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Article ; Online: Undergraduate Education in Transplantation.

Cobos, Marisa / Lausada, Natalia / Tanús, Roberto / Raimondi, J Clemente

2023 Volume 55, Issue 6, Page(s) 1466–1468

Abstract: Introduction: Training in the transplants of organs, tissues, and cells as a therapeutic modality of multiple pathologies is essential in undergraduate education. The medical aspects typical of the theme are associated with ethical, legal, religious, ... ...

Abstract	Introduction: Training in the transplants of organs, tissues, and cells as a therapeutic modality of multiple pathologies is essential in undergraduate education. The medical aspects typical of the theme are associated with ethical, legal, religious, and philosophical concerns, giving a holistic view of the process. We present a teaching model of the donation-transplant process with 15 years of experience. Methods: The subject of Organ, Tissue, and Cell Transplants began its activities in 2008. It is an elective, annual subject included in the last year of the medical career. Since its inception, it has established a continuous teaching methodology with a global approach to the donation and transplantation process. Results: During the last 15 years and until the moment of the presentation, 1057 students have registered for the subject, 80.6% (852) completed the requirements of approval of the course, 79.9% of the students presented for the final evaluation (681), and 96.4% (654) of the students passed the final assessment. The average final grade calculated was equal to 6.53 ± 2.9 points out of 10; 205 students (19.4%) still need to comply with the final evaluative instance. Conclusion: The available literature has different training modalities, but none resembles the model presented. It is concluded that, during these 15 years, the pedagogic expectations in the training of human resources have been exceeded.
MeSH term(s)	Humans ; Curriculum ; Education, Medical, Undergraduate ; Students, Medical
Language	English
Publishing date	2023-05-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	82046-5
ISSN	1873-2623 ; 0041-1345
ISSN (online)	1873-2623
ISSN	0041-1345
DOI	10.1016/j.transproceed.2023.03.078
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Article ; Online: When less is more: Experimental Bishop-Koop technique for reduction in the use of laboratory animals for intestinal pathophysiological studies.

Abate, Juan Cruz / Lausada, Natalia / Vecchio Dezillio, Leandro / Moreira, Jeremías / Marinoff, Ivana Ivanoff / Ferreyra Compagnucci, Maria Malena / Andrés Moreno, Ane Miren / Largo, Carlota / Rumbo, Martín / Hernández Oliveros, Francisco / Romanin, David / Stringa, Pablo

Laboratory animals

2023 Volume 57, Issue 4, Page(s) 443–454

Abstract: The use of animals to gain knowledge and understanding of diseases needs to be reduced and refined. In the field of intestinal research, because of the complexity of the gut immune system, living models testing is mandatory. Based on the 3Rs (replacement, ...

Abstract	The use of animals to gain knowledge and understanding of diseases needs to be reduced and refined. In the field of intestinal research, because of the complexity of the gut immune system, living models testing is mandatory. Based on the 3Rs (replacement, reduction and refinement) principles, we aimed to developed and apply the derived-intestinal surgical procedure described by Bishop and Koop (BK) in rats to refine experimental gastrointestinal procedures and reduce the number of animals used for research employing two models of intestinal inflammation: intestinal ischemia-reperfusion injury and chemical-induced colitis. Our results show the feasibility of the application of the BK technique in rodents, with good success after surgical procedure in both small and large intestine (100% survival, clinical recovery and weight regain). A considerable reduction in the use of the number of rats in both intestinal inflammation models (80% in case of intestinal ischemia-reperfusion damage and 66.6% in chemical-induced colitis in our experimental design) was achieved. Compared with conventional experimental models described by various research groups, we report excellent reproducibility of intestinal damage and functionality, survival rate and clinical status of the animals when BK is applied.
MeSH term(s)	Animals ; Rats ; Research Design ; Reproducibility of Results ; Animals, Laboratory ; Colitis ; Inflammation ; Reperfusion Injury
Language	English
Publishing date	2023-02-07
Publishing country	England
Document type	Journal Article
ZDB-ID	391008-8
ISSN	1758-1117 ; 0023-6772
ISSN (online)	1758-1117
ISSN	0023-6772
DOI	10.1177/00236772231151563
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Article ; Online: When less is more: Experimental Bishop–Koop technique for reduction in the use of laboratory animals for intestinal pathophysiological studies

