Department of Transplantation Immunology

Department of Transplantation Immunology

Department of Transplantation Immunology

 

 

Head: Prof. Vladimír Holáň, DrSc.

E-mail: holan@biomed.cas.cz
Tel.: +420 241 063 226

 

Research of this Department is focused on the isolation, characterization and cultivation of stem cells and their use for treatment of severe injuries or so far uncurable diseases. Stem cells are propagated in tissue cultures and using various nanofiber scaffolds transferred onto mechanically or chemically damaged ocular surface. The ability of transferred cells to inhibit a harmful inflammatory immune reaction ocurring in the site of injury and to support healing process is evaluated. The ultimate goal of the research is to get insights into the mechanisms of specific immune response after transplantation of stem cells with the aim to increase their antiinflammatory and therapeutic potential. The experience with the study of transplantation immunity and the combination of nanotechnologies with stem cell research enable to propose and test novel therapeutic approaches. The recent study is extended on the development of stem cell-based therapy for currently uncurable serious sight-threatening retinal diseases.

 

 

Research Scientists:

Prof. Vladimír Holáň, DrSc.

Assoc. Prof. Jitka Čejková, DrSc.

Alena Zajícová, PhD.

Eliška Javorková, PhD.

Čestmír Čejka, PhD.

Magdaléna Krulová, PhD.

 

PhD Students:

Michaela Hájková, MSc

Barbora Heřmánková, MSc

Pavla Boháčová, MSc

Jan Kössl, MSc

 

Undergraduate Students:

Kateřina Palacká

Sandra Žáčková

 

Technicans:

Lucie Holáňová

Jaroslava Knížová

Jana Herlová

Important results in 2015

 

1. Comparative study on the therapeutic potential of mesenchymal stem cells and tissue-specific limbal stem cells
The ability of mesenchymal stem cels (MSC) and limbal stem cells (LSC) to produce various immunoregulatory molecules and to modulate immune response was compared. The therapeutic potential of these stem cells was evaluated in the model of treatment of chemically damaged ocular surface in the rabbit. It was found that stem cells produce a number of immunoregulatory molecules and that MSC have comparable therapeutic properties as do tissue-specific LSC. These results bring the evidence that MSC can replace tissue-specific LSC in the cases when LSC are absent or difficult to obtain.

 

 


 

Characterization of MSC. The ability of MSC to express genes for immunomodulatory molecules was determined by real-time PCR. The cells were cultured untreated or were stimulated with LPS, IFN-γ with LPS plus IFN-γ

 

 
Therapeutic potential of MSC and LSC. Stem cells were transferred using nanofiber scaffold onto damaged ocular surface and their therapeutic potential was evaluated. The figure shows healthy rabbit eye, the eye shortly after chemical damage, damaged eye covered with nanofiber scaffold, untreated damaged eye two weeks after the injury and damaged eye treated with MSC or LSCC nebo LSC.

 

Publications:

Holáň V., Trošan P., Čejka Č., Javorková E., Zajícová A., Heřmánková B., Chudíčková M, Čejková J.: A comparative study of the therapeutic potential of mesenchymal stem cells and limbal epithelial stem cells for ocular surface reconstruction. Stem Cells Translat. Med. 4, 1052-1063, 2015. IF 5.709.

 

Heřmanková B., Zajícová A., Javorková E., Chudíčková M., Trošan P., Hájková M., Krulová M., Holáň V.: Suppression of IL-10 production by activated B cells via a cell contact-dependent cyclooxygenase-2 pathway upregulated in IFN-γ-treated mesenchymal stem cells. Immunobiology 221, 129-136, 2016. IF 3.044. 

 

 

Important results in 2014

 

1. The effects of cytokines on migration and therapeutic potential of mesenchymal stem cells in a model of local inflammatory reaction in damaged eye 

Using a well established model of chemically damaged ocular surface in mice we showed that systemically administered mesenchymal stem cells (MSCs) selectively migrate into the site of injury and inhibit infiltration with cells of of the immune system. The ability of MSCs to inhibit local inflammatory reaction can be modulated by their preincubation with cytokines which determine the development of T and B lymphocytes and regulate the secretory capacity of MSCs. The results thus show the possibilities to use cytokine-pretreated and systemically administered MSCs for the treatment of local inflammatory reaction.


