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Article ; Online: Protocol for the expansion of mouse immune effector cells for in vitro and in vivo studies.

Look, Thomas / Meister, Hanna / Weller, Michael / Weiss, Tobias

STAR protocols

2023 Volume 4, Issue 4, Page(s) 102700

Abstract: Reproducible and efficient expansion of different immune effector cells is required for pre-clinical studies investigating adoptive cell therapies against cancer. Here, we provide a protocol for the rapid expansion of mouse T cells, natural killer (NK) ... ...

Abstract	Reproducible and efficient expansion of different immune effector cells is required for pre-clinical studies investigating adoptive cell therapies against cancer. Here, we provide a protocol for the rapid expansion of mouse T cells, natural killer (NK) cells, and bone-marrow-derived macrophages (BMDMs). We describe steps for αCD3/αCD8 plate coating, isolating splenocytes, and expanding T cells and NK cells. Further, we detail procedures for bone marrow isolation and BMDM differentiation.
MeSH term(s)	Mice ; Animals ; Killer Cells, Natural ; Neoplasms/therapy ; T-Lymphocytes ; Bone Marrow
Language	English
Publishing date	2023-11-09
Publishing country	United States
Document type	Journal Article
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2023.102700
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Article ; Online: Protocol for the expansion of mouse immune effector cells for in vitro and in vivo studies

Thomas Look / Hanna Meister / Michael Weller / Tobias Weiss

STAR Protocols, Vol 4, Iss 4, Pp 102700- (2023)

2023

Abstract: Summary: Reproducible and efficient expansion of different immune effector cells is required for pre-clinical studies investigating adoptive cell therapies against cancer. Here, we provide a protocol for the rapid expansion of mouse T cells, natural ... ...

Abstract	Summary: Reproducible and efficient expansion of different immune effector cells is required for pre-clinical studies investigating adoptive cell therapies against cancer. Here, we provide a protocol for the rapid expansion of mouse T cells, natural killer (NK) cells, and bone-marrow-derived macrophages (BMDMs). We describe steps for αCD3/αCD8 plate coating, isolating splenocytes, and expanding T cells and NK cells. Further, we detail procedures for bone marrow isolation and BMDM differentiation. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Keywords	Cell Biology ; Cell culture ; Cell isolation ; Flow Cytometry ; Cancer ; Immunology ; Science (General) ; Q1-390
Language	English
Publishing date	2023-12-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
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Article ; Online: A priori prediction of breast cancer response to neoadjuvant chemotherapy using quantitative ultrasound, texture derivative and molecular subtype.

Sannachi, Lakshmanan / Osapoetra, Laurentius O / DiCenzo, Daniel / Halstead, Schontal / Wright, Frances / Look-Hong, Nicole / Slodkowska, Elzbieta / Gandhi, Sonal / Curpen, Belinda / Kolios, Michael C / Oelze, Michael / Czarnota, Gregory J

Scientific reports

2023 Volume 13, Issue 1, Page(s) 22687

Abstract: The purpose of this study was to investigate the performances of the tumor response prediction prior to neoadjuvant chemotherapy based on quantitative ultrasound, tumour core-margin, texture derivative analyses, and molecular parameters in a large cohort ...

Abstract	The purpose of this study was to investigate the performances of the tumor response prediction prior to neoadjuvant chemotherapy based on quantitative ultrasound, tumour core-margin, texture derivative analyses, and molecular parameters in a large cohort of patients (n = 208) with locally advanced and earlier-stage breast cancer and combined them to best determine tumour responses with machine learning approach. Two multi-features response prediction algorithms using a k-nearest neighbour and support vector machine were developed with leave-one-out and hold-out cross-validation methods to evaluate the performance of the response prediction models. In a leave-one-out approach, the quantitative ultrasound-texture analysis based model attained good classification performance with 80% of accuracy and AUC of 0.83. Including molecular subtype in the model improved the performance to 83% of accuracy and 0.87 of AUC. Due to limited number of samples in the training process, a model developed with a hold-out approach exhibited a slightly higher bias error in classification performance. The most relevant features selected in predicting the response groups are core-to-margin, texture-derivative, and molecular subtype. These results imply that that baseline tumour-margin, texture derivative analysis methods combined with molecular subtype can potentially be used for the prediction of ultimate treatment response in patients prior to neoadjuvant chemotherapy.
MeSH term(s)	Humans ; Female ; Breast Neoplasms/diagnostic imaging ; Breast Neoplasms/drug therapy ; Breast Neoplasms/pathology ; Neoadjuvant Therapy/methods ; Chemotherapy, Adjuvant ; Ultrasonography ; Algorithms ; Retrospective Studies
Language	English
Publishing date	2023-12-19
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/s41598-023-49478-3
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Article: SARS-CoV-2-host cell surface interactions and potential antiviral therapies.

