BACKGROUND: Interleukin 10 (IL-10) is a homodimeric cytokine that shows considerable clinical promise. Adeno-associated virus (AAV) vectors appear increasingly useful for in vivo gene-transfer applications. METHODS: A recombinant AAV type 2 vector encoding human IL-10 (rAAVhIL10) was constructed by using an adenoviral-free, three-plasmid co-transfection. Cytokine production was measured by using an enzyme-linked immunosorbent assay. Endotoxic shock was induced by lipopolysaccharide (LPS) injection. RESULTS: As media from rAAVhIL10-infected COS cells caused a dose-dependent blockade of IL-12 secretion from spleen cells of IL-10 knockout (KO) mice challenged with Brucella abortus, it was clear that vector-derived hIL-10 was biologically active in vitro. Intravenous or intramuscular administration of relatively modest levels of rAAVhIL10 (10(10) genomes) to IL-10 KO mice resulted in hIL-10 secretion into the bloodstream, which, at 8 weeks, gave median serum levels of 0.9 and 0.45 pg/ml, respectively. Acute endotoxic shock led to a 33% mortality rate, and severe morbidity, in control IL-10 KO mice, whereas no mortality and little morbidity were seen in IL-10 KO mice given rAAVhIL10 7 weeks earlier. CONCLUSIONS: The findings demonstrate that a modest dose of rAAVhIL10 administered in vivo provides long-term protection against LPS-induced endotoxic shock in a murine model. Thus, this vector may be useful for clinical applications requiring sustained IL-10 expression, for example in the treatment of several autoimmune diseases.

Animals↗

PubMedDate not supplied

In situ fixation of the neonatal brain and spinal cord.

A delay in the autopsy can result in significant tissue autolysis, especially in the central nervous system. We have developed a rapid technique of in situ fixation that preserves central nervous system tissues until the formal autopsy can be performed. Through the lateral margin of the anterior fontanelle, Zamboni's solution is injected percutaneously into the lateral ventricles and allowed to exit via an intrathecal spinal needle. The choice of fixative allows a wide array of postmortem studies to be done.

Autopsy↗

PubMedDate not supplied

Case management of HIV-infected children in Missouri.

The early referral of HIV-exposed children and their mothers to coordinated medical and social services has become increasingly important. In July 1989, the Missouri Department of Health initiated the Service Coordination Program to provide individualized referral (case management) for Missouri residents who were reported to have acquired immunodeficiency syndrome (AIDS) or HIV infection. The purpose of the Service Coordination Program is to assist persons in accessing medical and social services. The authors describe the characteristics of the 36 children (18 enrolled in the Service Coordination Program, and 18 not enrolled) reported to the Missouri Department of Health through September 1992. Although more detailed evaluations are necessary, preliminary data suggest that opportunities for early intervention may be facilitated by the Service Coordination Program if the child's HIV status is recognized early.

Child↗