 

 

Phenotype characterization of MSCs (A), labeling of MSCs with vital dye PKH26 (B, C) and detection of labeled MSCs in the injured eye (D).

 

 

Collaboration: Faculty of Science Charles University Prague

 

Publication:

Javorková E, Trošan P, Zajícová A, Krulová M, Hajková M, Holáň V, (2014): Modulation of the early infl ammatory microenvironment in alkaliburned eye by systemically administered interferon treated mesenchymal stem cells. Stem Cells Dev. 23, 2490-2500. IF 4.202

Holáň V, Zajícová A, Javorková E, Trošan P, Chudičková M, Pavlíková M, Krulová M, (2014): Distinct cytokines balance the development of regulatory T cells and IL-10-producing regulatory B cells. Immunology 141, 577-586. IF 3.3735 

 

Important results in 2013

1. Regeneration of severely damaged ocular surface by stem cells

Limbal and mesenchymal stem cells were cultured in vitro on nanofiber scaffolds and transferred on the damaged ocular surface in experimental animal models. The therapeutic effects of stem cells were evaluated by histology and according to the ability to inhibit expresssion of genes for proinflammatory molecules IL-2, IFN-gama, IL-17 and iNOS. The results thus demonstrate the ability of stem cells and nanofiber scaffolds to regenerate ocular surface after the injury.

 

Collaboration: Evropská oční klinika Lexum, Praha.

 

 

 

A scheme of immunoregulatory and therapeutic effects of mesenchymal stem cells.
There are number of diferent mechanisms by which MSC contribute to the healing process. Differentiation into epithelial-like cells, immunomodulation and the production of growth and trophic factors are the most important mechanisms.

 

A structure of nanofiber scaffold (A) and the growth of mesenchymal stem cells on nanofiber scaffold (B, C).
(A) Scanning electron microscopy image of nanofibers from copolymer polyamid 6/12, the avarage nanofiber diameter is 380 nm, (B) Mouse MSC growing on nanofibers (red filaments – F-actin stained with phalloidin, blue – DAPI staining of the nuclei), (C) Scanning electron microscopy of mouse MSC growing on nanofibers.

 

 

 

 
Chemically damaged eye of the rabbit covered by a nanofiber scaffold with stem cells.
The application of a nanofiber scaffold seeded with MSC onto the damaged ocular surface of a rabbit.

 

 

Publications:

  • Holáň, V., Javorková, E.: (2013) Mesenchymal stem cells, nanofiber scaffolds and ocular surface reconstruction. Stem Cells Rev. Rep. 9(5), 609-619. IF 4.523
  • Čejková J., Tošan P., Čejka Č., Lenčová A., Zajícová A., Javorková E., Kubinová Š., Syková E., Holáň V.: (2013) Suppression of alkali-induced oxidative injury to the cornea by mesenchymal stem cells growing on nanofiber scaffolds and transferred onto the damaged corneal surface. Exp. Eye Res. 116, 312-323. IF 3.026
  • Holáň, V., Javorková, E., Trošan, P.: The growth and delivery of mesenchymal and limbal stem cells using copolymer polyamide 6/12 nanofiber scaffolds. In: Wright, B. and Connon, C. J. (eds), Corneal Regenerative Medicine, Methods Mol. Biol., Humana Press – Springer, New York, London 2013, S. 187-199. ISBN 978-1-62703-431-9.

 

 

2. Distinct roles of cytokines in the development of regulatory T cells and regulatory B cells


The role of cytokines in development of regulatory T (Treg) and B (Breg) cells was described. TGF-beta was shown as the main cytokine determining the development of Tregs, but this cytokine inhbited activation of IL-10-producing Breg. On the contrary, another two cytokines, IL-12 and IFN-gama, enhanced the development of Breg. These results show distinct roles of cytokines in development of Treg and Breg, and suggest new approaches for targeted regulation of the immune system.

 

Collaboration: Faculty of Science Charles University Prague

 

 

 

 (A) Characterization of B cell populations by flow cytometry. (B) Detection of cells producing IL-10 using technology ELISPOT.

