Improvement of bovine ß-lactoglobulin production and secretion by Lactococcus lactis

Nouaille, Sébastien; Bermúdez‐Humarán, Luis G.; Adel‐Patient, Karine; Commissaire, Jacqueline; Gruss, Alexandra; Wal, Jean‐Michel; Azevedo, Vasco; Langella, Philippe; Châtel, Jean-Marc

doi:10.1590/s0100-879x2005000300005

Cited by 16 publications

(29 citation statements)

References 18 publications

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“…The constructions of recombinant L. lactis NZ9000 strains expressing a secreted form of IL-12 (L. lactis IL-12) and BLG fused with the LEISSTCDA propeptide (L. lactis BLG) have been described previously (8,42). The control strain, i.e., L. lactis NZ9000, containing an empty vector and all derived recombinant strains were grown at 30°C without shaking in M17 medium (Difco) supplemented with 0.5% glucose and 10 g/ml chloramphenicol.…”

Section: Methodsmentioning

confidence: 99%

Intranasal Coadministration of Live Lactococci Producing Interleukin-12 and a Major Cow's Milk Allergen Inhibits Allergic Reaction in Mice

Cortes-Perez

Ah‐Leung

Bermúdez‐Humarán

et al. 2007

Clin Vaccine Immunol

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The Th1/Th2 balance deregulation toward a Th2 immune response plays a central role in allergy. We previously demonstrated that administration of recombinant Lactococcus lactis strains expressing bovine ␤-lactoglobulin (BLG), a major cow's milk allergen, partially prevents mice from sensitization. In the present study, we aimed to improve this preventive effect by coadministration of L. lactis BLG and a second recombinant L. lactis strain producing biologically active interleukin-12 (IL-12). This L. lactis strain producing IL-12 was previously used to enhance the Food allergies, i.e., immediate-type, immunoglobulin E (IgE)-mediated immune responses, affect 2 to 2.5% of the general populations of Western countries (22,24,25). It results from the activation of the Th2-type helper lymphocytes. In mice, this response is mainly characterized by the production of interleukin-4 (IL-4) and IL-13, which induce the production of IgE and IgG1, and of IL-5, which attracts eosinophils. In contrast, the Th1 response is characterized by the induction of gamma interferon (IFN-␥) and IL-2 production and the stimulation of cellular immunity. Th1 and Th2 cells regulate their own development via the cytokines produced: IFN-␥ suppresses Th2-cell proliferation and promotes Th1-cell proliferation, whereas IL-4 promotes additional Th2-cell proliferation and inhibits Th1-cell development (39,43,45).A recent study suggests that an early allergen-specific induction of Th1 cells before allergy sensitization could not efficiently prevent the development of atopic disorders: Th1 priming was abolished in the presence of allergen-specific Th2 cells, whereas Th1 cells could not inhibit subsequent priming of Th2 cells (58). The use of adjuvants that increase the proliferation of Th1 cells and that render them more efficient in inhibiting Th2 cells would therefore be of great interest in the management of allergic diseases. These observations prompted us to explore the adjuvant effect of IL-12 in a mouse model of allergic reaction to bovine ␤-lactoglobulin (BLG), a major milk allergen (2). IL-12 is a heterodimeric cytokine produced by antigen-presenting cells that promotes the development of naïve Th cells into Th1 effector cells and that induces IFN-␥ production (51,52,54). IL-12 also inhibits Th2 class switching by repression of the IL-4 cytokine (15,17,46,54,55). Moreover, treatment with recombinant IL-12 has been shown to have positive effects on bronchial hyperresponsiveness and the eosinophilic response (11). Unfortunately, despite the therapeutic potential of this molecule, the toxicity of systemic IL-12 treatment observed during clinical trials has limited its use (18,31,32,33,40). This toxicity correlates with increased IFN-␥ levels, decreased glucose levels, and altered histological responses in the spleen and duodenum. This has motivated several recent investigations demonstrating that intranasal delivery of IL-12 is a less toxic route of inoculation compared to the commonly used systemic one (6,7,26).The gram-positive and nonpathogenic...

