A comparison of intrauterine hemopoietic cell transplantation and lentiviral gene transfer for the correction of severe β-thalassemia in a HbbTh3/+ murine model

Dighe, Niraja; Tan, Kang Wei; Tan, Lei; Shaw, Steven S. W.; Buckley, Suzanne M.K.; Sandikin, Dedy; Johana, Nuryanti; Tan, Yi-Wan; Biswas, Arijit; Choolani, Mahesh; Waddington, Simon N.; Antoniou, Michael; Chan, Jerry Ky; Mattar, Citra Nurfarah Zaini

doi:10.1016/j.exphem.2018.03.006

Cited by 11 publications

(10 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…14,15 Recipients not meeting the initial threshold chimerism were difficult to boost with serial postnatal booster transplants, even with chemoradiation for immunosuppression and to increase hemopoietic niche availability. 19,23,36,37 In these studies, allogenic IUT comprised the transplantation of BALB/c cells into purebred B6 fetuses and only ~30% of mice were chimeras after IUT. 14,15,23,37,38 Transplanted doses and HSC source ranged from 2.5E+6 fetal liver MNC/ pup to 20E+6 BM cells/pup resulting in dose-dependent chimerism.…”

Section: Discussionmentioning

confidence: 99%

“…Initial chimerism above this threshold promotes antigen‐specific NK downregulation and increased Treg activity in the host (through NK receptor calibration and thymic clearance of alloreactive effector T cells), inducing adaptive immune tolerance needed for stable long‐term engraftment, while sub‐threshold chimerism was predictive of engraftment loss via reactive effector T and NK cells 14,15 . Recipients not meeting the initial threshold chimerism were difficult to boost with serial postnatal booster transplants, even with chemoradiation for immunosuppression and to increase hemopoietic niche availability 19,23,36,37 . In these studies, allogenic IUT comprised the transplantation of BALB/c cells into purebred B6 fetuses and only ~30% of mice were chimeras after IUT 14,15,23,37,38 .…”

Section: Discussionmentioning

confidence: 99%

“…PB and BM-MNC were prepared by density gradient centrifugation using Ficoll-Paque PLUS density gradient media (GE Healthcare, NJ, USA) as described previously. 19 Liver MNC were prepared as described 17 : digestion by collagenase IV (0.04%, 10 minutes at 37°C), passing through 70 µm nylon mesh, centrifugation (150g × 5 minutes) to remove debris, pelleting of cells (400g × 5 minutes), resuspension of cell pellet in 40% of Percoll solution (overlaid on 70% Percoll), gradient centrifugation (800g × 25 minutes), and harvesting MNC from the Percoll interface. Spleen was minced and mashed through a 70 µm cell strainer producing a single-cell suspension flow-through which was centrifuged (400g × 5 minutes), the cell pellet was subjected to RBC lysis and washed with PBS.…”

Section: Isolation Of Mnc and Facs Analysismentioning

confidence: 99%

See 2 more Smart Citations

Maternal microchimerism and cell‐mediated immune‐modulation enhance engraftment following semi‐allogenic intrauterine transplantation

et al. 2021

Self Cite

View full text Add to dashboard Cite

Successful intrauterine hematopoietic cell transplantation (IUT) for congenital hemoglobinopathies is hampered by maternal alloresponsiveness. We investigate these interactions in semi‐allogenic murine IUT. E14 fetuses (B6 females × BALB/c males) were each treated with 5E+6 maternal (B6) or paternal (BALB/c) bone marrow cells and serially monitored for chimerism (>1% engraftment), trafficked maternal immune cells, and immune responsiveness to donor cells. A total of 41.0% of maternal IUT recipients (mIUT) were chimeras (mean donor chimerism 3.0 ± 1.3%) versus 75.0% of paternal IUT recipients (pIUT, 3.6 ± 1.1%). Chimeras showed higher maternal microchimerism of CD4, CD8, and CD19 than non‐chimeras. These maternal cells showed minimal responsiveness to B6 or BALB/c stimulation. To interrogate tolerance, mIUT were injected postnatally with 5E+6 B6 cells/pup; pIUT received BALB/c cells. IUT‐treated pups showed no changes in trafficked maternal or fetal immune cell levels compared to controls. Donor‐specific IgM and IgG were expressed by 1%‐3% of recipients. mIUT splenocytes showed greater proliferation of regulatory T cells (Treg) upon BALB/c stimulation, while B6 stimulation upregulated the pro‐inflammatory cytokines more than BALB/c. pIUT splenocytes produced identical Treg and cytokine responses to BALB/c and B6 cells, with higher Treg activity and lower pro‐inflammatory cytokine expression upon exposure to BALB/c. In contrast, naïve fetal splenocytes demonstrated greater alloresponsiveness to BALB/c compared to B6 cells. Thus pIUT, associated with increased maternal cell trafficking, modulates fetal Treg, and cytokine responsiveness to donor cells more efficiently than mIUT, resulting in improved engraftment. Paternal donor cells may be considered alternatively to maternal donor cells for intrauterine and postnatal transplantation to induce tolerance and maintain engraftment.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Isolation Of Mnc and Facs Analysismentioning

confidence: 99%

See 1 more Smart Citation

Maternal microchimerism and cell‐mediated immune‐modulation enhance engraftment following semi‐allogenic intrauterine transplantation

