Analysis of immune cells draining from the abdominal cavity as a novel tool to study intestinal transplant immunobiology

Summary During intestinal transplant (ITx) operation, intestinal lymphatics are not reconstituted. Consequently, trafficking immune cells drain freely into the abdominal cavity. Our aim was to evaluate whether leucocytes migrating from a transplanted intestine could be recovered from the abdominal draining fluid collected by a peritoneal drainage system in the early post-ITx period, and to determine potential applications of the assessment of draining cellular populations. The cell composition of the abdominal draining fluid was analysed during the first 11 post-ITx days. Using flow cytometry, immune cells from blood and draining fluid samples obtained the same day showed an almost complete lymphopenia in peripheral blood, whereas CD3+CD4+CD8 -, CD3+CD4-CD8+ and human leucocyte antigen D-related (HLA-DR)+CD19+ lymphocytes were the main populations in the draining fluid. Non-complicated recipients evolved from a mixed leucocyte pattern including granulocytes, monocytes and lymphocytes to an exclusively lymphocytic pattern along the first post-ITx week. At days 1-2 post-Itx, analysis by short tandem repeats fingerprinting of CD3 +CD8+ sorted T cells from draining fluid indicated that 50% of cells were from graft origin, whereas by day 11 post-ITx this proportion decreased to fewer than 1%. Our results show for the first time that the abdominal drainage fluid contains mainly immune cells trafficking from the implanted intestine, providing the opportunity to sample lymphocytes draining from the grafted organ along the post-ITx period. Therefore, this analysis may provide information useful for understanding ITx immunobiology and eventually could also be of interest for clinical management.Laboratorio de Investigaciones del Sistema Inmun

Analysis of immune cells draining from the abdominal cavity as a novel tool to study intestinal transplant immunobiology

Summary During intestinal transplant (ITx) operation, intestinal lymphatics are not reconstituted. Consequently, trafficking immune cells drain freely into the abdominal cavity. Our aim was to evaluate whether leucocytes migrating from a transplanted intestine could be recovered from the abdominal draining fluid collected by a peritoneal drainage system in the early post-ITx period, and to determine potential applications of the assessment of draining cellular populations. The cell composition of the abdominal draining fluid was analysed during the first 11 post-ITx days. Using flow cytometry, immune cells from blood and draining fluid samples obtained the same day showed an almost complete lymphopenia in peripheral blood, whereas CD3+CD4+CD8 -, CD3+CD4-CD8+ and human leucocyte antigen D-related (HLA-DR)+CD19+ lymphocytes were the main populations in the draining fluid. Non-complicated recipients evolved from a mixed leucocyte pattern including granulocytes, monocytes and lymphocytes to an exclusively lymphocytic pattern along the first post-ITx week. At days 1-2 post-Itx, analysis by short tandem repeats fingerprinting of CD3 +CD8+ sorted T cells from draining fluid indicated that 50% of cells were from graft origin, whereas by day 11 post-ITx this proportion decreased to fewer than 1%. Our results show for the first time that the abdominal drainage fluid contains mainly immune cells trafficking from the implanted intestine, providing the opportunity to sample lymphocytes draining from the grafted organ along the post-ITx period. Therefore, this analysis may provide information useful for understanding ITx immunobiology and eventually could also be of interest for clinical management.Laboratorio de Investigaciones del Sistema Inmun

Induction Versus Maintenance Immunosuppression After Intestinal Transplant: Determining Which Treatment Most Impacts Long-Term Patient And Graft Survival

