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Increased susceptibility of 129SvEvBrd mice to IgE-Mast cell mediated anaphylaxis.

Arumugam M, Ahrens R, Osterfeld H, Kottyan LC, Shang X, Maclennan JA, Zimmermann N, Zheng Y, Finkelman FD, Hogan SP - BMC Immunol. (2011)

Bottom Line: In vitro analyses of BMMCs revealed no difference in FcεRI and c-Kit expression, however, 129S5 BMMCs possessed greater proliferative capacity and reduced caspase-3-mediated apoptosis.IgE-BMMC degranulation assays demonstrated no difference in degranulation efficiency.We conclude that 129S5 mice have increased susceptibility to anaphylaxis as compared to BALB/c strain and their increased susceptibility was associated with altered mast cell proliferation and homeostatic tissue levels and responsiveness to histamine.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Biochemistry, National Institute of Siddha, Chennai, India.

ABSTRACT

Background: Experimental analyses have identified strain-dependent factors that regulate susceptibility to anaphylaxis in mice. We assessed the susceptibility of the widely used 129SvEvBrd (also known as 129S5) mouse strain to IgE/mast cell-mediated anaphylaxis as compared to BALB/c. Mice were subjected to passive and oral Ovalbumin [OVA]-induced active anaphylaxis. Tissue mast cell, plasma histamine, total IgE and OVA-specific IgE levels and susceptibility to histamine i.v infusion were assessed. Bone marrow mast cell (BMMC)s were examined for FcεRI, c-kit, degranulation efficiency, proliferation, apoptosis and cytokine profile.

Results: 129S5 mice had significantly increased susceptibility to passive and oral OVA-induced active anaphylaxis. Increased susceptibility to anaphylaxis was associated with increased homeostatic mast cell levels but not OVA-specific IgE or IgG1 levels. In vitro analyses of BMMCs revealed no difference in FcεRI and c-Kit expression, however, 129S5 BMMCs possessed greater proliferative capacity and reduced caspase-3-mediated apoptosis. IgE-BMMC degranulation assays demonstrated no difference in degranulation efficiency. Furthermore, 129S5 mice possessed increased sensitivity to histamine-induced hypothermia.

Conclusions: We conclude that 129S5 mice have increased susceptibility to anaphylaxis as compared to BALB/c strain and their increased susceptibility was associated with altered mast cell proliferation and homeostatic tissue levels and responsiveness to histamine. Given the wide spread usage of the 129SvEvBrd strain of mice in experimental gene targeting methodology, these data have important implications for studying IgE-reactions in mouse systems.

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Increased proliferation and decreased caspase-3-mediated apoptosis in 129S5 BMMC. (a) Representative flow cytometry plot (day 8) of CFSE-labeled BALB/c and 129S5 BMMCs. Divisional index of 129S5 and BALB/c BMMCs cultured in IL3 (20ng/ml) and SCF (10ng/ml) and CFSE-fluoresence decay was measured on day 4 and day 8 and analyzed by proliferation tool of Flowjo software to determine division index. (b) Percentage of 7AAD- Annexin V+ and (c) active caspase-3+ 129S5 and BALB/c BMMCs (5-week old) cultured in IL-3 (20 ng/ml) and SCF (10 ng/ml). Data represent mean ± SEM and is representative of three separate experiments. (a) Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). Single and double asterisks indicate a P value < 0.05 and < 0.01, respectively. (b and c) data were analyzed using a Students T-test. Double and triple asterisks indicate a P value < 0.01 and < 0.001, respectively.
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Figure 7: Increased proliferation and decreased caspase-3-mediated apoptosis in 129S5 BMMC. (a) Representative flow cytometry plot (day 8) of CFSE-labeled BALB/c and 129S5 BMMCs. Divisional index of 129S5 and BALB/c BMMCs cultured in IL3 (20ng/ml) and SCF (10ng/ml) and CFSE-fluoresence decay was measured on day 4 and day 8 and analyzed by proliferation tool of Flowjo software to determine division index. (b) Percentage of 7AAD- Annexin V+ and (c) active caspase-3+ 129S5 and BALB/c BMMCs (5-week old) cultured in IL-3 (20 ng/ml) and SCF (10 ng/ml). Data represent mean ± SEM and is representative of three separate experiments. (a) Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). Single and double asterisks indicate a P value < 0.05 and < 0.01, respectively. (b and c) data were analyzed using a Students T-test. Double and triple asterisks indicate a P value < 0.01 and < 0.001, respectively.

Mentions: To define if the divergence in susceptibility to anaphylaxis in 129S5 and BALB/c mice was due to inherent differences in mast cell function, we assessed phenotypic and degranulation properties of BMMCs. Surface expression of FcЄRI and c-Kit on BMMCs from 129S5 and BALB/c mice were comparable (results not shown). Similarly, the level of IgE-mediated degranulation of BMMCs, as measured by β-hexosaminidase activity and cytokine (IL-6, IL-13 and TNFα) release, was also not significantly different between strains (Figure 6a and results not shown). Since we observed increased systemic mast cell levels in 129S5 compared to BALB/c mice, we examined BMMC proliferation rate and apoptosis. Proliferation by 129S5 BMMCs in the presence of IL-3 and SCF was significantly increased relative to BALB/c BMMC (Figure 7a). Furthermore, the level of 129S5 BMMC apoptosis was reduced compared to BALB/c. Specifically, the percentage of BMMCs entering early apoptosis (Annexin V+ 7AAD-) was significantly reduced in 129S5 BMMCs compared to BALB/c (Figure.7b). Consistent with this observation, active Caspase-3 expression was significantly decreased in 129S5 BMMCs when compared to BALB/c BMMCs (Figure 7c).


