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1.
Cytotherapy ; 22(5): 276-290, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-32238299

RESUMEN

BACKGROUND AIMS: Key obstacles in human iNKT cell translational research and immunotherapy include the lack of robust protocols for dependable expansion of human iNKT cells and the paucity of data on phenotypes in post-expanded cells. METHODS: We delineate expansion methods using interleukin (IL)-2, IL-7 and allogeneic feeder cells and anti-CD2/CD3/CD28 stimulation by which to dependably augment Th2 polarization and direct cytotoxicity of human peripheral blood CD3+Vα24+Vß11+ iNKT cells. RESULTS: Gene and protein expression profiling demonstrated augmented Th2 cytokine secretion (IL-4, IL-5, IL-13) in expanded iNKT cells stimulated with anti-CD2/CD3/CD28 antibodies. Cytotoxic effector molecules including granzyme B were increased in expanded iNKT cells after CD2/CD3/CD28 stimulation. Direct cytotoxicity assays using unstimulated expanded iNKT cell effectors revealed α-galactosyl ceramide (α-GalCer)-dependent killing of the T-ALL cell line Jurkat. Moreover, CD2/CD3/CD28 stimulation of expanded iNKT cells augmented their (α-GalCer-independent) killing of Jurkat cells. Co-culture of expanded iNKT cells with stimulated responder cells confirmed contact-dependent inhibition of activated CD4+ and CD8+ responder T cells. DISCUSSION: These data establish a robust protocol to expand and novel pathways to enhance Th2 cytokine secretion and direct cytotoxicity in human iNKT cells, findings with direct implications for autoimmunity, vaccine augmentation and anti-infective immunity, cancer immunotherapy and transplantation.


Asunto(s)
Antígenos CD2/inmunología , Antígenos CD28/inmunología , Complejo CD3/inmunología , Proliferación Celular/efectos de los fármacos , Citocinas/metabolismo , Células T Asesinas Naturales/inmunología , Células Th2/inmunología , Anticuerpos/farmacología , Apoptosis/efectos de los fármacos , Donantes de Sangre , Trasplante de Células/métodos , Células Cultivadas , Perfilación de la Expresión Génica , Humanos , Inmunoterapia/métodos , Células Jurkat , Células K562 , Activación de Linfocitos/inmunología
2.
Blood ; 129(22): 3017-3030, 2017 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-28351935

RESUMEN

Nonmyeloablative conditioning using total lymphoid irradiation (TLI) and rabbit antithymocyte serum (ATS) (the murine preclinical equivalent of antithymocyte globulin [ATG]) facilitates immune tolerance after bone marrow transplantation (BMT) across major histocompatibility complex (MHC) disparities and may be a useful strategy for nonmalignant disorders. We previously reported that donor effector T-cell function and graft-versus-host disease (GVHD) are regulated via recipient invariant natural killer T-cell (iNKT) interleukin-4-driven expansion of donor Foxp3+ naturally occurring regulatory T cells (Tregs). This occurs via recipient iNKT- and STAT6-dependent expansion of recipient myeloid dendritic cells (MDCs) that induce contact-dependent expansion of donor Treg through PD-1/PD ligand signaling. After TLI/ATS + BMT, Gr-1lowCD11c+ MDCs and Gr-1highCD11cneg myeloid-derived suppressor cells (MDSCs) were enriched in GVHD target organs. We now report that the recovery of both recipient MDSCs (P < .01) and MDCs (P < .01) is significantly increased when the alkylator cyclophosphamide (CTX) is added to TLI/ATS conditioning. In a BALB/c → B6 lethal GVHD model, adoptive transfer of MDSCs from TLI/ATS/CTX-conditioned recipients is associated with significantly improved GVHD colitis and survival (P < .001), conversion of MDSCs to PD ligand-expressing MDCs, and increased donor naturally occurring Treg recovery (P < .01) compared with control treatment. Using BALB/c donors and ß-thalassemic HW-80 recipients, we found significantly improved rates of engraftment and GVHD following TLI/ATS/CTX compared with TLI/ATS, lethal or sublethal total body irradiation/ATS/CTX, or CTX/ATS conditioning. These data provide preclinical support for trials of TLI/ATG/alkylator regimens for MHC-mismatched BMT for hemoglobinopathies. The data also delineate innate immune mechanisms by which TLI/ATS/CTX conditioning may augment transplantation tolerance.