Laboratory Animals. 2023 Aug., v. 57, no. 4 p.443-454

2023

Abstract	The use of animals to gain knowledge and understanding of diseases needs to be reduced and refined. In the field of intestinal research, because of the complexity of the gut immune system, living models testing is mandatory. Based on the 3Rs (replacement, reduction and refinement) principles, we aimed to developed and apply the derived-intestinal surgical procedure described by Bishop and Koop (BK) in rats to refine experimental gastrointestinal procedures and reduce the number of animals used for research employing two models of intestinal inflammation: intestinal ischemia-reperfusion injury and chemical-induced colitis. Our results show the feasibility of the application of the BK technique in rodents, with good success after surgical procedure in both small and large intestine (100% survival, clinical recovery and weight regain). A considerable reduction in the use of the number of rats in both intestinal inflammation models (80% in case of intestinal ischemia-reperfusion damage and 66.6% in chemical-induced colitis in our experimental design) was achieved. Compared with conventional experimental models described by various research groups, we report excellent reproducibility of intestinal damage and functionality, survival rate and clinical status of the animals when BK is applied.
Keywords	animal use alternatives ; colitis ; immune system ; inflammation ; laboratories ; large intestine ; surgery ; survival rate ; Animal model ; animal use ; ethics and welfare ; organisms and models ; 3Rs ; reduction ; techniques
Language	English
Dates of publication	2023-08
Size	p. 443-454.
Publishing place	SAGE Publications
Document type	Article ; Online
ZDB-ID	391008-8
ISSN	1758-1117 ; 0023-6772
ISSN (online)	1758-1117
ISSN	0023-6772
DOI	10.1177/00236772231151563
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Article ; Online: Experimental Assessment of Intestinal Damage in Controlled Donation After Circulatory Death for Visceral Transplantation.

Stringa, Pablo / Vecchio Dezillio, Leandro Emmanuel / Talayero, Paloma / Serradilla, Javier / Errea, Agustina / Machuca, Mariana / Papa-Gobbi, Rodrigo / Camps Ortega, Onys / Pucci Molineris, Melisa / Lausada, Natalia / Andres Moreno, Ane Miren / Rumbo, Martin / Hernández Oliveros, Francisco

Transplant international : official journal of the European Society for Organ Transplantation

2023 Volume 36, Page(s) 10803

Abstract: There is an urgent need to address the shortage of potential multivisceral grafts in order to reduce the average time in waiting list. Since donation after circulatory death (DCD) has been successfully employed for other solid organs, a thorough ... ...

Abstract	There is an urgent need to address the shortage of potential multivisceral grafts in order to reduce the average time in waiting list. Since donation after circulatory death (DCD) has been successfully employed for other solid organs, a thorough evaluation of the use of intestinal grafts from DCD is warranted. Here, we have generated a model of Maastricht III DCD in rodents, focusing on the viability of intestinal and multivisceral grafts at five (DCD5) and twenty (DCD20) minutes of cardiac arrest compared to living and brain death donors. DCD groups exhibited time-dependent damage. DCD20 generated substantial intestinal mucosal injury and decreased number of Goblet cells whereas grafts from DCD5 closely resemble those of brain death and living donors groups in terms intestinal morphology, expression of tight junction proteins and number of Paneth and Globet cells. Upon transplantation, intestines from DCD5 showed increased ischemia/reperfusion damage compared to living donor grafts, however mucosal integrity was recovered 48 h after transplantation. No differences in terms of graft rejection, gene expression and absorptive function between DCD5 and living donor were observed at 7 post-transplant days. Collectively, our results highlight DCD as a possible strategy to increase multivisceral donation and transplantation procedures.
MeSH term(s)	Humans ; Brain Death ; Tissue and Organ Procurement ; Tissue Donors ; Liver Transplantation/methods ; Intestines ; Death ; Graft Survival ; Retrospective Studies
Language	English
Publishing date	2023-01-12
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	639435-8
ISSN	1432-2277 ; 0934-0874
ISSN (online)	1432-2277
ISSN	0934-0874
DOI	10.3389/ti.2023.10803
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Article ; Online: Experimental study to assess the impact of vasopressors administered during maintenance of the brain-dead donation in the quality of the intestinal graft.

