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Article ; Online: A step forward on the in vitro and in vivo assessment of a novel nanomedicine against melanoma.

Pinho, Jacinta O / Matias, Mariana / Godinho-Santos, Ana / Amaral, Joana D / Mendes, Eduarda / Perry, Maria Jesus / Francisco, Ana Paula / Rodrigues, Cecília M P / Gaspar, M Manuela

International journal of pharmaceutics

2023 Volume 640, Page(s) 123011

Abstract: Melanoma is the most aggressive form of skin cancer, with increasing incidence and mortality rates. To overcome current treatment limitations, a hybrid molecule (HM) combining a triazene and a sulfur L-tyrosine analogue, was recently synthesized, ... ...

Abstract	Melanoma is the most aggressive form of skin cancer, with increasing incidence and mortality rates. To overcome current treatment limitations, a hybrid molecule (HM) combining a triazene and a sulfur L-tyrosine analogue, was recently synthesized, incorporated in long blood circulating liposomes (LIP HM) and validated in an immunocompetent melanoma model. The present work constitutes a step forward in the therapeutic assessment of HM formulations. Here, human melanoma cells, A375 and MNT-1, were used and dacarbazine (DTIC), a triazene drug clinically available as first-line treatment for melanoma, constituted the positive control. In cell cycle analysis, A375 cells, after 24-h incubation with HM (60 μM) and DTIC (70 μM), resulted in a 1.2 fold increase (related to control) in the percentage of cells in G0/G1 phase. The therapeutic activity was evaluated in a human murine melanoma model (subcutaneously injected with A375 cells) to most closely resemble the human pathology. Animals treated with LIP HM exhibited the highest antimelanoma effect resulting in a 6-, 5- and 4-fold reduction on tumor volume compared to negative control, Free HM and DTIC groups, respectively. No toxic side effects were detected. Overall, these results constitute another step forward in the validation of the antimelanoma activity of LIP HM, using a murine model that more accurately simulates the pathology that occurs in human patients.
MeSH term(s)	Humans ; Animals ; Mice ; Nanomedicine ; Melanoma/metabolism ; Dacarbazine ; Skin Neoplasms/pathology ; Cell Line, Tumor ; Apoptosis
Chemical Substances	Dacarbazine (7GR28W0FJI)
Language	English
Publishing date	2023-05-03
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	428962-6
ISSN	1873-3476 ; 0378-5173
ISSN (online)	1873-3476
ISSN	0378-5173
DOI	10.1016/j.ijpharm.2023.123011
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Article ; Online: Towards personalized antibody cancer therapy: development of a microfluidic cell culture device for antibody selection.

Condelipes, Pedro G M / Fontes, Pedro Mendes / Godinho-Santos, Ana / Brás, Eduardo J S / Marques, Vanda / Afonso, Marta B / Rodrigues, Cecília M P / Chu, Virginia / Gonçalves, João / Conde, João Pedro

Lab on a chip

2022 Volume 22, Issue 23, Page(s) 4717–4728

Abstract: Antibody therapy has been one of the most successful therapies for a wide range of diseases, including cancer. One way of expediting antibody therapy development is through phage display technology. Here, by screening thousands of randomly assembled ... ...

Abstract	Antibody therapy has been one of the most successful therapies for a wide range of diseases, including cancer. One way of expediting antibody therapy development is through phage display technology. Here, by screening thousands of randomly assembled peptide sequences, it is possible to identify potential therapeutic candidates. Conventional screening technologies do not accommodate perfusion through the system, as is the case of standard plate-based cultures. This leads to a poor translation of the experimental results obtained
MeSH term(s)	Animals ; Humans ; Lab-On-A-Chip Devices ; Microfluidics/methods ; Receptors, CXCR4 ; Cell Culture Techniques ; Antibodies ; Bacteriophages ; Mammals ; Neoplasms/drug therapy
Chemical Substances	Receptors, CXCR4 ; Antibodies
Language	English
Publishing date	2022-11-22
Publishing country	England
Document type	Journal Article
ZDB-ID	2056646-3
ISSN	1473-0189 ; 1473-0197
ISSN (online)	1473-0189
ISSN	1473-0197
DOI	10.1039/d2lc00918h
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Article ; Online: Phenotype of BTK-lacking myeloid cells during prolonged COVID-19 and upon convalescent plasma.

