Inhibition of lactate transport by MCT-1 blockade improves chimeric antigen receptor T-cell therapy against B-cell malignancies.

BACKGROUND: Chimeric antigen receptor (CAR) T cells have shown remarkable results against B-cell malignancies, but only a minority of patients have long-term remission. The metabolic requirements of both tumor cells and activated T cells result in production of lactate. The export of lactate is facilitated by expression of monocarboxylate transporter (MCTs). CAR T cells express high levels of MCT-1 and MCT-4 on activation, while certain tumors predominantly express MCT-1. METHODS: Here, we studied the combination of CD19-specific CAR T-cell therapy with pharmacological blockade of MCT-1 against B-cell lymphoma. RESULTS: MCT-1 inhibition with small molecules AZD3965 or AR-C155858 induced CAR T-cell metabolic rewiring but their effector function and phenotype remained unchanged, suggesting CAR T cells are insensitive to MCT-1 inhibition. Moreover, improved cytotoxicity in vitro and antitumoral control on mouse models was found with the combination of CAR T cells and MCT-1 blockade. CONCLUSION: This work highlights the potential of selective targeting of lactate metabolism via MCT-1 in combination with CAR T cells therapies against B-cell malignancies.

Preclinical evaluation of chimeric antigen receptor T cells targeting the carcinoembryonic antigen as a potential immunotherapy for gallbladder cancer.

Gallbladder cancer (GBC) is commonly diagnosed at late stages when conventional treatments achieve only modest clinical benefit. Therefore, effective treatments for advanced GBC are needed. In this context, the administration of T cells genetically engineered with chimeric antigen receptors (CAR) has shown remarkable results in hematological cancers and is being extensively studied for solid tumors. Interestingly, GBC tumors express canonical tumor-associated antigens, including the carcinoembryonic antigen (CEA). However, the potential of CEA as a relevant antigen in GBC to be targeted by CAR-T cell-based immunotherapy has not been addressed. Here we show that CEA was expressed in 88% of GBC tumors, with higher levels associated with advanced disease stages. CAR-T cells specifically recognized plate-bound CEA as evidenced by up-regulation of 4-1BB, CD69 and PD-1, and production of effector cytokines IFN-γ and TNF-α. In addition, CD8+ CAR-T cells up-regulated the cytotoxic molecules granzyme B and perforin. Interestingly, CAR-T cell activation occurred even in the presence of PD-L1. Consistent with these results, CAR-T cells efficiently recognized GBC cell lines expressing CEA and PD-L1, but not a CEA-negative cell line. Furthermore, CAR-T cells exhibited in vitro cytotoxicity and reduced in vivo tumor growth of GB-d1 cells. In summary, we demonstrate that CEA represents a relevant antigen for GBC that can be targeted by CAR-T cells at the preclinical level. This study warrants further development of the adoptive transfer of CEA-specific CAR-T cells as a potential immunotherapy for GBC.

Comportamiento de indicadores relacionados con la asistencia a pacientes con hemorragia subaracnoidea espontánea en la provincia de Cienfuegos

Fundamento: la importancia sanitaria de la hemorragia subaracnoidea espontánea, como problema de salud, es un hecho reconocido. Objetivo determinar el comportamiento de algunos indicadores relacionados con la asistencia médica a pacientes con hemorragia subaracnoidea espontánea, en el contexto provincial. Métodos estudio observacional, descriptivo-correlacional y retrospectivo, de 96 pacientes con hemorragia subaracnoidea espontánea entre 2016 y 2021. Se analizó el comportamiento de indicadores seleccionados, en su relación con diversas variables. En el análisis estadístico se utilizó como estadígrafo el Odds Ratio y su intervalo de confianza. Resultados el 22 % de los pacientes fue diagnosticado pasadas las primeras 48 horas del inicio de los síntomas, mientras que 13 enfermos (14 %) requirieron más de una asistencia para el diagnóstico. La ocurrencia de diagnóstico tardío cuando no se identificó el sangramiento en la primera consulta fue significativa (OR 21,8[5,1;91,8]). Doce pacientes fueron admitidos fuera de unidades especializadas; esta situación se observó más en pacientes menores de 60 años (21% vs 4 %; OR 5,7[1,1;27,9]), y en quienes el diagnóstico se realizó después de las 48 horas del inicio de las manifestaciones (29 % vs 8 %; OR 4,6[1,3;16,2]). De los 35 pacientes trasladados a otra institución para tratamiento neuroquirúrgico solo cuatro (12 %) fueron evacuados en las primeras 72 horas. Conclusiones se identifican brechas en la atención al paciente con hemorragia subaracnoidea en el contexto provincial; se destacan el ingreso de enfermos fuera de unidades especializadas y el traslado tardío a instituciones con servicio de cirugía neurovascular.