Vecchio Dezillio, Leandro Emmanuel / Romanin, David Emmanuel / Ivanoff Marinoff, Ivana Mariel / Vernengo, Julieta / Abate Zárate, Juan Cruz / Machuca, Mariana Alejandra / Gondolesi, Gabriel Eduardo / Lausada, Natalia Raquel / Stringa, Pablo Luis / Rumbo, Martín

The journal of trauma and acute care surgery

2022 Volume 92, Issue 2, Page(s) 380–387

Abstract: Background: The hemodynamic maintenance of brain-dead donors will influence the quality of the organs procured for transplantation, including the intestine. Although norepinephrine (NE) and dopamine (DA) are commonly used to sustain mean arterial ... ...

Abstract	Background: The hemodynamic maintenance of brain-dead donors will influence the quality of the organs procured for transplantation, including the intestine. Although norepinephrine (NE) and dopamine (DA) are commonly used to sustain mean arterial pressure in humans, there are no standardized protocols for their use during maintenance of brain-dead donors. Our aim was to compare the effects of each drug, in the intestinal graft quality using a rat brain-dead donation model. Methods: Wistar rats (N = 17) underwent brain death (BD) for 2 hours with NE (NE group) or with DA (DA group) administration; the control group was mechanically ventilated for 2 hours without BD. Jejunum biopsies were obtained at the end of the maintenance period. Histological damage was evaluated using Park-Chiu scale. Villi/crypt ratio, mucosal thickness, Goblet cell count, and villi density were evaluated using ImageJ software (US National Institutes of Health, Bethesda, MD). Barrier damage was assessed by bacterial translocation culture counting on liver samples. The inflammatory status of the intestine was evaluated by CD3+ counting by immunohistochemistry and gene expression analysis of interleukin (IL)-6, IL-22, and CXCL10. Results: Norepinephrine-treated donors had higher focal ischemic injury in the intestinal mucosa without a substantial modification of morphometrical parameters compared with DA-treated donors. CD3+ mucosal infiltration was greater in intestines procured from brain-dead donors, being highest in NE (p ˂ 0.001). Local inflammatory mediators were affected in BD: DA and NE groups showed a trend to lower expression of IL-22, whereas CXCL10 expression was higher in NE versus control group. Brain death promoted intestinal bacterial translocation, but the use of NE resulted in the highest bacterial counting in the liver (p ˂ 0.01). Conclusion: Our results favor the use of DA instead of NE as main vasoactive drug to manage BD-associated hemodynamic instability. Dopamine may contribute to improve the quality of the intestinal graft, by better preserving barrier function and lowering immune cell infiltration.
MeSH term(s)	Animals ; Brain Death ; Chemokine CXCL10/metabolism ; Disease Models, Animal ; Dopamine/pharmacology ; Hemodynamics/drug effects ; Interleukin-6/metabolism ; Interleukins/metabolism ; Intestines/blood supply ; Intestines/drug effects ; Intestines/transplantation ; Male ; Norepinephrine/pharmacology ; Rats ; Rats, Wistar ; Interleukin-22
Chemical Substances	Chemokine CXCL10 ; Cxcl10 protein, rat ; Interleukin-6 ; Interleukins ; Dopamine (VTD58H1Z2X) ; Norepinephrine (X4W3ENH1CV)
Language	Spanish
Publishing date	2022-03-02
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2651070-4
ISSN	2163-0763 ; 2163-0755
ISSN (online)	2163-0763
ISSN	2163-0755
DOI	10.1097/TA.0000000000003473
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Article ; Online: The Novel N,N-bis-2-Hydroxyethyl-2-Aminoethanesulfonic Acid-Gluconate-Polyethylene Glycol-Hypothermic Machine Perfusion Solution Improves Static Cold Storage and Reduces Ischemia/Reperfusion Injury in Rat Liver Transplant.

