Substrate Rigidity Regulates Human T Cell Activation and Proliferation

O’Connor, Roddy S.; Hao, Xueli; Shen, K. Robert; Bashour, Keenan; Akimova, Tatiana; Hancock, Wayne W.; Kam, Lance C.; Milone, Michael C.

doi:10.4049/jimmunol.1102757

Cited by 254 publications

(288 citation statements)

References 51 publications

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“…Based on this information, it is reasonable to speculate that the BCR might have mechanosensing ability toward the substrate presenting the Ags that may allow it to regulate these molecular events in the cascades of B cell activation. This speculation is also supported by two recent reports on T cell mechanosensing capability (20,21).…”

supporting

confidence: 77%

“…However, for substrate stiffness .100 kPa, the activation of T cells did not seem to be enhanced, but was instead somewhat impaired (20). These results and speculations were further supported by a recent study by Milone and colleagues (21). Using polydimethylsiloxane (PDMS) substrates, they were able to produce substrate stiffnesses ranging from 100 to 8000 kPa.…”

Section: Discussionmentioning

confidence: 57%

“…We simply tethered anti-IgM surrogate Ags or NP8-BSA Ags to the surface of polyacrylamide gels. Our experimental system is very similar to the system used by Kam and colleagues (20) in which IgG anti-CD3 and anti-CD28 surrogate Ags were tethered to polyacrylamide gels, or to the system used by Milone and colleagues (21) in which IgG anti-CD3 and anti-CD28 surrogate Ags were tethered to PDMS gels. In their systems, no adhesive ligands were used, but in both cases, T cells similarly showed strong mechanosensing abilities.…”

Section: Discussionmentioning

confidence: 99%

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B Cell Activation Is Regulated by the Stiffness Properties of the Substrate Presenting the Antigens

Wan

Zhang

Fan

et al. 2013

The Journal of Immunology

109

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B lymphocytes are activated upon Ag sensing by BCRs. The substrate presenting the Ag can show different degrees of stiffness. It is not clear whether B cells can respond to changes in substrate stiffness. In this study we use high-resolution, high-speed live cell imaging techniques to capture the molecular events in B cell activation after the recognition of Ags tethered to polyacrylamide gel substrates with variable degrees of stiffness as quantified by Young’s modulus (2.6–22.1 kPa). We show that the initiation of B cell activation is extremely sensitive to substrate stiffness. B cells exhibit much stronger activation responses when encountering Ags tethered to substrates with a high degree of stiffness as measured by the accumulation of BCR, phospho-spleen tyrosine kinase, and phosphotyrosine molecules into the B cell immunological synapse. Ags tethered to stiff substrates induce the formation of more prominent BCR and phospho-spleen tyrosine kinase microclusters with significantly enhanced colocalization as compared with Ags tethered to soft substrates. Moreover, the expression of the B cell activation marker CD69 is enhanced in B cells encountering Ags on stiffer substrates. Through time-lapse live cell imaging, we find that the different responses of B cells to substrate stiffness are only demonstrated 5 min after BCR and Ag recognition. Using a series of cytoskeleton inhibitors, we determine that the mechanosensing ability of B cells is dependent on microtubules, and only mildly linked to the actin cytoskeleton. These results suggest the importance of the mechanical properties mediated by substrate stiffness in B cell activation.

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supporting

confidence: 77%

Section: Discussionmentioning

confidence: 57%

Section: Discussionmentioning

confidence: 99%

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B Cell Activation Is Regulated by the Stiffness Properties of the Substrate Presenting the Antigens

