Assessing the<i>In Vitro</i>Suppressive Capacity of Regulatory T Cells

Brusko, Todd M.; Hulme, Maigan A.; Myhr, Courtney; Haller, Michael J.; Atkinson, Mark A.

doi:10.1080/08820130701790368

Cited by 53 publications

(55 citation statements)

References 41 publications

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“…Effector T cells were analyzed for a CFSE profile on a FACSCalibur. The percentage suppression was calculated using the precursor frequency (PF) method described by Brusko et al (61). Briefly, percentage suppression = 100 × (1 -[PF (Treg + Teff) / PF (Teff)]).…”

Section: Methodsmentioning

confidence: 99%

Maternal alloantibodies induce a postnatal immune response that limits engraftment following in utero hematopoietic cell transplantation in mice

Merianos¹,

Tiblad²,

Santore³

et al. 2009

J. Clin. Invest.

114

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The lack of fetal immune responses to foreign antigens, i.e., fetal immunologic tolerance, is the most compelling rationale for prenatal stem cell and gene therapy. However, the frequency of engraftment following in utero hematopoietic cell transplantation (IUHCT) in the murine model is reduced in allogeneic, compared with congenic, recipients. This observation supports the existence of an immune barrier to fetal transplantation and challenges the classic assumptions of fetal tolerance. Here, we present evidence that supports the presence of an adaptive immune response in murine recipients of IUHCT that failed to maintain engraftment. However, when IUHCT recipients were fostered by surrogate mothers, they all maintained long-term chimerism. Furthermore, we have demonstrated that the cells responsible for rejection of the graft were recipient in origin. Our observations suggest a mechanism by which IUHCT-dependent sensitization of the maternal immune system and the subsequent transmission of maternal alloantibodies to pups through breast milk induces a postnatal adaptive immune response in the recipient, which, in turn, results in the ablation of engraftment after IUHCT. Finally, we showed that non-fostered pups that maintained their chimerism had higher levels of Tregs as well as a more suppressive Treg phenotype than their non-chimeric, non-fostered siblings. This study resolves the apparent contradiction of induction of an adaptive immune response in the pre-immune fetus and confirms the potential of actively acquired tolerance to facilitate prenatal therapeutic applications.

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Section: Methodsmentioning

confidence: 99%

Maternal alloantibodies induce a postnatal immune response that limits engraftment following in utero hematopoietic cell transplantation in mice

Merianos¹,

Tiblad²,

Santore³

et al. 2009

J. Clin. Invest.

114

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“…Plates were harvested on a Packard FilterMate Harvester and read on a Packard TopCount Scintillation and Luminescence Counter (Perkin Elmer, Waltham, MA). For both assays, percent suppression was calculated as previously described (17).…”

Section: Conclusion-thementioning

confidence: 99%

Expansion of Human Regulatory T-Cells From Patients With Type 1 Diabetes

et al. 2009

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OBJECTIVE— Regulatory T-cells (Tregs) have catalyzed the field of immune regulation. However, translating Treg-based therapies from animal models of autoimmunity to human clinical trials requires robust methods for the isolation and expansion of these cells—a need forming the basis for these studies. RESEARCH DESIGN AND METHODS— Tregs from recent-onset type 1 diabetic patients and healthy control subjects were isolated by fluorescence-activated cell sorting and compared for their capacity to expand in vitro in response to anti-CD3–anti-CD28–coated microbeads and IL-2. Expanded cells were examined for suppressive function, lineage markers and FOXP3, and cytokine production. RESULTS— Both CD4 + CD127 lo/− and CD4 + CD127 lo/− CD25 + T-cells could be expanded and used as Tregs. However, expansion of CD4 + CD127 lo/− cells required the addition of rapamycin to maintain lineage purity. In contrast, expansion of CD4 + CD127 lo/− CD25 + T-cells, especially the CD45RA + subset, resulted in high yield, functional Tregs that maintained higher FOXP3 expression in the absence of rapamycin. Tregs from type 1 diabetic patients and control subjects expanded similarly and were equally capable of suppressing T-cell proliferation. Regulatory cytokines were produced by Tregs after culture; however, a portion of FOXP3 + cells were capable of producing interferon (IFN)-γ after reactivation. IFN-γ production was observed from both CD45RO + and CD45RA + Treg populations. CONCLUSIONS— The results support the feasibility of isolating Tregs for in vitro expansion. Based on expansion capacity, FOXP3 stability, and functional properties, the CD4 + CD127 lo/− CD25 + T-cells represent a viable cell population for cellular therapy in this autoimmune disease.

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“…Regulatory T cells can act as active suppressor of lymphocyte response to tumor antigens in malignant neoplasms and many recent studies have explored the cellular and molecular mechanisms of this suppression [25][26][27]. In this study, we evaluated CTLA-4 and FOXP3 transcripts, as acceptable indicators of Tregs, in the peripheral blood from women with breast cancer and found that these transcripts significantly increase even in early stages of breast cancer.…”

Section: Discussionmentioning

confidence: 98%

Increased CTLA-4 and FOXP3 Transcripts in Peripheral Blood Mononuclear Cells of Patients with Breast Cancer

Jaberipour

Habibagahi

Hosseini

et al. 2010

Pathol. Oncol. Res.

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Generation of Regulatory T cells (Tregs) is known to play a major role in progression and modulation of the immune escape mechanisms in cancer. These cells express Forkhead/winged helix transcription factor (FOXP3) and also Cytotoxic T-lymphocyte antigen-4 (CTLA-4), as a negative regulatory molecule which, is a potential target for immunotherapy. We, therefore, evaluated FOXP3 and CTLA-4 transcripts in the peripheral blood mononuclear cells from 55 women with histologically-confirmed infiltrating ductal carcinoma of the breast. Blood samples from 40 healthy volunteer women without a history of malignancies or autoimmune disorders were also obtained as a control. The abundance of FOXP3 and CTLA-4 gene transcripts was determined by quantitative real-time PCR (qRT-PCR). Compared to healthy individuals, significantly higher amounts of these transcripts were found in the mononuclear cells from breast cancer patients. Also, a significant correlation was found between CTLA-4 and FOXP3 expressions in a group of patients. Among patients with early stage, nonmetastatic or low-grade disease, the relative expression of CTLA-4 was about 10-fold as much as in the control group. These patients also showed a significant increase, more than 10 fold, in mean relative FOXP3 expression. The results of this investigation point to functional activity of Treg cells in early stages of breast cancer, a finding which emphasizes the significance of Tregs as an imminent target for breast cancer immunotherapy.

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Assessing theIn VitroSuppressive Capacity of Regulatory T Cells

Cited by 53 publications

References 41 publications

Maternal alloantibodies induce a postnatal immune response that limits engraftment following in utero hematopoietic cell transplantation in mice

Maternal alloantibodies induce a postnatal immune response that limits engraftment following in utero hematopoietic cell transplantation in mice

Expansion of Human Regulatory T-Cells From Patients With Type 1 Diabetes

Increased CTLA-4 and FOXP3 Transcripts in Peripheral Blood Mononuclear Cells of Patients with Breast Cancer

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