Kendall O. Smith

Cryopreservation of varicella-zoster virions without loss of structural integrity or infectivity.

Varicella-zosterer virions present in infected cells or in a cell-free state were freeze-dried without loss of structural integrity of infectivity. Generally, yields of greater than 5 log10 foci/ml (infected cells) or greater than 4 log10 PFU/ml (cell-free virus) were recovered from varicella-zoster virus-infected human melanoma cells both before and after lyophilization in phosphate-buffered media containing 0.1-1.0 M sucrose. Virus frozen in solutions lacking sugar had little or no residual infectivity after vacuum sublimation was completed. Visualization by electron microscopy demonstrated large numbers of enveloped virions in the virus preparations lyophilized in media containing sucrose; in marked contrast, virus subjected to freeze-drying in buffered solutions without sugar consisted mainly of naked nucleocapsids. Water analyses by Karl Fischer titration suggested that residual moisture retained by sugar prevented disenvelopment of the varicella-zost virion.

Low-temperature storage of surface-attached living cell cultures

A process is described in which surface attached living cell cultures are treated, cooled, stored and revived in such a manner as to retain their viability and their adherence to culture surfaces.

Evidence for chronic viral infections in human arteries.

Summary Evidence is presented that measlesvirus antibody and herpesvirus antibody are sometimes present in human aortic tissues in a state not readily dissociable by extensive washing with buffered physiologic saline. This antibody is readily released from the tissues by high molarity potassium iodide solution, a treatment known to dissociate antigen-antibody complexes. We postulate: (1) that viral antigen-antibody complexes are sometimes present in aortic tissues; and (2) that a significant cause of vascular tissue injury in humans may be viral infection of the blood vessels themselves and/or deposit of viral antigen-antibody complexes in the vascular tissues.

Radioimmunoassay procedure for quantitating bacterial antibody in human sera

A RIA system was developed to detect antibodies in human sera against bacteria. Sonicates of Escherichia coli and Fusobacterium polymorphum were used as antigens to sensitize plastic-coated beads; antibodies to these antigens were detected with 125I-labeled antihuman globulin. Serum antibody titers against E. coli were determined by the serial dilution method; from the results the standard curve principle was applied in determining the relative amounts of antibodies against E. coli in serum samples tested at a single dilution. The coefficient of variation of the RIA procedure was less than 10%. Serum titers obtained by the RIA and indirect immunofluorescence test were compared; RIA was more sensitive, quantitative and objective. Absorption studies, using E. coli and F. polymorphum absorbentes against E. coli and F. polymorphum. This RIA procedure offers a combination of desirable advantages; it is sensitive, specific, objective, quantitative, and easy to perform.

Enzyme-linked immunosorbent assay (ELISA) for HIV antibody by a glass slide technique

An enzyme-linked immunosorbent assay (ELISA) technique is described which utilizes a commercially available glass microscope slide coated with hydrophobic teflon in such a pattern as to give 30 small circular wells, each of which has a glass bottom. Each well serves as a solid phase, analogous to a microtiter well for adsorption of purified human immunodeficiency virus (HIV) antigens. Since only 5-10 microliter volumes of reagents are used and rinsing processing is simple, the cost per test is much less than most other ELISA technologies. HIV antigen is stable for over 1 year at 37 degrees C when dried on the glass slides. The sensitivity and specificity of the micro slide immunoenzymatic assay (Micro-SIA) was studied by testing randomly selected, known HIV-seropositive and seronegative plasma. Results compare well with microtiter and Western blot assays. A simple vertical-beam colorimeter is described (useful in the Micro-SIA) which can be easily assembled by the user from commonly available components.

A Serological Study of Herpes simplex Virus Type 1 Antibody over a 13-Year Period

HSV-1 serum antibody titers determined by 50% plaque reduction and confirmed by radioimmunoassay in 11 samples from an individual over a 13-year period indicated a significant increase between the first sample and a sample taken 9 years later. This increase did not seem to reflect loss of antibody in the low titered serum sample due to storage.

Neutralizing Antibody Against Infectious Canine Hepatitis Virus in Veterinary Workers’Sera (ICHV)

Summary A high prevalence of neutralizing antibody to ICHV was found in the sera of veterinary and nonveterinary workers. When sera from veterinary and nonveterinary workers were paired according to age, sex and ethnic classification, veterinary workers were found to have significantly higher antibody titers than nonveterinary controls.