most... (Review)
Review
A review of literature on snakebites was undertaken to evaluate the various recommended and practised methods of treatment used over the years, and to choose the most effective and least mutilating therapy for snakebite victims. Descriptions and photographs of pit vipers indigenous to Canada are included for identification purposes. The only proved adequate treatment of snakebite was shown to be Antivenin administration. A complete description of the use of this substance, which is prepared from horse serum, is given, as well as precautions to be taken prior to its administration, in the event of sensitivity to the serum. Supporting therapy is described and brief comments are made on less satisfactory means of treatment.
Topics: Animals; Antivenins; Canada; Humans; Immune Sera; Skin Tests; Snake Bites; Snakes; Therapeutics; Viperidae
PubMed: 14158556
DOI: No ID Found -
The Journal of Experimental Medicine Apr 19651. The effects of normal rabbit serum and of rabbit antiserum to whole foetal mouse tissues, on the isolated limb bones of late foetal mice were studied in organ...
1. The effects of normal rabbit serum and of rabbit antiserum to whole foetal mouse tissues, on the isolated limb bones of late foetal mice were studied in organ culture, and the influence of hydrocortisone on these effects was investigated. 2. Unheated normal serum caused slight loss of metachromatic material from the cartilage matrix, and some resorption of both cartilage and bone. 3. In unheated antiserum to foetal mouse tissues, the terminal cartilage was smaller and less metachromatic than in paired controls in normal serum, while osteoclasis was so intense that in many explants the bone had almost disappeared. The amount of necrosis varied with different batches of antiserum. 4. The changes produced by normal serum and antiserum could be largely prevented by heating the sera to 57 degrees C for 45 minutes. 5. The effects could also be inhibited by the addition of hydrocortisone to the unheated sera; as little as 0.1 microg hydrocortisone per ml of medium had a well marked protective action. 6. It is suggested that (a) unheated antiserum causes a release of lysosomal enzymes with consequent breakdown of intercellular material, (b) this release is due to an indirect action on the lysosome via an increased permeability of the cell membrane, (c) hydrocortisone does not affect the antigen-antibody reaction, but inhibits the autolytic changes that normally follow this reaction, possibly by stabilising both the lysosomal and cell membranes.
Topics: Animals; Bone and Bones; Cell Membrane; Cell Membrane Permeability; Fetus; Hot Temperature; Hydrocortisone; Immune Sera; Lysosomes; Mice; Necrosis; Organ Culture Techniques; Pathology; Pharmacology; Research; Tissue Culture Techniques
PubMed: 14276776
DOI: 10.1084/jem.121.4.551 -
Microbiological Research Feb 2016Chlamydia psittaci (C. psittaci), an obligate intracellular agent of psittacosis, causes an atypical pneumonia in humans. The transmembrane head proteins (TMH) of C....
Chlamydia psittaci (C. psittaci), an obligate intracellular agent of psittacosis, causes an atypical pneumonia in humans. The transmembrane head proteins (TMH) of C. psittaci, putatively belong to the Inc family and presumably play similar roles. CPSIT_0844 and CPSIT_0846 were the putative TMH proteins of C. psittaci. To identify these two proteins, antisera were raised with fusion proteins which were prokaryotic expressed in Escherichia coli and purified. By immunofluorescence assay, CPSIT_0844 and CPSIT_0846 were localized in the inclusion membrane of C. psittaci-infected cells. By RT-PCR and western blot analysis to detect the temporal expression, CPSIT_0844 and CPSIT_0846 were detected as early as 12h post-infection (p.i.) and 6h p.i., separately; meanwhile, in secretions monitored with immunofluorescence assay, these proteins were observed in the inclusion membrane at 18h p.i. and remained in the inclusion membrane throughout the growth cycle. CPSIT_0844 and CPSIT_0846 could specifically be recognized by the antiserum of C. psittaci but failed to react with the antiserums of Chlamydia trachomatis and Chlamydia pneumoniae, which is consistent with the fact that they had no significant orthologs in C. trachomatis and C. pneumoniae. These results revealed that CPSIT_0844 and CPSIT_0846, the putative TMH family proteins, might be unique to C. psittaci and could be used to diagnose the infection caused by C. psittaci. Moreover, CPSIT_0844 and CPSIT_0846 could induce the expression of the inflammatory cytokines IL-1β, IL-6 and TNF-α in THP-1 cells, which might contribute to chlamydia-induced inflammatory pathologies.
