This Article Abstract Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wang, Y. Articles by Rose, N. R. Search for Related Content PubMed PubMed Citation Articles by Wang, Y. Articles by Rose, N. R.

Nasal administration of cardiac myosin suppresses autoimmune myocarditis in mice

^a Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, Maryland, USA, 21205
^b Department of Molecular Microbiology and Immunology, The Johns Hopkins Medical Institutions, Baltimore, Maryland, USA, 21205

View larger version (55K): [in a new window]   Figure 1 Nasal administration of CM reduced the severity of autoimmune myocarditis. (A) Female A/J mice were immunized subcutaneously twice on days 0 and 7 with porcine or murine CM (PCM or MCM) in complete Freund’s adjuvant for the induction of myocarditis. Three days prior to the first subcutaneous immunization with PCM (day –3), one group of mice (n = 9, open triangles) received a single nasal instillation of PCM, and the positive control group (n = 14, filled triangles) received equal volume of vehicle buffer. Three days prior to the first subcutaneous immunization with MCM, one group of mice (n = 9, open circles) received a single nasal administration of MCM, and the positive control group (n = 10, filled circles) received vehicle buffer only. All mice were sacrificed on day 21 when the heart sections were examined for evidence of myocarditis using the grading system described in the Methods section. The difference in severity between mice that received MCM intranasally and those that received vehicle buffer intranasally was statistically significant (p = 0.048). (B) Heart section from a PCM-immunized positive control mouse that received buffer intranasally and developed moderate myocarditis (grade 2.5) (24 x magnification). The inset on the top right corner is a closeup image (240 x magnification) of the cardiac infiltrate. (C) Heart section from the only mouse that developed cardiac infiltration in the group that received intranasal instillation of PCM (24 x magnification).

View larger version (25K): [in a new window]   Figure 2 Nasal administration of CM suppressed production of IL-2, TNF-, and IL-1ß by splenocytes in response to CM. Upon sacrifice on day 21, the spleens were collected from mice that received cardiac myosin intranasally (right bars, n = 9) and those that received vehicle buffer (left bars, n = 14). The splenocytes were cultured in vitro in the presence of either CM (10 µg/ml) or concanavalin A (ConA). The levels of cytokines produced in the culture supernatant were measured after 48 h and presented as the mean of each group ± SEM. The levels of IL-2 (p = 0.048), TNF- (p = 0.04), and IL-1ß (p = 0.02) produced in response to CM were significantly decreased in mice that received CM intranasally, compared to those that received buffer intranasally. The levels of IL-2 produced in response to ConA were not reduced after nasal instillation of myosin.

View larger version (14K): [in a new window]   Figure 3 Nasal administration of CM did not reduce the production of TGF-ß by splenocytes in response to CM. The splenocytes from mice that received either CM (right bar, n = 4) or vehicle buffer (left bar, n = 3) intranasally were cultured in vitro in the presence of CM (10 µg/ml). The levels of TGF-ß produced in the culture supernatant were measured and presented as the mean of each group ± SEM. There was no decrease in the levels of TGF-ß observed in mice that received CM nasally as compared to those that received buffer nasally.

View larger version (13K): [in a new window]   Figure 4 Nasal administration of CM reduced the serum levels of total IgE antibodies. Sera were collected on days 10 and 21 from mice that received CM intranasally (right bars, n = 9) and those that received vehicle buffer (left bars, n = 14). The serum levels of total IgE antibodies (ng/ml) were measured by a capture ELISA and presented as the mean of each group ± SEM. The levels of IgE were lower in mice that received CM intranasally on both days 10 and 21, compared to those that received buffer. The decrease in IgE levels on day 10 was statistically significant (p = 0.01).

View larger version (15K): [in a new window]   Figure 5 Nasal administration of CM reduced the serum levels of myosin-specific IgG1 antibodies. The levels of anti-myosin antibodies of the IgG1 subclass were measured in serum samples collected on both days 10 and 21 from mice that received CM nasally (right bars, n = 9) and from those that received buffer (left bars, n = 14). The antibody levels are presented as the mean of adjusted optical densities (adjusted ODs, see Methods section) of each group ± SEM. The levels of IgG1 anti-myosin antibodies were significantly reduced in mice that received CM intranasally on both day 10 (p = 0.04) and day 21 (p = 0.04), compared to the control mice that received buffer nasally.