Asunto(s)
Trasplante de Médula Ósea/métodos , Tolerancia Inmunológica , Acondicionamiento Pretrasplante/métodos , Talasemia beta/inmunología , Talasemia beta/terapia , Traslado Adoptivo , Animales , Suero Antilinfocítico/uso terapéutico , Ciclofosfamida/uso terapéutico , Modelos Animales de Enfermedad , Supervivencia de Injerto , Enfermedad Injerto contra Huésped/prevención & control , Irradiación Linfática , Complejo Mayor de Histocompatibilidad , Ratones , Ratones Endogámicos BALB C , Ratones Mutantes , Modelos Inmunológicos , Células Supresoras de Origen Mieloide/inmunología
3.
Vet Pathol ; 55(1): 76-97, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-28494703

RESUMEN

Only 2 major mast cell (MC) subtypes are commonly recognized in the mouse: the large connective tissue mast cells (CTMCs) and the mucosal mast cells (MMCs). Interepithelial mucosal inflammatory cells, most commonly identified as globule leukocytes (GLs), represent a third MC subtype in mice, which we term interepithelial mucosal mast cells (ieMMCs). This term clearly distinguishes ieMMCs from lamina proprial MMCs (lpMMCs) while clearly communicating their common MC lineage. Both lpMMCs and ieMMCs are rare in normal mouse intestinal mucosa, but increased numbers of ieMMCs are seen as part of type 2 immune responses to intestinal helminth infections and in food allergies. Interestingly, we found that increased ieMMCs were consistently associated with decreased mucosal inflammation and damage, suggesting that they might have a role in controlling helminth-induced immunopathology. We also found that ieMMC hyperplasia can develop in the absence of helminth infections, for example, in Treg-deficient mice, Arf null mice, some nude mice, and certain graft-vs-host responses. Since tuft cell hyperplasia plays a critical role in type 2 immune responses to intestinal helminths, we looked for (but did not find) any direct relationship between ieMMC and tuft cell numbers in the intestinal mucosa. Much remains to be learned about the differing functions of ieMMCs and lpMMCs in the intestinal mucosa, but an essential step in deciphering their roles in mucosal immune responses will be to apply immunohistochemistry methods to consistently and accurately identify them in tissue sections.


Asunto(s)
Intestinos/citología , Leucocitos/citología , Mastocitos/citología , Animales , Modelos Animales de Enfermedad , Helmintiasis Animal/inmunología , Helmintiasis Animal/patología , Mucosa Intestinal/citología , Mucosa Intestinal/patología , Intestinos/patología , Leucocitos/patología , Mastocitos/patología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL
6.
J Immunol ; 191(11): 5764-76, 2013 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-24190658