Gomes, André M C / Farias, Guilherme B / Trombetta, Amelia C / Godinho-Santos, Ana / Parreira, Inês / Gonçalves, Hélder Diogo / Simões, Mariana Lessa / Aguiar, Patrício / Deveza, Maria Manuel / Inácio, João / Sousa, Ana E / da Silva, Susana Lopes

European journal of haematology

2022 Volume 110, Issue 2, Page(s) 209–212

Abstract: XLA patient with 7-month course of COVID-19 with persistent plasma SARS-CoV-2 load revealed a sustained non-inflammatory profile of myeloid cells in association with contained severity of disease, arguing in favor of the use of BTK inhibitors in SARS-COV- ...

Abstract	XLA patient with 7-month course of COVID-19 with persistent plasma SARS-CoV-2 load revealed a sustained non-inflammatory profile of myeloid cells in association with contained severity of disease, arguing in favor of the use of BTK inhibitors in SARS-COV-2 infection.
MeSH term(s)	Humans ; Protein-Tyrosine Kinases ; Agammaglobulinaemia Tyrosine Kinase/genetics ; COVID-19 ; SARS-CoV-2 ; COVID-19 Serotherapy ; Myeloid Cells ; Phenotype ; Genetic Diseases, X-Linked
Chemical Substances	Protein-Tyrosine Kinases (EC 2.7.10.1) ; Agammaglobulinaemia Tyrosine Kinase (EC 2.7.10.2)
Language	English
Publishing date	2022-10-19
Publishing country	England
Document type	Case Reports
ZDB-ID	392482-8
ISSN	1600-0609 ; 0902-4441
ISSN (online)	1600-0609
ISSN	0902-4441
DOI	10.1111/ejh.13881
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Article ; Online: Early ART in Acute HIV-1 Infection: Impact on the B-Cell Compartment.

Badura, Robert / Foxall, Russell B / Ligeiro, Dario / Rocha, Miguel / Godinho-Santos, Ana / Trombetta, Amelia C / Sousa, Ana E

Frontiers in cellular and infection microbiology

2020 Volume 10, Page(s) 347

Abstract: HIV-1 infection induces B cell defects, not fully recovered upon antiretroviral therapy (ART). Acute infection and the early start of ART provide unique settings to address the impact of HIV on the B cell compartment. We took advantage of a cohort of 21 ... ...

Abstract	HIV-1 infection induces B cell defects, not fully recovered upon antiretroviral therapy (ART). Acute infection and the early start of ART provide unique settings to address the impact of HIV on the B cell compartment. We took advantage of a cohort of 21 seroconverters, grouped according to the presence of severe manifestations likely mediated by antibodies or immune complexes, such as Guillain-Barré syndrome and autoimmune thrombocytopenic purpura, with a follow-up of 8 weeks upon effective ART. We combined B and T cell phenotyping with serum immunoglobulin level measurement and quantification of sj-KRECs and ΔB to estimate bone marrow output and peripheral proliferative history of B cells, respectively. We observed marked B cell disturbances, notably a significant expansion of cells expressing low levels of CD21, in parallel with markers of both impaired bone marrow output and increased peripheral B cell proliferation. This B cell dysregulation is likely to contribute to the severe immune-mediated conditions, as attested by the higher serum IgG and the reduced levels of sj-KRECs with increased ΔB in these individuals as compared to those patients with mild disease. Nevertheless, upon starting ART, the dynamic of B cell recovery was not distinct in the two groups, featuring both persistent alterations by week 8. Overall, we showed for the first time that acute HIV-1 infection is associated with decreased bone marrow B cell output assessed by sj-KRECs. Our study emphasizes the need to intervene in both bone marrow and peripheral responses to facilitate B cell recovery during acute HIV-1 infection.
MeSH term(s)	B-Lymphocytes ; CD4-Positive T-Lymphocytes ; Cohort Studies ; HIV Infections/drug therapy ; HIV-1 ; Humans ; Lymphocyte Activation
Language	English
Publishing date	2020-07-16
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2619676-1
ISSN	2235-2988 ; 2235-2988
ISSN (online)	2235-2988
ISSN	2235-2988
DOI	10.3389/fcimb.2020.00347
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Article ; Online: CIB1 and CIB2 are HIV-1 helper factors involved in viral entry.

