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Sellberg, Felix
Publications (8 of 8) Show all publications
Watanabe, M., Kumagai-Braesch, M., Yao, M., Thunberg, S., Berglund, D., Sellberg, F., . . . Ericzon, B.-G. (2018). Ex Vivo Generation of Donor Antigen-Specific Immunomodulatory Cells A Comparison Study of Anti-CD80/86 mAbs and CTLA4-lg Costimulatory Blockade. Cell Transplantation, 27(11), 1692-1704
Open this publication in new window or tab >>Ex Vivo Generation of Donor Antigen-Specific Immunomodulatory Cells A Comparison Study of Anti-CD80/86 mAbs and CTLA4-lg Costimulatory Blockade
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2018 (English)In: Cell Transplantation, ISSN 0963-6897, E-ISSN 1555-3892, Vol. 27, no 11, p. 1692-1704Article in journal (Refereed) Published
Abstract [en]

Adoptive transfer of alloantigen-specific immunomodulatory cells generated ex vivo with anti-CD80/CD86 mAbs (2D10.4/IT2.2) holds promise for operational tolerance after transplantation. However, good manufacturing practice is required to allow widespread clinical application. Belatacept, a clinically approved cytotoxic T-lymphocyte antigen 4-immunoglobulin that also binds CD80/CD86, could be an alternative agent for 2D10.4/IT2.2. With the goal of generating an optimal cell treatment with clinically approved reagents, we evaluated the donor-specific immunomodulatory effects of belatacept- and 2D10.4/IT2.2-generated immunomodulatory cells. Immunomodulatory cells were generated by coculturing responder human peripheral blood mononuclear cells (PBMCs) (50 x 10(6) cells) with irradiated donor PBMCs (20 x 10(6) cells) from eight human leukocyte antigen-mismatched responder-donor pairs in the presence of either 2D10.4/IT2.2 (3 mu g/10(6) cells) or belatacept (40 mu g/10(6) cells). After 14 days of coculture, the frequencies of CD4(+) T cells, CD8(+) T cells, and natural killer cells as well as interferon gamma (IFN-gamma) production in the 2D10.4/IT2.2- and belatacept-treated groups were lower than those in the control group. The percentage of CD19(+) B cells was higher in the 2D10.4/IT2.2- and belatacept-treated groups than in the control group. The frequency of CD4(+)CD25(+)CD127(low)FOXP3(+) T cells increased from 4.1 +/- 1.0% (preculture) to 7.1 +/- 2.6% and 7.3 +/- 2.6% (day 14) in the 2D10.4/IT2.2- and belatacept-treated groups, respectively (p<0.05). Concurrently, delta-2 FOXP3 mRNA expression increased significantly. Compared with cells derived from the no-antibody treated control group, cells generated from both the 2D10.4/IT2.2- and belatacept-treated groups produced lower IFN-gamma and higher interleukin-10 levels in response to donor-antigens, as detected by enzyme-linked immunospot. Most importantly, 2D10.4/IT2.2- and belatacept-generated cells effectively impeded the proliferative responses of freshly isolated responder PBMCs against donor-antigens. Our results indicate that belatacept-generated donor-specific immunomodulatory cells possess comparable phenotypes and immunomodulatory efficacies to those generated with 2D10.4/IT2.2. We suggest that belatacept could be used for ex vivo generation of clinical grade alloantigen-specific immunomodulatory cells for tolerance induction after transplantation.

Keywords
Immunomodulatory cells, anti-CD80/86 antibodies, CTLA4-Ig, donor antigen specificity, ex vivo generation, FOXP3
National Category
Immunology Immunology in the medical area
Identifiers
urn:nbn:se:uu:diva-371403 (URN)10.1177/0963689718794642 (DOI)000450357000010 ()30261751 (PubMedID)
Funder
The Karolinska Institutet's Research Foundation
Available from: 2018-12-20 Created: 2018-12-20 Last updated: 2019-01-07Bibliographically approved
Berglund, E., Sellberg, F. & Berglund, D. (2017). Assessing the purity of regulatory T cells: A humble reminder [Letter to the editor]. Cytotherapy, 19(2), 329-332
Open this publication in new window or tab >>Assessing the purity of regulatory T cells: A humble reminder
2017 (English)In: Cytotherapy, ISSN 1465-3249, E-ISSN 1477-2566, Vol. 19, no 2, p. 329-332Article in journal, Letter (Refereed) Published
Keywords
cell therapy, regulatory T cells, FOXP3, flow cytometry
National Category
Cell and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-317095 (URN)10.1016/j.jcyt.2016.10.012 (DOI)000392890500017 ()27884702 (PubMedID)
Available from: 2017-03-10 Created: 2017-03-10 Last updated: 2018-01-13Bibliographically approved
Strandberg, G., Sellberg, F., Sommar, P., Ronaghi, M., Lubenow, N., Knutson, F. & Berglund, D. (2017). Standardizing the freeze-thaw preparation of growth factors from platelet lysate. Transfusion, 57(4), 1058-1065
Open this publication in new window or tab >>Standardizing the freeze-thaw preparation of growth factors from platelet lysate
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2017 (English)In: Transfusion, ISSN 0041-1132, E-ISSN 1537-2995, Vol. 57, no 4, p. 1058-1065Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Over the past decades, the focus on the regenerative properties of platelets (PLTs) has intensified and many PLT-derived growth factors are readily used in medical settings. A general lack of standardization in the preparation of these growth factors remains, however, and this study therefore examines the dynamics of growth factors throughout the freeze-thaw procedure.

