ABSTRACT
An increasing amount of research has been conducted on immunoglobulin Y (IgY) because the use of IgY offers several advantages with respect to diagnostic testing, including its easy accessibility, low cost and translatability to large-scale production, in addition to the fact that it can be ethically produced. In a previous work, immunoglobulin was produced and purified from egg yolks (IgY) reactive to hepatitis A virus (HAV) antigens. In the present work, this anti-HAV-specific IgY was used in an indirect immunofluorescence assay to detect viral antigens in liver biopsies that were obtained from experimentally infected cynomolgus monkeys. Fields that were positive for HAV antigen were detected in liver sections using confocal microscopy. In conclusion, egg yolks from immunised hens may be a reliable source for antibody production, which can be employed for immunological studies.
Subject(s)
Animals , Hepatitis A virus/immunology , Hepatitis A/diagnosis , Immunoglobulins/analysis , Liver/virology , Disease Models, Animal , Fluorescent Antibody Technique, Indirect , Hepatitis A Antibodies/immunology , Hepatitis A Antigens/immunology , Hepatitis A/immunology , Macaca fascicularis , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To find a suitable cell line for hepatitis A antigen expressed by vaccinia virus vector and to find a way of inactivation and preservation of the HAV recombinant antigen. Methods Series of cell lines such as K4,143, HEL, Hep-2 and Vero were inoculated with vaccinia virus that can express HAV recombinant antigen. ELISA was used to determine the contents of expression antigen. The characterization of the HAV antigen expressed by vaccinia virus was then analyzed after it was treated with different methods.</p><p><b>RESULTS</b>The expression of HAV recombinant antigen in K4,143 and HEL cell lines was a little more than expression in Hep-2 and Vero cell lines. The antigenicity is obviously higher when HAV recombinant antigen was inactivated by beta-propiolactone other than it was inactivated by formalin. It was best to preserve the prepared HAV recombinant antigen under -40 degrees C condition.</p><p><b>CONCLUSIONS</b>The application of vaccinia virus vector in hepatitis A antigen preparation was very useful and promising.</p>
Subject(s)
Animals , Humans , Cell Line , Enzyme-Linked Immunosorbent Assay , Formaldehyde , Pharmacology , Genetic Vectors , Hepatitis A Antigens , Genetics , Allergy and Immunology , Hepatitis A Vaccines , Allergy and Immunology , Propiolactone , Pharmacology , Recombinant Proteins , Allergy and Immunology , Vaccines, Synthetic , Allergy and Immunology , Vaccinia virus , GeneticsABSTRACT
Polyarteritis nodosa [PAN] first described by Kussmaul and Maier in 1866, is a multisystem necrotizing vasculitis of small and middle-sized muscular arteries. The presence of hepatitis B antigenemia [Hbs Ag] in approximately 30% of patients with PAN as well as immune complexes of Hbs Ag-Immunoglobulins and complement in the blood vessel walls strongly suggest the role of immunologic phenomena. The extremely poor prognosis of classic PAN has been modified by corticosteroid treatment with boluses of cyclophosphamide, and plasmapheresis. We report a case of PAN with renal, cardiac, central and peripheral nervous system involvement associated with active hepatitis B that got a total remission with corticosteroids, lamivudine and boluses of cyclophosphamide without plasmapheresis
Subject(s)
Humans , Male , Hepatitis A Antigens , Immunosuppressive Agents , Antiviral Agents , Cyclophosphamide , Lamivudine , PlasmapheresisABSTRACT
Studies were carried out to analyse the ultrastructural changes and the distribution of hepatitis A virus (HAV)/antigens at subcellular level in buffalo green monkey kidney (BGMK) cells persistently infected with HM-175 strain of HAV. HAV infected BGMK cells showed distinct abnormalities in the endoplasmic reticulum and cytoplasmic membrane as compared to uninfected cells. The abnormalities were characterized by wavy arrays, structures like myelin, annulate lamellae, cytoplasmic inclusion bodies and vesicles. The wavy arrays within the cytoplasm of the host cells appeared to represent degenerating membranes. A complex myelin like body was found in close association with a group of virus like particles. Annulate lamellae like structures involving single paired membrane were detected infrequently whereas the cytoplasmic vesicles were numerous in these cells. An indirect immunogold technique was utilized to localize the HAV antigenin infected cells. A high density immunogold label for HIV like particles was predominantly detected in cytoplasmic vesicles. These results suggest a strong association of membrane substructure in vesicle forms with the compartmentalized replication of HAV within persistently infected host cells.
Subject(s)
Animals , Antigens, Viral/analysis , Cell Line , Fluorescent Antibody Technique, Indirect/methods , Hepatitis A Antigens , Hepatovirus/isolation & purification , Immunohistochemistry/methods , Kidney/cytology , Microscopy, ElectronABSTRACT
Sera from healthy donors and four groups of subjects with acute viral hepatitis were tested for anti-HAV IgM by ELISA with hepatitis A virus antigen grown in tissue culture. The results were compared with those obtained by formalin inactivated HAV from 'HAVAB-M' test kit (Abbott Laboratories, USA). With both preparations of antigens, sera from healthy donors and patients suffering from acute hepatitis B or non-A, non-B did not show anti-HAV IgM antibodies whereas acute phase sera from hepatitis A patients showed strong reactions indicating the presence of anti-HAV IgM antibodies. The results indicate similarity in the specificity of both preparations of HAV against anti-HAV antibodies and encourage the use of tissue culture derived HAV for serological diagnosis of hepatitis A.