Godinho-Santos, Ana / Hance, Allan J / Gonçalves, João / Mammano, Fabrizio

Scientific reports

2016 Volume 6, Page(s) 30927

Abstract: HIV-1 relies on the host-cell machinery to accomplish its replication cycle, and characterization of these helper factors contributes to a better understanding of HIV-host interactions and can identify potential novel antiviral targets. Here we explored ... ...

Abstract	HIV-1 relies on the host-cell machinery to accomplish its replication cycle, and characterization of these helper factors contributes to a better understanding of HIV-host interactions and can identify potential novel antiviral targets. Here we explored the contribution of CIB2, previously identified by RNAi screening as a potential helper factor, and its homolog, CIB1. Knockdown of either CIB1 or CIB2 strongly impaired viral replication in Jurkat cells and in primary CD4+ T-lymphocytes, identifying these proteins as non-redundant helper factors. Knockdown of CIB1 and CIB2 impaired envelope-mediated viral entry for both for X4- and R5-tropic HIV-1, and both cell-free and cell-associated entry pathways were affected. In contrast, the level of CIB1 and CIB2 expression did not influence cell viability, cell proliferation, receptor-independent viral binding to the cell surface, or later steps in the viral replication cycle. CIB1 and CIB2 knockdown was found to reduce the expression of surface molecules implicated in HIV-1 infection, including CXCR4, CCR5 and integrin α4β7, suggesting at least one mechanism through which these proteins promote viral infection. Thus, this study identifies CIB1 and CIB2 as host helper factors for HIV-1 replication that are required for optimal receptor-mediated viral entry.
Language	English
Publishing date	2016-08-04
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/srep30927
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Article: Acute HIV-1 and SARS-CoV-2 Infections Share Slan+ Monocyte Depletion—Evidence from an Hyperacute HIV-1 Case Report

Farias, Guilherme B. / Badura, Robert / Conceição, Carolina M. / Gomes, André M. C. / Godinho-Santos, Ana / Laia, Joel / Rosmaninho, Pedro / Santos, Diana F. / Mota, Catarina / Almeida, Afonso R. M. / Fernandes, Susana M. / Trombetta, Amelia C. / Sousa, Ana E.

Viruses. 2021 Sept. 10, v. 13, no. 9

2021

Abstract: Monocytes are key modulators in acute viral infections, determining both inflammation and development of specific B- and T-cell responses. Recently, these cells were shown to be associated to different SARS-CoV-2 infection outcome. However, their role in ...