Wan

Zhang

Fan

et al. 2013

The Journal of Immunology

109

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“…Lastly, it is an intriguing question: how the stiffness feature of the substrates presenting the antigens will eventually affect the antibody responses in vivo? To the best of our knowledge, there is a lack of answers for all these questions.In this report, we tried to address these questions using substrates with softer (20 kPa) or stiffer (1100 kPa) stiffness that were made from polydimethylsiloxane (PDMS) following a protocol as recently reported for the mechanosensing studies on T cells [22,23]. PDMS is a biocompatible organosilicon elastomer that has been used as an anticaking or antibloating agent.…”

mentioning

confidence: 99%

“…Indeed, B1-8-BCR-specific hapten antigen 4-hydroxy-3-nitrophenyl (NP), Alexa 488 conjugated NP8-BSA, can stably bind to PDMS surface ( Fig. 1A and B) after an overnight incubation at 4°C following a published procedure [23]. 3D confocal microscopy revealed a lateral distribution of Alexa 488 conjugated NP8-BSA that was both planar and highly uniform (Fig.…”

mentioning

confidence: 99%

Substrate stiffness regulates B‐cell activation, proliferation, class switch, and T‐cell‐independent antibody responses in vivo

Zeng

Wan

et al. 2015

Eur J Immunol

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B cells use B-cell receptors (BCRs) to sense antigens that are usually presented on substrates with different stiffness. However, it is not known how substrate stiffness affects B-cell proliferation, class switch, and in vivo antibody responses. We addressed these questions using polydimethylsiloxane (PDMS) substrates with different stiffness (20 or 1100 kPa). Live cell imaging experiments suggested that antigens on stiffer substrates more efficiently trigger the synaptic accumulation of BCR and phospho-Syk molecules compared with antigens on softer substrates. In vitro expansion of mouse primary B cells shows different preferences for substrate stiffness when stimulated by different expansion stimuli. LPS equally drives B-cell proliferation on stiffer or softer substrates. Anti-CD40 antibodies enhance B-cell proliferation on stiffer substrates, while antigens enhance B-cell proliferation on softer substrates through a mechanism involving the enhanced phosphorylation of PI3K, Akt, and FoxO1. In vitro class switch differentiation of B cells prefers softer substrates. Lastly, NP67-Ficoll on softer substrates accounted for an enhanced antibody response in vivo. Thus, substrate stiffness regulates B-cell activation, proliferation, class switch, and T cell independent antibody responses in vivo, suggesting its broad application in manipulating the fate of B cells in vitro and in vivo. Eur. J. Immunol. 2015Immunol. . 45: 1621Immunol. -1634 Ig (mIg) and a heterodimer of Igα and Igβ in a 1 mIg:1 Igα-Igβ heterodimer stoichiometry [1,2]. Upon BCR engagement, the B cell initiates its activation that is followed by proliferation and differentiation into plasma cells and memory B cells [3]. It is appreciated that the antigens that encounter with B cells in vivo show a various degree of diversity based on their presentation forms, including antigen density [4,5], antigen affinity [4,5], antigen valency [6][7][8], and antigen mobility [9]. The early studies of ours and others showed that the initiation of B-cell activation is sensitive to antigen presentation forms, suggesting the sophisticated capability of B cells to sense the biophysical features of the antigens. For a long time, it has been neglected that the antigens encountered by B cells in vivo are actually presented on substrates with various stiffness features [10]. Stiffness usually represents the rigidity of an object, showing the extent that such an object resists deformation changes in response to an applied force. Young's modulus is usually used to describe the stiffness of an elastic substrate. For instance, the Young's modulus value of different virus particles is about 45-1000 MPa [11], while most of the mammalian cells show medium stiffness features from 0.01 to 1000 kPa [12]. Secreted soluble pathogen antigens in our plasma only exhibit very low stiffness of no more than 100 Pa [13]. Thus, substrates presenting the antigens on rigid virus capsid vesicles exhibit high stiffness features; those on the membrane of the virus-infected host cells show medium le...

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Microenvironmental modulation of the developing tumour: an immune‐stromal dialogue

2020

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Substrate Rigidity Regulates Human T Cell Activation and Proliferation

Cited by 254 publications

References 51 publications

B Cell Activation Is Regulated by the Stiffness Properties of the Substrate Presenting the Antigens

B Cell Activation Is Regulated by the Stiffness Properties of the Substrate Presenting the Antigens

Substrate stiffness regulates B‐cell activation, proliferation, class switch, and T‐cell‐independent antibody responses in vivo

Microenvironmental modulation of the developing tumour: an immune‐stromal dialogue

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