Ultrastructure of otic capsule sclerosis in Palmerston North autoimmune mice.

PURPOSE: Numerous temporal bone studies have reported a correlation between systemic autoimmune disease and osteogenic lesions within the inner ear. However, little is known of the cellular mechanisms that relate these two disease processes. The Palmerston North (PN) autoimmune strain mouse exhibits both spontaneous systemic autoimmune disease and otic capsule sclerotic lesions that are similar in many ways to those reported in humans. This suggests the PN mouse is a potential model in which to study the cellular events responsible for immune-related otic capsule lesions. Therefore, an evaluation of the fine structure of the PN modiolus was conducted to better understand these matrix changes of the inner ear. MATERIALS AND METHODS: Inner ears were collected from 15 PN mice at ages from 17 to 24 months and prepared for electron microscopy. The ears were ultrastructurally evaluated to characterize the lesions and their associated cytoarchitecture. RESULTS: The sclerotic lesions consisted of an electron-dense mass that appeared lobulated or layered, usually adjacent to the modiolar bone and blood vessels. Immediately surrounding the lesions were activated fibroblasts and fine fibrillar material in the extracellular space between them. The sclerotic foci often were apposed to normal modiolar bone that never appeared degraded. CONCLUSIONS: The similarities between these bony lesions and known human otic capsule diseases suggests parallel processes are involved. Thus, further study of the PN inner ear may provide insight into the cellular events that underlie otic capsule and other temporal bone alterations in systemic autoimmune diseases.

Inner ear basic fibroblast growth factor in CBA/J, C3H/HeJ, and autoimmune Palmerston north mice.

Basic fibroblast growth factor (bFGF) has a mitogenic effect on fibroblasts and osteoblasts for matrix proliferation and on endothelial cells for neovascularization. Because otic capsule osteogenesis in autoimmune disease subjects often involves abnormal matrix and vascular changes, bFGF may serve as a potential mediator for such bone disorders. To investigate this relationship, bFGF was evaluated in the Palmerston North autoimmune strain mouse, which develops otic capsule sclerotic lesions during the progression of its systemic disease. Inner ears from PN mice, along with control CBA/J and C3H/HeJ mice, were immunohistochemically stained with antibodies against bFGF to identify its presence and possible role in otic capsule disease. Although cells reactive for bFGF were observed along the lining of the otic capsule in all three strains, a significantly higher frequency was observed in the PN mice. Other sites of staining included connective tissue around the tensor tympani muscle and the geniculate ganglion. This identification of bFGF in the otic capsule raises the possibility that it may play some role in normal bone maintenance, as well as abnormal bone or connective tissue remodeling in autoimmune disease.

DNA binding to mouse cells is mediated by cell-surface molecules: the role of these DNA-binding molecules as target antigens in murine lupus.

Autoimmunity to a 28-29-kDa cell-surface DNA-binding molecule has previously been described in patients with systemic lupus erythematosus and related autoimmune diseases. This report describes experiments that implicate a similar antigen-antibody system in the evolution of autoimmunity in lupus-prone mice. DNA binding to murine spleen cells was found to be a saturable phenomenon that was inhibited by excess cold DNA and trypsinization. The role of autoimmunity to murine cell-surface DNA-binding molecules in lupus-prone mice (MRL lpr/lpr, MRL +/+, BXSB) was compared to normal mice (BALB/c, C3H.SW) by means of an assay that measured the inhibition of cell-surface DNA binding. Only sera from lupus strains had inhibitory activity and this component was shown to be an IgM autoantibody. Furthermore, we isolated a spontaneously occurring IgM monoclonal antibody from the spleen of an MRL/lpr mouse, which inhibited DNA binding to mouse cells. Time-course studies indicated that young female MRL/lpr mice lacked detectable activity against cell-surface DNA-binding molecules; however, by 8-10 weeks maximal inhibitory activity was observed. This response occurred prior to the development of significant antinuclear antibody activity. With the appearance of overt disease and anti-DNA antibodies, inhibition of DNA-binding activity became undetectable. These findings mirror previous studies on autoimmunity to a cell-surface DNA-binding molecule on human leucocytes, but have the added advantage of permitting the study of the temporal evolution of this inhibitory activity in relation to disease expression.