STUDY DESIGN AND METHODS: Plateletpheresis (PA) and PLT-poor plasma (PPP) samples were collected from 10 healthy donors. PA was lysed to produce PLT lysate (PL) for 1, 3, 5, 10, and 30 freeze-thaw cycles. The resulting growth factor and cytokine concentrations from PPP, PA, and PL of different cycles were analyzed and compared using enzyme-linked immunosorbent assay and multiplex bead assays.

RESULTS: PL produced by the freeze-thaw procedure resulted in approximately four-to 10-fold enrichment of transforming growth factor-b1, epidermal growth factor, PLT-derived growth factor (PDGF)-AB/BB, PLT factor-4, and fibroblast growth factor-2. The increase in concentrations plateaued at Cycles 3 and 5 and in some cases declined with further cycles. The concentrations of insulin-like growth factor-1, hepatocyte growth factor, vascular endothelial growth factor, and bone morphogenetic protein-2 in PL were essentially comparable to those in PPP.

CONCLUSION: Using the freeze-thaw method, optimal preparation of PL with regard to the concentration of growth factors was achieved at Cycles 3 to 5. Based on our findings, the clinical significance of using a greater number of cycles is likely limited.

Place, publisher, year, edition, pages
WILEY, 2017
National Category
Hematology
Identifiers
urn:nbn:se:uu:diva-328740 (URN)10.1111/trf.13998 (DOI)000402864500029 ()28182293 (PubMedID)
Available from: 2017-08-30 Created: 2017-08-30 Last updated: 2017-08-30Bibliographically approved
Fredriksson, F., Sellberg, F., Bowden, T., Engstrand, T., Berglund, D. & Engstrand Lilja, H. (2017). Sutures impregnated with carbazate-activated polyvinyl alcohol reduce intraperitoneal adhesions. Journal of Pediatric Surgery, 52(11), 1853-1858
Open this publication in new window or tab >>Sutures impregnated with carbazate-activated polyvinyl alcohol reduce intraperitoneal adhesions
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2017 (English)In: Journal of Pediatric Surgery, ISSN 0022-3468, E-ISSN 1531-5037, Vol. 52, no 11, p. 1853-1858Article in journal (Refereed) Published
Abstract [en]

Background: Intraperitoneal adhesions cause significant morbidity. They occur after peritoneal trauma, which induces oxidative stress with production of inflammatory cytokines, peroxidized proteins (carbonyls) and lipids (aldehydes). This study aimed to investigate if carbazate-activated polyvinyl alcohol (PVAC), an aldehyde-carbonyl inhibitor, can reduce intraperitoneal adhesions in an experimental model.

Material and methods: Male Sprague-Dawley rats (n = 110) underwent laparotomy, cecal abrasion and construction of a small bowel anastomosis. They either were treated with intraperitoneal instillation of PVAC or were sutured with PVAC-impregnated sutures. Thromboelastography analysis was performed using human blood and PVAC. The lipid peroxidation product malondialdehyde (MDA) and inflammatory cytokines IL-1 beta and IL-6 were quantified in peritoneal fluid. At day 7, bursting pressure of the anastomosis was measured and adhesions were blindly scored.

Results: PVAC in human blood decreased the production of the fibrin-thrombocyte mesh without affecting the coagulation cascade. MDA, IL-1 beta and IL-6 were increased after 6 h without significant difference between the groups. PVAC-impregnated sutures reduced intraperitoneal adhesions compared to controls (p = 0.0406) while intraperitoneal instillation of PVAC had no effect. Anastomotic bursting pressure was unchanged.

Conclusions: Intervention with an aldehyde-carbonyl inhibitor locally in the wound by PVAC-impregnated sutures might be a new strategy to reduce intraperitoneal adhesions.

Keywords
Intraperitoneal adhesion prevention, Peroxidation products, Experimental adhesion model
National Category
Surgery
Identifiers
urn:nbn:se:uu:diva-342398 (URN)10.1016/j.jpedsurg.2017.01.058 (DOI)000415328600030 ()28196659 (PubMedID)
Available from: 2018-02-21 Created: 2018-02-21 Last updated: 2018-02-21Bibliographically approved
Sellberg, F., Berglund, E., Ronaghi, M., Strandberg, G., Löf, H., Sommar, P., . . . Berglund, D. (2016). Composition of growth factors and cytokines in lysates obtained from fresh versus stored pathogen-inactivated platelet units. Transfusion and apheresis science, 55(3), 333-337
Open this publication in new window or tab >>Composition of growth factors and cytokines in lysates obtained from fresh versus stored pathogen-inactivated platelet units
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2016 (English)In: Transfusion and apheresis science, ISSN 1473-0502, E-ISSN 1878-1683, Vol. 55, no 3, p. 333-337Article in journal (Refereed) Published
Abstract [en]

Background: Platelet lysate is a readily available source of growth factors, and other mediators, which has been used in a variety of clinical applications. However, the product remains poorly standardized and the present investigation evaluates the composition of platelet lysate obtained from either fresh or stored pathogen-inactivated platelet units.