Abstract	Monocytes are key modulators in acute viral infections, determining both inflammation and development of specific B- and T-cell responses. Recently, these cells were shown to be associated to different SARS-CoV-2 infection outcome. However, their role in acute HIV-1 infection remains unclear. We had the opportunity to evaluate the mononuclear cell compartment in an early hyper-acute HIV-1 patient in comparison with an untreated chronic HIV-1 and a cohort of SARS-CoV-2 infected patients, by high dimensional flow cytometry using an unsupervised approach. A distinct polarization of the monocyte phenotype was observed in the two viral infections, with maintenance of pro-inflammatory M1-like profile in HIV-1, in contrast to the M2-like immunosuppressive shift in SARS-CoV-2. Noticeably, both acute infections had reduced CD14ˡᵒʷ/⁻CD16⁺ non-classical monocytes, with depletion of the population expressing Slan (6-sulfo LacNac), which is thought to contribute to immune surveillance through pro-inflammatory properties. This depletion indicates a potential role of these cells in acute viral infection, which has not previously been explored. The inflammatory state accompanied by the depletion of Slan+ monocytes may provide new insights on the critical events that determine the rate of viral set-point in acute HIV-1 infection and subsequent impact on transmission and reservoir establishment.
Keywords	HIV infections ; Severe acute respiratory syndrome coronavirus 2 ; T-lymphocytes ; case studies ; flow cytometry ; immunosuppression ; inflammation ; monitoring ; monocytes ; patients ; phenotype
Language	English
Dates of publication	2021-0910
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13091805
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Follicular Helper T Cells Are Major Human Immunodeficiency Virus-2 Reservoirs and Support Productive Infection.

Godinho-Santos, Ana / Foxall, Russell B / Antão, Ana V / Tavares, Bárbara / Ferreira, Tiago / Serra-Caetano, Ana / Matoso, Paula / Sousa, Ana E

The Journal of infectious diseases

2019 Volume 221, Issue 1, Page(s) 122–126

Abstract: Follicular helper T cells (Tfh), CD4 lymphocytes critical for efficient antibody responses, have been shown to be key human immunodeficiency virus (HIV)-1 reservoirs. Human immunodeficiency virus-2 infection represents a unique naturally occurring model ... ...

Abstract	Follicular helper T cells (Tfh), CD4 lymphocytes critical for efficient antibody responses, have been shown to be key human immunodeficiency virus (HIV)-1 reservoirs. Human immunodeficiency virus-2 infection represents a unique naturally occurring model for investigating Tfh role in HIV/acquired immune deficiency syndrome, given its slow rate of CD4 decline, low to undetectable viremia, and high neutralizing antibody titers throughout the disease course. In this study, we investigated, for the first time, Tfh susceptibility to HIV-2 infection by combining in vitro infection of tonsillar Tfh with the ex vivo study of circulating Tfh from HIV-2-infected patients. We reveal that Tfh support productive HIV-2 infection and are preferential viral targets in HIV-2-infected individuals.
MeSH term(s)	DNA, Viral/metabolism ; Female ; HIV Infections/immunology ; HIV Infections/virology ; HIV-1/physiology ; HIV-2/physiology ; Humans ; Middle Aged ; Palatine Tonsil/immunology ; Palatine Tonsil/pathology ; Primary Cell Culture ; RNA, Messenger/metabolism ; Receptors, CCR5/metabolism ; Receptors, CXCR4/metabolism ; T-Lymphocytes, Helper-Inducer/metabolism ; T-Lymphocytes, Helper-Inducer/virology ; Viral Tropism ; gag Gene Products, Human Immunodeficiency Virus/genetics
Chemical Substances	CCR5 protein, human ; CXCR4 protein, human ; DNA, Viral ; RNA, Messenger ; Receptors, CCR5 ; Receptors, CXCR4 ; gag Gene Products, Human Immunodeficiency Virus
Language	English
Publishing date	2019-09-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3019-3
ISSN	1537-6613 ; 0022-1899
ISSN (online)	1537-6613
ISSN	0022-1899
DOI	10.1093/infdis/jiz431
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Article ; Online: Acute HIV-1 and SARS-CoV-2 Infections Share Slan+ Monocyte Depletion-Evidence from an Hyperacute HIV-1 Case Report.

