Characterization of Four New Monoclonal Antibodies that Recognize Mouse Natural Killer Activation Receptors

Kung, Sam K. P.; Su, Ruey-Chyi; Shannon, James A.; Miller, Richard G.

doi:10.1089/02724570152057580

Cited by 3 publications

(2 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The lymphocyte population was gated, and two‐colour FCM performed. The percentages of CD3 – CD161 + cells (NK cells) and CD3 + CD161 + double‐positive cells (NK T cells) [21,22] in gated events were measured. These data were analysed using CellQuest software (Becton Dickinson).…”

Section: Methodsmentioning

confidence: 99%

Mushroom plant workers experience a shift towards a T helper type 2 dominant state: contribution of innate immunity to spore antigen

Saikai

Tanaka

Sato

et al. 2003

Clinical and Experimental Immunology

View full text Add to dashboard Cite

SUMMARYContemporary mushroom factories are places where there is a substantial risk of the occurrence of respiratory allergy. The aims of this investigation were to estimate its causative agents and to evaluate the contribution of innate immune response in mushroom workers who cultivate Hypsizigus marmoreus (Bunashimeji). Cross-sectional and follow-up studies were performed in the factory. We investigated CD1b, CD3, CD4, CD8, CD14, CD45RO, CD62L and CD161 expression in peripheral blood mononuclear cells (PBMC) by flow cytometry, and serum levels of interleukin (IL-2), IL-4, granulocytemacrophage colony stimulating factor (GM-CSF), IL-13 and interferon (IFN)-g by enzyme-linked immunosorbent assay (ELISA). Co-culture experiments of PBMC with spore extracts were also performed. Percentages of CD1b + monocytes, natural killer (NK), NK T and CD4 + T cells were increased in the workers compared with controls. Increases in Th2 type cells, Th2/Th1 ratio and serum IL-13 and decreased IFN-g were detected, indicating a Th2-biased status of the workers. The follow-up study showed that monocytes and NK cells increased soon after employment while CD4 + T, Th2 and NK T cells increased gradually as employment time lengthened. Serum precipitating antibody to the mushroom antigen could be detected at a later stage. Co-cultivation of PBMC with the spore extracts induced much higher CD1b expression, and suppressed secretion of Th1 cytokine in culture supernatants. These results indicate that the mushroom antigen contains highly immunogenic substances which stimulate PBMC into a Th2-biased in vivo status, and innate immune cells might also play a critical role in developing respiratory allergy in mushroom workers.

show abstract

Section: Methodsmentioning

confidence: 99%

Mushroom plant workers experience a shift towards a T helper type 2 dominant state: contribution of innate immunity to spore antigen

Saikai

Tanaka

Sato

et al. 2003

Clinical and Experimental Immunology

View full text Add to dashboard Cite

show abstract

“…Briefly, these cells secrete mouse IgG2a monoclonal antibody, which reacts with mouse natural killer cells. These hybridomas were obtained by fusion between SP2 myeloma cells and spleen cells (C3H × BALB/c) immunized with mouse spleen cells (Koo and Peppard, 1984;Kung et al, 2001). The cells were cultured and prepared using phosphatebuffered saline with pH 7.4 (PBS), bovine serum albumin (BSA), Tween 20, fluorescein diacetate, dimethylsulfoxide (DMSO), Iscove's Modified Dulbecco's Medium (Gibco, Invitrogen, Switzerland), foetal calf serum, normal mouse serum, hyaluronidase, and propidium iodine.…”