 

 

 

Publications:

  • Holáň, V., Zajícová, A., Javorková, E., Trošan, P., Chudičková, M., Pavlíková, M., Krulová, M.: (2013) Distinct cytokines balance the development of regulatory T cells and IL-10-producing regulatory B cells. Immunology. In press. IF 3.705
  • Kubera M., Curzytek K., Duda W., Leskiewicz M., Basta-Kaim A., Budziszewska B., Roman A., Zajícová A., Holáň V., Szczesny E., Lason W., Maes M.:. (2013) A new animal model of (chronic) depression induced by repeated and intermittent lipopolysaccharide administration for 4 months. Brain Behav. Immun. 31, 96-104. IF 5.612
  • Holáň, V., Krulová, M.: (2013) Common and small molecules as the ultimate regulatory and effector mediators of antigen-specific transplantation reactions. World J. Transplant. 3(4), 54-61. (new jurnal recently without IF)

GA ČR, 14-125805 – Treatment of severe ocular surface injuries using limbal and mesenchymal stem cells, 2014-2016.
 

GA UK, No. 80815 - The therapeutic potential of mesenchymal stem cells in diabetic retinopathy, 2015-2017.
 

TAČR GAMA - Commercionalization of VaV results in IEM AS CR, 2015-2017. 
 

MŠMT, NPU I: LO1309 - Center of cell therapy and tissue regeneration, 2014-2019.
 

MŠMT, NPU-I: LO1508 - Center of genomic and proteomic, 2015-2020.

2016

 

Ardan, T., Němcová, L., Bohuslavová, B., Klezlová, A., Popelka, Š., Studenovská, H.3, Hrnčiarová, E., Čejková, J., Motlík, J.: (2016) Reduced Levels of Tissue Inhibitors of Metalloproteinases (TIMPs) in UVB Irradiated Corneal Epithelium. Photochem. Photobiol., IN PRESS IF 2.008

Bílková, B., Albrecht, T., Chudičková, M., Holáň, V., Piálek, J., Vinkler, M.: (2016) Application of Concanavalin A during immune responsiveness skin-swelling tests facilitates measurement interpretation in mammalian ecology. Ecology and Evolution, 6 (13): 4551-4564. IF 2.537

Čejka, Č., Čejková, J., Trošan, P., Zajícová, A., Syková, E., Holáň, V.: (2016) Transfer of mesenchymal stem cells and cyclosporine A on alkali-injured rabbit cornea using nanofiber scaffolds strongly reduces corneal neovascularization and scar formation. Histol. Histopath., 31: 969-980. IF 1.875

Čejka, Č., Holáň, V., Trošan, P., Zajicová, A., Javorková E., Čejková, J.: (2016) The Favorable Effect of Mesenchymal Stem Cell Treatment on the Antioxidant Protective Mechanism in the Corneal Epithelium and Renewal of Corneal Optical Properties Changed after Alkali Burns. Oxidative Med. Cell. Longev., 5843809. IF 4.492

Čejková, J., Čejka, Č., Trošan, P., Zajícová, A., Syková, E., Holáň, V.: (2016) Treatment of alkali-injured cornea by cyclosporine A-loaded electrospun nanofibers - An alternative mode of therapy. Exp. Eye Res., 147: 128-137. IF 2.998

Hájková, M., Javorková, E., Zajicová, A., Trošan, P., Holáň, V., Krulová, M.: (2015) A local application of mesenchymal stem cells and cyclosporine A attenuates immune response by a switch in macrophage phenotype. J. Tissue Eng. Regen. Med., IN PRESS IF 4.710

Heřmánková, B., Holáň, V.: (2016) Perspectives of the Cell Therapy in Ophthalmology.2. The Potential of Stem Cells for the Retinal Diseases Treatment. Cesk. Slov. Oftalmol., 72(1): 272-275.

