Sanford H. Stone

Chronic relapsing experimental allergic encephalomyelitis: Identification and dynamics of T and B cells within the central nervous system

Abstract Using a monoclonal antibody against guinea pig T cells and anti-guinea pig immunoglobulins, T- and B-cell dynamics were studied by immunofluorescence in situ in the central nervous system (CNS) of animals with untreated and treated chronic relapsing experimental allergic encephalomyelitis (EAE). Treated animals were given a series of injections of either myelin basic protein (MBP) in incomplete Freunds adjuvant (IFA) or MBP and galactocerebroside in IFA. Within the CNS, T and B cells showed distinct distribution patterns in untreated chronic relapsing EAE, similar to that recently described in acute EAE. T cells were predominantly localized within the CNS parenchyma and B cells were mainly found in perivascular areas. B-cell infiltrates were more extensive than in acute EAE and, although most were centered around blood vessels, some were also detectable in the parenchyma. IgG, C 3 , and albumin deposits were common. These observations suggest an age-dependent difference in the immune response. In treated chronic EAE, the disease process was apparently arrested and T- and B-cell infiltrates in the white matter were negligible. Therefore, it appears that the present treatment protocol prevents lymphocytes from entering the CNS parenchyma.

Chronic relapsing experimental autoimmune encephalomyelitis ☆: Treatment with combinations of myelin components promotes clinical and structural recovery

Preliminary results are presented on the treatment of Strain 13 guinea pigs with chronic relapsing experimental autoimmune (allergic) encephalomyelitis (EAE) induced by a single sensitisation with whole spinal cord. Animals were treated at different stages of the disease with injection containing either myelin basic protein (MBP) alone in incomplete Freunds adjuvant (IFA), or MBP in combination with a lipid hapten of myelin, galactocerebroside (GC) in IFA. The rationale for this treatment stemmed from previous work which suggested that MBP was responsible for T cell sensitisation in EAE and that GC was important in producing demyelinating antibodies and that both myelin components were needed in the induction of disease. Although treatment with MBP alone caused some initial stabilisation of the disease process, subsequent relapses occurred in all animals. However, in animals given MBP and GC together, either early or late in the course of the disease, marked clinical improvement has been noted with little or no development of relapses over an observation period of more than one year post-treatment. In addition, evidence of extensive remyelination and oligodendroglial proliferation in CNS lesions has been found in MBP-GC-treated animals suggesting that this therapy might be beneficial for CNS repair and relevant to multiple sclerosis.

Association of hereditary cataracts in strain guinea-pigs with mutation of the gene for ζ-crystallin

Abstract Congenital nuclear cataracts transmitted by an autosomal dominant gene are present in a line of strain 13 N guinea-pigs. Studies on the lens proteins from these animals demonstrate changes in both the composition and structure of the crystallins relative to normal controls. The most prominent difference is in the ζ-crystallin, a taxon-specific crystallin which has been shown to be related to the alcohol dehydrogenases. In animals homozygous for the cataract phenotype the normal ζ-crystallin polypeptide is absent from the lens. Quantitation is difficult in the cataractous lenses from heterozygotes because of protein changes secondary to opacification: however in liver and kidney which have catalytic levels of the protein, the concentrations are approximately half that present in tissue from normal control animals. These findings suggest that in the cataractous animals a mutation has occurred in the gene for ζ-crystallin. In addition, a novel protein which is very similar to ζ-crystallin is synthesized only in the lenses of animals with cataract. This protein appears to be the product of the mutant gene for ζ-crystallin. These data support the hypothesis that this hereditary congenital cataract results from a specific mutation in the ζ-crystallin gene.

Acute experimental autoimmune encephalomyelitis: T- and B-cell distribution within the target organ

Abstract The sequence of T-cell events in the central nervous system (CNS) has been further studied during acute experimental autoimmune (allergic) encephalomyelitis (EAE). T cells were labeled by indirect fluorescence using a monoclonal antibody against guinea pig T cells. In addition, B cells, IgG, C 3 , and albumin were localized by direct fluorescence. T cells were demonstrable as early as 5 days postinoculation (PI), well before the onset of signs and were predominantly found within the CNS parenchyma. B cells and macrophages appeared later (10 days PI) and were restricted to the meninges and perivascular spaces. IgG and albumin were found from day 5 PI onwards and were probably indicative of damage to the blood-brain barrier. C 3 was observed later and was distributed similar to IgG. These findings (a) further underline the primary involvement of T cells in this autoimmune condition, (b) indicate a difference in the migratory activity of T and B cells in the CNS, (c) suggest that suppression of clinical EAE with myelin basic protein can be correlated with a diminution of inflammatory cells within the CNS, and (d) represent an unambiguous demonstration of T cells in situ in the target organ in EAE.

