Immune reconstitution after allogenic stem cell transplantation: An observational study in pediatric patients

Abstract Introduction The immune reconstitution (IR) after the allogenic hematopoietic stem cell transplantation (allo-HSCT) is a progressive process intrinsically correlated to the therapeutic success. It is essential to understand the interfering factors in IR to prevent the HSCT-related mortality. Methods We retrospectively evaluated the clinical outcomes, absolute lymphocyte counts (ALCs) and lymphocyte subtypes at different time-points of 111 pediatric patients with allogeneic HSCT for malignant and non-malignant diseases from 2013 to 2018. Results The ALCs gradually increased on D+30, D+100, and D+180 (medians 634/μL, 1022/μL and 1541/μL, respectively). On D+100, the CD3+CD8+ achieved the highest recovery rate (68%), followed by the CD16+CD56+ (47%), CD3+CD4+ (39%) and CD19+ (8%). The adequate ALC recovery was associated with age < 8 years, bone marrow grafts, myeloablative conditioning, non-use of serotherapy and non-haploidentical donors. The ALC and CD3+CD8+ on D+100 counts were higher in patients with the cytomegalovirus infection. The CD3+CD4+ recovery was associated with an age < 8 years, a non-malignant disease and a lower incidence of acute graft-versus-host disease ≥ grade 2. Furthermore, the ALC recovery on D+100 resulted in a higher overall survival, regardless of the disease type (HR 3.65, 1.05 - 12.71, p= 0.04). Conclusion Several factors influenced the IR after the allo-HSCT. The ALC ≥ 500/μL on D+100 was a simple IR predictor of survival, easily available to resource-limited centers.

Biomodulina T® como alternativa de tratamiento en el shock séptico en pediatría / Biomodulina T® as an alternative to the treatment of septic shock in pediatrics

Introducción: La sepsis y el shock séptico se encuentran entre las principales causas de morbilidad y mortalidad en la población pediátrica a nivel mundial. Encontrar soluciones alternativas para combatirlas, mediante el desarrollo de agentes inmunomoduladores, ha atraído el interés de investigadores en los últimos 20 años; Cuba cuenta con Biomodulina T®, un potente inmunomodulador. Objetivo: Demostrar que existe evidencia científica que avale la realización de ensayos clínicos controlados para la incorporación de la Biomodulina T® en las pautas de tratamientos de la sepsis en las terapias intensivas pediátricas. Material y Métodos: Se realizó una búsqueda en las bases de datos Medline, PubMed, SciELO, Lilacs, Cochrane Library y Web of Science, entre marzo de 2019 y marzo de 2020; se seleccionaron los 47 artículos de mayor relevancia para esta investigación. Desarrollo: La inmunopatogenia del shock se centra en un fenotipo complejo y alteraciones funcionales, tanto del sistema inmune innato como del sistema adaptativo, con disminución del número de células efectoras, aumento de subpoblaciones de linfocitos inmunosupresores y agotamiento de células T. Biomodulina T® estimula la producción de linfocitos T y robustece la diferenciación de las células linfoblastoides del timo. La práctica médica sugiere que su administración podría ser una estrategia prometedora para la restauración inmune en pacientes pediátricos con shock séptico. Conclusiones: Existe evidencia científica que respalda el uso de Biomodulina T® en pacientes con shock séptico, lo cual sustenta la fiabilidad de realizar ensayos clínicos controlados en población pediátrica para su posterior incorporación en las pautas de tratamientos en las terapias intensivas(AU)

Introduction: Sepsis and septic shock are among the main causes of morbidity and mortality in the pediatric population worldwide. Finding alternative solutions to combat them through the development of immunomodulatory agents has attracted the interest of researchers in the last 20 years; Cuba has Biomodulina T®, a powerful immunomodulator. Objective: To demonstrate that there is scientific evidence that supports the conduction of controlled clinical trials for the incorporation of Biomodulina T® into the treatment guidelines for sepsis in pediatric intensive care therapies. Material and Methods: A search was carried out in the Medline, PubMed, SciELO, Lilacs, the Cochrane Library and the Web of Science databases between March 2019 and March 2020; the 47 most relevant articlesfor this research were selected. Development: The immunopathogenesis of shock focuses on a complex phenotype and functional alterations of both the innate and adaptive immune systems with a decrease in the number of effector cells, an increase in subpopulations of immunosuppressive lymphocytes, and depletion of T cells. Biomodulina T® stimulates the production of T lymphocytes and strengthens the differentiation of lymphoblastoid cells of the thymus; medical practice suggests that its administration could be a promising strategy for immune restoration in pediatric patients with septic shock. Conclusions: There is scientific evidence that supports the use of Biomodulina T® in patients with septic shock, which supports the reliability of conducting controlled clinical trials in the pediatric population for its subsequent incorporation into treatment guidelines in intensive care therapies(AU)