Increased susceptibility of 129SvEvBrd mice to IgE-Mast cell mediated anaphylaxis.

Arumugam M, Ahrens R, Osterfeld H, Kottyan LC, Shang X, Maclennan JA, Zimmermann N, Zheng Y, Finkelman FD, Hogan SP - BMC Immunol. (2011)

Increased proliferation and decreased caspase-3-mediated apoptosis in 129S5 BMMC. (a) Representative flow cytometry plot (day 8) of CFSE-labeled BALB/c and 129S5 BMMCs. Divisional index of 129S5 and BALB/c BMMCs cultured in IL3 (20ng/ml) and SCF (10ng/ml) and CFSE-fluoresence decay was measured on day 4 and day 8 and analyzed by proliferation tool of Flowjo software to determine division index. (b) Percentage of 7AAD- Annexin V+ and (c) active caspase-3+ 129S5 and BALB/c BMMCs (5-week old) cultured in IL-3 (20 ng/ml) and SCF (10 ng/ml). Data represent mean ± SEM and is representative of three separate experiments. (a) Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). Single and double asterisks indicate a P value < 0.05 and < 0.01, respectively. (b and c) data were analyzed using a Students T-test. Double and triple asterisks indicate a P value < 0.01 and < 0.001, respectively.
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Figure 7: Increased proliferation and decreased caspase-3-mediated apoptosis in 129S5 BMMC. (a) Representative flow cytometry plot (day 8) of CFSE-labeled BALB/c and 129S5 BMMCs. Divisional index of 129S5 and BALB/c BMMCs cultured in IL3 (20ng/ml) and SCF (10ng/ml) and CFSE-fluoresence decay was measured on day 4 and day 8 and analyzed by proliferation tool of Flowjo software to determine division index. (b) Percentage of 7AAD- Annexin V+ and (c) active caspase-3+ 129S5 and BALB/c BMMCs (5-week old) cultured in IL-3 (20 ng/ml) and SCF (10 ng/ml). Data represent mean ± SEM and is representative of three separate experiments. (a) Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). Single and double asterisks indicate a P value < 0.05 and < 0.01, respectively. (b and c) data were analyzed using a Students T-test. Double and triple asterisks indicate a P value < 0.01 and < 0.001, respectively.
Mentions: To define if the divergence in susceptibility to anaphylaxis in 129S5 and BALB/c mice was due to inherent differences in mast cell function, we assessed phenotypic and degranulation properties of BMMCs. Surface expression of FcЄRI and c-Kit on BMMCs from 129S5 and BALB/c mice were comparable (results not shown). Similarly, the level of IgE-mediated degranulation of BMMCs, as measured by β-hexosaminidase activity and cytokine (IL-6, IL-13 and TNFα) release, was also not significantly different between strains (Figure 6a and results not shown). Since we observed increased systemic mast cell levels in 129S5 compared to BALB/c mice, we examined BMMC proliferation rate and apoptosis. Proliferation by 129S5 BMMCs in the presence of IL-3 and SCF was significantly increased relative to BALB/c BMMC (Figure 7a). Furthermore, the level of 129S5 BMMC apoptosis was reduced compared to BALB/c. Specifically, the percentage of BMMCs entering early apoptosis (Annexin V+ 7AAD-) was significantly reduced in 129S5 BMMCs compared to BALB/c (Figure.7b). Consistent with this observation, active Caspase-3 expression was significantly decreased in 129S5 BMMCs when compared to BALB/c BMMCs (Figure 7c).

Bottom Line: In vitro analyses of BMMCs revealed no difference in FcεRI and c-Kit expression, however, 129S5 BMMCs possessed greater proliferative capacity and reduced caspase-3-mediated apoptosis.IgE-BMMC degranulation assays demonstrated no difference in degranulation efficiency.We conclude that 129S5 mice have increased susceptibility to anaphylaxis as compared to BALB/c strain and their increased susceptibility was associated with altered mast cell proliferation and homeostatic tissue levels and responsiveness to histamine.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Biochemistry, National Institute of Siddha, Chennai, India.

ABSTRACT

Background: Experimental analyses have identified strain-dependent factors that regulate susceptibility to anaphylaxis in mice. We assessed the susceptibility of the widely used 129SvEvBrd (also known as 129S5) mouse strain to IgE/mast cell-mediated anaphylaxis as compared to BALB/c. Mice were subjected to passive and oral Ovalbumin [OVA]-induced active anaphylaxis. Tissue mast cell, plasma histamine, total IgE and OVA-specific IgE levels and susceptibility to histamine i.v infusion were assessed. Bone marrow mast cell (BMMC)s were examined for FcεRI, c-kit, degranulation efficiency, proliferation, apoptosis and cytokine profile.

Results: 129S5 mice had significantly increased susceptibility to passive and oral OVA-induced active anaphylaxis. Increased susceptibility to anaphylaxis was associated with increased homeostatic mast cell levels but not OVA-specific IgE or IgG1 levels. In vitro analyses of BMMCs revealed no difference in FcεRI and c-Kit expression, however, 129S5 BMMCs possessed greater proliferative capacity and reduced caspase-3-mediated apoptosis. IgE-BMMC degranulation assays demonstrated no difference in degranulation efficiency. Furthermore, 129S5 mice possessed increased sensitivity to histamine-induced hypothermia.

Conclusions: We conclude that 129S5 mice have increased susceptibility to anaphylaxis as compared to BALB/c strain and their increased susceptibility was associated with altered mast cell proliferation and homeostatic tissue levels and responsiveness to histamine. Given the wide spread usage of the 129SvEvBrd strain of mice in experimental gene targeting methodology, these data have important implications for studying IgE-reactions in mouse systems.

Show MeSH
Related in: MedlinePlus