Materials and Methods: Platelet pooled units (n = 10) were obtained from healthy blood donors and tested according to standard procedures. All units were pathogen inactivated using amotosalen hydrochloride and UVA exposure. Platelet lysate was subsequently produced at two separate time-points, either from fresh platelet units or after 5 days of storage, by repeated freeze-thaw cycles. The following mediators were determined at each time point: EGF, FGF-2, VEGF, IGF-1, PDGF-AB/BB, BMP-2, PF4, TGF-beta isoform 1, IL-1(i, IL-2, IL-6, IL-10, IL-12p70, 1L-17A, TNF-alpha, and IFN-gamma.

Results: The concentration of growth factors and cytokines was affected by time in storage. Notably, TGF-beta, PDGF-AB/BB, and PF4 showed an increase of 27.2% (p < 0.0001), 29.5% (p = 0.04) and 8.2% (p = 0.0004), respectively. A decrease was seen in the levels of IGF-1 and FGF-2 with 22% (p = 0.041) and 11% (p = 0.01), respectively. Cytokines were present only in very low concentrations and all other growth factors remained stable with time in storage.

Conclusion: The composition of mediators in platelet lysate obtained from pathogen inactivated platelet units differs when produced from fresh and stored platelet units, respectively. This underscores the need for further standardization and optimization of this important product, which potentially may influence the clinical effects.

Keywords
Platelet lysate, Pathogen-inactivation, Growth factor, Cytokine
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-314425 (URN)10.1016/j.transci.2016.08.004 (DOI)000390964500014 ()
Available from: 2017-02-02 Created: 2017-02-02 Last updated: 2017-11-29Bibliographically approved
Watanabe, M., Kumagai-Braesch, M., Thunberg, S., Henrikson, J., Sellberg, F., Lundgren, T., . . . Ericzon, B.-G. (2016). Ex vivo generation of alloantigen-specific T regulatory cells using selective T-cell costimulation blockade. Paper presented at International Congress of Immunology (ICI), AUG 21-26, 2016, Melbourne, AUSTRALIA. European Journal of Immunology, 46, 125-125
Open this publication in new window or tab >>Ex vivo generation of alloantigen-specific T regulatory cells using selective T-cell costimulation blockade
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2016 (English)In: European Journal of Immunology, ISSN 0014-2980, E-ISSN 1521-4141, Vol. 46, p. 125-125Article in journal, Meeting abstract (Other academic) Published
National Category
Immunology in the medical area
Identifiers
urn:nbn:se:uu:diva-308494 (URN)000383610400250 ()
Conference
International Congress of Immunology (ICI), AUG 21-26, 2016, Melbourne, AUSTRALIA
Available from: 2016-11-28 Created: 2016-11-28 Last updated: 2018-01-13Bibliographically approved
Sellberg, F., Ronaghi, M., Knutson, F. & Berglund, D. (2016). Inflammatory cytokines in serum samples of healthy blood donors [Letter to the editor]. Transfusion and apheresis science, 55(2), 246-247
Open this publication in new window or tab >>Inflammatory cytokines in serum samples of healthy blood donors
2016 (English)In: Transfusion and apheresis science, ISSN 1473-0502, E-ISSN 1878-1683, Vol. 55, no 2, p. 246-247Article in journal, Letter (Refereed) Published
National Category
Hematology
Identifiers
urn:nbn:se:uu:diva-313620 (URN)10.1016/j.transci.2016.07.020 (DOI)000386744900019 ()27475802 (PubMedID)
Available from: 2017-01-23 Created: 2017-01-23 Last updated: 2017-11-29Bibliographically approved
Watanabe, M., Kumagai-Braesch, M., Berglund, D., Thunberg, S., Jorns, C., Henriksson, J., . . . Ericzon, B.-G. (2015). Ex-Vivo Generation Of Alloantigen-Specific Regulatory T Cells Using Selective T-Cell Costimulation Blockade: A Comparison Study Between Anti-Cd80/86 Mabs And CTLA4-Ig. Transplant International, 28, 219-219
Open this publication in new window or tab >>Ex-Vivo Generation Of Alloantigen-Specific Regulatory T Cells Using Selective T-Cell Costimulation Blockade: A Comparison Study Between Anti-Cd80/86 Mabs And CTLA4-Ig
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2015 (English)In: Transplant International, ISSN 0934-0874, E-ISSN 1432-2277, Vol. 28, p. 219-219Article in journal, Meeting abstract (Other academic) Published
National Category
Surgery
Identifiers
urn:nbn:se:uu:diva-281010 (URN)000367726701255 ()
Available from: 2016-03-16 Created: 2016-03-16 Last updated: 2017-11-30Bibliographically approved
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