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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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Histamine-induces enhanced hypothermia in 129S5 mice. a) Rectal temperature 0-60 minutes and (b). Maximal rectal temperature change at 30 minutes following i.v. injection of histamine (5 mg/200 μl saline). Data presented as means ± SEM (n = 6-8) and is representative of two separate experiments. Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). (a) Single and double asterisks indicate a P value < 0.05 and < 0.01 compared with BALB/c + histamine. (b) Single asterisk indicate a P value less than < 0.05.
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Figure 3: Histamine-induces enhanced hypothermia in 129S5 mice. a) Rectal temperature 0-60 minutes and (b). Maximal rectal temperature change at 30 minutes following i.v. injection of histamine (5 mg/200 μl saline). Data presented as means ± SEM (n = 6-8) and is representative of two separate experiments. Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). (a) Single and double asterisks indicate a P value < 0.05 and < 0.01 compared with BALB/c + histamine. (b) Single asterisk indicate a P value less than < 0.05.

Mentions: Recently we and others have performed experimental analyses employing murine models of anaphylaxis and mice deficient in specific genes implicated in the regulation of IgE and mast cell function to identify the relative contribution of these molecules to the manifestations of IgE/mast cell-mediated anaphylaxis [15,19,25-32]. These analyses often involve usage of mice of different or mixed strains, in particularly 129S5, a mouse strain frequently used in gene targeting analyses, and the BALB/c, a "Th2-skewed" strain that develop robust allergic phenotypes. We were interested in assessing for the presence of strain-dependent anaphylactic phenotypes between these commonly utilized mouse strains to eliminate possible contribution of strain-dependent factors in studies employing gene targeted mice. To assess this, we initially examined passive IgE-mediated anaphylaxis in 129S5 and BALB/c mice. Intravenous (i.v.) infusion of anti-mouse IgE (EM95) resulted in anaphylaxis as evidenced by hypothermia (Figure 1). Notably, hypothermia was significantly more severe in the 129S5 mice compared to BALB/c (Figure 1a and 1b). To ascertain whether the increased hypothermia was associated with increased mast cell degranulation, we examined plasma histamine levels. Notably, we found significantly elevated plasma histamine levels in 129S5 mice compared to BALB/c (Figure 1c). Importantly, 129S5 mice were not significantly more susceptible to IgG mediated anaphylaxis (Figure 1d and 1e) indicating that this phenotype is specific to IgE/mast cell-mediated reactions. We have previously demonstrated that oral antigen triggered anaphylaxis is mast cell and IgE dependent [18,27]. To assess whether 129S5 mice had increased susceptibility to oral antigen triggered anaphylaxis, OVA-primed 129S5 and BALB/c mice were orally gavaged with OVA and body temperature measured; notably, hypothermia in the 129S5 mice was more severe than that observed in BALB/c mice (Figure 2a). To determine whether oral OVA-triggered hypothermia was histamine-dependent, mice received histamine receptor type-1 and -2 receptor antagonists (Triprolidine and Cimetidine) 30 minutes prior to the OVA challenge. H1 and H2 receptor antagonism blocked the OVA-induced hypothermia in both 129S5 and in BALB/c mice indicating that anaphylaxis was mast cell/histamine dependent (Figure 2b). To assess whether the increased susceptibility to IgE-reactions in 129S5 mice was due to increased sensitivity to mast cell mediators, mice were i.v. injected with histamine and hypothermia was assessed. Injection (i.v) of histamine induced a significant temperature decrease in both 129S5 mice and BALB/c mice, however the decrease in 129S5 mice was significantly greater compared to that in BALB/c mice (Figure 3a and 3b). These studies indicated that 129S5 mice have increased susceptibility to IgE/mast cell/histamine-mediated hypothermia relative to BALB/c mice.


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)

Histamine-induces enhanced hypothermia in 129S5 mice. a) Rectal temperature 0-60 minutes and (b). Maximal rectal temperature change at 30 minutes following i.v. injection of histamine (5 mg/200 μl saline). Data presented as means ± SEM (n = 6-8) and is representative of two separate experiments. Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). (a) Single and double asterisks indicate a P value < 0.05 and < 0.01 compared with BALB/c + histamine. (b) Single asterisk indicate a P value less than < 0.05.
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Figure 3: Histamine-induces enhanced hypothermia in 129S5 mice. a) Rectal temperature 0-60 minutes and (b). Maximal rectal temperature change at 30 minutes following i.v. injection of histamine (5 mg/200 μl saline). Data presented as means ± SEM (n = 6-8) and is representative of two separate experiments. Data were analyzed by one-way analysis of variance (ANOVA) and a post-hoc comparison test (Tukey-Kramer). (a) Single and double asterisks indicate a P value < 0.05 and < 0.01 compared with BALB/c + histamine. (b) Single asterisk indicate a P value less than < 0.05.
Mentions: Recently we and others have performed experimental analyses employing murine models of anaphylaxis and mice deficient in specific genes implicated in the regulation of IgE and mast cell function to identify the relative contribution of these molecules to the manifestations of IgE/mast cell-mediated anaphylaxis [15,19,25-32]. These analyses often involve usage of mice of different or mixed strains, in particularly 129S5, a mouse strain frequently used in gene targeting analyses, and the BALB/c, a "Th2-skewed" strain that develop robust allergic phenotypes. We were interested in assessing for the presence of strain-dependent anaphylactic phenotypes between these commonly utilized mouse strains to eliminate possible contribution of strain-dependent factors in studies employing gene targeted mice. To assess this, we initially examined passive IgE-mediated anaphylaxis in 129S5 and BALB/c mice. Intravenous (i.v.) infusion of anti-mouse IgE (EM95) resulted in anaphylaxis as evidenced by hypothermia (Figure 1). Notably, hypothermia was significantly more severe in the 129S5 mice compared to BALB/c (Figure 1a and 1b). To ascertain whether the increased hypothermia was associated with increased mast cell degranulation, we examined plasma histamine levels. Notably, we found significantly elevated plasma histamine levels in 129S5 mice compared to BALB/c (Figure 1c). Importantly, 129S5 mice were not significantly more susceptible to IgG mediated anaphylaxis (Figure 1d and 1e) indicating that this phenotype is specific to IgE/mast cell-mediated reactions. We have previously demonstrated that oral antigen triggered anaphylaxis is mast cell and IgE dependent [18,27]. To assess whether 129S5 mice had increased susceptibility to oral antigen triggered anaphylaxis, OVA-primed 129S5 and BALB/c mice were orally gavaged with OVA and body temperature measured; notably, hypothermia in the 129S5 mice was more severe than that observed in BALB/c mice (Figure 2a). To determine whether oral OVA-triggered hypothermia was histamine-dependent, mice received histamine receptor type-1 and -2 receptor antagonists (Triprolidine and Cimetidine) 30 minutes prior to the OVA challenge. H1 and H2 receptor antagonism blocked the OVA-induced hypothermia in both 129S5 and in BALB/c mice indicating that anaphylaxis was mast cell/histamine dependent (Figure 2b). To assess whether the increased susceptibility to IgE-reactions in 129S5 mice was due to increased sensitivity to mast cell mediators, mice were i.v. injected with histamine and hypothermia was assessed. Injection (i.v) of histamine induced a significant temperature decrease in both 129S5 mice and BALB/c mice, however the decrease in 129S5 mice was significantly greater compared to that in BALB/c mice (Figure 3a and 3b). These studies indicated that 129S5 mice have increased susceptibility to IgE/mast cell/histamine-mediated hypothermia relative to BALB/c mice.

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