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…In utero hematopoietic stem cell transplantation (IUT) represents potential cure for congenital hematological disorders with advantages over conventional postnatal transplantation, particularly avoidance of myeloablation, 1 but immune barriers cause poor engraftment, hampering clinical application. [2][3][4][5] Conventional dendritic cells (cDC) activate CD8 and CD4 T-cells via subtypes cDC1 and cDC2 respectively, [6][7][8] mediating antigen-specific tolerance by altering helper T-cell (Th) balance 9,10 and cytokine expression. [11][12][13] Active maternal-fetal trafficking occurs throughout pregnancy, 14,15 increasing after invasive procedures like IUT.…”

Section: Discussionmentioning

confidence: 99%

Maternal Dendritic Cells Influence Fetal Allograft Response and Tolerance

Mattar

Kandasamy

Johana

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Intrauterine hematopoietic cell transplantation (IUT), a promising therapy for congenital haematological disease, is limited by subtherapeutic donor cell chimerism (DCC). Microchimerism of maternal immune cells (MMc) trafficked into fetal IUT recipients may directly influence immune-mediated donor cell clearance. We investigated if maternal dendritic cell (DC) suppression reduces recipient alloresponsiveness to donor cells, improving DCC. IUT was performed at E14 in pregnancies resulting from crossing CD11c.DTR(B6) females and Balb/c males, with semiallogenic Balb/c or C57BL/6, or fully allogenic C3H donor cells, 24h after administering diphtheria toxin to the dam, transiently suppressing maternal DC. This resulted in reduced MMc in recipient fetuses, greatest after Balb/c transplantation, also associated with the highest DCC, and lowest with C3H. Maternal-derived T-cell receptor (TCR) clonotypes were enriched in IUT recipients and displayed substantially reduced diversity. Recipient pups with reduced MMc increased expression of regulatory T-cell subtypes, reduced expression of proinflammatory cytokines, and demonstrated enhanced TCR clonotype diversity. DCC was primarily related to donor cell type and not influenced by MMc. Our data indicate that donor cell origin and MMc are distinct factors determining IUT effectiveness. MMc independently influences DCC and recipient tolerance to donor cells and may present novel therapeutic targets to improve transplantation outcomes.

show abstract

“…www.nature.com/scientificreports www.nature.com/scientificreports/ in need. However, many trials have encountered difficulties in genetically manipulating HSCs efficiently [45][46][47] . Understanding the resistance mechanisms and developing methods to overcome them will help many researchers and impact the broader perspectives of stem cell utilization in regenerative medicine.…”

Section: Csh-enhanced Transduction Does Not Limit Transplantation Int...mentioning

confidence: 99%

Cyclosporine H Improves the Multi-Vector Lentiviral Transduction of Murine Haematopoietic Progenitors and Stem Cells

Olender

Bujanover

Sharabi

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Haematopoietic stem cells (HSCs) have the potential for lifetime production of blood and immune cells. The introduction of transgenes into HSCs is important for basic research, as well as for multiple clinical applications, because HSC transplantation is an already established procedure. Recently, a major advancement has been reported in the use of cyclosporine H (CsH), which can significantly enhance the lentivirus (LV) transduction of human haematopoietic stem and progenitor cells (HSPCs). In this study, we employed CsH for LV transduction of murine HSCs and defined haematopoietic progenitors, confirming previous findings in more specific subsets of primitive haematopoietic cells. Our data confirm increased efficiencies, in agreement with the published data. We further experimented with the transduction with the simultaneous use of several vectors. The use of CsH yielded an even more robust increase in rates of multi-vector infection than the increase for a single-vector. CsH was reported to reduce the innate resistance mechanism against LV infection. We indeed found that additional pretreatment could increase the efficiency of transduction, in agreement with the originally reported results. Our data also suggest that CsH does not reduce the efficiency of transplantation into immune-competent hosts or the differentiation of HSCs while enhancing stable long-term expression in vivo. This new additive will surely help many studies in animal models and might be very useful for the development of novel HSC gene therapy approaches.

show abstract

A comparison of intrauterine hemopoietic cell transplantation and lentiviral gene transfer for the correction of severe β-thalassemia in a HbbTh3/+ murine model

Cited by 11 publications

References 62 publications

Maternal microchimerism and cell‐mediated immune‐modulation enhance engraftment following semi‐allogenic intrauterine transplantation

Maternal microchimerism and cell‐mediated immune‐modulation enhance engraftment following semi‐allogenic intrauterine transplantation

Maternal Dendritic Cells Influence Fetal Allograft Response and Tolerance

Cyclosporine H Improves the Multi-Vector Lentiviral Transduction of Murine Haematopoietic Progenitors and Stem Cells

Contact Info

Product

Resources

About