Carnevale, Matías E / Lausada, Natalia / Juan de Paz, Leonardo / Stringa, Pablo / Machuca, Mariana / Rumbo, Martin / Guibert, Edgardo E / Tiribelli, Claudio / Gondolesi, Gabriel E / Rodriguez, Joaquin V

Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society

2019 Volume 25, Issue 9, Page(s) 1375–1386

Abstract: Organ transplantation is the treatment of choice against terminal and irreversible organ failure. Optimal preservation of the graft is crucial to counteract cold ischemia effects. As we developed an N,N-bis-2-hydroxyethyl-2-aminoethanesulfonic acid- ... ...

Abstract	Organ transplantation is the treatment of choice against terminal and irreversible organ failure. Optimal preservation of the graft is crucial to counteract cold ischemia effects. As we developed an N,N-bis-2-hydroxyethyl-2-aminoethanesulfonic acid-gluconate-polyethylene glycol (BGP)-based solution (hypothermic machine perfusion [HMP]), we aimed to analyze the use of this solution on static cold storage (SCS) of rat livers for transplantation as compared with the histidine tryptophan ketoglutarate (HTK) preservation solution. Livers procured from adult male Sprague Dawley rats were preserved with BGP-HMP or HTK solutions. Liver total water content and metabolites were measured during the SCS at 0°C for 24 hours. The function and viability of the preserved rat livers were first assessed ex vivo after rewarming (90 minutes at 37°C) and in vivo using the experimental model of reduced-size heterotopic liver transplantation. After SCS, the water and glycogen content in both groups remained unchanged as well as the tissue glutathione concentration. In the ex vivo studies, livers preserved with the BGP-HMP solution were hemodynamically more efficient and the O
MeSH term(s)	Alkanesulfonic Acids/chemistry ; Allografts/blood supply ; Allografts/pathology ; Animals ; Cold Ischemia/adverse effects ; Disease Models, Animal ; Gluconates/administration & dosage ; Gluconates/chemistry ; Glucose/administration & dosage ; Humans ; Liver/blood supply ; Liver/pathology ; Liver Transplantation/adverse effects ; Liver Transplantation/methods ; Male ; Mannitol/administration & dosage ; Organ Preservation/methods ; Organ Preservation Solutions/administration & dosage ; Organ Preservation Solutions/chemistry ; Perfusion/methods ; Polyethylene Glycols/administration & dosage ; Polyethylene Glycols/chemistry ; Potassium Chloride/administration & dosage ; Procaine/administration & dosage ; Rats ; Reperfusion Injury/diagnosis ; Reperfusion Injury/etiology ; Reperfusion Injury/pathology ; Reperfusion Injury/prevention & control ; Time Factors
Chemical Substances	Alkanesulfonic Acids ; Bretschneider cardioplegic solution ; Gluconates ; Organ Preservation Solutions ; BES (10191-18-1) ; Mannitol (3OWL53L36A) ; Polyethylene Glycols (3WJQ0SDW1A) ; Procaine (4Z8Y51M438) ; Potassium Chloride (660YQ98I10) ; Glucose (IY9XDZ35W2) ; gluconic acid (R4R8J0Q44B)
Language	English
Publishing date	2019-07-04
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2006866-9
ISSN	1527-6473 ; 1527-6465
ISSN (online)	1527-6473
ISSN	1527-6465
DOI	10.1002/lt.25573
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Article: Inmunosupresión en donantes renales. Expresión de citoquinas proinflamatorias.

