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1.
J Leukoc Biol ; 2024 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-39213186

RESUMEN

Eosinophils are granulocytes that can accumulate in increased numbers in tissues and/or peripheral blood in disease. Phenotyping of eosinophils in health and disease has the potential to improve the precision of diagnosis and choice of therapies for eosinophilic-associated diseases. Transcriptional profiling of eosinophils has been plagued by cell fragility and difficulty isolating high quality RNA. With several technological advances, single-cell RNA sequencing (scRNA-seq) has become possible with eosinophils, at least from mice, while bulk RNA sequencing and microarrays have been performed in both murine and human samples. Anticipating more eosinophil transcriptional profiles in the coming years, we provide a summary of prior studies conducted on mouse and human eosinophils in blood and tissue, with a discussion of the advantages and potential pitfalls of various approaches. Common technical standards in studying eosinophil biology would help advance the field and make cross-study comparisons possible. Knowledge gaps and opportunities include identifying a minimal set of genes that define the eosinophil lineage, comparative studies between active disease and remission vs. homeostasis or development, especially in humans, and a comprehensive comparison between murine and human eosinophils at the transcriptional level. Characterizing such transcriptional patterns will be important to understanding the complex and diverse roles of eosinophils in both health and disease.

3.
Allergy Asthma Proc ; 42(6): 515-521, 2021 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-34871159

RESUMEN

Background: Acute allergic reactions to messenger RNA (mRNA) vaccines are rare but may limit public health immunization efforts. Objectives: To characterize suspected allergic reactions to the first dose of coronavirus disease 2019 (COVID-19) mRNA vaccine and to assess the safety and utility of a two-step graded-dose protocol for the second dose of the Pfizer-BioNTech vaccine in patients with a history of low suspicion of anaphylaxis to their first dose. Methods: This was a retrospective evaluation of referrals to the allergy and immunology clinic for a presumed allergic reaction to the first dose of the COVID-19 mRNA vaccine (Pfizer-BioNTech or Moderna) between December 17, 2020, and February 28, 2021. Recommendations for the second dose and outcomes were evaluated by trained board-certified allergists. Results: Seventy-seven patients presented with a Pfizer-BioNTech reaction (56 [72.7%]) or with a Moderna reaction (21 [27.3%]). Most patients (69.7%) had symptom onset within 4 hours. Most commonly reported symptoms were cutaneous (51.9%), cardiovascular (48.1%), and respiratory (33.8%) symptoms. Recommendations included to proceed with the single dose (70.1%), two-step graded dose (19.5%), or deferral (10.4%). Twelve of 15 patients completed the second dose with a graded-dose protocol. Of these patients, five reported at least one or more similar symptoms as experienced with their first dose. Conclusion: Of the patients with presumed allergic reactions to their first dose of COVID-19 mRNA vaccine, most were able to safely receive the second dose. For those with a low suspicion of anaphylaxis, the two-step graded protocol with the Pfizer-BioNTech vaccine was well tolerated. A graded-dose protocol could be an effective strategy for second-dose vaccination in those who may otherwise defer the second dose.


Asunto(s)
Anafilaxia/inducido químicamente , Vacunas contra la COVID-19/efectos adversos , COVID-19/prevención & control , Hipersensibilidad , Vacunas Sintéticas/efectos adversos , Adulto , Anciano , COVID-19/epidemiología , Vacunas contra la COVID-19/administración & dosificación , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , SARS-CoV-2 , Vacunas Sintéticas/administración & dosificación , Vacunas de ARNm
4.
Ann Allergy Asthma Immunol ; 127(1): 70-75.e2, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33753218