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Section: Methodsmentioning

confidence: 99%

Intranasal Coadministration of Live Lactococci Producing Interleukin-12 and a Major Cow's Milk Allergen Inhibits Allergic Reaction in Mice

Cortes-Perez

Ah‐Leung

Bermúdez‐Humarán

et al. 2007

Clin Vaccine Immunol

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“…The PCR fragment was digested with NcoI and BstBI and fused to pIL-Ter cut by NcoI and ClaI. The resulting plasmid, pLacTd-Ter, cut by BglII and SalI, was then fused to BamHI-HindIII and HindIII-XhoI fragments from pSEC:Nuc-BLG or pSEC: LEISS-Nuc-BLG (28). The sequence encoding the Nuc protein was removed by NsiI digestion, resulting in plasmids pLacTd:BLG and pLacTd:LEISS-BLG.…”

Section: Methodsmentioning

confidence: 99%

Constitutive Delivery of Bovine β-Lactoglobulin to the Digestive Tracts of Gnotobiotic Mice by EngineeredLactobacillus casei

Hazebrouck¹,

Oozeer

Adel‐Patient³

et al. 2006

Appl Environ Microbiol

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The gut microbiota is critical for maturation of the immune system. Recent evidence suggests that early establishment of lactobacilli in the intestinal microbiota, during neonatal colonization or by probiotic supplementation, could prevent the development of allergic disorders. Postnatal maturation of the gut immune system with allergen-producing lactobacilli colonizing the digestive tract could then affect the development of further allergic sensitization. In this paper, we describe construction of a recombinant Lactobacillus casei strain that can constitutively deliver bovine ␤-lactoglobulin (BLG), a major cow's milk allergen, to the guts of gnotobiotic mice. The blg gene was inserted into the L. casei chromosome downstream of an endogenous promoter. BLG production was improved by fusing the propeptide LEISSTCDA (LEISS) to the BLG mature moiety. This led to a 10-fold increase in LEISS-BLG production compared to the production obtained without the propeptide and also led to enhanced secretion corresponding to 5% of the total production. After inoculation into germfree C3H/HeN mice, the genetic stability of the recombinant strain and in vivo BLG production were confirmed for at least 10 weeks. BLG stimulation of spleen cells from mice monoassociated with the BLG-producing lactobacilli induced secretion of the Th1 cytokine gamma interferon and, to a lesser extent, the Th2 cytokine interleukin-5. No BLG-specific immunoglobulin G1 (IgG1), IgG2a, or IgA was detected in sera or in fecal samples. These results suggest that gut colonization with allergen-producing lactobacilli could provide a useful model for studying the modulation of allergic disorders.Food allergy affects 1 to 2% of adults and 5 to 8% of children in Western countries (25). With an incidence of 1.9 to 2.8%, cow's milk allergy is the most common allergy in early infancy (13). Patients may be sensitized to various proteins, mainly ␤-lactoglobulin (BLG) and caseins (39). Food allergy generally corresponds to an inappropriate immune response characterized by disruption of the Th1/Th2 balance toward a Th2 profile that results in the production of immunoglobulins E (IgE) specific for food antigens. Th2 cells produce interleukin-4 (IL-4), IL-5, and IL-13, whereas the Th1 response is characterized by gamma interferon (IFN-␥) and IL-12 synthesis. The Th1 and Th2 responses inhibit each other's development and function via the cytokines produced (26).Development of allergy is multifactorial, and it includes genetic factors and also different environmental factors, such as lifestyle and the intestinal microbiota. The intestinal microbiota seems to be critical because of its role in the postnatal maturation of the immune system. At birth, the digestive tract is sterile and the neonatal immune response is characterized by a polarized Th2 cytokine profile. During postnatal gut colonization, the gut immune system is exposed to a wide range of bacterial antigens, which apparently play a major role in driving the initial Th2-skewed immune response toward a more finely...

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“…Total protein was extracted from cultures of wild-type MG1363, MG1363(pLIG1), and MG1363 . We checked that BLG was produced only by our positive control, a nisininduced culture of L. lactis NZ900(pLEISS-BLG) (31). Lactococcus lactis is able to transfer genetic material to Caco-2 cells.…”

Section: Blg Expression In Cos-7 Cells Is Higher With Plig:blg1mentioning

confidence: 99%

Use of Native Lactococci as Vehicles for Delivery of DNA into Mammalian Epithelial Cells

Guimarães

Innocentin

Lefèvre

et al. 2006

Appl Environ Microbiol

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Improvement of bovine ß-lactoglobulin production and secretion by Lactococcus lactis

Cited by 16 publications

References 18 publications

Intranasal Coadministration of Live Lactococci Producing Interleukin-12 and a Major Cow's Milk Allergen Inhibits Allergic Reaction in Mice

Intranasal Coadministration of Live Lactococci Producing Interleukin-12 and a Major Cow's Milk Allergen Inhibits Allergic Reaction in Mice

Constitutive Delivery of Bovine β-Lactoglobulin to the Digestive Tracts of Gnotobiotic Mice by EngineeredLactobacillus casei

Use of Native Lactococci as Vehicles for Delivery of DNA into Mammalian Epithelial Cells

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