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1.
J Immunol Methods ; 531: 113715, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38936465

RESUMEN

Antibody-dependent cellular phagocytosis (ADCP) is a cellular process by which antibody-opsonized targets (pathogens or cells) activate the Fc receptors on the surface of phagocytes to induce phagocytosis, resulting in internalization and degradation of pathogens or target cells through phagosome acidification. Besides NK cells-mediated antibody-dependent cellular cytotoxicity (ADCC), tumor-infiltrated monocytes and macrophages can directly kill tumor cells in the presence of tumor antigen-specific antibodies through ADCP, representing another attractive strategy for cancer immunotherapy. Even though several methods have been developed to measure ADCP, an automated and high-throughput quantitative assay should offer highly desirable advantages for drug discovery. In this study we established a new ADCP assay to identify therapeutical monoclonal antibodies (mAbs) that facilitate macrophages phagocytosis of live target cells. We used Incucyte, an imaging system for live cell analysis. By labeling the live target cells with a pH sensitive dye (pHrodo), we successfully monitored the ADCP in real time. We demonstrated that our image-based assay is robust and quantitative, suitable for screening and characterization of therapeutical mAbs that directly kill target cells through ADCP. Furthermore, we found different subtypes of macrophages have distinct ADCP activities using both mouse and human primary macrophages differentiated in vitro. By studying various mAbs with mutations in their Fc regions using our assay, we showed that the variants with increased binding to Fc gamma receptors (FcγRs) have enhanced ADCP activities.


Asunto(s)
Anticuerpos Monoclonales , Macrófagos , Fagocitosis , Humanos , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacología , Macrófagos/inmunología , Macrófagos/metabolismo , Citotoxicidad Celular Dependiente de Anticuerpos , Animales , Ratones , Línea Celular Tumoral
2.
Medicine (Baltimore) ; 103(9): e37199, 2024 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-38428902

RESUMEN

BACKGROUND: The goal of anesthesia in patients with hypertrophic obstructive cardiomyopathy (HOCM) is to reduce the risk of left ventricular outflow tract obstruction triggered by anesthetics. Remimazolam is a newly developed anesthetic that has been reported to have superior hemodynamic stability. There have been no reports on the completion of non-cardiac surgery with remimazolam in patients with HOCM. METHODS: Here we report the case of a 49-year-old man diagnosed with hypertrophic obstructive cardiomyopathy who underwent resection of colon cancer with remimazolam and remifentanil anesthesia. A bolus 0.3 mg/kg remimazolam was administered for anesthesia induction, and then adjusted to 2 mg/kg/h to maintain anesthesia. Set the pain threshold index to 50 to auto-control the infusion speed of remifentanil. RESULTS: No hypotension occurred during anesthesia, and norepinephrine was not administered. After conversion to open surgery, the patient's blood pressure elevated and reduced with urapidil and esmolol. CONCLUSION: In this patient with HOCM, remimazolam and remifentanil provided adequate anesthesia for induction and maintenance to complete the right hemicolectomy.


Asunto(s)
Anestésicos , Benzodiazepinas , Cardiomiopatía Hipertrófica , Neoplasias Colorrectales , Masculino , Humanos , Persona de Mediana Edad , Remifentanilo , Anestesia General , Cardiomiopatía Hipertrófica/complicaciones , Cardiomiopatía Hipertrófica/cirugía , Neoplasias Colorrectales/complicaciones , Neoplasias Colorrectales/cirugía
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