Background: the spontaneous subarachnoid hemorrhage's health importance, it's recognized as a health problem. Objective: to determine the behavior of some indicators related to medical care for patients with spontaneous subarachnoid hemorrhage, in the provincial context. Methods: Observational, descriptive-correlational and retrospective study of 96 patients with spontaneous subarachnoid hemorrhage between 2016 and 2021. The behavior of selected indicators was analyzed in relation to various variables. In the statistical analysis, the Odds Ratio and its confidence interval were used as statisticians. Results: 22% of the patients were diagnosed after the first 48 hours after the onset of symptoms, while 13 patients (14%) required more than one assistance for diagnosis. The occurrence of late diagnosis when bleeding was not identified at the first visit was significant (OR 21.8[5.1;91.8]). Twelve patients were admitted outside of specialized units; this situation was observed more in patients under 60 years of age (21% vs 4%; OR 5.7[1.1;27.9]), and in whom the diagnosis was made 48 hours after the onset of manifestations (29% vs 8%; OR 4.6[1.3;16.2]). Of the 35 patients transferred to another institution for neurosurgical treatment, only four (12%) were evacuated in the first 72 hours. Conclusions: gaps are identified in the patients' care with subarachnoid hemorrhage in the provincial context; The admission of patients outside specialized units and the late transfer to institutions with neurovascular surgery service stand out.

Identification of Compound Heterozygous EVC2 Gene Variants in Two Mexican Families with Ellis-van Creveld Syndrome.

BACKGROUND: Ellis-van Creveld syndrome (EvCS) is an autosomal recessive ciliopathy with a disproportionate short stature, polydactyly, dystrophic nails, oral defects, and cardiac anomalies. It is caused by pathogenic variants in the EVC or EVC2 genes. To obtain further insight into the genetics of EvCS, we identified the genetic defect for the EVC2 gene in two Mexican patients. METHODS: Two Mexican families were enrolled in this study. Exome sequencing was applied in the probands to screen potential genetic variant(s), and then Sanger sequencing was used to identify the variant in the parents. Finally, a prediction of the three-dimensional structure of the mutant proteins was made. RESULTS: One patient has a compound heterozygous EVC2 mutation: a novel heterozygous variant c.519_519 + 1delinsT inherited from her mother, and a heterozygous variant c.2161delC (p.L721fs) inherited from her father. The second patient has a previously reported compound heterozygous EVC2 mutation: nonsense mutation c.645G > A (p.W215*) in exon 5 inherited from her mother, and c.273dup (p.K92fs) in exon 2 inherited from her father. In both cases, the diagnostic was Ellis-van Creveld syndrome. Three-dimensional modeling of the EVC2 protein showed that truncated proteins are produced in both patients due to the generation of premature stop codons. CONCLUSION: The identified novel heterozygous EVC2 variants, c.2161delC and c.519_519 + 1delinsT, were responsible for the Ellis-van Creveld syndrome in one of the Mexican patients. In the second Mexican patient, we identified a compound heterozygous variant, c.645G > A and c.273dup, responsible for EvCS. The findings in this study extend the EVC2 mutation spectrum and may provide new insights into the EVC2 causation and diagnosis with implications for genetic counseling and clinical management.

Dopaminergic Signalling Enhances IL-2 Production and Strengthens Anti-Tumour Response Exerted by Cytotoxic T Lymphocytes in a Melanoma Mouse Model.

Dopamine has emerged as an important regulator of immunity. Recent evidence has shown that signalling through low-affinity dopamine receptors exerts anti-inflammatory effects, whilst stimulation of high-affinity dopamine receptors potentiates immunity in different models. However, the dopaminergic regulation of CD8+ T-cells in anti-tumour immunity remains poorly explored. Here, we studied the role of dopamine receptor D3 (DRD3), which displays the highest affinity for dopamine, in the function of CD8+ T-cells and its consequences in the anti-tumour immune response. We observed that the deficiency of Drd3 (the gene encoding DRD3) in CD8+ T-cells limits their in vivo expansion, leading to an impaired anti-tumour response in a mouse melanoma model. Mechanistic analyses suggest that DRD3 stimulation favours the production of interleukin 2 (IL-2) and the surface expression of CD25, the α-chain IL-2 receptor, which are required for expansion and effector differentiation of CD8+ T-cells. Thus, our results provide genetic and pharmacologic evidence indicating that DRD3 favours the production of IL-2 by CD8+ T-cells, which is associated with higher expansion and acquisition of effector function of these cells, promoting a more potent anti-tumour response in a melanoma mouse model. These findings contribute to understanding how dopaminergic signalling affects the cellular immune response and represent an opportunity to improve melanoma therapy.

Linfangiomatosis esplénica en un paciente adulto: Reporte de caso

Los linfangiomas son malformaciones benignas del sistema linfático que afectan con mayor frecuencia a los niños y en raras ocasiones a adultos. Presentamos el caso de una mujer de 36 años, con cuadro clínico de 6 meses de dolor abdominal difuso sin irritación peritoneal, y asociado a adinamia. Como parte de los estudios, se solicitó una tomografía axial computarizada con contraste de abdomen, que reveló múltiples lesiones esplénicas hipodensas hipovasculares, con sospecha diagnóstica de lesiones linfoproliferativas. Por tal motivo se solicitó una esplenectomía diagnóstica, que finalmente mostró hallazgos macroscópicos, microscópicos y de inmunohistoquímica (positividad de las lesiones quísticas para CD31 y D2-40) compatibles con linfagiomatosis esplénica difusa.