RESUMEN

BACKGROUND: Immunoglobulin E-mediated food allergy (FA) affects children and adults with variable age of onset. Phenotype and quality of life (QoL) differences between childhood-onset FA (COFA) and adult-onset FA (AOFA) are not known. OBJECTIVE: To identify phenotypic and QoL differences between AOFA and COFA. METHODS: A cross-sectional study of adults (≥18 years old) seen at Northwestern Memorial HealthCare clinics between 2002 and 2017 with an International Classification of Diseases ninth and tenth revision diagnosis of FA. Subjects completed a FA history survey and a FA QoL questionnaire. FA characteristics and QoL scores were compared between groups. RESULTS: Among 294 consented subjects, 202 had a clinical history consistent with labeled immunoglobulin E-mediated FA. The onset of FA symptoms occurred before age 18 years (COFA) in 80 subjects and after age 18 years in 122 (AOFA) subjects. Shellfish reactions were most common in AOFA-labeled subjects (28%), whereas tree nut reactions were the most common in COFA-labeled subjects (55%) compared with other triggers. Hives (68% vs 52%, P = .03), facial swelling (69% vs 50%, P = .009), wheezing (56% vs 29%, P < .001), and vomiting (41% vs 22%, P = .005) were more often observed in COFA compared with AOFA. Total QoL was significantly reduced in COFA compared with AOFA (3.6 vs 3.0, P = .003) along with specific domains related to the following: allergen avoidance and dietary restriction (3.7 vs 3.1, P = .006), emotional impact (3.9 vs 3.2, P = .003), and risk of accidental exposure (3.6 vs 2.8, P = .001). CONCLUSION: There are differences in specific food triggers and symptoms in adult-onset and childhood-onset labeled FA. Adults labeled with childhood-onset FA have reduced QoL.


Asunto(s)
Hipersensibilidad a los Alimentos/psicología , Calidad de Vida , Adulto , Edad de Inicio , Niño , Estudios Transversales , Femenino , Hipersensibilidad a los Alimentos/inmunología , Humanos , Inmunoglobulina E/inmunología , Masculino , Persona de Mediana Edad , Encuestas y Cuestionarios
5.
J Allergy Clin Immunol ; 146(2): 307-314.e4, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-32554082

RESUMEN

BACKGROUND: The Centers for Disease Control and Prevention advises that patients with moderate to severe asthma belong to a high-risk group that is susceptible to severe coronavirus disease 2019 (COVID-19). However, the association between asthma and COVID-19 has not been well-established. OBJECTIVE: The primary objective was to determine the prevalence of asthma among patients with COVID-19 in a major US health system. We assessed the clinical characteristics and comorbidities in asthmatic and nonasthmatic patients with COVID-19. We also determined the risk of hospitalization associated with asthma and/or inhaled corticosteroid use. METHODS: Medical records of patients with COVID-19 were searched by a computer algorithm (March 1 to April 15, 2020), and chart review was used to validate the diagnosis of asthma and medications prescribed for asthma. All patients had PCR-confirmed COVID-19. Demographic and clinical features were characterized. Regression models were used to assess the associations between asthma and corticosteroid use and the risk of COVID-19-related hospitalization. RESULTS: Of 1526 patients identified with COVID-19, 220 (14%) were classified as having asthma. Asthma was not associated with an increased risk of hospitalization (relative risk, 0.96; 95% CI, 0.77-1.19) after adjusting for age, sex, and comorbidities. The ongoing use of inhaled corticosteroids did not increase the risk of hospitalization in a similar adjusted model (relative risk, 1.39; 95% CI, 0.90-2.15). CONCLUSIONS: Despite a substantial prevalence of asthma in our COVID-19 cohort, asthma was not associated with an increased risk of hospitalization. Similarly, the use of inhaled corticosteroids with or without systemic corticosteroids was not associated with COVID-19-related hospitalization.


