Toll-like receptors 4 and 9 are responsible for the maintenance of the inflammatory reaction in canine steroid-responsive meningitis-arteritis, a large animal model for neutrophilic meningitis

Abstract

Background: Steroid-responsive meningitis-arteritis (SRMA) is a systemic inflammatory disease affecting young adult dogs and a potential large animal model for neutrophilic meningitis. Similarities between SRMA and infectious central nervous system (CNS) diseases in lymphocyte subsets suggest an infectious origin.Toll-like receptors (TLRs) are pattern recognition receptors playing an important role in innate immunity. Due to their ability to recognize both self and non-self antigens, we hypothesize that TLRs are among the key factors for the induction of the inflammatory process in SRMA and provide an indirect hint on the etiology of the disease.

Methods: The expression profile of cell surface TLRs (TLR2, TLR4 and TLR5) and intracellular TLRs (TLR3 and TLR9) of canine leukocytes was analyzed by immunophenotyping and subsequent flow cytometric measurements. Experiments were performed on cerebrospinal fluid (CSF) and peripheral blood (PB) samples of dogs affected with SRMA during the acute phase (n = 14) as well as during treatment (n = 23) and compared with those of dogs with bacterial meningitis (n = 3), meningoencephalitis of unknown etiology (n = 6), neoplasia of the central nervous system (n = 6) and a group of dogs with miscellaneous neurological diseases (n = 9). Two additional control groups consisted of dogs with pyogenic infections (n = 13) and of healthy dogs (n = 6).

Results: All examined groups showed a high percentage of TLR2, TLR4 and TLR5 positive PB polymorphonuclear cells (PMNs) in comparison to healthy dogs. Very high values of TLR9 positive PB PMNs were detected in acute SRMA. Only a few similarities were found between SRMA patients and dogs with pyogenic infections, both groups were characterized by high expression of TLR4 positive PB monocytes. Glucocorticosteroid therapy reduced TLR2, TLR4 and TLR9 expression in PB monocytes.

Conclusions: A relatively high expression of TLR4 and TLR9 in acute SRMA suggests that these two receptors might be involved in the inflammatory process in SRMA, enhancing the autoimmune reaction. Systematic CSF cell analysis for TLRs can be performed in future treatment studies in larger animals, such as dogs.

Figure 1

Population of CSF leukocytes: granulocytes (R1, green), lymphocytes (R2, pink) and monocytes (R3, red).

Figure 2

Percentage of TLR positive PMNs in different disease categories. Boxes contain values from the 1^st to the 3^rd quartile, lines inside boxes indicate median values, endpoints of vertical lines display the 5^th to 95^th percentile and · represent the outliers. Asterisks indicate statistically significant differences (* P <0.05; ** P <0.01; *** P <0.005). BM, bacterial meningitis; CSF, cerebrospinal fluid; Healthy, healthy dogs; Mix, miscellaneous diseases of the nervous system; MUE, meningoencephalitis of unknown etiology; PB, peripheral blood; PMNs, polymorphonuclear cells; Pyo, pyogenic infection; SRMA, steroid-responsive meningitis-arteritis; SRMA A, SRMA Acute; SRMA Th, SRMA under therapy; TLR, Toll-like receptor.

Figure 3

Precentage of TLR positive monocytes in different disease categories. Boxes contain values from the 1^st to the 3^rd quartile, lines inside boxes indicate median values, endpoints of vertical lines display the 5^th to 95^th percentile and · represents the outliers. Asterisks indicate statistically significant differences (* P <0.05; ** P <0.01; *** P <0.005). BM, bacterial meningitis; CSF, cerebrospinal fluid; Healthy, healthy dogs; Mix, miscellaneous diseases of the nervous system; MUE, meningoencephalitis of unknown etiology; PB, peripheral blood; PMNs, polymorphonuclear cells; Pyo, pyogenic infection; SRMA, steroid-responsive meningitis-arteritis; SRMA A, SRMA Acute; SRMA Th, SRMA under therapy; TLR, Toll-like receptor.

Figure 4

Percentage of TLR positive lymphocytes in different disease categories. Boxes contain values from the 1^st to the 3^rd quartile, lines inside boxes indicate median values, endpoints of vertical lines display the 5^th to 95^th percentile and · represents the outliers. Asterisks indicate statistically significant differences (* P <0.05; ** P <0.01; *** P <0.005) BM, bacterial meningitis; CSF, cerebrospinal fluid; Healthy, healthy dogs; Mix, miscellaneous diseases of the nervous system; MUE, meningoencephalitis of unknown etiology; PB, peripheral blood; PMNs, polymorphonuclear cells; Pyo, pyogenic infection; SRMA, steroid-responsive meningitis-arteritis; SRMA A, SRMA Acute; SRMA Th, SRMA under therapy; TLR, Toll-like receptor.