Butnariu, Aura-Bianca / Look, Alex / Grillo, Marta / Tabish, Tanveer A / McGarvey, Michael J / Pranjol, Md Zahidul I

Interface focus

2021 Volume 12, Issue 1, Page(s) 20200081

Abstract: In this review, we reveal the latest developments at the interface between SARS-CoV-2 and the host cell surface. In particular, we evaluate the current and potential mechanisms of binding, fusion and the conformational changes of the spike (S) protein to ...

Abstract	In this review, we reveal the latest developments at the interface between SARS-CoV-2 and the host cell surface. In particular, we evaluate the current and potential mechanisms of binding, fusion and the conformational changes of the spike (S) protein to host cell surface receptors, especially the human angiotensin-converting enzyme 2 (ACE2) receptor. For instance, upon the initial attachment, the receptor binding domain of the S protein forms primarily hydrogen bonds with the protease domain of ACE2 resulting in conformational changes within the secondary structure. These surface interactions are of paramount importance and have been therapeutically exploited for antiviral design, such as monoclonal antibodies. Additionally, we provide an insight into novel therapies that target viral non-structural proteins, such as viral RNA polymerase. An example of which is remdesivir which has now been approved for use in COVID-19 patients by the US Food and Drug Administration. Establishing further understanding of the molecular details at the cell surface will undoubtably aid the development of more efficacious and selectively targeted therapies to reduce the burden of COVID-19.
Language	English
Publishing date	2021-12-10
Publishing country	England
Document type	Journal Article ; Review
ISSN	2042-8898
ISSN	2042-8898
DOI	10.1098/rsfs.2020.0081
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Article: Laparoscopic Treatment of an Incarcerated Meckel's Diverticulum in a Femoral Hernia.

Paasch, Christoph / De Santo, Gianluca / Look, Peter / Boettge, Katherina / Hünerbein, Michael

Case reports in surgery

2019 Volume 2019, Page(s) 3140706

Abstract: Meckel's diverticulum (MD) is the persistence of the omphalomesenteric duct. It is usually asymptomatic but may present with bleeding, infections, and intestinal obstruction. It also may be a content of a hernia sac, a so-called Littre hernia. Herein, we ...

Abstract	Meckel's diverticulum (MD) is the persistence of the omphalomesenteric duct. It is usually asymptomatic but may present with bleeding, infections, and intestinal obstruction. It also may be a content of a hernia sac, a so-called Littre hernia. Herein, we will present the case of a 75-year-old female, who suffered from a painful swelling of the right inguinal region. Ultrasound imaging detected an inguinal hernia with incarcerated blind ending small bowel. Immediately, a laparoscopy was conducted. We diagnosed a right femoral hernia with an incarcerated MD. A TAPP (transabdominal preperitoneal) procedure was performed and the MD tangential stapled. Due to an uneventful postoperative course, the patient left the hospital after two days. An incarceration of a MD in a femoral hernia is rare. Tangential resection of the MD with simultaneous hernia repair in a TAPP technique seems to be a sufficient approach, when it is conducted by an experienced surgeon.
Keywords	covid19
Language	English
Publishing date	2019-08-08
Publishing country	United States
Document type	Case Reports
ZDB-ID	2657697-1
ISSN	2090-6919 ; 2090-6900
ISSN (online)	2090-6919
ISSN	2090-6900
DOI	10.1155/2019/3140706
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Article ; Online: Targeted delivery of tumor necrosis factor in combination with CCNU induces a T cell-dependent regression of glioblastoma.

Look, Thomas / Puca, Emanuele / Bühler, Marcel / Kirschenbaum, Daniel / De Luca, Roberto / Stucchi, Riccardo / Ravazza, Domenico / Di Nitto, Cesare / Roth, Patrick / Katzenelenbogen, Yonatan / Weiner, Assaf / Rindlisbacher, Lukas / Becher, Burkhard / Amit, Ido / Weller, Michael / Neri, Dario / Hemmerle, Teresa / Weiss, Tobias

Science translational medicine

2023 Volume 15, Issue 697, Page(s) eadf2281

Abstract: Glioblastoma is the most aggressive primary brain tumor with an unmet need for more effective therapies. Here, we investigated combination therapies based on L19TNF, an antibody-cytokine fusion protein based on tumor necrosis factor that selectively ... ...