Asunto(s)
Asma/epidemiología , Betacoronavirus/patogenicidad , Enfermedad de la Arteria Coronaria/epidemiología , Infecciones por Coronavirus/epidemiología , Diabetes Mellitus/epidemiología , Hipertensión/epidemiología , Obesidad/epidemiología , Neumonía Viral/epidemiología , Administración por Inhalación , Corticoesteroides/uso terapéutico , Adulto , Factores de Edad , Anciano , Antiasmáticos/uso terapéutico , Asma/diagnóstico , Asma/tratamiento farmacológico , Asma/fisiopatología , COVID-19 , Prueba de COVID-19 , Técnicas de Laboratorio Clínico/métodos , Comorbilidad , Enfermedad de la Arteria Coronaria/diagnóstico , Enfermedad de la Arteria Coronaria/fisiopatología , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/fisiopatología , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/fisiopatología , Femenino , Hospitalización/estadística & datos numéricos , Humanos , Hipertensión/diagnóstico , Hipertensión/fisiopatología , Illinois/epidemiología , Masculino , Persona de Mediana Edad , Modelos Estadísticos , Obesidad/diagnóstico , Obesidad/fisiopatología , Pandemias , Neumonía Viral/diagnóstico , Neumonía Viral/fisiopatología , Prevalencia , Estudios Retrospectivos , Factores de Riesgo , SARS-CoV-2
6.
J Clin Invest ; 130(9): 4759-4770, 2020 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-32484802

RESUMEN

No known therapies can prevent anaphylaxis. Bruton's tyrosine kinase (BTK) is an enzyme thought to be essential for high-affinity IgE receptor (FcεRI) signaling in human cells. We tested the hypothesis that FDA-approved BTK inhibitors (BTKis) would prevent IgE-mediated responses including anaphylaxis. We showed that irreversible BTKis broadly prevented IgE-mediated degranulation and cytokine production in primary human mast cells and blocked allergen-induced contraction of isolated human bronchi. To address their efficacy in vivo, we created and used what we believe to be a novel humanized mouse model of anaphylaxis that does not require marrow ablation or human tissue implantation. After a single intravenous injection of human CD34+ cells, NSG-SGM3 mice supported the population of mature human tissue-resident mast cells and basophils. These mice showed excellent responses during passive systemic anaphylaxis using human IgE to selectively evoke human mast cell and basophil activation, and response severity was controllable by alteration of the amount of allergen used for challenge. Remarkably, pretreatment with just 2 oral doses of the BTKi acalabrutinib completely prevented moderate IgE-mediated anaphylaxis in these mice and also significantly protected against death during severe anaphylaxis. Our data suggest that BTKis may be able to prevent anaphylaxis in humans by inhibiting FcεRI-mediated signaling.


Asunto(s)
Agammaglobulinemia Tirosina Quinasa/antagonistas & inhibidores , Anafilaxia/prevención & control , Benzamidas/farmacología , Inmunoglobulina E/inmunología , Inhibidores de Proteínas Quinasas/farmacología , Pirazinas/farmacología , Agammaglobulinemia Tirosina Quinasa/inmunología , Anafilaxia/inmunología , Anafilaxia/patología , Animales , Humanos , Ratones , Ratones Endogámicos NOD , Ratones SCID , Receptores de IgE/inmunología
7.
PLoS One ; 15(1): e0226701, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31940364

RESUMEN

IgE-primed mast cells in peripheral tissues, including the skin, lung, and intestine, are key initiators of allergen-triggered edema and inflammation. Particularly in severe forms of allergy, this inflammation becomes strongly neutrophil dominated, and yet how mast cells coordinate this type of response is unknown. We and others have reported that activated mast cells--a hematopoietic cell type--can produce IL-33, a cytokine known to participate in allergic responses but generally considered as being of epithelial origin and driving Type 2 immune responses (e.g., ILC2 and eosinophil activation). Using models of skin anaphylaxis, our data reveal that mast cell-derived IL-33 also initiates neutrophilic inflammation. We demonstrate a cellular crosstalk mechanism whereby activated mast cells crosstalk to IL-33 receptor-bearing basophils, driving these basophils to adopt a unique response signature rich in neutrophil-associated molecules. We further establish that basophil expression of CXCL1 is necessary for IgE-driven neutrophilic inflammation. Our findings thus unearth a new mechanism by which mast cells initiate local inflammation after antigen triggering and might explain the complex inflammatory phenotypes observed in severe allergic diseases. Moreover, our findings (i) establish a functional link from IL-33 to neutrophilic inflammation that extends IL-33-mediated biology well beyond that of Type 2 immunity, and (ii) demonstrate the functional importance of hematopoietic cell-derived IL-33 in allergic pathogenesis.