Cicora, Federico / Roberti, Javier / Lausada, Natalia / González, Pedro / Guerrieri, Diego / Stringa, Pablo / Raimondi, Clemente

Medicina

2012 Volume 72, Issue 1, Page(s) 3–9

Abstract: The ischemia-reperfusion injury (IRI) remains a major problem in transplantation. The objective of this study was to evaluate the effects of preconditioning a donor group with rapamycin and another donor group with tacrolimus to prevent IRI. Twelve hours ...

Title translation	Immunosuppression in kidney donors with rapamycin and tacrolimus. Proinflammatory cytokine expression.
Abstract	The ischemia-reperfusion injury (IRI) remains a major problem in transplantation. The objective of this study was to evaluate the effects of preconditioning a donor group with rapamycin and another donor group with tacrolimus to prevent IRI. Twelve hours before nephrectomy, donor Wistar rats received immunosuppressive drugs. The sample was divided into four experimental groups: a sham group, an untreated control group, a group treated with rapamycin (2 mg/kg) and a group treated with tacrolimus (0.3 mg/kg). Left kidneys were removed and, after three hours of cold ischemia, grafts were transplanted. Twenty-four hours later, the transplanted organs were recovered for histological analysis and evaluation of cytokine expression. The pre-conditioning treatment with rapamycin or tacrolimus significantly reduced donor blood urea nitrogen and creatinine levels compared with control group (BUN: p < 0.001 vs. control and creatinine: p < 0.001 vs. control). Acute tubular necrosis was significantly lower in donors treated with immunosuppressant drugs compared with the control group (p < 0.001). Finally, inflammatory cytokines such as TNF-a, IL-6 and rIL-21 showed lower levels in the graft of pre-treated animals. This exploratory experimental study shows that preconditioning donors with rapamycin and tacrolimus in different groups improves clinical outcome and pathology in recipients and reduces in situ pro-inflammatory cytokines associated with Th17 differentiation, creating a favorable environment for the differentiation of regulatory T cells (Tregs).
MeSH term(s)	Animals ; Cytokines/biosynthesis ; Disease Models, Animal ; Immunosuppression ; Immunosuppressive Agents/therapeutic use ; Inflammation/metabolism ; Inflammation Mediators/metabolism ; Kidney Transplantation/immunology ; Living Donors ; Male ; Rats ; Rats, Wistar ; Reperfusion Injury/pathology ; Reperfusion Injury/prevention & control ; Sirolimus/therapeutic use ; Tacrolimus/therapeutic use ; Transplantation Conditioning/methods ; Tumor Necrosis Factor-alpha/biosynthesis
Chemical Substances	Cytokines ; Immunosuppressive Agents ; Inflammation Mediators ; Tumor Necrosis Factor-alpha ; Sirolimus (W36ZG6FT64) ; Tacrolimus (WM0HAQ4WNM)
Language	Spanish
Publishing date	2012
Publishing country	Argentina
Document type	English Abstract ; Journal Article
ZDB-ID	411586-7
ISSN	1669-9106 ; 0025-7680 ; 0325-951X
ISSN (online)	1669-9106
ISSN	0025-7680 ; 0325-951X
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Article ; Online: CD4+ T cell-derived IL-22 enhances liver metastasis by promoting angiogenesis.

Zhang, Tao / Wahib, Ramez / Zazara, Dimitra E / Lücke, Jöran / Shiri, Ahmad Mustafa / Kempski, Jan / Zhao, Lilan / Agalioti, Theodora / Machicote, Andres Pablo / Giannou, Olympia / Belios, Ioannis / Jia, Rongrong / Zhang, Siwen / Tintelnot, Joseph / Seese, Hannes / Grass, Julia Kristin / Mercanoglu, Baris / Stern, Louisa / Scognamiglio, Pasquale /