RESUMEN

We showed previously that nonmyeloablative total lymphoid irradiation/rabbit anti-thymocyte serum (TLI/ATS) conditioning facilitates potent donor-recipient immune tolerance following bone marrow transplantation (BMT) across MHC barriers via recipient invariant NKT (iNKT) cell-derived IL-4-dependent expansion of donor Foxp3(+) naturally occurring regulatory T cells (nTregs). In this study, we report a more specific mechanism. Wild-type (WT) BALB/c (H-2(d)) hosts were administered TLI/ATS and BMT from WT or STAT6(-/-) C57BL/6 (H-2(b)) donors. Following STAT6(-/-) BMT, donor nTregs demonstrated no loss of proliferation in vivo, indicating that an IL-4-responsive population in the recipient, rather than the donor, drives donor nTreg proliferation. In graft-versus-host disease (GVHD) target organs, three recipient CD11b(+) cell subsets (Gr-1(high)CD11c(-), Gr-1(int)CD11c(-), and Gr-1(low)CD11c(+)) were enriched early after TLI/ATS + BMT versus total body irradiation/ATS + BMT. Gr-1(low)CD11c(+) cells induced potent H-2K(b+)CD4(+)Foxp3(+) nTreg proliferation in vitro in 72-h MLRs. Gr-1(low)CD11c(+) cells were reduced significantly in STAT6(-/-) and iNKT cell-deficient Jα18(-/-) BALB/c recipients after TLI/ATS + BMT. Depletion of CD11b(+) cells resulted in severe acute GVHD, and adoptive transfer of WT Gr-1(low)CD11c(+) cells to Jα18(-/-) BALB/c recipients of TLI/ATS + BMT restored day-6 donor Foxp3(+) nTreg proliferation and protection from CD8 effector T cell-mediated GVHD. Blockade of programmed death ligand 1 and 2, but not CD40, TGF-ß signaling, arginase 1, or iNOS, inhibited nTreg proliferation in cocultures of recipient-derived Gr-1(low)CD11c(+) cells with donor nTregs. Through iNKT-dependent Th2 polarization, myeloid-derived immunomodulatory dendritic cells are expanded after nonmyeloablative TLI/ATS conditioning and allogeneic BMT, induce PD-1 ligand-dependent donor nTreg proliferation, and maintain potent graft-versus-host immune tolerance.


Asunto(s)
Trasplante de Médula Ósea , Células Dendríticas/inmunología , Factores de Transcripción Forkhead/metabolismo , Células Mieloides/inmunología , Linfocitos T Reguladores/inmunología , Tolerancia al Trasplante/inmunología , Animales , Antígeno CD11c , Antígenos CD4/metabolismo , Proliferación Celular , Factores de Transcripción Forkhead/genética , Inmunomodulación , Irradiación Linfática , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Receptor de Muerte Celular Programada 1/metabolismo , Factor de Transcripción STAT6/genética , Donantes de Tejidos
7.
Biol Blood Marrow Transplant ; 20(8): 1224-8, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24769325

RESUMEN

The safety, pharmacokinetics, and biological effect of plerixafor in children as part of a conditioning regimen for chemo-sensitization in allogeneic hematopoietic stem cell transplantation (HSCT) have not been studied. This is a phase I study of plerixafor designed to evaluate its tolerability at dose of .24 mg/kg given intravenously on day -4 (level 1); day -4 and day -3 (level 2); or day -4, day -3, and day -2 (level 3) in combination with fludarabine, thiotepa, melphalan, and rabbit antithymocytic globulin for a second allogeneic HSCT in children with refractory or relapsed leukemia. Immunophenotype analysis was performed on blood and bone marrow before and after plerixafor administration. Twelve patients were enrolled. Plerixafor at all 3 levels was well tolerated without dose-limiting toxicity. Transient gastrointestinal side effects of National Cancer Institute-grade 1 or 2 in severity were the most common adverse events. The area under the concentration-time curve increased proportionally to the dose level. Plerixafor clearance was higher in males and increased linearly with body weight and glomerular filtration rate. The clearance decreased and the elimination half-life increased significantly from dose level 1 to 3 (P < .001). Biologically, the proportion of CXCR4(+) blasts and lymphocytes both in the bone marrow and peripheral blood increased after plerixafor administration.


Asunto(s)
Compuestos Heterocíclicos/farmacocinética , Compuestos Heterocíclicos/uso terapéutico , Leucemia/etiología , Bencilaminas , Biomarcadores Farmacológicos , Ciclamas , Femenino , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Compuestos Heterocíclicos/administración & dosificación , Humanos , Inmunofenotipificación , Leucemia/tratamiento farmacológico , Masculino , Recurrencia , Acondicionamiento Pretrasplante , Trasplante Homólogo , Resultado del Tratamiento
8.
Blood ; 120(2): 468-72, 2012 Jul 12.
Artículo en Inglés | MEDLINE | ID: mdl-22517895