Interleukin-12 over-expression in malignant melanoma B16 cells reduces programmed death-1 expression on T cells in mice with immune reconstitution / 南方医科大学学报

OBJECTIVE@#To investigate whether interleukin-12 (IL-12) over-expression in malignant melanoma B16 cells affects the expression level of programmed death-1 (PD-1) on T cells in mice during immune microenvironment reconstruction.@*METHODS@#B16 cells were transfected with an IL-12 expression lentiviral vector, and IL-12 over-expression in the cells was verified qPCR and ELISA. Plate cloning assay was used to compare the cell proliferation activity between B16 cells and B16/IL-12 cells. The expression of IL-12 protein in B16/IL-12 cells-derived tumor tissue were detected by ELISA. C57BL/6 mice were inoculated with B16 cells or B16/IL-12 cells, and 14 days later the proportion of T cells with high expression of PD-1 in the tumor-draining lymph nodes was detected by flow cytometry. Mouse models of immune reconstitution established by 650 cGy X-ray radiation were inoculated with B16 (B16+RT group) or B16/IL-12 (B16/IL-12+RT group) cells, with the mice without X-ray radiation prior to B16 cell inoculation as controls. Tumor growth in the mice was recorded at different time points, and on day 14, flow cytometry was performed to detect the proportion of T cells with high PD-1 expression in the tumor-draining lymph nodes and in the tumor tissue.@*RESULTS@#B16 cells infected with the IL-12-overexpressing lentiviral vector showed significantly increased mRNA and protein levels of IL-12 ( < 0.001) without obvious changes in cell viability (>0.05). B16/IL-12 cells expressed higher levels of IL-12 than B16 cells ( < 0.01). In the tumor-bearing mouse models, the proportion of CD4 PD-1 T cells was significantly lower in B16/IL-12 group than in B16 group ( < 0.01). In the mice with X-ray radiation-induced immune reconstitution, PD-1 expressions on CD4 T cells ( < 0.05) and CD8+ T cells ( < 0.01) were significantly higher in B16+ RT group than in the control mice and in B16/IL-12+RT group ( < 0.01 or 0.001); the tumors grew more slowly in B16/IL-12+RT group than in B16 + RT group ( < 0.001).@*CONCLUSIONS@#During immune microenvironment reconstruction, overexpression IL-12 in the tumor microenvironment can reduce the percentage of PD-1 T cells and suppress the growth of malignant melanoma in mice.

Advances in newborn screening and immune system reconstitution of severe combined immunodeficiency / 浙江大学学报·医学版

Severe combined immunodeficiency disease (SCID) is a group of rare congenital diseases characterized by severe deficiencies in T lymphocyte counts and/or function. The recurrent, persistent and severe infections are its clinical manifestations. Neonatal screening and immune system reconstruction would improve the prognosis of SCID children. Newborn screening programs based on T-cell receptor excision circles (TRECs) quantitative detection have been carried out in clinical practice, however, the methods still have some limitations. Other new methods such as mass spectrometry and T lymphocyte-specific biomarker assays are still under investigation. Hematopoietic stem cell transplantation and gene therapy are the two main methods for reconstructing immune function in SCID children. Through improving the success rate of transplantation and the long-term safety and stability of viral vectors, some achievements have been made by many centers already. However, large-scale prospective studies are needed for evaluation of the long-term efficacy. In this article, the recent progress in newborn screening and immune reconstitution of SCID is reviewed.