Fard-Aghaie, Mohammad / Seeger, Philipp / Wakker, Jonas / Kemper, Marius / Brunswig, Benjamin / Duprée, Anna / Lykoudis, Panagis M / Pikouli, Anastasia / Giorgakis, Emmanouil / Stringa, Pablo / Lausada, Natalia / Gentilini, Maria Virginia / Gondolesi, Gabriel E / Bachmann, Kai / Busch, Philipp / Grotelüschen, Rainer / Maroulis, Ioannis C / Arck, Petra C / Nakano, Ryosuke / Thomson, Angus W / Ghadban, Tarik / Tachezy, Michael / Melling, Nathaniel / Achilles, Eike-Gert / Puelles, Victor G / Nickel, Felix / Hackert, Thilo / Mann, Oliver / Izbicki, Jakob R / Li, Jun / Gagliani, Nicola / Huber, Samuel / Giannou, Anastasios D

Oncoimmunology

2023 Volume 12, Issue 1, Page(s) 2269634

Abstract: Metastasis is a cancer-related systemic disease and is responsible for the greatest mortality rate among cancer patients. Interestingly, the interaction between the immune system and cancer cells seems to play a key role in metastasis formation in the ... ...

Abstract	Metastasis is a cancer-related systemic disease and is responsible for the greatest mortality rate among cancer patients. Interestingly, the interaction between the immune system and cancer cells seems to play a key role in metastasis formation in the target organ. However, this complex network is only partially understood. We previously found that IL-22 produced by tissue resident iNKT17 cells promotes cancer cell extravasation, the early step of metastasis. Based on these data, we aimed here to decipher the role of IL-22 in the last step of metastasis formation. We found that IL-22 levels were increased in established metastatic sites in both human and mouse. We also found that Th22 cells were the key source of IL-22 in established metastasis sites, and that deletion of IL-22 in CD4+ T cells was protective in liver metastasis formation. Accordingly, the administration of a murine IL-22 neutralizing antibody in the establishment of metastasis formation significantly reduced the metastatic burden in a mouse model. Mechanistically, IL-22-producing Th22 cells promoted angiogenesis in established metastasis sites. In conclusion, our findings highlight that IL-22 is equally as important in contributing to metastasis formation at late metastatic stages, and thus, identify it as a novel therapeutic target in established metastasis.
MeSH term(s)	Humans ; Animals ; Mice ; CD4-Positive T-Lymphocytes ; Interleukins ; Liver Neoplasms ; Interleukin-22
Chemical Substances	Interleukins
Language	English
Publishing date	2023-10-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2645309-5
ISSN	2162-402X ; 2162-402X
ISSN (online)	2162-402X
ISSN	2162-402X
DOI	10.1080/2162402X.2023.2269634
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Article: Gut Permeability and Glucose Absorption Are Affected at Early Stages of Graft Rejection in a Small Bowel Transplant Rat Model.

Stringa, Pablo / Romanin, David / Lausada, Natalia / Papa Gobbi, Rodrigo / Zanuzzi, Carolina / Martín, Pedro / Abate, Juan Cruz / Cabanne, Ana / Arnal, Nathalie / Vecchio, Leandro / Milesi, Verónica / Portiansky, Enrique / Gondolesi, Gabriel / Rumbo, Martin

Transplantation direct

2017 Volume 3, Issue 11, Page(s) e220

Abstract: Background: Intestinal transplantation (ITx) faces many challenges due to the complexity of surgery and to the multiple immunological reactions that lead to the necessity of rigorous follow-up for early detection of acute cellular rejection (ACR). Our ... ...