Chronic relapsing experimental allergic encephalomyelitis ☆: Correlation of circulating lymphocyte fluctuations with disease activity in suppressed and unsuppressed animals

Groups of juvenile Strain 13 guinea pigs sensitized for chronic relapsing experimental allergic encephalomyelitis (EAE) with isogeneic central nervous system (CNS) tissue in complete Freunds adjuvant (CFA) were either left to develop late-onset chronic EAE (unsuppressed), or given a series of injections of bovine myelin basic protein (MBP) in incomplete Freunds adjuvant (IFA) to suppress the disease. All unsuppressed animals developed disease and all suppressed animals remained healthy over a 27-month period of study. some unsuppressed and suppressed animals were rechallenged with CNS tissue in CFA 12 or 26 months post-inoculation (PI). Unsuppressed animals all became sick 2-4 weeks after rechallenge, while rechallenged, suppressed animals were protected, indicating that the suppression was permanent. Pathologic findings in the CNS complemented the clinical changes. Circulating lymphocyte studies were performed on animals from all groups. Early (active, high-affinity rosetting) T cell levels in unsuppressed animals showed significant decreases during exacerbations (P less than 0.01) and normal values during remissions. After rechallenge, circulating early T cells decreased in unsuppressed animals with the development of signs. In suppressed animals, early T cells showed significant elevations during, and for a short time after, the period of suppressive injections, and normal values afterwards. These levels did not change significantly after rechallenge. Late (total, 24 hour rosetting) T cell and B cell values showed minor fluctuations only which did not correlate with disease activity. These results indicate that chronic relapsing EAE can be successfully suppressed with MBP in IFA, that this suppression is permanent and that the immunologic findings presented correlate well with the clinical and pathologic facets of the disease. the findings are presented in terms of their relevance to multiple sclerosis.

Suppression of acute and chronic experimental allergic encephalomyelitis in strain 13 guinea pigs: A clinical and pathological study☆

Adult inbred Strain 13 guinea pigs develop an acute, fatal form of experimental allergic encephalomyelitis (EAE) about 2 weeks after a single injection of isologous spinal cord in complete Freunds adjuvant (CFA), but similarly injected juveniles develop a delayed, rarely fatal chronic form. Thirty-seven sensitised adult Strain 13 animals were separated into 2 groups. One group was permitted to develop acute EAE. The other group was injected intramuscularly with 1 mg of guinea pig or bovine myelin basic protein (MBP) in incomplete Freunds adjuvant (IFA) on day 2, 7 or 10 post-inoculation (PI) followed by 0.2 mg in IFA every third day for a total of 10 doses. Animals in the unsuppressed group succumbed to acute EAE 13-16 days post-sensitisation. No animal in the suppressed group died during this period. Animals treated with MBP beginning 2 days PI showed no clinical signs, but mild clinical manifestations occurred in animals suppressed from days 7 and 10 PI. These signs remitted by 21 days post-sensitisation. One suppressed animal (out of 21) died during the fourth week postsensitisation. The other 20 suppressed animals appeared clinically normal towards the end of the course of MBP injections and remained so for the 6 months of study. Morphological examination revealed that CNS lesions occurred in all animals. In animals suppressed with MBP beginning on day 2 PI, lesions consisted only of a few meningeal inflammatory cells. Animals given MBP beginning on day 7 or 10 PI and sampled 1-2 weeks later, had lesions which could not be distinguished from those occurring in the non-suppressed acute EAE group. In time, the suppressed animals developed lesions which were typical of chronic EAE with remyelination as a predominant feature. Preliminary experiments on the suppression of chronic EAE in 5 juvenile Strain 13 guinea pigs have revealed that 3 MBP-injected animals failed to develop clinical disease over a 28-week period of study although lesions typical of chronic EAE were present. Simultaneously, 2 non-suppressed juvenile animals developed clinical signs by 12 weeks. These were associated with both acute inflammation and demyelination superimposed upon regions of chronic demyelinative activity.

Acute and chronic autoimmune encephalomyelitis: age, strain, and sex dependency. The importance of the source of antigen.

Summary A variety of conditions favoring the production of chronic forms of AE existed between the optimal conditions for acute AE (guinea pig antigen in adult Strain 13/N) and for strong resistance to AE (rabbit antigen in newborn Strain 2/N). The resistance of Strain 2/N guinea pigs to autoimmune encephalomyelitis is not absolute. Older Strain 2/N female guinea pigs regularly develop severe or lethal AE when sensitized with guinea pig spinal cord in Freunds complete adjuvant. Guinea pig spinal cord is a more potent encephalitogen than rabbit spinal cord in Strains 2/N and 13/N guinea pigs. Sensitized Strain 2/N guinea pigs of an age group showing no overt clinical signs of AE nevertheless showed striking histological lesions of AE in the central nervous system.

Adoptive Autoimmune Encephalomyelitis in Inbred Guinea Pigs: Immunological and Histological Aspects

Major variables which determine the induction and severity of adoptive autoimmune encephalomyelitis are the age and strain of the animal, and the amount of killed mycobacteria in the adjuvant. Control of these factors results in consistent production of this disease in high incidence and in severe form. The pathologic changes in the central nervous system can be correlated with the clinical disease. Maturity of the target tissues in the central nervous system of the newborn appears to be an important factor which distinguishes the response of the guinea pig from that of other species.

Autoimmune Encephalomyelitis and Ocular Lesions in Monkeys Sensitized during the Neonatal Period

The neonatal rhesus monkey is susceptible to the induction of autoimmune encephalomyelitis. The disease has been produced regularly by injection of neonatal animals with guinea pig spinal cord antigen in complete Freunds adjuvant. The onset of the disease, as compared with onset in adults, is delayed and is most often heralded by intrinsic eye lesions, notably widespread retinal hemorrhages.

Autoimmune Chorioretinitis in Rhesus Monkeys

Monkeys injected with monkey retinal tissue incorporated in Freunlds complete adjuvant developed ocular lesions characterized by choroiditic patches in the fundus periphery and sheathing of retinal vessels. Bovine retina, monkey choroid plexus, and guinea pig kidney were ineffective in this respect.