Lymphangiomas are benign malformations of the lymphatic system. They most often affect children and rarely adults. We present the case of a 36-year-old woman with a 6-month clinical picture of diffuse abdominal pain without peritoneal irritation associated with adynamia. As part of the studies, a computerized axial tomography with abdominal contrast was requested, which revealed multiple hypovascular hypodense splenic lesions with suspected diagnosis of lymphoproliferative lesions. For this reason, a diagnostic splenectomy was requested, which showed macroscopic, microscopic and immunohistochemical findings (positivity of cystic lesions for CD31 and D2-40) compatible with diffuse splenic lymphagiomatosis

Comportamiento y manejo actual de la infección por virus de la hepatitis B / Behavior and current management of hepatitis B virus infection

RESUMEN Introducción: Las hepatitis virales son un problema de salud a nivel mundial, provocan elevada mortalidad y morbilidad. A pesar de los avances en la prevención, diagnóstico y tratamiento de la hepatitis B, esta enfermedad es todavía preocupación de las autoridades sanitarias. Objetivo: Realizar una actualización acerca del comportamiento y manejo actual de la infección por el virus de la hepatitis B. Métodos: Se realizó una revisión bibliográfica durante abril y mayo de 2021, se consultaron 211 artículos en idioma inglés y español en las bases de datos como SciELO, PubMed, Lilacs y Medigraphic, de ellos se utilizaron 30 citas. Se empleó la combinación de términos y operadores booleanos y métodos teóricos como: análisis-síntesis, inducción-deducción e histórico-lógico. Resultados: Se abordó el comportamiento de la infección causada por el virus de la hepatitis B, se hizo hincapié en su epidemiología, características morfológicas y funcionales que permiten su infectividad, evolución y manifestaciones clínicas, complicaciones, tratamiento y perspectivas actuales. Conclusiones: La historia natural de la infección es dinámica y las manifestaciones clínicas dependen de diversos factores. La infección por el virus de la hepatitis B puede causar complicaciones como: la cirrosis hepática y el carcinoma hepatocelular y su principal vía de transmisión es la parenteral. El diagnóstico se realiza a través de técnicas serológicas. Es imprescindible la atención diferenciada a los grupos con riesgo y, aunque se cuenta con una vacuna eficaz, aún existen casos y se han desarrollado tratamientos con resultados positivos.

ABSTRACT Introduction: Viral hepatitis is a global health problem, causing high mortality and morbidity. Despite advances in the prevention, diagnosis and treatment of hepatitis B, this disease is still a concern of health authorities. Objective: Toupdate on current behavior and management of hepatitis B virus infection. Methods: A bibliographic review was carried out during April and May 2021, consulting 211 articles in English and Spanish from databases such as SciELO, PubMed, Lilacs and Medigraphic, of which 30 citations were used. The combination of Boolean terms and operators and theoretical methods such as: analysis-synthesis, induction-deduction and historical-logical were used. Results: The behavior of the infection caused by the hepatitis B virus was addressed, emphasizing its epidemiology, morphological and functional characteristics that allow its infectivity, evolution and clinical manifestations, complications, treatment and current perspectives. Conclusions: The natural history of infection is dynamic, and clinical manifestations depend on various factors. Hepatitis B virus infection can cause complications such as liver cirrhosis and hepatocellular carcinoma, and its main route of transmission is parenterally. The diagnosis is made mainly through serological techniques. Differentiated care for groups at risk is essential and, although there is an effective vaccine, there are still cases and treatments have been developed with positive results.

Actualización sobre hepatitis A

Introducción: La hepatitis es una enfermedad inflamatoria que afecta al hígado. El virus de la hepatitis A produce un cuadro infeccioso agudo por lo general autolimitado en el ser humano, para el cual no existe tratamiento específico. Objetivo: Realizar una actualización acerca del comportamiento y manejo actual de la infección por el virus de la hepatitis A. Métodos: Se realizó una revisión bibliográfica durante septiembre y octubre de 2021, se consultaron 156 artículos en idioma inglés y español de bases de datos como SciELO, PubMed, LILACS y Medigraphic, de ellos se utilizaron 30 citas. Se empleó la combinación de términos y operadores booleanos y métodos teóricos como: análisis-síntesis, inducción-deducción e histórico-lógico. Resultados: Se abordó el comportamiento de la infección causada por el virus de la hepatitis A, se hizo hincapié en su epidemiología, características morfológicas y funcionales que permiten su infectividad, evolución y manifestaciones clínicas, complicaciones, tratamiento y perspectivas actuales. Conclusiones: El virus de la hepatitis A produce un cuadro infeccioso agudo. Se presenta tanto de forma esporádica como epidémica. Es la forma más común de hepatitis viral aguda. La transmisión es de persona a persona por vía fecal oral. Los síntomas relacionados con las hepatitis virales agudas son variables e inespecíficos. El diagnóstico de certeza se realiza mediante las pruebas serológicas y técnicas de biología molecular para la identificación del virus. No existe tratamiento específico para esta infección.