Lacrimal and salivary gland inflammation in the C3H/Ipr autoimmune strain mouse: a potential mode for Sjögren's syndrome.

Sjögren's syndrome is an autoimmune inflammatory disease that affects the lacrimal and salivary glands. To identify a potential animal model for study of Sjögren's syndrome, an evaluation was made of lacrimal and salivary glands in the C3H/Ipr autoimmune strain mouse at ages before (2 months) and after (5 months) systemic autoimmune disease onset at 3 to 4 months. Quantitative and qualitative analyses of C3H/Ipr lacrimal and salivary (parotid, submandibular, and sublingual) gland histopathology were performed using age-matched C3H/HeJ nonautoimmune mice to control for inflammation of nonautoimmune origin. No lacrimal or salivary gland inflammation was seen in either of the strains at 2 months of age and measures of systemic autoimmune disease were negative. At 5 months of age, the nonautoimmune C3H/HeJ controls showed a slight increase in lacrimal gland inflammation, but this was not significantly different from the 2 month old controls. A significant increase in lacrimal gland inflammation was found in the 5 month old C3H/Ipr autoimmune mice in a histologic pattern similar to that of Sjögren's syndrome in human beings. Furthermore, the degree of inflammation was positively correlated with serum immune complexes and spleen weight. Sporadic inflammation of the submandibular gland was seen in both autoimmune and control mice, but this was neither statistically significant nor correlated with measures of autoimmunity. No significant inflammation was seen in the parotid or sublingual glands.

Cochlear IgG in the C3H/lpr autoimmune strain mouse.

The inner ear of the C3H/lpr autoimmune strain mouse was evaluated to identify potential mechanisms by which systemic autoimmune disease interferes with auditory function. The inner ears were immunohistochemically stained for IgG at ages before (2 months) and after (6-10 months) autoimmune disease onset and compared to age-matched nonautoimmune C3H/HeJ controls. Immunoreactivity for IgG was not seen in the 2 month C3H/lpr autoimmune mice or in either age group of the C3H/HeJ controls. On the other hand, all older C3H/lpr mice showed reaction product in the vessels of the cochlea, particularly the stria vascularis and bony capsule. Less frequent sites of staining were the geniculate ganglion, marrow cavities of the bony capsule, tensor tympani muscle, and on one occasion, a hair cell of the organ of Corti. These findings indicate that IgG is widespread within the cochlea and its vessels during systemic autoimmune disease and not directed against any specific sensorineural structure. This suggests a generalized or indirect mechanism whereby such systemic disease affects the inner ear.

Stria vascularis ultrastructural pathology in the C3H/lpr autoimmune strain mouse: a potential mechanism for immune-related hearing loss.

The stria vascularis in the C3H/lpr autoimmune strain mouse was ultrastructurally examined in order to better understand the potential mechanisms by which systemic autoimmune disease affects the ear. The inner ear from C3H/lpr mice before disease onset and C3H/HeJ controls showed no apparent pathology. However, the stria vascularis from older C3H/lpr mice after systemic autoimmune disease onset showed considerable intercellular edema around the stria capillaries and thickening of the capillary basement membrane, compared to controls. These observations suggest that perivascular abnormalities, which are the hallmark of systemic autoimmune diseases, may underlie the stria dysfunction and hearing loss seen in autoimmune diseases in humans.

Immunohistochemical analysis of otic capsule osteogenesis in the Palmerston North autoimmune mouse.

Autoimmunity and the immune complex disease associated with it have been hypothesized to be the cause of several idiopathic diseases of the inner ear--including the new bone formation associated with otic capsule osteogenesis and otosclerosis. The Palmerston North (PN) autoimmune mouse strain, which exhibits both spontaneous systemic autoimmune disease and otic capsule bone formation, has been proposed as a model relating these two disease processes. To investigate the potential role of immunopathologic processes in PN otic capsule lesion formation, inner ears from PN mice were immunostained for the presence of IgG and complement (C3), two immunologic markers involved in the development of the vascular and perivascular changes associated with immune complex deposition. Both systemic autoimmune disease and otic capsule bony lesions were confirmed in all animals. However, immunohistochemical analyses did not establish a direct relationship between the two conditions as complement was absent in all lesions and IgG stained positive in only one instance. These results suggest that immune complex deposition is not directly involved in the otic capsule lesions of the PN mouse, and alternate mechanisms relating autoimmune disease and otic capsule osteogenesis must be explored.