Heřmánková, B., Zajícová, A., Javorková, E., Chudíčková, M., Trošan, P., Hájková, M., Krulová, M., Holáň, V.: (2016) Suppression of IL-10 production by activated B cells via a cell contact-dependent cyclooxygenase-2 pathway upregulated in IFN-γ-treated mesenchymal stem cells. Immunobiology, 221(2): 129-136. IF 2.795

Holáň, V., Heřmánková, B., Boháčová, P., Kössl, J., Chudičková, M., Hajková, M., Krulová, M., Zajícová, A., Javorková, E.: (2016) Distinct Immunoregulatory Mechanisms in Mesenchymal Stem Cells: Role of the Cytokine Environment. Stem Cell Reviews and Reports, IN PRESS IF 3.111

Chudičková, M., Brůža, P., Zajícová, A., Trošan, P., Svobodová, L., Javorková, E., Kubinová, Š., Holáň, V.: (2015) Targeted neural differentiation of murine mesenchymal stem cells by a protocol simulating the inflammatory site of neural injury. J. Tissue Eng. Regen. Med., IN PRESS IF 4.710

Javorková, E., Holáň, V.: (2016) Perspectives of the Cell Therapy in Ophthalmology1. The Application of Stem Cells in the Regeneration of Damaged Surface of the Eye. Cesk. Slov. Oftalmol., 72(1): 268-271.

Trošan, P., Javorková, E., Zajícová, A., Hájková, M., Heřmánková, B., Kössl, J., Krulová, M., Holáň, V.: (2016) The Supportive Role of Insulin-Like Growth Factor-I in the Differentiation of Murine Mesenchymal Stem Cells into Corneal-Like Cells. Stem Cells Dev., IN PRESS IF 3.777

 

 

 

2015

 

Holan V., Trosan P., Cejka C., Javorkova E., Zajicova A., Hermankova B., Chudickova M, Cejkova J.: A comparative study of the therapeutic potential of mesenchymal stem cells and limbal epithelial stem cells for ocular surface reconstruction. Stem Cells Translat. Med. 4, 1052-1063, 2015.

Hajkova M., Javorkova E., Zajicova A., Trosan P., Holan V., Krulova M.: A local application of MSCs and Cyclosporine A attenuates immune response by a switch in macrophage phenotype. J. Tissue Eng. Regen. Med., 2015 Jun 29. doi: 10.1002/term.2044. [Epub ahead of print]. 2015.

Chudickova M., Bruza P., Zajicova A.., Trosan P., Svobodova L., Javorkova E.,Kubinova K., Holan V.: Targeted neural differentiation of murine mesenchymal stem cells by a protocol simulating the inflammatory site of neural injury. J. Tissue Eng. Regen. Med., 2015 Jun 29. doi: 10.1002/term.2044. [Epub ahead of print]. 2015.

Cejka C., Cejkova J.: Oxidative stress to the cornea, changes in corneal optical properties, and advances in treatment of corneal oxidative injuries. Oxid. Med. Cell. Longev. 2015;2015:591530. doi: 10.1155/2015/591530. Epub 2015.

Cejkova J., Cejka C.: The role of oxidative stress in corneal diseases and injuries. Histol. Histopathol. 30, 893-900, 2015.

Holan V., Hermankova B., Zajicova A.: Nanofiber scaffolds for the growth and transfer of stem cells for the treatment of ocular surface damage. In: From Functionalized Nanostructures Towards Engineered Macrostructions, Eds C. Caruana, J. Hornik, Zittau, J. Hradec, pp. 74-81, 2015.

Holan V., Bohacova P., Kossl J., Javorkova E.: Immunosuppressive drug-loaded nanofibers for the local inhibition of harmful inflammatory reactions. In: From Functionalized Nanostructures Towards Engineered Macrostructions, Eds C. Caruana, J. Hornik, Zittau, J. Hradec, pp. 17-23, 2015.

Kubinova S., Zajicova A., Holan V., Sykova A.: Nanofibers as scaffolds for cell transfer in   tissue regeneration In: From Functionalized Nanostructures Towards Engineered Macrostructions, Eds C. Caruana, J. Hornik, Zittau, J. Hradec, pp. 82-98, 2015.

Striz I., Holan V.: Cytokines in Clinical Medicine, Maxdorf Jessenius Press, Prague, 2015.