RESUMEN

In patients with acute leukemia, detection of minimal residual disease (MRD) before allogeneic hematopoietic cell transplantation (HCT) correlates with risk of relapse. However, the level of MRD that is most likely to preclude cure by HCT is unclear, and the benefit of further chemotherapy to reduce MRD before HCT is unknown. In 122 children with very-high-risk acute lymphoblastic leukemia (ALL; n = 64) or acute myeloid leukemia (AML, n = 58), higher MRD levels at the time of HCT predicted a poorer survival after HCT (P = .0019); MRD was an independent prognostic factor in a multivariate analysis (P = .0035). However, the increase in risk of death associated with a similar increment of MRD was greater in ALL than in AML, suggesting that a pretransplantation reduction of leukemia burden would have a higher impact in ALL. At any given MRD level, survival rates were higher for patients treated in recent protocols: the 5-year overall survival for patients with ALL was 49% if MRD was detectable and 88% if it was not and the corresponding rates for patients with AML were 67% and 80%, respectively. Although MRD before HCT is a strong prognostic factor, its impact has diminished and should not be regarded as a contraindication for HCT.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas , Leucemia Mieloide Aguda/patología , Leucemia Mieloide Aguda/terapia , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Niño , Estudios de Cohortes , Contraindicaciones , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Neoplasia Residual , Pronóstico , Factores de Riesgo , Trasplante Homólogo
9.
Chemotherapy ; 60(1): 24-36, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25341697

RESUMEN

BACKGROUND: Chemotherapy-induced mucositis (CIM) complicates cancer therapy and limits maximum tolerated doses and efficacy. Rodent models do not reproducibly mimic clinical CIM, so alternative models are needed. METHODS: CIM severity was assessed after weaned pigs were treated with doxorubicin (5 and 3.75 mg/kg) using clinical observations, laboratory parameters and gastrointestinal structure and functions. Bovine colostrum was provided as an experimental intervention to the pigs treated receiving the 3.75 mg/kg dose. RESULTS: Doxorubin at 3.75 mg/kg decreased food intake and weight gain (p < 0.05) and caused diarrhea and vomiting that coincided with damage to the small intestine mucosa based on histological scoring (p < 0.05). It resulted in higher serum TNF-α concentrations, increased chloride secretion and reduced brush border membrane disaccharidase activities and carrier-mediated glucose uptake (all p < 0.05). The gastrointestinal damage and dysfunction resemble the clinical and laboratory features of CIM in humans; these can be partially prevented by providing cow colostrum. CONCLUSION: The weaned pig is a relevant large animal for studying CIM and evaluating existing and experimental interventions for mucositis.


Asunto(s)
Antibióticos Antineoplásicos/toxicidad , Doxorrubicina/toxicidad , Mucosa Intestinal/efectos de los fármacos , Mucositis/inducido químicamente , Animales , Modelos Animales de Enfermedad , Ingestión de Alimentos/efectos de los fármacos , Glucosa/metabolismo , Interleucina-10/sangre , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Mucositis/patología , Porcinos , Factor de Necrosis Tumoral alfa/sangre , Aumento de Peso/efectos de los fármacos
10.
Sci Immunol ; 9(92): eado2161, 2024 Feb 02.
Artículo en Inglés | MEDLINE | ID: mdl-38306415

RESUMEN

Lineage-specific effects of upstream promoters affect ST2 expression and effector function in TH1cells.


Asunto(s)
Proteína 1 Similar al Receptor de Interleucina-1 , Proteína 1 Similar al Receptor de Interleucina-1/genética , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Regiones Promotoras Genéticas
11.
Sci Immunol ; 9(97): eadr2965, 2024 Jul 05.
Artículo en Inglés | MEDLINE | ID: mdl-38968340

RESUMEN

Lineage-specific effects of upstream promoters affect ST2 expression and effector function in TH1 cells.