Inner ear pathology in the Palmerston North autoimmune strain mouse.

The Palmerston North autoimmune strain mouse is a model for spontaneous systemic lupus erythematosus. Inner ear structure and function were examined during the onset and progression of systemic autoimmune disease to identify potentially correlated auditory system pathology. The onset of systemic disease occurred at 4 to 5 months of age and was characterized by elevated serum immune complexes, cryoglobulins, and antinuclear antibodies. Coincident with the onset of autoimmune disease was degeneration of the apical turn stria vascularis and outer hair cells. These cochlear changes progressed basalward. At 10 months of age, auditory brainstem response thresholds were elevated and the stria vascularis area was measurably smaller throughout the cochlea. Immunohistochemical staining showed immunoglobulin G deposits within the organ of Corti, the vas spirale of the basilar membrane, the scala tympani, and marrow cavities of the bony otic capsule. These results suggest that cochlear pathology may be immune mediated in this mouse, which would make the strain suitable for the study of the mechanisms relating inner ear abnormalities and autoimmune disease.

Autoimmune disease and cochlear pathology in the C3H/lpr strain mouse.

The C3H/lpr autoimmune strain mouse is a model for spontaneous systemic lupus erythematosus. Inner ear structure and function were examined during systemic autoimmune disease progression to identify correlated auditory system pathology. Onset of the systemic disease occurred at 2-3 months of age and was characterized by elevated serum immune complexes, cryoglobulins, and antinuclear antibodies. Coincident with the onset of autoimmune disease was degeneration of the stria vascularis. Early edema of the stria occurred in the apex and progressed basalward with duration of the disease. By 10 months of age, stria vascularis area was smaller and auditory brainstem response thresholds were elevated. No degeneration of hair cells was seen at any age, suggesting that the stria vascularis may be the primary anatomic site of autoimmune auditory damage in this mouse model.

Cultured mesenteric vascular smooth muscle cells express dopamine DA1-receptors.

Incubation of cultured mesenteric vascular smooth muscle cells with dopamine, in the presence of propranolol, caused an increase in cyclic AMP formation in a concentration-dependent manner (Ka apparent 6.8 +/- 0.5 microM). This effect of dopamine was inhibited by the DA1-receptor antagonist SCH 23390 (Ki = 1 nM). These results suggest that cultured mesenteric vascular smooth muscle cells express DA1-receptors linked to adenylate cyclase.

Effects of oral dimethyl sulfoxide and dimethyl sulfone on murine autoimmune lymphoproliferative disease.

The results from several studies examining the effects of DMSO on autoimmune phenomena have been inconclusive, possibly because of differences in experimental models, treatment regimens and doses employed. In the present investigation, autoimmune strain MRL/lpr, C3H/lpr, and male BXSB mice were placed on a continuous treatment regimen with 3% DMSO or 3% DMSO2 in the drinking water, ad libitum, commencing at 1 to 2 months of age, before spontaneous disease development could be detected. This represented doses of 8-10 g/kg/day of DMSO and 6-8 g/kg/day of DMSO2. Both compounds were observed to extend the mean life span of MRL/lpr mice from 5 1/2 months to over 10 months of age. All strains showed decreased antinuclear antibody responses and significant diminution of lymphadenopathy, splenomegaly, and anemia development. Serum IgG levels and spleen IgM antibody plaque formation, however, did not differ from control values. There was no indication of involvement of systemic immunosuppressive or antiproliferative effects, and treated animals were observed to remain healthy and vigorous with no signs of toxicity. These results demonstrate that high doses of both DMSO and its major in vivo metabolite, DMSO2, provide significant protection against the development of murine autoimmune lymphoproliferative disease. Possible mechanisms of protection are discussed.

Vitamin C and immunity: influence of ascorbate on prostaglandin E2 synthesis and implications for natural killer cell activity.

A regimen of vitamin C in the drinking water of several strains of inbred mice has been observed previously to be without substantial effect on natural killer (NK) cell activity. In the present study, macrophage cultures derived from bone marrow of a number of these strains showed, on the average, a 90-100% increase in prostaglandin E2 (PGE2) production when ascorbate was added. This latter finding might suggest an explanation for the apparent failure of ascorbate to augment NK activity while enhancing interferon production.