Abstract	Glioblastoma is the most aggressive primary brain tumor with an unmet need for more effective therapies. Here, we investigated combination therapies based on L19TNF, an antibody-cytokine fusion protein based on tumor necrosis factor that selectively localizes to cancer neovasculature. Using immunocompetent orthotopic glioma mouse models, we identified strong anti-glioma activity of L19TNF in combination with the alkylating agent CCNU, which cured the majority of tumor-bearing mice, whereas monotherapies only had limited efficacy. In situ and ex vivo immunophenotypic and molecular profiling in the mouse models revealed that L19TNF and CCNU induced tumor DNA damage and treatment-associated tumor necrosis. In addition, this combination also up-regulated tumor endothelial cell adhesion molecules, promoted the infiltration of immune cells into the tumor, induced immunostimulatory pathways, and decreased immunosuppression pathways. MHC immunopeptidomics demonstrated that L19TNF and CCNU increased antigen presentation on MHC class I molecules. The antitumor activity was T cell dependent and completely abrogated in immunodeficient mouse models. On the basis of these encouraging results, we translated this treatment combination to patients with glioblastoma. The clinical translation is ongoing but already shows objective responses in three of five patients in the first recurrent glioblastoma patient cohort treated with L19TNF in combination with CCNU (NCT04573192).
MeSH term(s)	Animals ; Mice ; Glioblastoma/drug therapy ; T-Lymphocytes ; Neoplasm Recurrence, Local ; Tumor Necrosis Factor-alpha ; Disease Models, Animal ; Lomustine
Chemical Substances	Tumor Necrosis Factor-alpha ; Lomustine (7BRF0Z81KG)
Language	English
Publishing date	2023-05-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2518854-9
ISSN	1946-6242 ; 1946-6234
ISSN (online)	1946-6242
ISSN	1946-6234
DOI	10.1126/scitranslmed.adf2281
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Zs.A 6941: Show issues

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Article ; Online: Multifunctional mRNA-Based CAR T Cells Display Promising Antitumor Activity Against Glioblastoma.

Meister, Hanna / Look, Thomas / Roth, Patrick / Pascolo, Steve / Sahin, Ugur / Lee, Sohyon / Hale, Benjamin D / Snijder, Berend / Regli, Luca / Ravi, Vidhya M / Heiland, Dieter Henrik / Sentman, Charles L / Weller, Michael / Weiss, Tobias

Clinical cancer research : an official journal of the American Association for Cancer Research

2022 Volume 28, Issue 21, Page(s) 4747–4756

Abstract: Purpose: Most chimeric antigen receptor (CAR) T-cell strategies against glioblastoma have demonstrated only modest therapeutic activity and are based on persistent gene modification strategies that have limited transgene capacity, long manufacturing ... ...

Abstract	Purpose: Most chimeric antigen receptor (CAR) T-cell strategies against glioblastoma have demonstrated only modest therapeutic activity and are based on persistent gene modification strategies that have limited transgene capacity, long manufacturing processes, and the risk for uncontrollable off-tumor toxicities. mRNA-based T-cell modifications are an emerging safe, rapid, and cost-effective alternative to overcome these challenges, but are underexplored against glioblastoma. Experimental design: We generated mouse and human mRNA-based multifunctional T cells coexpressing a multitargeting CAR based on the natural killer group 2D (NKG2D) receptor and the proinflammatory cytokines IL12 and IFNα2 and assessed their antiglioma activity in vitro and in vivo. Results: Compared with T cells that either expressed the CAR or cytokines alone, multifunctional CAR T cells demonstrated increased antiglioma activity in vitro and in vivo in three orthotopic immunocompetent mouse glioma models without signs of toxicity. Mechanistically, the coexpression of IL12 and IFNα2 in addition to the CAR promoted a proinflammatory tumor microenvironment and reduced T-cell exhaustion as demonstrated by ex vivo immune phenotyping, cytokine profiling, and RNA sequencing. The translational potential was demonstrated by image-based single-cell analyses of mRNA-modified T cells in patient glioblastoma samples with a complex cellular microenvironment. This revealed strong antiglioma activity of human mRNA-based multifunctional NKG2D CAR T cells coexpressing IL12 and IFNα2 whereas T cells that expressed either the CAR or cytokines alone did not demonstrate comparable antiglioma activity. Conclusions: These data provide a robust rationale for future clinical studies with mRNA-based multifunctional CAR T cells to treat malignant brain tumors.
MeSH term(s)	Humans ; Mice ; Animals ; Glioblastoma/genetics ; Glioblastoma/therapy ; Glioblastoma/pathology ; Immunotherapy, Adoptive ; NK Cell Lectin-Like Receptor Subfamily K/genetics ; RNA, Messenger/genetics ; Xenograft Model Antitumor Assays ; Cell Line, Tumor ; Receptors, Chimeric Antigen/genetics ; T-Lymphocytes ; Cytokines ; Interleukin-12 ; Tumor Microenvironment/genetics
Chemical Substances	NK Cell Lectin-Like Receptor Subfamily K ; RNA, Messenger ; Receptors, Chimeric Antigen ; Cytokines ; Interleukin-12 (187348-17-0)
Language	English
Publishing date	2022-08-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1225457-5
ISSN	1557-3265 ; 1078-0432
ISSN (online)	1557-3265
ISSN	1078-0432
DOI	10.1158/1078-0432.CCR-21-4384
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Zs.A 4223: Show issues