Asunto(s)
Células TH1 , Animales , Humanos , Células TH1/inmunología , Regiones Promotoras Genéticas/genética , Ratones
12.
Biol Blood Marrow Transplant ; 19(2): 291-7, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23063628

RESUMEN

Children with relapsed or refractory solid tumors face dismal prognoses, and novel therapies are desperately needed. Allogeneic hematopoietic cell transplantation (HCT) offers potential for cell-based therapy, but the toxicity of myeloablation limits this approach in heavily pretreated patients. We sought to determine the feasibility of HCT in a cohort of 24 children with incurable solid tumors using human leukocyte antigen-matched sibling or unrelated donors and a minimal conditioning regimen. Before stem cell infusion, all patients received 3 daily doses of 30 mg/m(2) fludarabine followed by 2 Gy of total body irradiation. Hematopoietic cell recovery was rapid and reliable. Median time to neutrophil engraftment was 13.5 days for sibling donors and 12 days for unrelated donors. Donor lymphocyte infusions were used safely in 4 patients, all of whom had either improved chimerism or apparent tumor response. Graft-versus-host disease was comparable across donor sources and did not affect survival. Relapse remains a substantial barrier, although objective graft-versus-tumor effect was observed in several patients. Four patients with detectable disease before HCT achieved a complete response for at least 30 days after HCT, and two remain long-term survivors. Three patients were in complete response before HCT and remained in remission for 3, 6, and 74 months after HCT. Early disease response was associated with improved survival. Allogeneic HCT using this conditioning regimen offers a potential platform for novel immunotherapies.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas/métodos , Neoplasias/tratamiento farmacológico , Neoplasias/cirugía , Acondicionamiento Pretrasplante/métodos , Adolescente , Adulto , Niño , Preescolar , Quimerismo , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Humanos , Neoplasias/radioterapia , Recurrencia , Inducción de Remisión , Acondicionamiento Pretrasplante/efectos adversos , Trasplante Homólogo , Vidarabina/administración & dosificación , Vidarabina/análogos & derivados , Irradiación Corporal Total , Adulto Joven
13.
Blood ; 118(2): 223-30, 2011 Jul 14.
Artículo en Inglés | MEDLINE | ID: mdl-21613256

RESUMEN

We evaluated 190 children with very high-risk leukemia, who underwent allogeneic hematopoietic cell transplantation in 2 sequential treatment eras, to determine whether those treated with contemporary protocols had a high risk of relapse or toxic death, and whether non-HLA-identical transplantations yielded poor outcomes. For the recent cohorts, the 5-year overall survival rates were 65% for the 37 patients with acute lymphoblastic leukemia and 74% for the 46 with acute myeloid leukemia; these rates compared favorably with those of earlier cohorts (28%, n = 57; and 34%, n = 50, respectively). Improvement in the recent cohorts was observed regardless of donor type (sibling, 70% vs 24%; unrelated, 61% vs 37%; and haploidentical, 88% vs 19%), attributable to less infection (hazard ratio [HR] = 0.12; P = .005), regimen-related toxicity (HR = 0.25; P = .002), and leukemia-related death (HR = 0.40; P = .01). Survival probability was dependent on leukemia status (first remission vs more advanced disease; HR = 0.63; P = .03) or minimal residual disease (positive vs negative; HR = 2.10; P = .01) at the time of transplantation. We concluded that transplantation has improved over time and should be considered for all children with very high-risk leukemia, regardless of matched donor availability.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas/estadística & datos numéricos , Leucemia/epidemiología , Leucemia/terapia , Niño , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Humanos , Leucemia/mortalidad , Leucemia/patología , Masculino , Estadificación de Neoplasias , Neoplasia Residual/epidemiología , Estudios Retrospectivos , Factores de Riesgo , Análisis de Supervivencia , Donantes de Tejidos , Resultado del Tratamiento
14.
Sci Immunol ; 8(82): eadi0472, 2023 04 14.
Artículo en Inglés | MEDLINE | ID: mdl-37027482

RESUMEN

CD169+ macrophage-intrinsic IL-10 production mitigates mortality from sepsis.


Asunto(s)
Interleucina-10 , Macrófagos , Sepsis , Humanos , Interleucina-10/metabolismo
15.
Sci Immunol ; 8(87): eadk4486, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37656779

RESUMEN

In successful melanoma immunotherapy, clonal TCRs can recognize multiple tumor-specific antigens simultaneously through cross-reactivity.