Topics: Bacterial Outer Membrane Proteins; Bacterial Proteins; Chlamydia Infections; Chlamydophila psittaci; Cloning, Molecular; Cytokines; Escherichia coli; Fluorescent Antibody Technique; Genes, Bacterial; HeLa Cells; Humans; Immune Sera; Psittacosis; Recombinant Proteins; Tumor Necrosis Factor-alpha
PubMed: 26805615
DOI: 10.1016/j.micres.2015.11.005 -
The Journal of Clinical Investigation Jul 1983The relationship between group B streptococcal (GBS) type-specific antisera and the type II-specific polysaccharide is evaluated from a structural and immunologic...
The relationship between group B streptococcal (GBS) type-specific antisera and the type II-specific polysaccharide is evaluated from a structural and immunologic viewpoint. Although all GBS type-specific polysaccharides are composed of the same monosaccharides, the type II antigen is more complex structurally and contains these sugars in a molar ratio different from the other antigens. Type II polysaccharide has two side chains. One contains only sialic acid and is less susceptible to acid cleavage than sialic acid residues found on types III, Ia, and Ib polysaccharides. The other side chain is composed of galactose as the only sugar. Immunochemical studies demonstrate that the type II polysaccharide has several immunodeterminants. One of these determinants is likely to be the side-chain galactose, while sialic acid appears to comprise part of another immunodeterminant, more complex than sialic acid alone. A series of cross-reactions is demonstrated between the type II native antigen and antisera to serotypes Ia, III, and Ib by a sensitive radioactive antigen-binding assay, which account for additional, complex immunodeterminants. The strongest of these cross-reactions is with type Ia antiserum and the weakest with Ib antiserum. Since Ia and Ib polysaccharides differ in only one linkage, these findings suggest that the trisaccharide beta D-N-acetyl-glucosamine-p(1 leads to 3) beta D-galactose-p(1 leads to 4) beta D-glucose-p [[beta D-GlcNAcp(1 leads to 3) beta D-Galp(1 leads to 4)beta D-Glcap]] is the likely common site responsible for the interaction of the type II native polysaccharide and type Ia antiserum. Another cross-reaction is observed between type III antiserum and type II native antigen. Inhibition studies indicate that the most likely cross-reactive determinant in this case is [beta D-Galp(1 leads to 4)beta D-GlcNAcp]. Type II polysaccharide has been utilized in a human vaccine trial to test safety and immunogenicity. The polysaccharide is highly immunogenic, inducing an antibody response in 95% of recipients, and nontoxic, with side-effects confined to minimal local reactions. Despite the cross-reactions observed between type-specific antigens and antibody prepared by immunization of rabbits with whole bacteria, which suggest shared immunodeterminants, similar cross-reactions were not detected in human sera after immunization with purified type II polysaccharide.
Topics: Adult; Animals; Antibodies; Antibody Specificity; Bacterial Capsules; Epitopes; Humans; Immune Sera; Immunization; Immunologic Techniques; Polysaccharides, Bacterial; Rabbits
PubMed: 6192144
DOI: 10.1172/jci110965 -
The Indian Journal of Medical Research Jul 2012Phospholipase A2 (PLA2 ) is one of the major constituents of krait venom associated with several pathophysiological actions like myotoxicity, cardiotoxicity,... (Comparative Study)
Comparative Study
BACKGROUND & OBJECTIVES
Phospholipase A2 (PLA2 ) is one of the major constituents of krait venom associated with several pathophysiological actions like myotoxicity, cardiotoxicity, neurotoxicity, etc. As there was no specific antiserum available against Bungarus fasciatus venom, this study was done with synthetic herbal compounds, anti PLA2 rabbit antiserum and commercial polyvalent snake venom antiserum to neutralize the PLA2 induced toxicities in experimental models.
METHODS
B. fasciatus venom phospholipase A2 fraction 38 (BF-38) was isolated by ion exchange chromatography, molecular weight was determined by mass spectrometry and its N terminal amino acid sequence was identified. Monospecific rabbit antiserum was raised against the PLA2 in presence of Freund complete adjuvant. The neutralization of PLA2 induced toxicities was done in in vitro and in in vivo models using synthetic herbal compounds, anti PLA2 rabbit antiserum and commercial polyvalent snake venom antiserum.