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Article ; Online: Use of protective antigen of Bacillus anthracis as a model recombinant antigen to evaluate toll-like receptors 2, 3, 4, 7 and 9 agonists in mice using established functional antibody assays, antigen-specific antibody assays and cellular assays.

Inglefield, Jon / Catania, Jason / Harris, Andrea / Hickey, Thomas / Ma, Zhidong / Minang, Jacob / Baranji, Katalin / Spangler, Tarl / Look, Jee / Ruiz, Christian / Lu, Hang / Alleva, David / Reece, Joshua J / Lacy, Michael J

Vaccine

2022 Volume 40, Issue 38, Page(s) 5544–5555

Abstract: Toll-like receptor (TLR) agonists can act as immune stimulants alone or as part of alum or oil formulations. Humoral and cellular immune responses were utilized to assess quantitative and qualitative immune response enhancement by TLR agonists using ... ...

Abstract	Toll-like receptor (TLR) agonists can act as immune stimulants alone or as part of alum or oil formulations. Humoral and cellular immune responses were utilized to assess quantitative and qualitative immune response enhancement by TLR agonists using recombinant protective antigen (rPA) of B. anthracis as a model antigen. To rPA, combined with aluminum hydroxide (Alhydrogel; Al(OH)3) or squalene (AddaVax™), was added one of 7 TLR agonists: TLR2 agonist Pam3CysSK4 (PamS), TLR3 agonist double stranded polyinosinic:polycytidylic acid (PolyIC), TLR4 agonists Monophosphoryl lipid A (MPLA) or glucopyranosyl lipid A (GLA), TLR7-8 agonists 3M-052 or Resiquimod (Resiq), or TLR9 agonist CPG 7909 (CPG). CD-1 or BALB/c mice received two intraperitoneal or intramuscular immunizations 14 days apart, followed by serum or spleen sampling 14 days later. All TLR agonists except PamS induced high levels of B. anthracis lethal toxin-neutralizing antibodies and immunoglobulin G (IgG) anti-PA. Some responses were >100-fold higher than those without a TLR agonist, and IP delivery (0.5 mL) induced higher TLR-mediated antibody response increases compared to IM delivery (0.05 mL). TLR7-8 and TLR9 agonists induced profound shifts of IgG anti-PA response to IgG2a or IgG2b. Compared to the 14-day immunization schedule, use of a shortened immunization schedule of only 7 days between prime and boost found that TLR9 agonist CPG in a squalene formulation maintained higher interferon-γ-positive cells than TLR4 agonist GLA. Variability in antibody responses was lower in BALB/c mice than CD-1 mice but antibody responses were higher in CD-1 mice. Lower serum 50% effective concentration (EC50) values were found for rPA-agonist formulations and squalene formulations compared to Al(OH)3 formulations. Lower EC50 values also were associated with low frequency detection of linear peptide epitopes. In summary, TLR agonists elicited cellular immune responses and markedly boosted humoral responses.
MeSH term(s)	Adjuvants, Immunologic ; Aluminum Hydroxide ; Animals ; Antigens ; Bacillus anthracis ; Immunoglobulin G ; Mice ; Mice, Inbred BALB C ; Squalene ; Toll-Like Receptor 2 ; Toll-Like Receptor 4/agonists ; Toll-Like Receptor 7/agonists ; Toll-Like Receptor 9/agonists
Chemical Substances	Adjuvants, Immunologic ; Antigens ; Immunoglobulin G ; Toll-Like Receptor 2 ; Toll-Like Receptor 4 ; Toll-Like Receptor 7 ; Toll-Like Receptor 9 ; Aluminum Hydroxide (5QB0T2IUN0) ; Squalene (7QWM220FJH)
Language	English
Publishing date	2022-06-27
Publishing country	Netherlands
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2022.06.017
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Zs.A 1930: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 458: Show issues

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Article ; Online: Effects of Tibiofibular and Ankle Joint Manipulation on Hip Strength and Muscle Activation.