Asunto(s)
Basófilos/patología , Hipersensibilidad/inmunología , Hipersensibilidad/metabolismo , Interleucina-33/metabolismo , Mastocitos/patología , Animales , Comunicación Celular , Quimiocina CXCL1/metabolismo , Regulación de la Expresión Génica/inmunología , Hipersensibilidad/complicaciones , Inflamación/complicaciones , Ratones , Infiltración Neutrófila
8.
J Am Acad Dermatol ; 83(3): 737-744, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31862404

RESUMEN

BACKGROUND: Previous studies found conflicting results about whether hidradenitis suppurativa (HS) is associated with depression or anxiety. OBJECTIVES: To determine the relationship of HS with depression and anxiety. METHODS: A systematic review was performed of published observational studies in MEDLINE, PubMed, Embase, Global Resource for Eczema Trials (GREAT), Latin American and Caribbean Health Sciences Literature (LILACS), Cochrane, Scopus, and PsychInfo that analyzed depression or anxiety in HS. Two reviewers performed title/abstract review and data extraction. Meta-analysis was performed with random-effects weighting. RESULTS: Thirty-eight studies met inclusion criteria; 27 had sufficient data for meta-analysis. The prevalences of depression (26.5% vs 6.6%) and anxiety (18.1% vs 7.1%) were higher in persons with versus without HS. Patients with HS had higher odds of depression in 12 of 13 studies and pooled analysis (odds ratio, 2.54; 95% confidence interval, 2.15-3.01), and anxiety in 6 of 6 studies and pooled analysis (odds ratio, 2.00; 95% confidence interval, 1.66-2.42). Similar results were found in sensitivity analyses for different methods of HS diagnosis (physician diagnosed and chart review) and control groups (healthy and dermatologic control individuals). HS was associated with higher antidepressant and anxiolytic use and with suicidality, but not mean depression and anxiety scale scores. LIMITATIONS: Individual-level data were unavailable. CONCLUSIONS: Patients with HS have higher odds of depression, anxiety, and suicidality.


Asunto(s)
Ansiedad/epidemiología , Depresión/epidemiología , Hidradenitis Supurativa/complicaciones , Ideación Suicida , Ansiolíticos/uso terapéutico , Antidepresivos/uso terapéutico , Ansiedad/diagnóstico , Ansiedad/tratamiento farmacológico , Ansiedad/etiología , Depresión/diagnóstico , Depresión/tratamiento farmacológico , Depresión/etiología , Prescripciones de Medicamentos/estadística & datos numéricos , Hidradenitis Supurativa/psicología , Hidradenitis Supurativa/rehabilitación , Humanos , Cuestionario de Salud del Paciente/estadística & datos numéricos , Prevalencia
9.
Int J Mol Sci ; 20(1)2018 Dec 21.
Artículo en Inglés | MEDLINE | ID: mdl-30577572

