Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed

Wright, Kandis; Dziuk, Rachel; Mital, Payal; Kaur, Gurvinder; Dufour, Jannette M.

doi:10.3727/096368916x692032

Cited by 18 publications

(28 citation statements)

References 34 publications

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“…NPSC and NPI were isolated as described previously 35 . Prior to transplantation the number of cells was determined by measuring the total cellular DNA content using the Quant-iT PicoGreen dsDNA quantification assay (Invitrogen, Carlsbad, CA, USA) 35 . Aliquots consisting of 11 × 10 6 cells were gently placed under the kidney capsule of isofluorane-anesthetized Lewis rats 35 .…”

Section: Methodsmentioning

confidence: 99%

Neonatal Pig Sertoli Cells Survive Xenotransplantation by Creating an Immune Modulatory Environment Involving CD4 and CD8 Regulatory T Cells

et al. 2020

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The acute cell-mediated immune response presents a significant barrier to xenotransplantation. Immune-privileged Sertoli cells (SC) can prolong the survival of co-transplanted cells including xenogeneic islets, hepatocytes, and neurons by protecting them from immune rejection. Additionally, SC survive as allo- and xenografts without the use of any immunosuppressive drugs suggesting elucidating the survival mechanism(s) of SC could be used to improve survival of xenografts. In this study, the survival and immune response generated toward neonatal pig SC (NPSC) or neonatal pig islets (NPI), nonimmune-privileged controls, was compared after xenotransplantation into naïve Lewis rats without immune suppression. The NPSC survived throughout the study, while NPI were rejected within 9 days. Analysis of the grafts revealed that macrophages and T cells were the main immune cells infiltrating the NPSC and NPI grafts. Further characterization of the T cells within the grafts indicated that the NPSC grafts contained significantly more cluster of differentiation 4 (CD4) and cluster of differentiation 8 (CD8) regulatory T cells (Tregs) at early time points than the NPI grafts. Additionally, the presence of increased amounts of interleukin 10 (IL-10) and transforming growth factor (TGF) β and decreased levels of tumor necrosis factor (TNF) α and apoptosis in the NPSC grafts compared to NPI grafts suggests the presence of regulatory immune cells in the NPSC grafts. The NPSC expressed several immunoregulatory factors such as TGFβ, thrombospondin-1 (THBS1), indoleamine-pyrrole 2,3-dioxygenase, and galectin-1, which could promote the recruitment of these regulatory immune cells to the NPSC grafts. In contrast, NPI grafts had fewer Tregs and increased apoptosis and inflammation (increased TNFα, decreased IL-10 and TGFβ) suggestive of cytotoxic immune cells that contribute to their early rejection. Collectively, our data suggest that a regulatory graft environment with regulatory immune cells including CD4 and CD8 Tregs in NPSC grafts could be attributed to the prolonged survival of the NPSC xenografts.

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Section: Methodsmentioning

confidence: 99%

Neonatal Pig Sertoli Cells Survive Xenotransplantation by Creating an Immune Modulatory Environment Involving CD4 and CD8 Regulatory T Cells

et al. 2020

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“…Although xenotransplantation has vast clinical potential, it is limited by the problem of immune responses against xenogeneic tissue (Wright et al 2016 ). Additionally, xenotransplanted cells, including vascularized organ xenografts, show loss of function within a short time of transplantation in dissonant species combinations.…”

Section: Introductionmentioning

confidence: 99%

GM1 Induced the inflammatory response related to the Raf-1/MEK1/2/ERK1/2 pathway in co-culture of pig mesenchymal stem cells with RAW264.7

Kwak

Seo

Lee

et al. 2018

Animal Cells and Systems

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Pig-human xenotransplantation can trigger cell-mediated immune responses. We explored the role of gangliosides in inflammation related to immune rejection in xenotransplantation. Co-culture of xenogeneic cells (pig-MSCs and RAW264.7) was used to emulate xenotransplantation conditions. MTT assay results indicated that cell viability was significantly decreased in pADMSCs co-cultured with RAW264.7 cells. GM1 and GM3 were highly expressed in pADMSCs co-cultured with RAW264.7 cells. pADMSCs co-cultured with RAW264.7 cells strongly expressed pro-inflammatory proteins such as COX-2, iNOS, p50, p65, pIκBα, and TNF-α. GM1-knockdown pADMSCs co-cultured with RAW 264.7 cells did not show significantly altered cell viability, but pro-inflammatory proteins were markedly inhibited. Co-culture of pADMSCs with RAW264.7 cells induced significant phosphorylation (p) of JNK1/2 and pERK1/2. However, pERK1/2 and pJNK1/2 were decreased and MEK1/2 and Raf1 were suppressed in GM1-knockdown pADMSCs co-cultured with RAW264.7 cells. Thus, the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways were significantly upregulated in response to increases of GM1 in co-cultured xenogeneic cells. However, the inflammatory response was suppressed in co-culture of GM1-knockdown pADMSCs with RAW264.7 cells via down-regulation of the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways. Therefore, the ganglioside GM1 appears to play a major role in the inflammatory response in xenotransplantation via the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways.