Lawrence, Michael A / Raymond, Jamie T / Look, Amy E / Woodard, Nicholas M / Schicker, Christina M / Swanson, Brian T

Journal of manipulative and physiological therapeutics

2020 Volume 43, Issue 5, Page(s) 406–417

Abstract: Objectives: The purpose of this study was to determine whether high-velocity, low-amplitude ankle region manipulations could increase force output and muscle activation of hip musculature in individuals with a history of ankle sprain and unilateral ... ...

Abstract	Objectives: The purpose of this study was to determine whether high-velocity, low-amplitude ankle region manipulations could increase force output and muscle activation of hip musculature in individuals with a history of ankle sprain and unilateral tensor fascia latae (TFL) weakness during muscle testing. Methods: This investigation used a single-arm repeated measures design. Twenty-five participants' force outputs were tested at three time points (before manipulation, immediately after manipulation, and 48 hours after manipulation), and muscle activation of the rectus femoris, gluteus medius, and TFL was measured before and immediately after manipulation. Manipulations were applied to the talocrural, subtalar, proximal, and distal tibiofibular joints of the weaker limb. No contralateral manipulations were applied. Two-way repeated measures analysis of variance was used to compare maximal and average force production for each limb. In addition, paired t tests were used to compare muscle activation before and after manipulations. Results: There was a significant limb × time interaction. The involved limb average force increased from before manipulation (65.7 N) to 48 hours after manipulation (77.8 N; P = .014), maximal force increased (76.9 N) 48 hours after manipulation (87.8 N; P = .030), and gluteus medius activation increased (9.8% maximum, 12.2% average) immediately after manipulation. No significant differences were found in the uninvolved limb. Conclusion: The results of this study suggest that high-velocity, low-amplitude ankle region manipulations might improve hip abductor strength in individuals with a history of ankle sprain and unilateral weakness during a TFL muscle test.
MeSH term(s)	Adult ; Ankle ; Ankle Injuries/therapy ; Ankle Joint/physiology ; Electromyography/methods ; Exercise Therapy/methods ; Female ; Hip Joint/physiology ; Humans ; Isometric Contraction ; Male ; Manipulation, Orthopedic/methods ; Muscle, Skeletal/physiology ; Range of Motion, Articular
Language	English
Publishing date	2020-07-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	761054-3
ISSN	1532-6586 ; 0161-4754
ISSN (online)	1532-6586
ISSN	0161-4754
DOI	10.1016/j.jmpt.2019.10.005
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Zs.A 1859: Show issues

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Article ; Online: The long-term memory benefits of a daytime nap compared with cramming.

Cousins, James N / Wong, Kian F / Raghunath, Bindiya L / Look, Carol / Chee, Michael W L

Sleep

2018 Volume 42, Issue 1

Abstract: Study objectives: Daytime naps benefit long-term memory relative to taking a break and remaining awake. However, the use of naps as a practical way to improve learning has not been examined, in particular, how memory following a nap compares with ... ...

Abstract	Study objectives: Daytime naps benefit long-term memory relative to taking a break and remaining awake. However, the use of naps as a practical way to improve learning has not been examined, in particular, how memory following a nap compares with spending the equivalent amount of time cramming. Methods: Young adults learned detailed factual knowledge in sessions that flanked 1 hr spent napping (n = 27), taking a break (n = 27), or cramming that information (n = 30). Recall was examined 30 min and 1 week after learning. Results: When tested 30 min after learning, cramming and napping led to significantly better memory than taking a break. After a week, napping maintained this significant advantage, but cramming did not. Conclusions: These findings demonstrate the longer-term benefits of napping for retention of memoranda akin to what students encounter daily and encourage more widespread adoption of napping in education.
MeSH term(s)	Adult ; Female ; Humans ; Learning/physiology ; Male ; Memory, Long-Term/physiology ; Mental Recall/physiology ; Sleep/physiology ; Wakefulness/physiology ; Young Adult
Language	English
Publishing date	2018-12-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	424441-2
ISSN	1550-9109 ; 0161-8105
ISSN (online)	1550-9109
ISSN	0161-8105
DOI	10.1093/sleep/zsy207
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