RESUMEN

Sialic acid-binding Ig-like lectin 8 (Siglec-8) is expressed on the surface of human eosinophils, mast cells, and basophils-cells that participate in allergic and other diseases. Ligation of Siglec-8 by specific glycan ligands or antibodies triggers eosinophil death and inhibits mast cell degranulation; consequences that could be leveraged as treatment. However, Siglec-8 is not expressed in murine and most other species, thus limiting preclinical studies in vivo. Based on a ROSA26 knock-in vector, a construct was generated that contains the CAG promoter, a LoxP-floxed-Neo-STOP fragment, and full-length Siglec-8 cDNA. Through homologous recombination, this Siglec-8 construct was targeted into the mouse genome of C57BL/6 embryonic stem (ES) cells, and chimeric mice carrying the ROSA26-Siglec-8 gene were generated. After cross-breeding to mast cell-selective Cre-recombinase transgenic lines (CPA3-Cre, and Mcpt5-Cre), the expression of Siglec-8 in different cell types was determined by RT-PCR and flow cytometry. Peritoneal mast cells (dual FcεRI⁺ and c-Kit⁺) showed the strongest levels of surface Siglec-8 expression by multicolor flow cytometry compared to expression levels on tissue-derived mast cells. Siglec-8 was seen on a small percentage of peritoneal basophils, but not other leukocytes from CPA3-Siglec-8 mice. Siglec-8 mRNA and surface protein were also detected on bone marrow-derived mast cells. Transgenic expression of Siglec-8 in mice did not affect endogenous numbers of mast cells when quantified from multiple tissues. Thus, we generated two novel mouse strains, in which human Siglec-8 is selectively expressed on mast cells. These mice may enable the study of Siglec-8 biology in mast cells and its therapeutic targeting in vivo.


Asunto(s)
Antígenos CD/genética , Antígenos de Diferenciación de Linfocitos B/genética , Regulación de la Expresión Génica , Lectinas/genética , Mastocitos/metabolismo , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos B/metabolismo , Línea Celular , Técnicas de Sustitución del Gen , Marcación de Gen , Humanos , Hipersensibilidad/genética , Hipersensibilidad/inmunología , Lectinas/metabolismo , Mastocitos/inmunología , Ratones , Ratones Transgénicos , Especificidad de Órganos/genética
10.
Cancer Metab ; 6: 10, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30140438

RESUMEN

BACKGROUND: T cells and cancer cells utilize glycolysis for proliferation. The hexokinase (1-4) family of enzymes catalyze the first step of glycolysis. Hexokinase 2 (HK2) is one of the most highly upregulated metabolic enzymes in both cancer and activated T cells. HK2 is required for the development and/or growth of cancer in several cancer models, but the necessity of HK2 in T cells is not fully understood. The clinical applicability of HK2 inhibition in cancer may be significantly limited by any potential negative effects of HK2 inhibition on T cells. Therefore, we investigated the necessity of HK2 for T cell function. In order to identify additional therapeutic cancer targets, we performed RNA-seq to compare in vivo proliferating T cells to T cell leukemia. METHODS: HK2 was genetically ablated in mouse T cells using a floxed Hk2 allele crossed to CD4-Cre. CD4+ and CD8+ cells from mice were characterized metabolically and tested in vitro. T cell function in vivo was tested in a mouse model of colitis, Th2-mediated lung inflammation, and viral infection. Treg function was tested by crossing Hk2-floxed mice to FoxP3-Cre mice. Hematopoietic function was tested by deleting HK2 from bone marrow with Vav1-iCre. RNA-seq was used to compare T cells proliferating in response to virus with primary T-ALL leukemia induced with mutant Notch1 expression. RESULTS: We unexpectedly report that HK2 is largely dispensable for in vitro T cell activation, proliferation, and differentiation. Loss of HK2 does not impair in vivo viral immunity and causes only a small impairment in the development of pathological inflammation. HK2 is not required for Treg function or hematopoiesis in vivo. One hundred sixty-seven metabolic genes were identified as being differentially expressed between T cells and leukemia. CONCLUSIONS: HK2 is a highly upregulated enzyme in cancer and in T cells. The requirement for HK2 in various cancer models has been described previously. Our finding that T cells are able to withstand the loss of HK2 indicates that HK2 may be a promising candidate for cancer therapy. Furthermore, we identify several other potential metabolic targets in T-ALL leukemia that could spare T cell function.