Objectives: Immunosuppressive strategies for intestinal transplant have changed over time. However, specific intestinal transplant-oriented protocols and reports on long-term maintenance regimens are scarce. Our objective was to evaluate the impact of 2 different initial immunosuppressive protocols based on thymoglobulin (group A) and basiliximab (anti-interleukin 2 antibody) (group B) and of changes to maintenance immunosuppression over long-term follow-up in intestinal transplant recipients. Materials and Methods: We performed a retrospective analysis of a prospectively established protocol for intestinal transplant immunosuppression, conducted between May 2006 and December 2020. We analyzed 51 intestinal transplant recipients, with 6 patients excluded because of early death or graft loss. Acute cellular rejection frequency and grade, number of acute cellular rejection episodes, time to the first acute cellular rejection episode, response to treatment, number of patients who progressed to chronic allograft rejection, kidney function, infections, incidence of posttransplant lymphoproliferative disorder and graft-versus-host disease, and patient and graft survival were analyzed. Results: In the study groups, there were 87 acute cellular rejection episodes in 45 patients (33 in group A and 54 in group B). We found degree of acute cellular rejection to be mild in 45 patients, moderate in 18, and severe in 24 (not significant between groups). Our comparison of induction therapy (thymoglobulin [group A] vs interleukin 2 antibody [group B]) did not show any statistical difference during clinical followup. Long-term review showed that all patients were on tacrolimus. Five-year patient and graft survival rates were 62% and 45% for group A and 54% and 46% for group B, respectively (not significant). Conclusions: Long-term patient and graft outcomes reflected the use of an individualized follow-up with adjustments and changes in immunosuppressive medications according to the patient’s clinical course and complications rather than based on the induction immunosuppressive protocol used.Fil: Gentilini, María Virginia. Universidad Favaloro; Argentina. Fundación Favaloro; ArgentinaFil: Perez Illidge, Luis. Fundación Favaloro; Argentina. Universidad Favaloro; ArgentinaFil: Pedraza, Néstor. Colombiana de Trasplantes; ColombiaFil: Nemirovsky, Sergio Ivan. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Fernandez, María Florencia. Fundación Favaloro; ArgentinaFil: Ramisch, Diego. Fundación Favaloro; ArgentinaFil: Solar, Héctor. Fundación Favaloro; ArgentinaFil: Rumbo, Martín. Facultad de Ciencias Exactas, Universidad Nacional de la Plata; ArgentinaFil: Rumbo, Carolina. Fundación Favaloro; ArgentinaFil: Gondolesi, Gabriel Eduardo. Universidad Favaloro; Argentina. Fundación Favaloro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Analysis of immune cells draining from the abdominal cavity as a novel tool to study intestinal transplant immunobiology

Summary During intestinal transplant (ITx) operation, intestinal lymphatics are not reconstituted. Consequently, trafficking immune cells drain freely into the abdominal cavity. Our aim was to evaluate whether leucocytes migrating from a transplanted intestine could be recovered from the abdominal draining fluid collected by a peritoneal drainage system in the early post-ITx period, and to determine potential applications of the assessment of draining cellular populations. The cell composition of the abdominal draining fluid was analysed during the first 11 post-ITx days. Using flow cytometry, immune cells from blood and draining fluid samples obtained the same day showed an almost complete lymphopenia in peripheral blood, whereas CD3+CD4+CD8 -, CD3+CD4-CD8+ and human leucocyte antigen D-related (HLA-DR)+CD19+ lymphocytes were the main populations in the draining fluid. Non-complicated recipients evolved from a mixed leucocyte pattern including granulocytes, monocytes and lymphocytes to an exclusively lymphocytic pattern along the first post-ITx week. At days 1-2 post-Itx, analysis by short tandem repeats fingerprinting of CD3 +CD8+ sorted T cells from draining fluid indicated that 50% of cells were from graft origin, whereas by day 11 post-ITx this proportion decreased to fewer than 1%. Our results show for the first time that the abdominal drainage fluid contains mainly immune cells trafficking from the implanted intestine, providing the opportunity to sample lymphocytes draining from the grafted organ along the post-ITx period. Therefore, this analysis may provide information useful for understanding ITx immunobiology and eventually could also be of interest for clinical management.Laboratorio de Investigaciones del Sistema Inmun

Neutrophil extracellular traps stimulate proinflammatory responses in human airway epithelial cells