Effects of High Cytomegalovirus DNA Load on Immune Reconstitution and Clinical Outcomes after Single Unrelated Cord Blood Transplantation / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of cytomegalovirus (CMV) DNA load on immune reconstitution and clinical outcomes of patients after unrelated cord blood transplantation (UCBT).@*METHODS@#Eight-color flow cytometry was used to dynamically monitor the changes of peripheral blood lymphocyte subsets of 41 patients at one year after UCBT, and 10 healthy volunteers were enroled as controls. Patients were divided into two groups according to the DNA load of CMV (DNA copies <1000/ml and DNA copies ≥1000/ml). Comparative analyse of the effect of CMV DNA load on lymphocyte subsets and transplantation outcomes were carried out after transplantation.@*RESULTS@#The high CMV DNA load group showed a faster and expanded T cell reconstitution, and the differences between the two groups were statistically significant at one and nine months after transplantation (0.38×10 /L vs 0.25×10 /L, P=0.015 and 2.53×10 /L vs 1.36×10 /L, P=0.006, respectively). Further analysis of T cell subsets suggested that CD8 T cells presented a higher and faster recovery in the high DNA load group, and the differences between the two groups were statistically significant at one and nine months after transplantation (0.20×10 /L vs 0.10×10 /L, P=0.038 and 1.62×10 /L vs 0.68×10 /L, P=0.003, respectively). In addition, there were no significant differences in levels of B cells, regulatory B cells and NK cells between the two groups. Outcomes after one- and a-half-year transplantation showed that there were no significant difference in relapse, non-relapse mortality and overall survival between the high and the low DNA load groups (7.7% vs 7.5%) (P=0.900) (15.4% vs 21.4%) (P=0.686) and (76.9% vs 78.6%) (P=0.889) respectively.@*CONCLUSION@#The high CMV DNA load induces a faster and long-lasting expansion of T cells, mainly as the expansion of CD8 T cells after UCBT. Besides, under the current pre-emptive treatment of CMV, the high CMV DNA load does not affect the early survival of patients with acute myeloid leukemia after UCBT.

Comparison of reference values for immune recovery between event-free patients receiving haploidentical allografts and those receiving human leukocyte antigen-matched sibling donor allografts / 医学前沿

To establish optimal reference values for recovered immune cell subsets, we prospectively investigated post-transplant immune reconstitution (IR) in 144 patients who received allogeneic stem cell transplantation (allo- SCT) and without showing any of the following events: poor graft function, grades II‒IV acute graft-versus-host disease (GVHD), serious chronic GVHD, serious bacterial infection, invasive fungal infection, or relapse or death in the first year after transplantation. IR was rapid in monocytes, intermediate in lymphocytes, CD3 Tcells, CD8 T cells, and CD19 B cells, and very slow in CD4 T cells in the entire patient cohort. Immune recovery was generally faster under HLA-matched sibling donor transplantation than under haploidentical transplantation. Results suggest that patients with an IR comparable to the reference values display superior survival, and the levels of recovery in immune cells need not reach those in healthy donor in the first year after transplantation.We suggest that data from this recipient cohort should be used as reference values for post-transplant immune cell counts in patients receiving HSCT.

Síndrome de reconstitución inmune en un paciente de sida con una tuberculosis diseminada / Immune reconstitution syndrome in an AIDS patient with disseminated tuberculosis

La tuberculosis es un factor de riesgo en los pacientes con sida, ya que una vez iniciado el tratamiento antirretroviral pueden de desarrollar un síndrome de reconstitución inmune, lo que favorecería el deterioro del su estado clínico. Se presenta el caso de un paciente masculino, de 24 años de edad, diagnosticado de sida hace 4 años, y tratamiento irregular con antirretrovirales. Acudió al Hospital Universitario Clínico Quirúrgico "Comandante Faustino Pérez Hernández" con fiebre elevada, acompañado de cuadro general, manifestaciones respiratorias y dolor inguinal derecho. En el examen físico se constató un cuadro adénico generalizado, fue hospitalizado para estudio y tratamiento. Se diagnosticó un síndrome de reconstitución inmune en un paciente de sida con una tuberculosis diseminada, el cual fallece a pesar de la terapéutica impuesta. Este síndrome se caracteriza por una restauración gradual de la inmunidad patógeno-específica, donde el sistema inmune es capaz de reconocer patógenos presentes pero clínicamente ocultos. Se asocia a otros factores de riesgo y puede ser letal; de ahí que el reconocimiento oportuno de los pacientes con alto riesgo de contraerlo, así como un adecuado manejo sobre cuándo iniciar la terapia antirretroviral en cada caso específico, es quizá la única forma de prevenir su desarrollo (AU).

Tuberculosis is a risk factor in patients with AIDS, because once the retroviral treatment begins they can develop an immune reconstitution syndrome that would favor the deterioration of their clinical status. The case of a male patient, aged 24 years is presented. He was diagnosed with AIDS four years ago, and was irregularly treated with antiretroviral. The patient assisted the Clinic-surgical University Hospital "Comandante Faustino Pérez Hernández" with high fever accompanied by general characteristics, respiratory manifestations and right inguinal pain. At the physical examination, generalized adenic characteristics were found. A syndrome of immune reconstitution was diagnosed in an AIDS patient with disseminated tuberculosis; the patient died in spite of the imposed therapy. This syndrome is characterized by the gradual restoration of the pathogen-specific immunity, where the immune system is able of recognizing the pathogens that are present but clinically hidden. It is associated to other risk facts and may be lethal; therefore the timely recognition of the patients at high risk of suffering it, and also an adequate management about when to begin the anti-retroviral therapy in each specific case, is the unique way of preventing its development (AU).