11.
J Immunol ; 198(12): 4868-4878, 2017 06 15.
Artículo en Inglés | MEDLINE | ID: mdl-28476932

RESUMEN

Mast cells and basophils are developmentally related cells whose activation is a hallmark of allergy. Functionally, mast cells and basophils overlap in their ability to produce several mediators, including histamine and granule proteases, but studies have increasingly demonstrated nonredundant roles. To characterize the transcriptional heterogeneity of mast cells and basophils upon their activation, we performed large-scale comparative microarrays of murine bone marrow-derived mast cells and bone marrow-derived basophils (BMBs) at rest, upon an adaptive-type activation (IgE cross-linking), or upon an innate-type activation (IL-33 stimulation). Hierarchical clustering demonstrated that bone marrow-derived mast cells and BMBs shared specific activation-associated transcriptional signatures but differed in other signatures both between cell type and between activation mode. In bone marrow-derived mast cells, IgE cross-linking upregulated 785 genes, including Egr2, Ccl1, and Fxyd6, whereas IL-33 stimulation induced 823 genes, including Ccl1, Egr2, and Il1b. Focused bioinformatics pathway analysis demonstrated that IgE activation aligned with processes such as oxidative phosphorylation, angiogenesis, and the p53 pathway. The IL-33-activated transcriptome was enriched in genes commonly altered by NF-κB in response to TNF, by IL-6 via STAT3, and in response to IFN-γ. Furthermore, BMBs activated via IgE cross-linking selectively induced immune response genes Ccl1, Il3, and Il2 compared with IL-33-stimulated BMBs. Principal-component analysis revealed key cell- and activation-specific clustering. Overall, our data demonstrate that mast cells and basophils have cell- and activation-specific transcriptional responses and suggest that context-specific gene networks and pathways may shape how the immune system responds to allergens and innate cytokines.


Asunto(s)
Basófilos/inmunología , Biología Computacional , Mastocitos/inmunología , Transcripción Genética , Alérgenos/metabolismo , Animales , Basófilos/metabolismo , Células de la Médula Ósea/inmunología , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Redes Reguladoras de Genes , Inmunoglobulina E/química , Inmunoglobulina E/inmunología , Interleucina-33/farmacología , Interleucinas/genética , Interleucinas/metabolismo , Mastocitos/metabolismo , Ratones , Receptores de IgE/química , Receptores de IgE/inmunología , Análisis de Matrices Tisulares
12.
J Immunol ; 197(9): 3445-3453, 2016 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-27683753

RESUMEN

Eosinophils are important in the pathogenesis of many diseases, including asthma, eosinophilic esophagitis, and eczema. Whereas IL-5 is crucial for supporting mature eosinophils (EoMs), the signals that support earlier eosinophil lineage events are less defined. The IL-33R, ST2, is expressed on several inflammatory cells, including eosinophils, and is best characterized for its role during the initiation of allergic responses in peripheral tissues. Recently, ST2 expression was described on hematopoietic progenitor subsets, where its function remains controversial. Our findings demonstrate that IL-33 is required for basal eosinophil homeostasis, because both IL-33- and ST2-deficient mice exhibited diminished peripheral blood eosinophil numbers at baseline. Exogenous IL-33 administration increased EoMs in both the bone marrow and the periphery in wild-type and IL-33-deficient, but not ST2-deficient, mice. Systemic IL-5 was also increased under this treatment, and blocking IL-5 with a neutralizing Ab ablated the IL-33-induced EoM expansion. The homeostatic hypereosinophilia seen in IL-5-transgenic mice was significantly lower with ST2 deficiency despite similar elevations in systemic IL-5. Finally, in vitro treatment of bone marrow cells with IL-33, but not IL-5, led to specific early expansion of IL-5Rα-expressing precursor cells. In summary, our findings establish a basal defect in eosinophilopoiesis in IL-33- and ST2-deficient mice and a mechanism whereby IL-33 supports EoMs by driving both systemic IL-5 production and the expansion of IL-5Rα-expressing precursor cells.


