Gender, age, and season at immunization uniquely influence the genetic control of susceptibility to histopathological lesions and clinical signs of experimental allergic encephalomyelitis: implications for the genetics of multiple sclerosis

Abstract

Multiple sclerosis (MS), the principal inflammatory demyelinating disease of the central nervous system (CNS), is believed to have an immunopathological etiology arising from gene-environment interactions. In this study, we examined the effect of sex, age, and season at immunization on the susceptibility of (B10.S x SJL/J) F(2) intercross mice to experimental allergic encephalomyelitis (EAE), the foremost animal model of MS. Results of logistic regression analyses suggest that female mice were more likely to exhibit CNS lesions than male mice [odds ratio (OR) = 2.28 for brain lesions; OR = 2.37 for spinal cord (SC) lesions]. Although statistically significant associations were seen between brain and SC lesions and age at the time of injection or month of injection when examined separately; these associations disappeared when controlling for sex in multiple logistic regression analyses. These results suggest that the sex of the mouse is more important in influencing the development of brain and SC lesions than was either age or month of immunization. When examining clinical disease as the endpoint, the OR for the age at immunization is 1.04, indicating that the odds of being affected increase by 4% for each increasing week of age. When controlled for age, the OR for injection in the summer months (July through September) is 1.90, suggesting that the odds of being clinically affected are 90% greater for F(2) intercross animals injected in the summercompared to those injected in the winter to spring months (February through May). In contrast to CNS lesions, the age and season at immunization significantly and independently influenced susceptibility to clinical EAE and did so equally in both males and females. Linkage analysis to eae5, the H2-linked locus controlling susceptibility to clinical disease, was performed using 6- to 12- and >12-week-old cohorts as well as summer and winter/spring cohorts of F(2) mice. Significant linkage of clinical EAE to eae5 was observed with the 6- to 12-week-old and summer populations. In contrast, linkage of clinical EAE to eae5 was not detected with the >12-week-old and winter/spring populations. These results indicate that age and seasonal effects are capable of overriding eae5-dependent genetic control of susceptibility to clinical EAE and have significant implications for the genetics of MS.

Figure 1

Characteristic histopathological lesions in murine EAE. A: Normal SC white matter under the pia mater. B: Myelin stained (light blue) in normal SC. C: Pia mater and subpial white matter is infiltrated (perivascular pattern) by acute inflammatory cells consisting predominately of neutrophils and lesser numbers of chronic inflammatory cells (lymphocytes/macrophages). There is extensive destruction of the subpial white matter with swelling and fragmentation of axon sheaths (arrows) and reactive gliosis. D: There is extensive demyelination (loss of light blue staining) in the subpial white matter. E: Normal brain white matter (cerebrum, external capsule). F: Myelin stained (light blue) in normal external capsule of the cerebrum. G: The perivascular space in the external capsule is infiltrated by lymphocytes (arrowhead). H: Demyelination is absent or minimal in areas of chronic inflammation. Left, H&E stain; right, Luxol fast blue-periodic acid-Schiff stain. Scale bar, 50 μm.