ABSTRACT
By reversing the usual order of double-staining procedures, we obtained optimal conditions for simultaneous detection of guinea pig lymphotropic herpesvirus (GPHLV) and surface immunoglobulin G-positive (SIgG+) cells in paraffin-embedded tissue sections. Of the different fixatives tested, Bouin's solution gave the best preservation of morphology and cell surface IgG. Double immunolabeling was best achieved when avidin-biotin complex immunoperoxidase (ABC-IP) staining was performed first for detection of intracellular viral antigen, followed by immunogold-silver staining (IGSS) for localization of cell surface IgG.
Subject(s)
Antigens, Viral/analysis , Herpesviridae Infections/diagnosis , Herpesviridae/isolation & purification , Immunoglobulin G/analysis , Receptors, Antigen, B-Cell/analysis , Animals , Female , Guinea Pigs , Herpesviridae/immunology , Immunoenzyme Techniques , Immunohistochemistry , Lymph Nodes/immunology , Lymph Nodes/microbiology , Spleen/immunology , Spleen/microbiologyABSTRACT
Female guinea pigs pretreated for 2 days with cyclosporin A (CsA), were then inoculated intranasally (IN) or intraperitoneally (IP) with a lymphotropic herpesvirus (GPHLV) and followed by 4 additional daily doses of CsA. Immunosuppressed animals did not show lymphocytosis and virus infectivity titers in their spleen, cervical lymph node and blood mononuclear cells were lower than those of oil-treated controls. While IN-inoculated CsA-treated animals expressed higher virus infectivity titer in their lungs compared to oil-treated controls, this difference was not seen in the IP-inoculated groups. At histopathology, lymphoid depletion was seen in all CsA-treated animals, but lymphocytic interstitial pneumonia was observed only in the lungs of IN-inoculated CsA-treated guinea pigs. Thus, CsA immunosuppression altered the pathogenesis of primary GPHLV infection with some notable differences attributed to the route of virus inoculation.