CBA/N X-linked defect delays expression of the Y-linked accelerated autoimmune disease in BXSB mice.

BXSB male mice spontaneously develop progressive autoimmune disease characterized by high serum immunoglobulins, including anti-nuclear antibodies (ANA), enlarged spleen and lymph nodes, and diffuse proliferative glomerulonephritis. Females develop symptoms at a much slower rate. The mechanisms underlying the autoimmune disease and the nature of the Y-linked accelerating factor have not yet been elucidated. We found that the male progeny of the cross between the non-autoimmune strain CBA/Ca and BXSB (CBA/Ca X BXSB)F1 showed progressing signs of autoimmunity starting at 6 to 7 mo. In contrast, the male progeny that resulted from BXSB males crossed with immune-defective CBA/N females (Xid) were devoid of splenic B colonies, were nonresponsive to TNP-Ficoll, and were free of autoimmune disease for at least 10 mo. At 18 mo, some of the (CBA/N X BXSB)F1 mice developed weak antinuclear antibodies, but no spleen or lymph node enlargement was seen. The same mice had low anti-TNP Ficoll responses but did not produce B colonies in vitro. The role of the X chromosome in regulating expression of autoimmunity in young and old BXSB mice is discussed.

Vitamin C and immunity: natural killer (NK) cell factor.

The influence of a vitamin C regimen, 250 mg% in the drinking water, on natural killer (NK) cell activity was investigated in three highly inbred strains of mice. Spleen effector cells from these donors, both tap water control and experimental after 4-5 weeks, were tested against YAC-1 murine lymphoma target cells in a 4-hour 51Cr release assay. Ascorbate treatment was observed to be without effect on NK activity in the autoimmune- and lymphoma-prone NZB strain as well as in the normal and low cancer-incidence BALB/c and DBA/2. The relative levels of hemopoietic stem cells, purported to be decisive in determining NK levels, were of a similar order in these three strains as their relative NK activities. It appears that the established association of vitamin C with modulation of the immune response, as observed in activation of T cell-mediated immunity and the enhancement of interferon production, would not include alterations in natural cytotoxic reactivity.

Androgen sensitivity and autoimmune disease. I. Influence of sex and testosterone on the humoral immune response of autoimmune and non-autoimmune mouse strains to sheep erythrocytes.

Pre- and post-puberal NZB, DBA/2 and BALB/c mice showed no sex differences in primary IgM plaque-forming cell responses to sheep erythrocyte immunization. Orchiectomy increased and testosterone implants reduced antibody responses only if followed by sublethal irradiation suggesting that androgens may affect rapidly regenerating stem cells and/or their differentiating progeny. Strain differences in target organ sensitivity to androgen were not observed suggesting that NZB autoimmunity does not arise from a pathologic defect in androgen responsiveness.

Autoimmunity and tumor resistance. 3-Methylcholanthrene tumorigenesis in New Zealand Black mice.

Skin tumors induced by the subcutaneous injection of 3-methylcholanthrene (3-MC) in New Zealand Black (NZB) mice had a delayed development and lower frequency compared with BALB/c and C57BL mice. In the SJL/J strain, the incidence of tumors was lower than in the NZB, but with the same delayed development. Most of the tumors in the BALB/c, C57BL, and SJL/J strains were sarcomas; more than one third of the tumors in the NZB mice were squamous cell carcinomas. The greatest frequency and most rapid development of tumors in the NZB, as a function of age at the time of injection of 3-MC, occurred at 4 months. Young (3.5 and 7 weeks) and 12-month-old tumor incidence in the 4-month-old NZB was decreased by treatment with antithymocyte serum (ATS). Five hundred rad whole-body x-irradiation accelerated the onset of tumors but did not increase the final incidence. 3-MC injection and the presence of skin tumors had no influence on the development of glomerulonephritis or hematopoietic neoplasms in the NZB mice. Coombs' positive anemia was not influenced by 3-MC injection, but there was an earlier and increased incidence of positive Coombs' tests in tumor-bearing animals. Liver aryl hydrocarbon hydroxylase (AHH) specific activity was low in the young NZB, increased gradually with age, and was higher in the female mice.