Section: Chemicals and Cellsmentioning

confidence: 99%

Screening cell surface receptors using micromosaic immunoassays

Wolf

Zimmermann

Delamarche

et al. 2006

Biomed Microdevices

View full text Add to dashboard Cite

This report presents a general method for screening cell surface receptors using so-called micromosaic immunoassays. This method employs a microfluidic chip having n (n = 11) independent flow paths to move cells over m (m = 11) lines of surface-patterned antibodies for screening individual cells in a parallel, combinatorial, fast and flexible manner. The antibodies are patterned as 30-microm-wide lines on a poly(dimethylsiloxane) layer used to seal the area of the chip in which screening is being monitored. Mouse hybridoma cells having CD44 cell surface receptors and anti-CD44 antibodies were used to establish a proof-of-concept for this method. Both the capture antibodies and the cells were fluorescently labelled to allow the position of the cells to be accurately tracked over the binding sites using an inverted fluorescence microscope. The chips and cells were maintained at a constant temperature between 20 to 37 degrees C, and flow velocities of the cells over the capture areas were 100-280 microm s(-1), resulting in a approximately 0.1-0.3 s residency time of the cells on each of the eleven 30 x 30 microm s2 capture areas. Binding of the cells appeared to be specific to the capture areas, with a yield of 30% when the assay was performed at a temperature of 37 degrees C and with a slow flow velocity. We suggest that this proof-of-concept is broadly applicable to the screening of cells for medical/diagnostic purposes as well as for basic research on the interaction of cells with surfaces.

show abstract

High-dimensional single cell mass cytometry analysis of the murine hematopoietic system reveals signatures induced by ageing and physiological pathogen challenges

et al. 2021

View full text Add to dashboard Cite

Background Immune ageing is a result of repetitive microbial challenges along with cell intrinsic or systemic changes occurring during ageing. Mice under ‘specific-pathogen-free’ (SPF) conditions are frequently used to assess immune ageing in long-term experiments. However, physiological pathogenic challenges are reduced in SPF mice. The question arises to what extent murine experiments performed under SPF conditions are suited to analyze immune ageing in mice and serve as models for human immune ageing. Our previous comparisons of same aged mice with different microbial exposures, unambiguously identified distinct clusters of immune cells characteristic for numerous previous pathogen encounters in particular in pet shop mice. Results We here performed single cell mass cytometry assessing splenic as secondary and bone marrow as primary lymphoid organ-derived leukocytes isolated from young versus aged SPF mice in order to delineate alterations of the murine hematopoietic system induced during ageing. We then compared immune clusters from young and aged SPF mice to pet shop mice in order to delineate alterations of the murine hematopoietic system induced by physiological pathogenic challenges and those caused by cell intrinsic or systemic changes during ageing. Notably, distinct immune signatures were similarly altered in both pet shop and aged SPF mice in comparison to young SPF mice, including increased frequencies of memory T lymphocytes, effector-cytokine producing T cells, plasma cells and mature NK cells. However, elevated frequencies of CD4+ T cells, total NK cells, granulocytes, pDCs, cDCs and decreased frequencies of naïve B cells were specifically identified only in pet shop mice. In aged SPF mice specifically the frequencies of splenic IgM+ plasma cells, CD8+ T cells and CD4+ CD25+ Treg were increased as compared to pet shop mice and young mice. Conclusions Our study dissects firstly how ageing impacts both innate and adaptive immune cells in primary and secondary lymphoid organs. Secondly, it partly distinguishes murine intrinsic immune ageing alterations from those induced by physiological pathogen challenges highlighting the importance of designing mouse models for their use in preclinical research including vaccines and immunotherapies.

show abstract

Characterization of Four New Monoclonal Antibodies that Recognize Mouse Natural Killer Activation Receptors

Cited by 3 publications

References 26 publications

Mushroom plant workers experience a shift towards a T helper type 2 dominant state: contribution of innate immunity to spore antigen

Mushroom plant workers experience a shift towards a T helper type 2 dominant state: contribution of innate immunity to spore antigen

Screening cell surface receptors using micromosaic immunoassays

High-dimensional single cell mass cytometry analysis of the murine hematopoietic system reveals signatures induced by ageing and physiological pathogen challenges

Contact Info

Product

Resources

About