Farias, Guilherme B / Badura, Robert / Conceição, Carolina M / Gomes, André M C / Godinho-Santos, Ana / Laia, Joel / Rosmaninho, Pedro / Santos, Diana F / Mota, Catarina / Almeida, Afonso R M / Fernandes, Susana M / Trombetta, Amelia C / Sousa, Ana E

Viruses

2021 Volume 13, Issue 9

Abstract	Monocytes are key modulators in acute viral infections, determining both inflammation and development of specific B- and T-cell responses. Recently, these cells were shown to be associated to different SARS-CoV-2 infection outcome. However, their role in acute HIV-1 infection remains unclear. We had the opportunity to evaluate the mononuclear cell compartment in an early hyper-acute HIV-1 patient in comparison with an untreated chronic HIV-1 and a cohort of SARS-CoV-2 infected patients, by high dimensional flow cytometry using an unsupervised approach. A distinct polarization of the monocyte phenotype was observed in the two viral infections, with maintenance of pro-inflammatory M1-like profile in HIV-1, in contrast to the M2-like immunosuppressive shift in SARS-CoV-2. Noticeably, both acute infections had reduced CD14
MeSH term(s)	Adult ; Aged ; Amino Sugars/immunology ; COVID-19/immunology ; Cohort Studies ; Female ; HIV Infections/immunology ; HIV-1/immunology ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Monocytes/immunology ; Young Adult
Chemical Substances	6-sulfo-LacNac ; Amino Sugars
Language	English
Publishing date	2021-09-10
Publishing country	Switzerland
Document type	Case Reports ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13091805
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Article ; Online: Severe COVID-19 Recovery Is Associated with Timely Acquisition of a Myeloid Cell Immune-Regulatory Phenotype.

Trombetta, Amelia C / Farias, Guilherme B / Gomes, André M C / Godinho-Santos, Ana / Rosmaninho, Pedro / Conceição, Carolina M / Laia, Joel / Santos, Diana F / Almeida, Afonso R M / Mota, Catarina / Gomes, Andreia / Serrano, Marta / Veldhoen, Marc / Sousa, Ana E / Fernandes, Susana M

Frontiers in immunology

2021 Volume 12, Page(s) 691725

Abstract: After more than one year since the COVID-19 outbreak, patients with severe disease still constitute the bottleneck of the pandemic management. Aberrant inflammatory responses, ranging from cytokine storm to immune-suppression, were described in COVID-19 ... ...

Abstract	After more than one year since the COVID-19 outbreak, patients with severe disease still constitute the bottleneck of the pandemic management. Aberrant inflammatory responses, ranging from cytokine storm to immune-suppression, were described in COVID-19 and no treatment was demonstrated to change the prognosis significantly. Therefore, there is an urgent need for understanding the underlying pathogenic mechanisms to guide therapeutic interventions. This study was designed to assess myeloid cell activation and phenotype leading to recovery in patients surviving severe COVID-19. We evaluated longitudinally patients with COVID-19 related respiratory insufficiency, stratified according to the need of intensive care unit admission (ICU, n = 11, and No-ICU, n = 9), and age and sex matched healthy controls (HCs, n = 11), by flow cytometry and a wide array of serum inflammatory/immune-regulatory mediators. All patients featured systemic immune-regulatory myeloid cell phenotype as assessed by both unsupervised and supervised analysis of circulating monocyte and dendritic cell subsets. Specifically, we observed a reduction of CD14lowCD16+ monocytes, and reduced expression of CD80, CD86, and Slan. Moreover, mDCs, pDCs, and basophils were significantly reduced, in comparison to healthy subjects. Contemporaneously, both monocytes and DCs showed increased expression of CD163, CD204, CD206, and PD-L1 immune-regulatory markers. The expansion of M2-like monocytes was significantly higher at admission in patients featuring detectable SARS-CoV-2 plasma viral load and it was positively correlated with the levels of specific antibodies. In No-ICU patients, we observed a peak of the alterations at admission and a progressive regression to a phenotype similar to HCs at discharge. Interestingly, in ICU patients, the expression of immuno-suppressive markers progressively increased until discharge. Notably, an increase of M2-like HLA-DRhighPD-L1+ cells in CD14++CD16- monocytes and in dendritic cell subsets was observed at ICU discharge. Furthermore, IFN-γ and IL-12p40 showed a decline over time in ICU patients, while high values of IL1RA and IL-10 were maintained. In conclusion, these results support that timely acquisition of a myeloid cell immune-regulatory phenotype might contribute to recovery in severe systemic SARS-CoV-2 infection and suggest that therapeutic agents favoring an innate immune system regulatory shift may represent the best strategy to be implemented at this stage.
MeSH term(s)	Adult ; Aged ; COVID-19/immunology ; Cell Differentiation ; Critical Care ; Cytokines/metabolism ; Female ; Humans ; Immunomodulation ; Male ; Middle Aged ; Monocytes/immunology ; Myeloid-Derived Suppressor Cells/immunology ; Phenotype ; Respiratory Insufficiency ; SARS-CoV-2/physiology ; Severity of Illness Index ; Th2 Cells/immunology
Chemical Substances	Cytokines
Language	English
Publishing date	2021-06-23
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2021.691725
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Article ; Online: SARS-CoV2 pneumonia recovery is linked to expansion of innate lymphoid cells type 2 expressing CCR10.