Asunto(s)
Eosinófilos/fisiología , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Interleucina-33/metabolismo , Interleucina-5/metabolismo , Neutrófilos/fisiología , Animales , Células de la Médula Ósea/fisiología , Diferenciación Celular , Linaje de la Célula , Células Cultivadas , Regulación de la Expresión Génica , Hematopoyesis , Homeostasis , Humanos , Proteína 1 Similar al Receptor de Interleucina-1/genética , Interleucina-33/genética , Interleucina-5/genética , Subunidad alfa del Receptor de Interleucina-5/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados
13.
Immunotherapy ; 7(8): 913-22, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26268849

RESUMEN

Food allergy affects around 10% of the population. As the prevalence of food allergy continues to increase, disproportionately in children, new therapies for food allergy are being investigated. While there are no approved treatments for food allergy, immunotherapy facilitates significant desensitization and protection from accidental exposure. Nevertheless, current immunotherapies do not entirely nor permanently eliminate sensitivity to the food allergen. Since the rates of sustained unresponsiveness are significantly lower than desensitization, future therapies that enhance the rates of long-term tolerance in patients will catalyze progress in this field over the next 5-10 years.


Asunto(s)
Hipersensibilidad a los Alimentos/inmunología , Hipersensibilidad a los Alimentos/terapia , Tolerancia Inmunológica/inmunología , Inmunoterapia/métodos , Basófilos/inmunología , Niño , Desensibilización Inmunológica/métodos , Humanos , Inmunoterapia/tendencias , Mastocitos/inmunología , Inmunoterapia Sublingual/métodos , Linfocitos T/inmunología
14.
Nat Chem Biol ; 8(5): 434-6, 2012 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-22446837

RESUMEN

We performed bottom-up engineering of a synthetic pathway in Escherichia coli for the production of eukaryotic trimannosyl chitobiose glycans and the transfer of these glycans to specific asparagine residues in target proteins. The glycan biosynthesis was enabled by four eukaryotic glycosyltransferases, including the yeast uridine diphosphate-N-acetylglucosamine transferases Alg13 and Alg14 and the mannosyltransferases Alg1 and Alg2. By including the bacterial oligosaccharyltransferase PglB from Campylobacter jejuni, we successfully transferred glycans to eukaryotic proteins.


Asunto(s)
Disacáridos/biosíntesis , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Polisacáridos/biosíntesis , Ingeniería de Proteínas , Campylobacter jejuni/enzimología , Glicosilación , Hexosiltransferasas/metabolismo , Manosiltransferasas/metabolismo , Proteínas de la Membrana/metabolismo , N-Acetilglucosaminiltransferasas/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo
15.
Biotechnol Prog ; 28(2): 454-8, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22275211

RESUMEN

The most significant factor contributing to the presence of host cell protein (HCP) impurities in Protein A chromatography eluates is their association with the product monoclonal antibodies (mAbs) has been reported previously, and it has been suggested that more efficacious column washes may be developed by targeting the disruption of the mAbs-HCP interaction. However, characterization of this interaction is not straight forward as it is likely to involve multiple proteins and/or types of interaction. This work is an attempt to begin to understand the contribution of HCP subpopulations and/or mAb interaction propensity to the variability in HCP levels in the Protein A eluate. We performed a flowthrough (FT) recycling study with product respiking using two antibody molecules of apparently different HCP interaction propensities. In each case, the ELISA assay showed depletion of select subpopulations of HCP in Protein A eluates in subsequent column runs, while the feedstock HCP in the FTs remained unchanged from its native harvested cell culture fluid (HCCF) levels. In a separate study, the final FT from each molecule's recycling study was cross-spiked with various mAbs. In this case, Protein A eluate levels remained low for all but two molecules which were known as having high apparent HCP interaction propensity. The results of these studies suggest that mAbs may preferentially bind to select subsets of HCPs, and the degree of interaction and/or identity of the associated HCPs may vary depending on the mAb.


Asunto(s)
Anticuerpos Monoclonales/aislamiento & purificación , Inmunoglobulina G/aislamiento & purificación , Proteínas/aislamiento & purificación , Proteína Estafilocócica A/química , Animales , Anticuerpos Monoclonales/química , Células CHO , Cromatografía de Afinidad , Cricetinae , Inmunoglobulina G/química , Unión Proteica , Proteínas/química
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