RESULTS
A toxic PLA2 (BF-38) was purified from the B. fasciatus venom by CM-cellulose and HPLC, of 13.17 kDa and a minor band of 7.3 kDa using ESI-MS. The 13.17 kDa PLA2 sequence was NLYQFKNMIQC. The 7.3 kDa toxin sequence was RKCLTKYSQDNES and was found to be <10 per cent w/w. Anti PLA2 rabbit antiserum produced faint precipitant band in immunogel diffusion and showed low titre value. The commercial polyvalent snake venom antiserum, anti PLA2 rabbit antiserum and the synthetic herbal compounds neutralized the PLA2 induced toxicities at different intensities.
INTERPRETATION & CONCLUSIONS
Our results suggested that synthetic herbal compound (BA) along with antiserum might provide effective protection against PLA2 induced toxicities of B. fasciatus venom.
Topics: Amino Acid Sequence; Animals; Benzoates; Bungarotoxins; Bungarus; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Hydroxybenzoate Ethers; Immune Sera; Mass Spectrometry; Molecular Sequence Data; Phospholipase A2 Inhibitors; Phospholipases A2; Plant Extracts; Rabbits; Salicylic Acid; Toxicity Tests
PubMed: 22885262
DOI: No ID Found -
The Korean Journal of Parasitology Feb 2016The sodium hydrogen exchanger 1 (NHE1), which functions in maintaining the ratio of Na(+) and H(+) ions, is widely distributed in cell plasma membranes. It plays a...
The sodium hydrogen exchanger 1 (NHE1), which functions in maintaining the ratio of Na(+) and H(+) ions, is widely distributed in cell plasma membranes. It plays a prominent role in pH balancing, cell proliferation, differentiation, adhesion, and migration. However, its exact subcellular location and biological functions in Toxoplasma gondii are largely unclear. In this study, we cloned the C-terminal sequence of T. gondii NHE1 (TgNHE1) incorporating the C-terminal peptide of NHE1 (C-NHE1) into the pGEX4T-1 expression plasmid. The peptide sequence was predicted to have good antigenicity based on the information obtained from an immune epitope database. After induction of heterologous gene expression with isopropyl-b-D-thiogalactoside, the recombinant C-NHE1 protein successfully expressed in a soluble form was purified by glutathione sepharose beads as an immunogen for production of a rabbit polyclonal antiserum. The specificity of this antiserum was confirmed by western blotting and immunofluorescence. The antiserum could reduce T. gondii invasion into host cells, indicated by the decreased TgNHE1 expression in T. gondii parasites that were pre-incubated with antiserum in the process of cell entry. Furthermore, the antiserum reduced the virulence of T. gondii parasites to host cells in vitro, possibly by blocking the release of Ca(2+). In this regard, this antiserum has potential to be a valuable tool for further studies of TgNHE1.
Topics: Animals; Cell Line; Immune Sera; Male; Mice; Protozoan Proteins; Rabbits; Recombinant Proteins; Sheep; Sodium-Hydrogen Exchangers; Toxoplasma; Toxoplasmosis
PubMed: 26951975
DOI: 10.3347/kjp.2016.54.1.21 -
Poultry Science Feb 2019Contamination of avian attenuated vaccines by avian leukosis virus (ALV) is considered to be a specific horizontal transmission of ALV. Eradication of ALV contamination...
Contamination of avian attenuated vaccines by avian leukosis virus (ALV) is considered to be a specific horizontal transmission of ALV. Eradication of ALV contamination in vaccine virus seeds is thus a precondition of qualified vaccine production. In this study, we used the nucleoside reverse transcriptase (RT) inhibitor azidothymidine (AZT) together with monofactor antiserum against avian leukosis virus subgroup A (ALV-A) to remove ALV-A from vaccine virus seeds. Different doses of ALV-A were artificially added to the Newcastle disease virus (NDV) vaccine seeds, then the ALV-contaminated attenuated virus vaccine virus seeds were cultured in DF-1 cells. Single-drug treatment with 5 μg/mL AZT or 5% (v/v) as well as combined treatment with AZT and antiserum significantly suppressed ALV-A replication (P < 0.001) in the vaccine virus seeds. Complete absence of virus replication was observed in cells exposed to joint treatment with AZT and antiserum. The treated virus seeds met the requirements of the Chinese Ministry of Agriculture. Therefore, combined treatment with AZT and antiserum can be used to eradicate contaminating ALV-A from vaccine virus seeds, thus providing a new approach for improving vaccine safety.
Topics: Avian Leukosis Virus; Decontamination; Immune Sera; Immunologic Factors; Newcastle disease virus; Reverse Transcriptase Inhibitors; Viral Vaccines; Zidovudine
PubMed: 30544189
DOI: 10.3382/ps/pey257