Temporal expression of inflammatory mediators in brain basilar artery vasculitis and cerebrospinal fluid of rabbits with coccidioidal meningitis

Abstract

SummaryStrokes due to transmural vasculitis associated with coccidioidal meningitis result in significant morbidity and mortality. The immunological and inflammatory processes responsible are poorly understood. To determine the inflammatory mediators, i.e. cytokines, chemokines, iNOS, matrix metalloproteinase-9 (MMP-9), that possibly contribute to vasculitis, temporal mRNA expression in brain basilar artery samples and MMP-9 protein in the CSF of male NZW rabbits infected intracisternally with 6·5 × × × × 10 4 arthroconidia of Coccidioides immitis were assessed. Five infected and 3 sham-injected rabbits at each time point were euthanized 4, 9, 14 and 20 days post infection. All infected rabbits had neurological abnormalities and severe vasculitis in the basilar arteries on days 9-20. In basilar arteries of infected animals versus controls, mRNAs encoding for IL-6, iNOS, IFN-γ γ γ γ , IL-2, MCP-1, IL-1β β β β , IL-10, TNF-α α α α , CCR-1, MMP-9, TGF-β β β β , as well as MMP-9 protein in CSF, were found to be significantly up-regulated. Thus, this study identified inflammatory mediators associated with CNS vasculitis and meningitis due to C. immitis infection. Assessment of the individual contribution of each mediator to vasculitis may offer novel approaches to the treatment of coccidioidal CNS infection. This study also provides unique methodology for immunology studies in a rabbit model.

Expression change of interleukin-8 gene in rabbit basilar artery after subarachnoid hemorrhage

“…Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene served as an inner reference. Primers were determined according to the reported literatures [3,4] . The primer sequences for IL-8 were: forward: 5'-CAT GGA TCT GTC GTA GGG CT-3', reverse: 5'-CTG ACC AAC AGA CCA GGG TT-3' (with a desired product of 390-bp); and for GAPDH were: forward: 5'-TCA CCA TCT TCC AGG AGC GA-3', reverse: 5'-CAC AAT GCC GAA GTG GTC GT-3' (with a desired product of 293-bp).…”

Comparative Efficacies of Lipid-Complexed Amphotericin B and Liposomal Amphotericin B against Coccidioidal Meningitis in Rabbits

“…Dodge Animal Health, Ft. Dodge, IA), xylazine (The Butler Co., Dublin, OH), and acepromazine (Butler) and infected by direct cisternal puncture with 5 ϫ 10 4 Coccidioides posadasii strain Silveira arthroconidia (1,2,5,6,(12)(13)(14). All procedures in these studies were done with the approval of the Animal Care and Use Committee of the California Institute for Medical Research.…”

“…An experimental model of coccidioidal meningitis in 3-kg male New Zealand White rabbits (Myrtles Rabbitry, Inc., Thompson Station, TN) was established as described previously (1,2,5,6,(12)(13)(14). In brief, rabbits were anesthetized using a cocktail of ketamine (Ft.…”

Quantitation of rabbit cytokine mRNA by real-time RT-PCR

“…Similarly, the rabbit is an important model for tuberculosis due to its formation of pulmonary cavities, a distinct characteristic of human tuberculosis that other small animal models do not develop (Helke et al, 2005). The rabbit is also used to study a variety of other diseases such as enteropathogenic and enterohemorrhagic E. coli infection (Zhu et al, 2006), meningitis and vasculitis caused by Coccidioides immitis (Zucker et al, 2006), and various ocular diseases including those caused by herpes viruses (Weisbroth et al, 1974;Morishige et al, 2006).…”

“…This method has been supplanted by quantitative real time RT-PCR which offers advantages in terms of labor, sensitivity, and time. Zucker et al very recently reported RT-PCR assays for relative quantitation of a number of pro-inflammatory molecules as well as a few cytokines (Zucker et al, 2006). In this study, we describe the development of a quantitative real-time RT-PCR assay for measuring mRNA for rabbit cytokines IFN-γ, IL-2, IL-4, IL-10, and TNF-α using the LightCycler system (Roche Applied Science, Mannheim, Germany).…”