Introduction: Hepatitis is an inflammatory disease that affects the liver. The hepatitis A virus produces a generally self-limited acute infectious picture in humans, for which there is no specific treatment. Objective: To update on current behavior and management of hepatitis A virus infection. Methods: A bibliographic review was carried out during September and October 2021, consulting 156 articles in English and Spanish from databases such as SciELO, PubMed, Lilacs and Medigraphic, of which 30 citations were used. The combination of Boolean terms and operators and theoretical methods such as: analysis-synthesis, induction-deduction and historical-logical was used. Results: The behavior of the infection caused by the hepatitis A virus was addressed, emphasizing its epidemiology, morphological and functional characteristics that allow its infectivity, evolution and clinical manifestations, complications, treatment and current perspectives. Conclusions: The hepatitis A virus produces an acute infectious picture. It occurs both sporadically and epidemically. It is the most common form of acute viral hepatitis. Transmission is person-to-person by the fecal oral route. Symptoms related to acute viral hepatitis are variable and nonspecific. The diagnosis of certainty is carried out by means of serological tests and molecular biology techniques for the identification of the virus. There is no specific treatment for this infection.

El HeberNasvac®, una opción en el tratamiento de la hepatitis B crónica

Introducción: En Cuba se produce la vacuna HeberNasvac® para el tratamiento de la hepatitis B crónica. Su relevancia radica en lograr resultados de control virológico sostenido en una mayor proporción de pacientes. Objetivo: Evaluar seguridad y efectividad de la vacuna terapéutica HeberNasvac® en el tratamiento de la hepatitis B crónica, en la provincia Camagüey en el periodo comprendido de enero de 2019 a diciembre de 2020. Métodos: Se realizó un estudio cuasi experimental de intervención terapéutica, en pacientes atendidos en la Consulta Provincial de hepatitis virales crónicas. El universo estuvo constituido por 24 pacientes adultos, con carga viral detectable al inicio del estudio. La fuente primaria de la investigación estuvo dada por la historia clínica. Resultados: En el primer ciclo fueron más frecuentes la cefalea y los estornudos; y en el segundo ciclo la fiebre y el malestar general. Al concluir el tratamiento la mayoría mostraron mejoría de resultados de las pruebas de función hepática. Antes del tratamiento el mayor número de pacientes presentó una carga viral detectable por encima de 250 copias/mL y después de haber recibido tratamiento varios de los casos resultaron carga no detectable. El tratamiento se consideró con mediana seguridad en el mayor número de pacientes y la efectividad fue alta. Conclusiones: Se presentaron más eventos adversos en el segundo ciclo del tratamiento. Las pruebas de función hepática mostraron mejoría al concluir tratamiento. La carga viral después del tratamiento presentó un descenso. El tratamiento mostró mediana seguridad y efectividad alta.

Introduction: Cuba produces the HeberNasvac® vaccine for the treatment of chronic hepatitis B. Its relevance lies in achieving sustained virological control results in a greater proportion of patients. Objective: To evaluate the safety and effectiveness of the therapeutic vaccine HeberNasvac® in the treatment of chronic hepatitis B, in the province of Camagüey in the period from January 2019 to December 2020. Methods: A quasi-experimental study of therapeutic intervention was carried out in patients treated at the Provincial Consultation of Chronic Viral Hepatitis. The universe consisted of 24 adult patients, with detectable viral load at the beginning of the study. The primary source of the investigation was given by the medical history. Results: In the first cycle, headache and sneezing were more prevalent; and in the second cycle, fever and general malaise were more prevalent. At the end of the treatment, the majority showed improvement in the results of liver function tests. Before treatment, the largest number of patients had a detectable viral load above 250 copies / mL and after receiving treatment, several of the cases resulted in an undetectable load. The treatment was considered to be of medium safety in the largest number of patients and the effectiveness was high. Conclusions: There were more adverse events in the second cycle of treatment. Liver function tests showed improvement at the end of treatment. The viral load after treatment showed a decrease. The treatment showed medium safety and high effectiveness.

Comportamiento de las enfermedades inflamatorias del intestino en el Servicio Provincial de Coloproctología de Camagüey / Behavior of the inflammatory diseases of the intestine in Camagüey's provincial Colon-proctology service

RESUMEN Fundamento: la enfermedad inflamatoria del intestino se define como una afectación inflamatoria crónica del tubo digestivo de causa desconocida que evoluciona de modo recurrente con brotes, remisiones que pueden presentar diversas complicaciones y manifestaciones extra digestivas. Objetivo: describir el comportamiento de las enfermedades inflamatorias del intestino en el Servicio Provincial de Coloproctología del Hospital Universitario Manuel Ascunce Domenech. Métodos: se realizó un estudio descriptivo y transversal, desde abril de 2016 hasta abril de 2018. El universo de estudio estuvo constituido por los 100 pacientes diagnosticados por estudios de endoscopía y biopsia de enfermedad inflamatoria del intestino en el servicio ya mencionado. La fuente primaria de la investigación estuvo dada por un formulario diseñado por los autores. Resultados: los pacientes diagnosticados de colitis ulcerosa ocuparon el primer lugar con predominio de la edad de diagnóstico entre 30-39 años; casi la mitad de los pacientes estudiados presentaron antecedentes familiares positivos de enfermedad inflamatoria intestinal, la localización distal fue la más frecuente para la colitis ulcerosa, mientras que para el Crohn solo fue la perineal reportado con un solo caso; las manifestaciones clínicas intestinales más frecuentes correspondieron a las diarreas. Conclusiones: la colitis ulcerativa resultó ser más frecuente que la enfermedad de Crohn y que la colitis inespecífica con la edad de diagnóstico entre 30-39 años, con predominio de antecedentes familiares de primer orden de la enfermedad, en la colitis ulcerosa prevaleció la localización distal y en el Crohn fue perineal donde la diarrea fue el síntoma más frecuente en ambas.