 

2014

 

Holan V., Zajicova A., Javorkova E., Trosan P., Chudickova M., Pavlikova M., Krulova M.: Distinct cytokines balance the development of regulatory T cells and IL-10-producing regulatory B cells. Immunology 141, 577-586, 2014

Javorkova E., Trosan P., Zajicova A., Krulova M., Hajkova M., Holan V.: Modulation of the early inflammatory microenvironment in alkali-burned eye by systemically administered interferon-γ treated mesenchymal stem cells. Stem Cells Dev. 23, 2490-2500, 2014.

Cejkova J., Olmiere C., Cejka C., Trosan P., Holan V.: The healing of alkali-injured cornea is stimulated by a novel matrix regenerating agent (RGTA, CACICOL20) – a biopolymer mimicking heparan sulfates reducing proteolytic, oxidative and nitrosative damage. Histol. Histopathiol. 29, 457-478, 2014.

Zajicova A., Javorkova E., Trosan P., Chudickova M., Hajkova M., Krulova M., Holan V.: A low-molecular weight leukocyte dialysable extract selectively enhances the development of RORγt+CD4+ T cells and IL-17 production. Folia Biol. (Praha) 60, 253-260, 2014.

Koci Z., Turnovcova K., Dubsky M., Baranovicova L., Holan V., Chudickova M., Sykova E., Kubinova S.: Characterization of human adipose tissue-derived stromal cells isolated from diabetic patients´s distal limbs with critical limb ischemia. Cell. Biochem. Funct. 32, 597-604, 2014.

Zablotskii V., Lunov O., Novotná B., Churpita O., Trosan P., Holan V., Syková E., Dejneka A., Kubinová S.: Down-regulation of adipogenesis of mesenchymal stem cells by oscillating high-gradient magnetic fields and mechanical vibration. Applied Physics Letters 105, 103702, 2014.

Holáň V.: Kmenové buňky a imunita. Alergie 16, 233-239, 2014.

 

2013

 

Holáň V., Javorková E.: Mesenchymal stem cells, nanofiber scaffolds and ocular surface reconstruction. Stem Cells Rev. Rep. 9, 609-619, 2013.

Holáň V., Krulová M.: Common and small molecules as the ultimate regulatory and effector mediators of antigen-specific transplantation reactions. World J. Transplant.3, 54-61, 2013.

Čejková J., Tošan P., Čejka Č., Lenčová A., Zajícová A., Javorková E., Kubinová Š., Syková E., Holáň V.: Suppression of alkali-induced oxidative injury to the cornea by mesenchymal stem cells growing on nanofiber scaffolds and transferred onto the damaged corneal surface. Exp. Eye Res. 116, 312-323, 2013.

Holáň V., Javorková E., Trošan P.: The growth and delivery of mesenchymal and limbal stem cells using copolymer polyamide 6/12 nanofiber scaffolds. In: Corneal Regenerative Medicine, Methods Mol. Biol., Eds. B. Wright and C. J. Connon, Humana Press - Springer, 1014, 187-199, 2013.

Kubera M., Curzytek K., Duda W., Leskiewicz M., Basta-Kaim A., Budziszewska B., Roman A., Zajícová A., Holáň V., Szczesny E., Lason W., Maes M.:. A new animal model of (chronic) depression induced by repeated and intermittent lipopolysaccharide administration for 4 months. Brain Behav. Immun. 31, 96-104, 2013.

Vitova A., Kuffova L., Klaska I. P., Holáň V, Cornall R. J, Forrester J. V.: The high-risk corneal re-graft model: a justification for tissue matching in humans. Transplant Inter. Transplant Inter., 26, 453-461 2013.

Čejková J., Olmiere C., Čejka Č., Trošan P., Holáň V.: The healing of alkali-injured cornea is stimulated by a novel matrix regenerating agent (RGTA, CACICOL20) – a biopolymer mimicking heparan sulfates reducing proteolytic, oxidative and nitrosative damage. Histol. Histopathol., in press, 2013.

Holáň V., Zajícová A., Javorková E., Trošan P., Chudičková M., Pavlíková M., Krulová M.: Distinct cytokines balance the development of regulatory T cells and IL-10-producing regulatory B cells. Immunology, in press, 2013.

Department od Cell Biology, Faculty of Natural Science, Charles University, Prague

Department of Polymer Gels, Institute of Macromolecular Chemistry, Czech Academy of Sciences, Prague