Tissue injury leads to the release of uric acid (UA). At high local concentrations, UA can form monosodium urate crystals (MSU). MSU and UA stimulate neutrophils to release extracellular traps (NET). Here, we investigated whether these NET could be involved in the development of inflammation by stimulating cytokine release by airway epithelial cells. We found that NET significantly increased the secretion of CXCL8/IL-8 and IL-6 by alveolar and bronchial epithelial cells. These effects were not observed when NETosis was inhibited by Diphenyleneiodonium, elastase inhibitor, or Cl-amidine. Similar findings were made with NET induced by cigarette smoke extract, suggesting that NET proinflammatory capacity is independent of the inducing stimulus. Furthermore, NET affected neither the viability and morphology of epithelial cells nor the barrier integrity of polarized cells. The epithelial stimulatory capacity of NET was not affected by degradation of DNA with micrococcal nuclease, treatment with heparin, or inhibition of the elastase immobilized to DNA, but it was significantly reduced by pretreatment with an anti-HMGB-1 blocking antibody. Altogether, our findings indicate that NET exert direct proinflammatory effects on airway epithelial cells that might contribute in vivo to the further recruitment of neutrophils and the perpetuation of inflammation upon lung tissue damage.Instituto de Estudios Inmunológicos y Fisiopatológico

Non-conventional vascular accesses for the management of superior vena cava syndrome in patients with Intestinal Failure: case series and systematic review

Background: Type III Intestinal Failure (IF) is a devastating clinical condition.characterized by the inability of the gut to absorb necessary macronutrients, and/or water and electrolytes, requiring Parenteral Nutrition (PN) as chronic therapy. Long-term PN may lead to life-threatening complications; the loss of central venous access (LCVA) is the most frequent and challenging. To date, few studies in the literature have reported the relevance of Non-conventional Vascular Accesses (NCVA) in the management IF as part of the comprehensive multidisciplinary care. Methods: A retrospective analysis of a database collected from January 2006 to December 2019 was performed using SPSS v25.0 for statistical analysis, followed by a systematic review, using the PRISMA.methodology Results: From January 2006 to December 2019, 184 NCVA were placed in 71 patients with LCVA as IF-related complication; 173 were placed in 61 patients by interventional radiology (IR) and 11 NCVA were placed in 10 patients by the surgical team during the intestinal transplant (ITx) operation. From the 173 IR procedures 166 (95.9%) were successful with 3 ± 2.7 procedures/patient; average catheter permanence rate was 738.68 ± 997 days; complications related to the procedures occurred in 18/173 (10.4%), including two deaths. On the other hand, among the 11 NCVA implanted by the surgical team, 7 (64%) were successful and were safely withdrawn 30 days after ITx when were no longer needed; 2 (18%) catheters malfunctioned during the first week and could not be further used, and 1 was accidently removed; average catheter permanence rate was 26 ± 4 days. There was one complication (9%) requiring laparotomy; there was no mortality associated the procedure in this group. A systematic review was conducted to evaluate the success and safety of NCVA as part of the treatment of HPN-related complications; from 337,542 papers, 14 studies were included. A total of 28 HPN-patients with LCVA received NCVA; 34 procedures were successfully performed, while procedure-related complications were reported in 11.7%, as well as one death. Conclusions: The data analyzed show that NCVAs may be successfully placed by expert teams, allowing to sustain long-term PN, as well as increasing the Intestinal Transplantation applicability for candidates in the extreme need of vascular access.Fil: Pérez Illidge, Luis Carlos. Universidad Favaloro; ArgentinaFil: Ramisch, Diego. Universidad Favaloro; ArgentinaFil: Valdivieso, León. Universidad Favaloro; ArgentinaFil: Guzman, Carlos. Universidad Favaloro; ArgentinaFil: Antoni, Diego. Universidad Favaloro; ArgentinaFil: Rumbo, Carolina. Universidad Favaloro; ArgentinaFil: Trentadue, Julio. Universidad Favaloro; ArgentinaFil: Solar, Héctor. Universidad Favaloro; ArgentinaFil: Gentilini, Maria Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Medicina Traslacional, Trasplante y Bioingeniería. Fundación Favaloro. Instituto de Medicina Traslacional, Trasplante y Bioingeniería; ArgentinaFil: Gondolesi, Gabriel Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Medicina Traslacional, Trasplante y Bioingeniería. Fundación Favaloro. Instituto de Medicina Traslacional, Trasplante y Bioingeniería; Argentin