ABSTRACT Background: the inflammatory disease of the intestine defines like an inflammatory chronic affectation of the alimentary canal of unknown etiology that evolves of recurrent mode with sprouts and remissions and can present various complications and extra digestive manifestations. Objective: to describe the behavior of the inflammatory diseases of the intestine in Provincial Colon-proctology Service of the Teaching Hospital Manuel Ascunce Domenech. Methods: a descriptive cross-sectional study was carried out from April, 2016 to April, 2018. The universe of study was constituted for the 100 patients diagnosed by endoscopy's studies and biopsy of inflammatory disease of the intestine in the aforementioned service. The investigation's primary source was given for a fill-out form designed by the authors. Results: the patients diagnosed of ulcerous colitis occupied the first place with a predominance of age diagnosis between 30-39 years old, almost half of the studied patients presented familiar background of intestinal inflammatory disease, the distal localization showed the most frequent for the ulcerative idiopathic colitis, while for Crohn the most frequent localization was perinea localization with just a case; the clinical intestinal manifestation more frequent corresponded to the diarrheas. Conclusions: the ulcerative colitis turned out to be more frequent than the disease of Crohn and then unspecified colitis with an age diagnosis between 30-39 years old; and with predominance of first-rate family record of the disease; in the ulcerative colitis prevailed the location distal and in the Crohn it was perineal being the diarrhea the most frequent symptom in both.

The Heteromeric Complex Formed by Dopamine Receptor D5 and CCR9 Leads the Gut Homing of CD4+ T Cells Upon Inflammation.

BACKGROUND AND AIMS: CD4+ T cells constitute central players in inflammatory bowel diseases (IBDs), driving inflammation in the gut mucosa. Current evidence indicates that CCR9 and the integrin α4ß7 are necessary and sufficient to imprint colonic homing on CD4+ T cells upon inflammation. Interestingly, dopaminergic signaling has been previously involved in leukocyte homing. Despite dopamine levels are strongly reduced in the inflamed gut mucosa, the role of dopamine in the gut homing of T cells remains unknown. Here, we study how dopaminergic signaling affects T cells upon gut inflammation. METHODS: Gut inflammation was induced by transfer of naïve T cells into Rag1-/- mice or by administration of dextran sodium sulfate. T cell migration and differentiation were evaluated by adoptive transfer of congenic lymphocytes followed by flow cytometry analysis. Protein interaction was studied by bioluminescence resonance energy transfer analysis, bimolecular fluorescence complementation, and in situ proximity ligation assays. RESULTS: We show the surface receptor providing colonic tropism to effector CD4+ T cells upon inflammation is not CCR9 but the complex formed by CCR9 and the dopamine receptor D5 (DRD5). Assembly of the heteromeric complex was demonstrated in vitro and in vivo using samples from mouse and human origin. The CCR9:DRD5 heteroreceptor was upregulated in the intestinal mucosa of IBD patients. Signaling assays confirmed that complexes behave differently than individual receptors. Remarkably, the disruption of CCR9:DRD5 assembly attenuated the recruitment of CD4+ T cells into the colonic mucosa. CONCLUSIONS: Our findings describe a key homing receptor involved in gut inflammation and introduce a new cell surface module in immune cells: macromolecular complexes formed by G protein-coupled receptors integrating the sensing of multiple molecular cues.

Modulation of Cell Surface Receptor Expression by Modified Vaccinia Virus Ankara in Leukocytes of Healthy and HIV-Infected Individuals.

Viral vectors are increasingly used as delivery means to induce a specific immunity in humans and animals. However, they also impact the immune system, and it depends on the given context whether this is beneficial or not. The attenuated vaccinia virus strain modified vaccinia virus Ankara (MVA) has been used as a viral vector in clinical studies intended to treat and prevent cancer and infectious diseases. The adjuvant property of MVA is thought to be due to its capability to stimulate innate immunity. Here, we confirmed that MVA induces interleukin-8 (IL-8), and this chemokine was upregulated significantly more in monocytes and HLA-DRbright dendritic cells (DCs) of HIV-infected patients on combined antiretroviral therapy (ART) than in cells of healthy persons. The effect of MVA on cell surface receptors is mostly unknown. Using mass cytometry profiling, we investigated the expression of 17 cell surface receptors in leukocytes after ex vivo infection of human whole-blood samples with MVA. We found that MVA downregulates most of the characteristic cell surface markers in particular types of leukocytes. In contrast, C-X-C motif chemokine receptor 4 (CXCR4) was significantly upregulated in each leukocyte type of healthy persons. Additionally, we detected a relative higher cell surface expression of the HIV-1 co-receptors C-C motif chemokine receptor 5 (CCR5) and CXCR4 in leukocytes of HIV-ART patients than in healthy persons. Importantly, we showed that MVA infection significantly downregulated CCR5 in CD4+ T cells, CD8+ T cells, B cells, and three different DC populations. CD86, a costimulatory molecule for T cells, was significantly upregulated in HLA-DRbright DCs after MVA infection of whole blood from HIV-ART patients. However, MVA was unable to downregulate cell surface expression of CD11b and CD32 in monocytes and neutrophils of HIV-ART patients to the same extent as in monocytes and neutrophils of healthy persons. In summary, MVA modulates the expression of many different kinds of cell surface receptors in leukocytes, which can vary in cells originating from persons previously infected with other pathogens.