16.
Biol Blood Marrow Transplant ; 18(12): 1911-20, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22842333

RESUMEN

This study analyzes the hematopoietic cell transplantation experience in patients with immune deficiency at a single institution. The objective is to comprehensively evaluate the short-term and long-term outcomes with various preparative regimens, donor grafts, and ex vivo manipulations to identify transplantation approaches that most likely favor early donor immune competency without generating excessive toxicity. Clinical outcomes were evaluated in 52 consecutive patients with immune deficiencies. Thirty-seven of the 52 patients (71%) survived with attenuation of their underlying disease. The use of a melphalan-based reduced-intensity conditioning preparative regimen and immunomagnetic CD3(+) T cell depletion techniques (when T cell depletion was indicated) were associated with improved event-free survival. Survivors who received a preparative regimen other than a melphalan-based reduced-intensity regimen suffered from therapy-related morbidities or chronic/recurrent infections. Our findings indicate that melphalan-based reduced-intensity conditioning regimens and immunomagnetic CD3(+) T cell depletion limit therapy-related toxicity, and demonstrate promising results for the early establishment of donor immune competency.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas/métodos , Depleción Linfocítica/métodos , Inmunodeficiencia Combinada Grave/cirugía , Linfocitos T/inmunología , Adolescente , Adulto , Niño , Preescolar , Supervivencia sin Enfermedad , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Estudios Retrospectivos , Inmunodeficiencia Combinada Grave/inmunología , Linfocitos T/citología , Trasplante Homólogo , Adulto Joven
17.
Sci Immunol ; 7(77): eadf5127, 2022 11 11.
Artículo en Inglés | MEDLINE | ID: mdl-36332010

RESUMEN

ABCC1 is an ATP-dependent cGAMP exporter responsible for negatively regulating STING signaling.


Asunto(s)
Proteínas de la Membrana , Transducción de Señal , Proteínas de la Membrana/genética
18.
Sci Immunol ; 7(72): eadd2045, 2022 06 03.
Artículo en Inglés | MEDLINE | ID: mdl-35658011

RESUMEN

In a mouse model of pneumococcal meningitis, skull channels provide extravascular signaling to the skull marrow capable of initiating local marrow hematopoiesis.


Asunto(s)
Meningitis Neumocócica , Animales , Médula Ósea , Modelos Animales de Enfermedad , Hematopoyesis , Ratones , Cráneo
19.
Sci Immunol ; 7(67): eabn9190, 2022 Jan 07.
Artículo en Inglés | MEDLINE | ID: mdl-34995095

RESUMEN

CoA-driven mitochondrial metabolism enhances the anti-tumor properties of IL-22­producing CD8+ T cells.


Asunto(s)
Tutoría , Neoplasias , Humanos
20.
Front Immunol ; 13: 815828, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35493491

RESUMEN

Mass cytometry has revolutionized immunophenotyping, particularly in exploratory settings where simultaneous breadth and depth of characterization of immune populations is needed with limited samples such as in preclinical and clinical tumor immunotherapy. Mass cytometry is also a powerful tool for single-cell immunological assays, especially for complex and simultaneous characterization of diverse intratumoral immune subsets or immunotherapeutic cell populations. Through the elimination of spectral overlap seen in optical flow cytometry by replacement of fluorescent labels with metal isotopes, mass cytometry allows, on average, robust analysis of 60 individual parameters simultaneously. This is, however, associated with significantly increased complexity in the design, execution, and interpretation of mass cytometry experiments. To address the key pitfalls associated with the fragmentation, complexity, and analysis of data in mass cytometry for immunologists who are novices to these techniques, we have developed a comprehensive resource guide. Included in this review are experiment and panel design, antibody conjugations, sample staining, sample acquisition, and data pre-processing and analysis. Where feasible multiple resources for the same process are compared, allowing researchers experienced in flow cytometry but with minimal mass cytometry expertise to develop a data-driven and streamlined project workflow. It is our hope that this manuscript will prove a useful resource for both beginning and advanced users of mass cytometry.


Asunto(s)
Anticuerpos , Análisis de la Célula Individual , Citometría de Flujo/métodos , Inmunofenotipificación , Análisis de la Célula Individual/métodos , Coloración y Etiquetado
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