Neutrophil Extracellular Traps Stimulate Proinflammatory Responses in Human Airway Epithelial Cells

Tissue injury leads to the release of uric acid (UA). At high local concentrations, UA can form monosodium urate crystals (MSU). MSU and UA stimulate neutrophils to release extracellular traps (NET). Here, we investigated whether these NET could be involved in the development of inflammation by stimulating cytokine release by airway epithelial cells. We found that NET significantly increased the secretion of CXCL8/IL-8 and IL-6 by alveolar and bronchial epithelial cells. These effects were not observed when NETosis was inhibited by Diphenyleneiodonium, elastase inhibitor, or Cl-amidine. Similar findings were made with NET induced by cigarette smoke extract, suggesting that NET proinflammatory capacity is independent of the inducing stimulus. Furthermore, NET affected neither the viability and morphology of epithelial cells nor the barrier integrity of polarized cells. The epithelial stimulatory capacity of NET was not affected by degradation of DNA with micrococcal nuclease, treatment with heparin, or inhibition of the elastase immobilized to DNA, but it was significantly reduced by pretreatment with an anti-HMGB-1 blocking antibody. Altogether, our findings indicate that NET exert direct proinflammatory effects on airway epithelial cells that might contribute in vivo to the further recruitment of neutrophils and the perpetuation of inflammation upon lung tissue damage.Fil: Sabbione, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Keitelman, Irene Angélica. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Iula, Leonardo Jairo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Ferrero, Mariana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Giordano, Mirta Nilda. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; ArgentinaFil: Baldi, Pablo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Rumbo, Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Jancic, Carolina Cristina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Trevani, Analía Silvina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentin

Experience with the use of teduglutide in paediatric patients with intestinal failure in a center in Argentina

Introducción. El teduglutide es un análogo sintético del péptido símil glucagón tipo 2, aprobado recientemente en Europa y en los Estado Unidos para uso pediátrico, para promover la adaptación intestinal en casos de síndrome de intestino corto. La experiencia en niños es limitada. Objetivo. El objetivo de este trabajo es presentar la evolución de la primera serie pediátrica tratada con teduglutide en Argentina. Material y métodos. Se realizó un estudio prospectivo sobre registros de pacientes menores de 18 años tratados en una unidad de rehabilitación y trasplante de intestino desde 2017. De 62 niños con síndrome de intestino corto, 5 recibieron teduglutide 0,05 mg/kg/día subcutáneo. Se analizaron los diagnósticos, el tipo de anatomía y la evolución de los aportes de nutrición parenteral. Resultados. La etiología del síndrome de intestino corto fue igual a 3 malformaciones congénitas (2 atresias, 1 gastrosquisis); dos causas en niños mayores: 1 vólvulo y 1 trauma abdominal. La anatomía de los pacientes era tres tipo 2A, uno tipo 2B y uno tipo 3A. La longitud del intestino remanente (media y DS) fue de 25,7 ± 19 cm, 1 con válvula ileocecal y colon, 4 hemicolon izquierdo. La edad al inicio del aporte de nutrición parenteral fue 6,2 ± 0,4 años, el tiempo de aporte de nutrición parenteral previo fue de 7,1 ± 4 años. El tiempo de tratamiento con teduglutide fue de 52,2 ± 39 semanas. El aporte de nutrición parenteral se redujo en 56 ± 48%, en todos los pacientes, pudiendo suspenderse en 2 (a las semanas 29° y 24° del tratamiento). No se registró deterioro del z score de IMC/edad (0,16 inicial ± 0,3 vs. 0,14 ± 1,02 al corte), ni de la talla/edad (-2,01 ± 1,5 vs. -1,76 ± 1,42). Conclusión. Se llegó a la conclusión de que el uso de teduglutide como alternativa terapéutica en el síndrome de intestino corto en pediatría resultó efectiva y segura en este grupo de pacientes, permitiendo restaurar la suficiencia intestinal o reduciendo los aportes de nutrición parenteral.Introduction. Teduglutide is a synthetic analogue of the glucagon-like peptide type 2, recently approved in Europe and in the United States for paediatric use, to promote intestinal adaptation in short bowel syndrome cases. The experience in children is limited. Objective. The aim of this work is to present the evolution of the first paediatric series treated with teduglutide in Argentina. Material and methods. A prospective study was realized on patient records under 18 years treated in a rehabilitation and intestine transplant unit since 2017. Of 62 children with short bowel syndrome, 5 received teduglutide 0.05 mg/kg/subcutaneous day. Diagnostics, type of anatomy and evolution of parenteral nutrition requirements were realized . Results. Etiology of short bowel syndrome: 3 congenital malformations: 2 atresias, 1 gastroschisis, two causes in older children: 1 volvulus and 1 abdominal trauma. Anatomy. Three patients type 2A, one type 2B and one type 3A, the length of the remaining intestine (mean and DS) 25.7 ± 19 cm, 1 with ileocecal valve and colon, 4 left hemicolon. The age at the beginning of parenteral nutrition was 6.2 ± 0.4 years, time on previous parenteral nutrition was 7.1 ± 4 years. The treatment time with teduglutide was 52.2 ± 39 weeks. The parenteral nutrition requirements were reduced by 56 ± 48%, in all patients, and could be suspended in 2 (at the 29th and 24th weeks of treatment). There was no deterioration of the z score of BMI/age (initial 0.16 ± 0.3 vs. 0.14 ± 1.02), nor of the height/age (-2.01 ± 1.5 vs. -1.76 ± 1.42). Conclusion. It was concluded that the use of teduglutide as a therapeutic alternative in the short bowel syndrome in paediatrics was effective and safe in this group of patients, allowing the restoration of intestinal sufficiency or reducing the requirements of parenteral nutrition.Fil: Martinez, Maria Ines. Fundación Favaloro; ArgentinaFil: Rumbo, Carolina. Fundación Favaloro; ArgentinaFil: Garcia Hervá, Dolores. Fundación Favaloro; ArgentinaFil: Trentadeue, Julio. Fundación Favaloro; ArgentinaFil: Gondolesi, Gabriel Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Medicina Traslacional, Trasplante y Bioingeniería. Fundación Favaloro. Instituto de Medicina Traslacional, Trasplante y Bioingeniería; Argentina. Universidad Favaloro. Área de Investigación y Desarrollo; ArgentinaFil: Fernández, Adriana. Fundación Favaloro; Argentin