Antithrombin III Contributes to the Protective Effects of Fresh Frozen Plasma Following Hemorrhagic Shock by Preventing Syndecan-1 Shedding and Endothelial Barrier Disruption.

BACKGROUND: Endothelial dysfunction during hemorrhagic shock (HS) is associated with loss of cell-associated syndecan-1 (Sdc1) and hyperpermeability. Fresh frozen plasma (FFP) preserves Sdc1 and reduces permeability following HS, although the key mediators remain unknown. Antithrombin III (ATIII) is a plasma protein with potent anti-inflammatory and endothelial protective activity. We hypothesized that the protective effects of FFP on endothelial Sdc1 and permeability are mediated, in part, through ATIII. METHODS: ATIII and Sdc1 were measured in severely injured patients upon admission (N = 125) and hospital day 3 (N = 90) for correlation analysis. In vitro effects of ATIII on human lung microvascular endothelial cells (HLMVECs) were determined by pretreating cells with vehicle, FFP, ATIII-deficient FFP, or purified ATIII followed by TNFα stimulation. Sdc1 expression was measured by immunostaining and permeability by electrical impedance. To determine the role of ATIII in vivo, male mice were subjected to a fixed pressure exsanguination model of HS, followed by resuscitation with FFP, ATIII-deficient FFP, or ATIII-deficient FFP with ATIII repletion. Lung Sdc1 expression was assessed by immunostaining. RESULTS: Pearson correlation analysis showed a significant negative correlation between plasma levels of Sdc1 and ATIII (R = -0.62; P < 0.0001) in injured patients on hospital day 3. Also, in vitro, FFP and ATIII prevented TNFα-induced permeability (P < 0.05 vs TNFα) in HLMVECs. ATIII-deficient FFP had no effect; however, ATIII restoration reestablished its protective effects in a dose-dependent manner. Similarly, FFP and ATIII prevented TNFα-induced Sdc1 shedding in HLMVECs; however, ATIII-deficient FFP did not. In mice, Sdc1 expression was increased following FFP resuscitation (1.7 ±â€Š0.5, P < 0.01) vs. HS alone (1.0 ±â€Š0.3); however, no improvement was seen following ATIII-deficient FFP treatment (1.3 ±â€Š0.4, P = 0.3). ATIII restoration improved Sdc1 expression (1.5 ±â€Š0.9, P < 0.05) similar to that of FFP resuscitation. CONCLUSIONS: ATIII plays a role in FFP-mediated protection of endothelial Sdc1 expression and barrier function, making it a potential therapeutic target to mitigate HS-induced endothelial dysfunction. Further studies are needed to elucidate the mechanisms by which ATIII protects the endothelium.

Tissue-resident memory CD8+ T cells amplify anti-tumor immunity by triggering antigen spreading through dendritic cells.

Tissue-resident memory CD8+ T (Trm) cells mediate potent local innate and adaptive immune responses and play a central role against solid tumors. However, whether Trm cells cross-talk with dendritic cells (DCs) to support anti-tumor immunity remains unclear. Here we show that antigen-specific activation of skin Trm cells leads to maturation and migration to draining lymph nodes of cross-presenting dermal DCs. Tumor rejection mediated by Trm cells triggers the spread of cytotoxic CD8+ T cell responses against tumor-derived neo- and self-antigens via dermal DCs. These responses suppress the growth of intradermal tumors and disseminated melanoma lacking the Trm cell-targeted epitope. Moreover, analysis of RNA sequencing data from human melanoma tumors reveals that enrichment of a Trm cell gene signature associates with DC activation and improved survival. This work unveils the ability of Trm cells to amplify the breath of cytotoxic CD8+ T cell responses through DCs, thereby strengthening anti-tumor immunity.

Cystathionine-γ-lyase (CSE) deficiency increases erythropoiesis and promotes mitochondrial electron transport via the upregulation of coproporphyrinogen III oxidase and consequent stimulation of heme biosynthesis.