Abstract	Background: Intestinal transplantation (ITx) faces many challenges due to the complexity of surgery and to the multiple immunological reactions that lead to the necessity of rigorous follow-up for early detection of acute cellular rejection (ACR). Our aim was to determine the kinetics of ACR using an experimental ITx model, with emphasis in the characterization of the process using different approaches, including the use of functional assays of absorptive and barrier function. Methods: ITx in rats conducting serial sampling was performed. Clinical monitoring, graft histology, proinflammatory gene expression, and nitrosative stress determination were performed. Also, glucose absorption, barrier function using ovalbumin translocation, and contractile function were analyzed. Results: The model used reproduced the different stages of ACR. Allogeneic ITx recipients showed signs of rejection from postoperative day (POD) 5, with increasing severity until 12 POD. Histological evaluation showed mild rejection in early sampling and severe rejection at late stages, with alterations in all graft layers. IL-6, CXCL 10, IFNg, and nitrite plasmas levels showed behavior coincident with histopathology. Remarkably, allogeneic grafts showed a marked alteration of glucose absorptive capacity from POD 5 that was sustained until endpoint. Coincidently, barrier function alteration was evidenced by luminal ovalbumin translocation to serum. Contractile function was progressively impaired along ACR. Conclusions: Glucose absorption and barrier function are altered at early stages of ACR when histological alterations or gene expression changes were much subtle. This observation may provide simple evaluation tools that could be eventually translated to the clinics to contribute to early ACR diagnosis.
Language	English
Publishing date	2017-10-06
Publishing country	United States
Document type	Journal Article
ISSN	2373-8731
ISSN	2373-8731
DOI	10.1097/TXD.0000000000000718
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Article: Down-regulation of intestinal epithelial innate response by probiotic yeasts isolated from kefir

Romanin, David / Serradell, María / González Maciel, Dolores / Lausada, Natalia / Garrote, Graciela L / Rumbo, Martín

International journal of food microbiology. 2010 June 15, v. 140, no. 2-3

2010

Abstract: Kefir is obtained by milk fermentation with a complex microbial population included in a matrix of polysaccharide and proteins. Several health-promoting activities has been attributed to kefir consumption. The aim of this study was to select ... ...

Abstract	Kefir is obtained by milk fermentation with a complex microbial population included in a matrix of polysaccharide and proteins. Several health-promoting activities has been attributed to kefir consumption. The aim of this study was to select microorganisms from kefir able to down-regulate intestinal epithelial innate response and further characterize this activity. Caco-2 cells stably transfected with a human CCL20 promoter luciferase reporter were used to screen a collection of 24 yeast and 23 bacterial strains isolated from kefir. The Toll-like receptor 5 agonist, flagellin was used to activate the reporter cells, while pre-incubation with the selected strains was tested to identify strains with the capacity to inhibit cell activation. In this system, 21 yeast strains from the genera Saccharomyces, Kluyveromyces and Issatchenkia inhibited almost 100% of the flagellin-dependent activation, whereas only some lactobacilli strains showed a partial effect. K. marxianus CIDCA 8154 was selected for further characterization. Inhibitory activity was confirmed at transcriptional level on Caco-2/TC-7 and HT-29 cells upon flagellin stimulation. A similar effect was observed using other pro-inflammatory stimulation such as IL-1β and TNF-α. Pre-incubation with yeasts induced a down-regulation of NF-κB signalling in epithelial cells in vitro, as well as expression of other pro-inflammatory chemokines such as CXCL8 and CXCL2. Furthermore, modulation of CCL20 mRNA expression upon flagellin stimulation was evidenced in vivo, in a mouse ligated intestinal loop model. Results indicate kefir contains microorganisms able to abolish the intestinal epithelial inflammatory response that could explain some of the properties attributed to this fermented milk.
Keywords	intestinal mucosa ; gene expression regulation ; mucosal immunity ; probiotics ; yeasts ; isolation ; kefir ; human cell lines ; functional foods ; cell culture ; inflammation ; anti-inflammatory activity
Language	English
Dates of publication	2010-0615
Size	p. 102-108.
Publishing place	[Amsterdam; New York, NY]: Elsevier Science
Document type	Article
ZDB-ID	87122-9
ISSN	1879-3460 ; 0168-1605
ISSN (online)	1879-3460
ISSN	0168-1605
DOI	10.1016/j.ijfoodmicro.2010.04.014
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