Gomes, André M C / Farias, Guilherme B / Dias-Silva, Manuel / Laia, Joel / Trombetta, Amelia C / Godinho-Santos, Ana / Rosmaninho, Pedro / Santos, Diana F / Conceição, Carolina M / Costa-Reis, Renato / Adão-Serrano, Maria / Mota, Catarina / Almeida, Afonso R M / Sousa, Ana E / Fernandes, Susana M

European journal of immunology

2021 Volume 51, Issue 12, Page(s) 3194–3201

Abstract: Accelerate lung repair in SARS-CoV-2 pneumonia is essential for pandemic handling. Innate lymphoid cells (ILCs) are likely players, given their role in mucosal protection and tissue homeostasis. We studied ILC subpopulations at two time points in a ... ...

Abstract	Accelerate lung repair in SARS-CoV-2 pneumonia is essential for pandemic handling. Innate lymphoid cells (ILCs) are likely players, given their role in mucosal protection and tissue homeostasis. We studied ILC subpopulations at two time points in a cohort of patients admitted in the hospital due to SARS-CoV-2 infection. COVID-19 patients with moderate/severe respiratory failure featured profound depletion of circulating ILCs at hospital admission, in agreement with overall lymphocyte depletion. However, ILCs recovered in direct correlation with lung function improvement as measured by oxygenation index and in negative association with inflammatory and lung/endothelial damage markers like RAGE. While both ILC1 and ILC2 expanded, ILC2 showed the most striking phenotype changes, with CCR10 upregulation in strong correlation with these parameters. Overall, CCR10
MeSH term(s)	Adult ; Aged ; Antigens, Neoplasm/metabolism ; Biomarkers/metabolism ; COVID-19/immunology ; Cell Proliferation ; Cytokines/metabolism ; Female ; Humans ; Immunity, Innate ; Lung/pathology ; Lymphocytes/immunology ; Male ; Middle Aged ; Mitogen-Activated Protein Kinases/metabolism ; Pneumonia, Viral/immunology ; Receptors, CCR10/metabolism ; Recovery of Function ; SARS-CoV-2/physiology ; Th2 Cells/immunology ; Up-Regulation
Chemical Substances	Antigens, Neoplasm ; Biomarkers ; CCR10 protein, human ; Cytokines ; Receptors, CCR10 ; MOK protein, human (EC 2.7.11.22) ; Mitogen-Activated Protein Kinases (EC 2.7.11.24)
Language	English
Publishing date	2021-10-20
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	120108-6
ISSN	1521-4141 ; 0014-2980
ISSN (online)	1521-4141
ISSN	0014-2980
DOI	10.1002/eji.202149311
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