BACKGROUND: Hydrogen sulfide (H2S) is an endogenous gasotransmitter produced by mammalian cells. The current study investigated the potential role of H2S in the regulation of heme biosynthesis using mice deficient in cystathionine gamma-lyase (CSE), one of the three major mammalian H2S-producing enzymes. METHODS: Wild-type and global CSE-/- mice, as well as mitochondria prepared from their liver were used. In vivo, arterial and venous blood gases were measured, and survival of the mice to severe global hypoxia was monitored. Ex vivo, expression of various heme biosynthetic enzymes including coproporphyrinogen oxidase (CPOX) was measured, and mitochondrial function was evaluated using Extracellular Flux Analysis. Urine samples were collected to measure the oxidized porphyrinogen intermediates. The in vivo/ex vivo studies were complemented with mitochondrial bioenergetic studies in hepatocytes in vitro. Moreover, the potential effect of H2S on the CPOX promoter was studied in cells expressing a CPOX promoter construct system. RESULTS: The main findings are as follows: (1) CSE-/- mice exhibit elevated red blood cell counts and red blood cell mean corpuscular volumes compared to wild-type mice; (2) these changes are associated with elevated plasma and liver heme levels and (3) these alterations are likely due to an induction of CPOX (the sixth enzyme involved in heme biosynthesis) in CSE-/- mice. (4) Based on in vitro promoter data the promoter activation of CPOX is directly influenced by H2S, the product of CSE. With respect to the potential functional relevance of these findings, (5) the increased circulating red blood cell numbers do not correspond to any detectable alterations in blood gas parameters under resting conditions, (6) nor do they affect the hypoxia tolerance of the animals in an acute severe hypoxia model. However, there may be a functional interaction between the CSE system and the CPOX system in terms of mitochondrial bioenergetics: (7) CSE-/- hepatocytes and mitochondria isolated from them exhibit increased oxidative phosphorylation parameters, and (8) this increase is partially blunted after CPOX silencing. Although heme is essential for the biosynthesis of mitochondrial electron chain complexes, and CPOX is required for heme biosynthesis, (9) the observed functional mitochondrial alterations are not associated with detectable changes in mitochondrial electron transport chain protein expression. CONCLUSIONS: The CSE system regulates the expression of CPOX and consequent heme synthesis. These effects in turn, do not influence global oxygen transport parameters, but may regulate mitochondrial electron transport.

Regulation of endothelial cell permeability by platelet-derived extracellular vesicles.

BACKGROUND: Platelet (Plt)-derived extracellular vesicles (Plt-EVs) have hemostatic properties similar to Plts. In addition to hemostasis, Plts also function to stabilize the vasculature and maintain endothelial cell (EC) barrier integrity. We hypothesized that Plt-EVs would inhibit vascular EC permeability, similar to fresh Plts. To investigate this hypothesis, we used in vitro and in vivo models of vascular endothelial compromise and bleeding. METHODS: In the vitro model, Plt-EVs were isolated by ultracentrifugation and characterized for Plt markers and particle size distribution. Effects of Plts and Plt-EVs on endothelial barrier function were assessed by transendothelial electrical resistance measurements and histological analysis of endothelial junction proteins. Hemostatic potential of Plt-EVs and Plts was assessed by multiple electrode Plt aggregometry. Using an in vivo model, the effects of Plts and Plt-EVs on vascular permeability and bleeding were assessed in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice by an established Miles assay of vascular permeability and a tail snip bleeding assay. RESULTS: In the in vitro model, Plt-EVs displayed exosomal size distribution and expressed Plt-specific surface markers. Platelets and Plt-EVs decreased EC permeability and restored EC junctions after thrombin challenge. Multiplate aggregometry revealed that Plt-EVs enhanced thrombin receptor-activating peptide-mediated aggregation of whole blood, whereas Plts enhanced thrombin receptor-activating peptide-, arachidonic acid-, collagen-, and adenosine diphosphate-mediated aggregation. In the in vivo model, Plt-EVs are equivalent to Plts in attenuating vascular endothelial growth factor (VEGF)-A-induced vascular permeability and uncontrolled blood loss in a tail snip hemorrhage model. CONCLUSION: Our study is the first to report that Plt-EVs might provide a feasible product for transfusion in trauma patients to attenuate bleeding, inhibit vascular permeability, and mitigate the endotheliopathy of trauma.

La familia, la comunicación humana y el enfoque sistémico en su relación con la esquizofrenia / Family, human communication and systemic approach in their relation with schizophrenia

Para el análisis teórico y práctico de la familia es necesario conocer su funcionamiento, el concepto de sistema y sus diferentes teorías, la comunicación como elemento primordial en todo grupo social y las particularidades del doble vínculo en aquellas con un miembro esquizofrénico, ya que generalmente muestran problemas en la comunicación, no solo desde el comportamiento del enfermo, sino también desde los demás familiares. En ese sentido se efectuó una revisión bibliográfica relacionada con el tema que permite la selección del enfoque sistémico para la aplicación en la práctica diaria.

For the theoretical and practical analysis of the family it is necessary to know its way of functioning, the system concept and its different theories, communication as main element in all social group and the particularities of double binding in those with a schizophrenic member, since generally they show problems in communication, not only from the sick person's behavior, but also from the other family members behavior. In that sense, a literature review related to the topic was carried out which allows the selection of the systemic approach for the implementation in the daily practice.

Screening of north-east Mexico medicinal plants with activities against herpes simplex virus and human cancer cell line.

