The role of eosinophils and neutrophils in helminth-induced keratitis.

PURPOSE: Intrastromal injection of mice with antigens from the parasitic helminth that causes river blindness (Onchocerca volvulus) induces eosinophil recruitment to the corneal stroma at the time of maximum corneal opacification and neovascularization. The present study was conducted to examine the role of eosinophils and neutrophils in onchocercal keratitis in control C57Bl/6 mice and in interleukin-5 gene knockout (IL-5(-/-)) mice. METHODS: C57Bl/6 and IL-5(-/-) mice were immunized subcutaneously and injected intrastromally with soluble O. volvulus antigens. Mice were killed at various times thereafter. Development of keratitis was assessed by slit lamp examination, and inflammatory cells in the cornea were identified by immunohistochemistry. RESULTS: A biphasic recruitment of inflammatory cells was observed in C57Bl/6 mice; neutrophils predominated during the first 72 hours after intrastromal injection and subsequently declined, whereas eosinophil recruitment increased as time elapsed and comprised the majority (90%) of cells in the cornea by day 7. In contrast, neutrophils were the predominant inflammatory cells in IL-5(-/-) mice at early and late time points and were associated with extensive stromal damage and corneal opacification and neovascularization. Eosinophils were not detected in these mice at any time. CONCLUSIONS: In the absence of eosinophils, neutrophils can mediate keratitis induced by helminth antigens. Together with the early neutrophilic infiltrate in control animals, these observations indicate that neutrophils have an important role in onchocercal keratitis.

Occurrence, clinical manifestation and the epidemiological implications of naturally occurring canine trypanosomosis in western Kenya.

Domestic dogs were screened for Trypanosoma brucei infection using the haematocrit centrifugation technique as part of routine active surveillance exercises in the Busia and Teso districts of Kenya. The purpose was to assess the role of dogs as sentinels for the occurrence of human sleeping sickness. Out of 200 dogs screened, five were found to be infected at the various test sites. These five succumbed to the disease within four weeks, and exhibited a distinct and pronounced corneal opacity before death. Blood from two naturally infected dogs were tested for the presence of the serum resistance associated (SRA) gene and one tested positive, confirming it as human infective (T. brucei rhodesiense) prevalence (0.5%). It is considered that the occurrence of this clinical sign could be used as an early warning prediction of future outbreaks. This type of prediction could form an integral part of an indigenous technical knowledge set in areas lying at the edges of the tsetse (Glossina) belts where T. brucei is the main trypanosome species that affects dogs. The occurrence of corneal opacity in dogs could indicate a rise in the levels of T. brucei a proportion of which could be human infective T. b. rhodesiense circulating in the population early enough before disease outbreak occurs. It is thought that during sleeping sickness epidemics the domestic dog will be the first casualty rapidly succumbing to disease long before it is noticed in man. Prompt prediction of disease outbreaks would thus enable early interventions that would reduce the morbidity, mortality and the general economic losses associated with sleeping sickness to be instituted.

Immune mechanisms in Onchocerca volvulus-mediated corneal disease (river blindness).

Infection with the parasitic nematode Onchocerca volvulus can lead to severe visual impairment and blindness. In an effort to characterize the molecular basis for the inflammatory response in the cornea, we have developed a murine model for O. volvulus-mediated keratitis in which parasite antigens are injected into the corneal stroma of sensitized mice. This model reproduces the two main clinical features of human disease, corneal opacification and neovascularization. Histological analysis of corneas from these mice reveals a biphasic recruitment of neutrophils and eosinophils to the central cornea, along with a small, but persistent number of CD3+ cells. In this review, we present evidence that production of antigen-specific T cell and antibody responses are essential for development of O. volvulus keratitis, and we propose a sequence of molecular and cellular events that lead to migration of inflammatory cells to the cornea and to loss of corneal clarity.

The microfilarial load in the anterior segment of the eye. A parameter of intensity of onchocerciasis.

The presence of microfilariae of Onchocerca volvulus in the eye is associated with an increased risk of deterioration of existing eye lesions. An opthalmological and parasitological examination of 630 persons was carried out in a hyperendemic focus of onchocerciasis in northern Togo. The prevalence of microfilariae increased in the cornea as well as the anterior chamber up to the age of 40-50 years, then decreased. The prevalence of onchocercal punctate keratitis, on the other hand, showed a peak for the age group 10-20 years. In two-thirds of the cases microfilariae were present in the anterior chamber as well as in the cornea. The relative distribution of microfilariae between the anterior chamber and the cornea did not change with the development of severe anterior lesions but in cases with severe posterior lesions relatively more microfilariae were found in the anterior chamber than in the cornea. In all cases of severe ocular lesions the numbers of microfilariae both in the anterior chamber and in the cornea were increased. The average number of microfilariae in the eye can be used as a parameter to enumerate the severity of ocular onchocerciasis.

CXC chemokine receptor 2 but not C-C chemokine receptor 1 expression is essential for neutrophil recruitment to the cornea in helminth-mediated keratitis (river blindness).

Infiltration of neutrophils and eosinophils into the mammalian cornea can result in loss of corneal clarity and severe visual impairment. To identify mediators of granulocyte recruitment to the corneal stroma, we determined the relative contribution of chemokine receptors CXC chemokine receptor (CXCR)-2 (IL-8R homologue) and CCR1 using a murine model of ocular onchocerciasis (river blindness) in which neutrophils and eosinophils migrate from peripheral vessels to the central cornea. CXCR2(-/-) and CCR1(-/-) mice were immunized s.c. and injected into the corneal stroma with Ags from the parasitic helminth Onchocerca volvulus. We found that production of macrophage-inflammatory protein (MIP)-2, KC, and MIP-1 alpha was localized to the corneal stroma, rather than to the epithelium, which was consistent with the location of neutrophils in the cornea. CCR1 deficiency did not inhibit neutrophil or eosinophil infiltration to the cornea or development of corneal opacification. In marked contrast, neutrophil recruitment to the corneas of CXCR2(-/-) mice was significantly impaired (p < 0.0001 compared with control, BALB/c mice) with only occasional neutrophils detected in the central cornea. Furthermore, CXCR2(-/-) mice developed only mild corneal opacification compared with BALB/c mice. These differences were not due to impaired KC and MIP-2 production in the corneal stroma of CXCR2(-/-) mice, which was similar to BALB/c mice. Furthermore, although MIP-1 alpha production was lower in CXCR2(-/-) mice than BALB/c mice, eosinophil recruitment to the cornea was not impaired. These observations demonstrate the critical role for CXCR2 expression in neutrophil infiltration to the cornea and may indicate a target for immune intervention in neutrophil-mediated corneal inflammation.