Calidad bacteriológica de comidas listas para su consumo, preparadas y distribuidas en la isla de Margarita (Venezuela), período 1991-2009 / Bacteriological quality of ready to eat meals prepared and distributed in Margarita island (Venezuela) during 1991-2009

Se valoró la calidad bacteriológica de 696 comidas listas para su consumo preparadas desde 1991 a 2009. 454 muestras correspondían a alimentos tratados térmicamente y el resto a alimentos que llevaban ingredientes no so metidos a cocción. Las muestras se escogieron al final de su elaboración, y antes de distribuirlas, en diferentes establecimientos de res tauración (hoteles, empresas de catering, restaurantes, supermercados, panaderías, luncherías, areperas, ventas ambulantes, cantinas y comedores es colares y casas de alimentación). Los criterios considerados fueron NMP/g de Coliformes, Coliformes fecales y Escherichia coli; y recuentos (UFC/g) de aerobios mesófilos y Staphylo coccus aureus (coagulasa +), según normas COVENIN y FONDONORMA. Los resultados se compararon con requisitos internos establecidos por el Laboratorio de Microbiolo gía, Fundación La Salle de C.N. Al momento de la captación el promedio de la temperatura de las comidas servidas en frío fue 13,4 °C y el de las cocinadas 32,2 °C. Los menores porcentajes de cumplimiento a todos los requisitos los tuvieron las rebanadas de jamón y queso (28%), rellenos de arepas y empanadas (28%), pastas italianas (23%), postres horneados (13%) y ensaladas con ingredientes crudos (7%). En este tipo de ensaladas sólo cumplieron los requisitos de Coliformes, Coliformes fecales y re cuentos de aerobios mesófilos el 19%; 31% y 24% de las muestras respectivamente. El menor cumplimiento de S. aureus (72%) se obtuvo en las rebanadas de jamón y queso y de E. coli (45%) en los rellenos de arepas y empanadas. Los establecimientos que presentaron los mayores valores de conformidad fueron las casas de alimentación (66%), comedores escolares (51%) y catering (50%) y los menores, los kioscos y ventas ambulantes (24%), supermercados y panaderías (17%) y cantinas escolares (0%). Las causas por las cuales los resultados bacteriológicos no cumplieron los criterios internos del Laboratorio pudieran estar relacionadas, entre otras, con la calidad de la materia prima, fallas higiénicas en la preparación de los alimentos, temperatura de mantenimiento al igual que tiempo y temperatura de cocción.

The bacteriological quality of 696 samples of ready to eat (RTE) food prepared from 1991 to 2009 was assessed. 454 samples corresponded to food subject to thermal treatment and the remainder carried ingredients that were not subject to cooking. The samples were chosen at the end of their elabo ration and before distributing, from different dinning establishments (hotels, caterers, restaurants, supermarkets, baker shops, luncheon rooms, arepa shops, ambulatory sales, canteens, school lunchrooms, and feeding houses). Total and fecal coliforms, E. coli, aerobic plate count and S. aureus (coagulase +) according to COVENIN and FONDONORMA norms were the criteria used for the investigation. The results were compared with internal requirements established by the microbiology laboratory of La Salle Natural Sciences Foundation. At the moment of sampling, the average temperature of cool dishes was 13.4 °C and of hot dishes was 32.2 °C. The smallest percentage of observance to all requisites were: ham and cheese slices (28%); arepas and turnovers (28%); Italian pasta (23%), baked desserts (13%) and salad with raw ingredients (7%). In these last ones, the requirements of total coliforms, fecal coliforms and aerobic plate count norms were met only by the 19%, 31% and 24% of samples respectively. The lowest compliance with S. aureus (72%) was found in the ham and cheese slices, and with E. coli (45%) were the arepas and turnovers. The establishments that presented the highest compliance values were the feeding houses (66%), school lunchrooms (51%) and caterers (50%); while those with the lowest values were the ambulatory sales (24%), supermarkets and baker shops (17%), and school canteens (0%). The reasons for non compliance with the internal requirements can be related, among other things, with the quality of raw material, hygienic failure in food handling, maintenance temperatures, and time and temperature of cooking.