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“…The use of immune-privileged SCs as a vehicle for cell-based gene therapy could circumvent these issues. For instance, it has been demonstrated that SCs survive without the use of immunosuppressive drugs when transplanted as allo- and xenografts (Korbutt et al , 1997 ; Dufour et al , 2003 , 2004; Gores et al , 2003 ; Wright et al , 2016 ). Therefore, SC-based gene therapy is not likely to cause a cytokine storm.…”

Section: Discussionmentioning

confidence: 99%

“…Before transplantation, MSC-LV-mI or nontransduced MSC-1 cells were aggregated on nontissue culture-treated Petri dishes containing Ham's F10 medium (Thermo Fisher Scientific) with supplements and 10% FBS for 48 h at 37°C (Kaur et al , 2014b ; Wright et al , 2016 ). Six or 20 million MSC-LV-mI and 20 million nontransduced MSC-1 cells, calculated using a double-stranded DNA quantitation assay (Quant-iT PicoGreen ® ds DNA Assay Kit; Thermo Fisher Scientific), were transplanted into the left renal subcapsular space of isoflurane-anesthetized diabetic BALB/c or SCID-Beige mice as described previously (Halley et al , 2010 ; Kaur et al , 2014b ; Wright et al , 2016 ). Graft-bearing kidneys were collected over a span of 20–85 days posttransplantation and used for RT-qPCR, immunohistochemistry, or cell culture.…”

Section: Methodsmentioning

confidence: 99%

“…SCs create this environment by forming the blood–testis barrier and secreting several immunomodulatory factors (Mital et al , 2010 , 2011 ; Kaur et al , 2014a ). These immunoregulatory factors also allow SCs to survive when transplanted across immunological barriers as allo- and xenografts without the use of chronic immunosuppressive drugs (Korbutt et al , 1997 ; Dufour et al , 2003 , 2004 ; Gores et al , 2003 ; Wright et al , 2016 ). These unique properties of SCs led to the concept that SCs would make excellent targets as vehicles for the delivery of therapeutic proteins (i.e., cell-based gene therapy).…”

Section: Introductionmentioning

confidence: 99%

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Sertoli Cells Engineered to Express Insulin to Lower Blood Glucose in Diabetic Mice

Kaur

Thompson

Babcock

et al. 2018

DNA and Cell Biology

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Long-term survival of allo- and xenotransplanted immune-privileged Sertoli cells (SCs) is well documented suggesting that SCs can be used to deliver foreign proteins for cell-based gene therapy. The aim of this study was to use a lentivirus carrying proinsulin cDNA to achieve stable expression and lowering of blood glucose levels (BGLs). A SC line transduced with the lentivirus (MSC-LV-mI) maintained stable insulin expression in vitro. These MSC-LV-mI cells were transplanted and grafts were analyzed for cell survival, continued proinsulin mRNA, and insulin protein expression. All grafts contained MSC-LV-mI cells that expressed proinsulin mRNA and insulin protein. Transplantation of MSC-LV-mI cells into diabetic mice significantly lowered BGLs for 4 days after transplantation. Interestingly, in three transplanted SCID mice and one transplanted BALB/c mouse, the BGLs again significantly lowered by day 50 and 70, respectively. This is the first time SC transduced with a lentiviral vector was able to stably express insulin and lower BGLs. In conclusion, a SC line can be modified to stably express therapeutic proteins (e.g., insulin), thus taking us one step further in the use of SCs as an immune-privileged vehicle for cell-based gene therapy.

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Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed

Cited by 18 publications

References 34 publications

Neonatal Pig Sertoli Cells Survive Xenotransplantation by Creating an Immune Modulatory Environment Involving CD4 and CD8 Regulatory T Cells

Neonatal Pig Sertoli Cells Survive Xenotransplantation by Creating an Immune Modulatory Environment Involving CD4 and CD8 Regulatory T Cells

GM1 Induced the inflammatory response related to the Raf-1/MEK1/2/ERK1/2 pathway in co-culture of pig mesenchymal stem cells with RAW264.7

Sertoli Cells Engineered to Express Insulin to Lower Blood Glucose in Diabetic Mice

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