Intestinal transplantation in pediatrics: Analysis of the first recipient series in Argentina

El presente trabajo comunica la evolución posoperatoria inmediata y alejada de los pacientes pediátricos trasplantados en un único centro en la Argentina, desde marzo de 2006 hasta marzo de 2010, en variables demográficas, indicaciones, contraindicaciones, evolución y supervivencia de pacientes e injerto. Basándose en los resultados se puede concluir que el trasplante intestinal debe ser considerado como un tratamiento válido para todos aquellos pacientes que presenten insuficiencia intestinal, con complicaciones del soporte parenteral. Los resultados adquiridos en nuestro programa son similares a los comunicados internacionalmente y abren una nueva perspectiva para un grupo especial de niños que carecían de solución en nuestro medio.The present is a retrospective analysis of all pediatric patients that underwent intestinal transplant from march 2006 to march 2010, describing demographics, indications, contraindications, clinical follow up and survival in a single center in Argentina. Based on the results shown one can conclude that intestinal transplant should be considered as a valid treatment for patients with intestinal insufficiency and complications related to parenteral nutrition. The results of our program are similar to those reported in the international Intestinal Transplant Registry. This opens a new perspective to a special population that otherwise would not have any other therapeutic option.Fil: Trentadue, Julio. Fundación Favaloro; ArgentinaFil: Rumbo, Carolina. Fundación Favaloro; ArgentinaFil: Garcia Hervás, Dolores. Fundación Favaloro; ArgentinaFil: Saá, Gladys. Fundación Favaloro; ArgentinaFil: Martínez, María Inés. Fundación Favaloro; ArgentinaFil: Orce, Guillermo. Fundación Favaloro; ArgentinaFil: Fernández, Adriana. Fundación Favaloro; ArgentinaFil: Gondolesi, Gabriel Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Favaloro. Área de Investigación y Desarrollo; Argentin