Chicken anemia virus (CAV) is an important viral pathogen that causes anemia and severe immunodeficiency syndrome in chickens worldwide. In this study, a potential diagnostic monoclonal antibody against the CAV VP1 protein was developed which can precisely recognize the CAV antigen for diagnostic and virus recovery purposes. The VP1 gene of CAV encoding the N-terminus-deleted VP1 protein, VP1Nd129, was cloned into an Escherichia (E.) coli expression vector. After isopropyl-beta-D-thiogalactopyronoside induction, VP1Nd129 protein was shown to be successfully expressed in the E. coli. By performing an enzyme-linked immunoabsorbent assay using two coating antigens, purified VP1Nd129 and CAV-infected liver tissue lysate, E3 monoclonal antibody (mAb) was found to have higher reactivity against VP1 protein than the other positive clones according to the result of limiting dilution method from 64 clones. Using immunohistochemistry, the presence of the VP1-specific mAb, E3, was confirmed using CAV-infected liver and thymus tissues as positive-infected samples. Additionally, CAV particle purification was also performed using an immunoaffinity column containing E3 mAb. The monoclonal E3 mAb developed in this study will not only be very useful for detecting CAV infection and performing histopathology studies of infected chickens, but may also be used to purify CAV particles in the future.
Animals ; Antibodies, Monoclonal/biosynthesis/genetics/*immunology ; Antigens, Viral/analysis ; Capsid Proteins/genetics/*immunology ; Chicken anemia virus/genetics/*immunology ; *Chickens ; Circoviridae Infections/blood/immunology/*veterinary/virology ; Escherichia coli/genetics ; Immunohistochemistry/veterinary ; Liver/virology ; Mice ; Mice, Inbred BALB C ; Microscopy, Fluorescence/veterinary ; Poultry Diseases/blood/immunology/*virology ; Specific Pathogen-Free Organisms ; Thymus Gland/virology

OBJECTIVETo investigate clinicopathological features of DiGeorge syndrome (DGS).

METHODThe clinical features, histological and immunohistochemical findings were analyzed in 5 cases of DGS by autopsy.

RESULTSFive cases of DGS in male infants aged 4 days, 1 month, 7 months, 10 months, and 13 months respectively. Gross and microscopic observations revealed that thymic cortex was depleted of lymphocytes or showed few, dispersed lymphocytes. The thymic medulla showed predominantly epithelial cells with calcified Hassall bodies as well as lymphocyte depletion. T lymphocytes were also scarce in the tonsils, lymph nodes, spleen, and mucosa-associated lymphatic tissue of ileum. In addition, 3 of the 5 patients also showed parathyroid aplasia or dysplasia, and congenital hypertrophy of the ventricular septum.

CONCLUSIONSThe pathological changes indicate that clinicians should be aware of defects of immune system if the infants suffer from severe infections. Pathologists should recognize the importance of abnormalities of lymphohematopoietic tissues in the diagnosis of primary immunodeficiency diseases such as DGS.

Autopsy ; DiGeorge Syndrome ; immunology ; pathology ; virology ; Hepatitis, Viral, Human ; pathology ; Humans ; Hypertrophy, Left Ventricular ; pathology ; Infant ; Infant, Newborn ; Lymphocyte Count ; Male ; Parathyroid Glands ; pathology ; Pneumonia, Viral ; pathology ; T-Lymphocytes ; immunology ; pathology ; Thymus Gland ; pathology