Ex vivo testing of immune responses in precision-cut lung slices

Henjaković, Maja; Sewald, Katherina; Switalla, Simone; Kaiser, Daniel R.; Müller, Meike; Veres, Tibor Z.; Martin, Christian; Uhlig, Stefan; Krug, Norbert; Braun, Armin

doi:10.1016/j.taap.2008.04.003

Cited by 97 publications

(87 citation statements)

References 41 publications

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“…This is in contrast to other publications that report a limited viability of PCLS between 1-6 d in culture. 7,20,21 We further demonstrated that the viability of human PCLS, as shown by the LDH assay, remained above 80% for the full 14-day cultivation period. This type of long-term cultivation could increase the experimental window for using PCLS, maximizing data sets generated in this model.…”

Section: Discussionmentioning

confidence: 60%

“…Previous studies have shown increased production of TNF-a, IL1b and IL-6 as well as COX-2 induction by PCLS after in vitro stimulation with LPS. 2,7 In addition to these proteins, we also detected increases in IL-8, IL-10 and MIP-1b production. Unlike Switalla et al, we did not detect IL-12p40 or IFN-g production in culture supernatants, potentially due to a different duration of LPS stimulation (18 vs. 24 hours).…”

Section: Discussionmentioning

confidence: 76%

“…7,22 However, it was also shown that exposure to LPS could directly cause tissue damage by triggering apoptotic cell death in bronchial epithelial 23 and endothelial 24 cells. In addition, LPS was reported to induce apoptosis in human alveolar macrophages in a dosedependent manner.…”

Section: Discussionmentioning

confidence: 99%

“…Previous work on this technology have shown that PCLS can easily be prepared in abundance just from a single human lung lobe and can be prepared from human as well as different laboratory animal species, including mice, rats, guinea pigs, sheep and monkeys. [1][2][3][4][5] Multiple studies have focused on assessing various characteristics of cultured PCLS, such as measurement of bronchoconstriction [6][7][8] and vascular responses 9,10 induced by various stimuli. In addition, PCLS have been used to evaluate allergy, 11 asthma, 6 toxicology, [12][13][14] and infectious disease responses.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of inflammatory and immune responses in long-term cultured human precision-cut lung slices

Temann

Golovina

Neuhaus

et al. 2016

Human Vaccines & Immunotherapeutics

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The development of systems that are more accurate and time-efficient in predicting safety and efficacy of target products in humans are critically important in reducing the cost and duration of pharmaceutical development. To circumvent some of the limitations imposed by the use of animal models, ex vivo systems, such as precision-cut lung slices (PCLS), have been proposed as an alternative for evaluating safety, immunogenicity and efficacy of vaccines and pharmaceuticals. In this study, we have established a human PCLS system and methodology for PCLS cultivation that can provide long-term viability and functionality in culture. Using these techniques, we found that cultured PCLS remained viable for at least 14 d in culture and maintained normal metabolic activity, tissue homeostasis and structural integrity. To investigate whether cultured PCLS remained functional, lipopolysaccharide (LPS) was used as a target stimulating compound. We observed that after an 18-hour incubation with LPS, cultured PCLS produced a set of pro-inflammatory cytokines, including TNF-a, IL-1b, IL-6 and IL-10 as well as the enzyme COX-2. Furthermore, cultured PCLS were shown to be capable of generating re-call immune responses, characterized by cytokine production, against antigens commonly found in routine vaccinations against influenza virus and tetanus toxoid. Taken together, these results suggest that human PCLS have the potential to be used as an alternative, high-throughput, ex vivo system for evaluating the safety, and potentially immunogenicity, of vaccines and pharmaceuticals.

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Section: Discussionmentioning

confidence: 60%

Section: Discussionmentioning

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Evaluation of inflammatory and immune responses in long-term cultured human precision-cut lung slices

Temann

Golovina

Neuhaus

et al. 2016

Human Vaccines & Immunotherapeutics

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“…PCLSs retain the organizational features of the in vivo lung, while providing microscopic access for individual cells (3)(4)(5)(6). Despite these advantages, the use of hPCLSs is confounded by the unpredictable and limited availability of human tissue.…”

mentioning

confidence: 99%

Cryopreserved Human Precision-Cut Lung Slices as a Bioassay for Live Tissue Banking. A Viability Study of Bronchodilation with Bitter-Taste Receptor Agonists

Bai

Krishnamoorthy

Patel

et al. 2016

Am J Respir Cell Mol Biol

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Human precision-cut lung slices (hPCLSs) provide a unique ex vivo model for translational research. However, the limited and unpredictable availability of human lung tissue greatly impedes their use. Here, we demonstrate that cryopreservation of hPCLSs facilitates banking of live human lung tissue for routine use. Our results show that cryopreservation had little effect on overall cell viability and vital functions of immune cells, including phagocytes and T lymphocytes. In addition, airway contraction and relaxation in response to specific agonists and antagonists, respectively, were unchanged after cryopreservation. At the subcellular level, cryopreserved hPCLSs maintained Ca 21 -dependent regulatory mechanisms for the control of airway smooth muscle cell contractility. To exemplify the use of cryopreserved hPCLSs in smooth muscle research, we provide evidence that bitter-taste receptor (TAS2R) agonists relax airways by blocking Ca 21 oscillations in airway smooth muscle cells. In conclusion, the banking of cryopreserved hPCLSs provides a robust bioassay for translational research of lung physiology and disease.

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