The plants examined in this study have previous biological activity reports indicating the possibility of found activity against herpes and cancer cell. The aim of this contribution was to carry out a screening of Juglans mollis (Juglandaceae), Persea americana (Lauraceae), Hamelia patens (Rubiaceae), Salvia texana (Lamiaceae), Salvia ballotaeflora (Lamiaceae), Ceanothus coeruleus (Rhamnaceae), Chrysactinia mexicana (Asteraceae) y Clematis drummondii (Ranunculaceae), against HeLa cells, VHS-1 and VHS-2. The method MTT was used to determine the 50% cytotoxic concentration (CC50), in Vero and HeLa cell lines. To determine the 50% inhibitory concentration (IC50) against herpes, the plaque reduction method was used. Results showed that none of the plants exhibited activity against HeLa cells. About antiherpetic activity, J. mollis and S. ballotaeflora extracts present antiherpetic activity in terms of their SI, increasingly interest for further studies on the isolation of compounds with antiherpetic activity and about the mechanisms of action that produce this activity.

[Perfil de expresión de Ki67 en lesiones melanocíticas palmoplantares: estudio de casos y controles]. / Expression profile of Ki67 in palmoplantar melanocytic lesions: a case-control study.

INTRODUCCIÓN: El melanoma acral lentiginoso es una neoplasia maligna que afecta a población predominantemente no caucásica. Debido al diagnóstico tardío suele tener mal pronóstico, además de que se considera una neoplasia biológicamente más agresiva, incluso cuando se detecta tempranamente. OBJETIVO: Determinar la expresión de Ki67 en el melanoma acral lentiginoso invasor y compararla con los nevos acrales. MÉTODO: Estudio transversal, descriptivo, observacional. Se realizó inmunohistoquímica con marcador Ki67 en 17 biopsias de melanoma acral lentiginoso invasor (casos) y 17 biopsias de nevos palmoplantares (controles). Se determinó la expresión nuclear de Ki-67 y se comparó entre ambos grupos. RESULTADOS: La media de expresión de Ki67 fue del 8.5% en el grupo control y del 34% en el grupo de melanomas, siendo esta diferencia estadísticamente significativa (p < 0.0001). DISCUSIÓN: La expresión de Ki67 en los melanomas acrales es considerablemente mayor que en los nevos acrales. El valor pronóstico del marcador Ki67 sigue siendo considerado controversial. Sin embargo, hay estudios en los que en combinación con otros marcadores se refuerza su valor pronóstico. CONCLUSIONES: Por la gran diferencia en inmunorreactividad de Ki67 entre melanomas y nevos, la expresión de Ki67, referida como índice proliferativo, podría ser considerada como factor pronóstico incluso más objetivo que el índice mitótico. BACKGROUND: Acral lentiginous melanoma is a malignant neoplasm which appears in hands and feet. Acral lentiginous melanoma has an unclear etiology, and usually affects non-Caucasian population. Because it is frequently diagnosed lately, acral melanoma has bad prognosis; however, it is biologically more aggressive than other clinicopathological types of melanoma, even when diagnosed early. OBJECTIVE: To determine the expression of Ki67 in invasive lentiginous acral melanoma and to compare it with acral nevi. METHOD: Cross-sectional, descriptive, observational study. Immunohistochemistry with Ki67 marker was performed on 17 biopsies of invasive lentiginous acral melanoma (cases) and 17 biopsies of palmoplantar nevi (controls). Nuclear expression of Ki-67 was determined and both were compared between both groups. RESULTS: The mean expression of Ki67 was 8.5% in the control group, and 34% in the melanoma group, which was statistically significant (p < 0.0001). DISCUSSION: Ki67 expression in acral lentiginous melanomas is higher than in acral nevi. Prognostic value of Ki67 is still considered controversial. However, there are several studies where, in combination with other markers, their prognostic value is reinforced. CONCLUSIONS: Due to the wide gap in Ki67 expression between melanomas and nevi showed in this study, Ki67 expression, referred to as a proliferative index, could be considered as a prognostic factor even more objective than the mitotic index.

Vaccination-induced skin-resident memory CD8+ T cells mediate strong protection against cutaneous melanoma.

Memory CD8+ T cell responses have the potential to mediate long-lasting protection against cancers. Resident memory CD8+ T (Trm) cells stably reside in non-lymphoid tissues and mediate superior innate and adaptive immunity against pathogens. Emerging evidence indicates that Trm cells develop in human solid cancers and play a key role in controlling tumor growth. However, the specific contribution of Trm cells to anti-tumor immunity is incompletely understood. Moreover, clinically applicable vaccination strategies that efficiently establish Trm cell responses remain largely unexplored and are expected to strongly protect against tumors. Here we demonstrated that a single intradermal administration of gene- or protein-based vaccines efficiently induces specific Trm cell responses against models of tumor-specific and self-antigens, which accumulated in vaccinated and distant non-vaccinated skin. Vaccination-induced Trm cells were largely resistant to in vivo intravascular staining and antibody-dependent depletion. Intradermal, but not intraperitoneal vaccination, generated memory precursors expressing skin-homing molecules in circulation and Trm cells in skin. Interestingly, vaccination-induced Trm cell responses strongly suppressed the growth of B16F10 melanoma, independently of circulating memory CD8+ T cells, and were able to infiltrate tumors. This work highlights the therapeutic potential of vaccination-induced Trm cell responses to achieve potent protection against skin malignancies.