Chemokine gene expression during allograft rejection: Comparison of two quantitative PCR techniques

Carvalho-Gaspar, Manuela; Billing, J. Stephen; Spriewald, Bernd M.; Wood, Kathryn J.

doi:10.1016/j.jim.2005.03.003

Cited by 42 publications

(30 citation statements)

References 40 publications

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“…Skin grafts were harvested at different time points after transplantation and snap frozen. DNase I-treated total RNA was later isolated by using the Absolutely RNA Miniprep kit (Stratagene) and reverse-transcribed by the M-MLV reverse transcriptase (Invitrogen) as previously described (28). Real-time quantification was performed by using the PRISM 7700 sequence detection system (PE Applied Biosystem) using a fluorogenic probe as described previously (28).…”

Section: In Vivo Suppression Analysis On Proliferation Survival Andmentioning

confidence: 99%

“…DNase I-treated total RNA was later isolated by using the Absolutely RNA Miniprep kit (Stratagene) and reverse-transcribed by the M-MLV reverse transcriptase (Invitrogen) as previously described (28). Real-time quantification was performed by using the PRISM 7700 sequence detection system (PE Applied Biosystem) using a fluorogenic probe as described previously (28). Specific primers and probe for hypoxanthine phosphoribosyltransferase (HPRT) and foxp3 have been previously described (23,29).…”

Section: In Vivo Suppression Analysis On Proliferation Survival Andmentioning

confidence: 99%

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Allograft rejection mediated by memory T cells is resistant to regulation

Yang

Brook

Carvalho-Gaspar

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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198

157

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Alloreactive memory T cells may be refractory to many of the tolerance-inducing strategies that are effective against naive T cells and thus present a significant barrier to long-term allograft survival. Because CD4 ؉ CD25 ؉ regulatory T cells (Tregs) are critical elements of many approaches to successful induction/maintenance of transplantation tolerance, we used MHC class I and II alloreactive TCR-transgenic models to explore the ability of antigen-specific Tregs to control antigen-specific memory T cell responses. Upon coadoptive transfer into RAG-1 ؊/؊ mice, we found that Tregs effectively suppressed the ability of naive T cells to reject skin grafts, but neither antigen-unprimed nor antigen-primed Tregs suppressed rejection by memory T cells. Interestingly, different mechanisms appeared to be active in the ability of Tregs to control naive T cell-mediated graft rejection in the class II versus class I alloreactive models. In the former case, we observed decreased early expansion of effector cells in lymphoid tissue. In contrast, in the class I model, an effect of Tregs on early proliferation and expansion was not observed. However, at a late time point, significant differences in cell numbers were seen, suggesting effects on responding T cell survival. Overall, these data indicate that the relative resistance of both CD4 ؉ and CD8 ؉ alloreactive memory T cells to regulation may mediate resistance to tolerance induction seen in hosts with preexisting alloantigen-specific immunity and further indicate the multiplicity of mechanisms by which Tregs may control alloimmune responses in vivo.suppression ͉ tolerance ͉ transplantation

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Section: In Vivo Suppression Analysis On Proliferation Survival Andmentioning

confidence: 99%

Section: In Vivo Suppression Analysis On Proliferation Survival Andmentioning

confidence: 99%

Allograft rejection mediated by memory T cells is resistant to regulation

Yang

Brook

Carvalho-Gaspar

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

198

157

View full text Add to dashboard Cite

show abstract

“…Skin grafts were harvested at different time points after transplantation, snap frozen, and DNase I-treated total RNA was later isolated using the Absolutely RNA Miniprep kit (Stratagene) and reversed transcribed by the M-MLV reverse transcriptase (Invitrogen) (26). Real-time quantification was performed using the ABI Prism 7700 Sequence Detection System (PE Applied Biosystems) using either the fluorogenic probe (CD8␣ Taqman Gene Expression Assay from Applied Biosystems, hypoxanthine phosphoribosyl transferase (HPRT), CD3␥, IFN-␥, and perforin) or the SYBR Green technology (HPRT, CCL5, XCL1, CXCL9, and CXCL10) as described previously (26).…”

Section: Real-time Pcrmentioning

confidence: 99%

“…Real-time quantification was performed using the ABI Prism 7700 Sequence Detection System (PE Applied Biosystems) using either the fluorogenic probe (CD8␣ Taqman Gene Expression Assay from Applied Biosystems, hypoxanthine phosphoribosyl transferase (HPRT), CD3␥, IFN-␥, and perforin) or the SYBR Green technology (HPRT, CCL5, XCL1, CXCL9, and CXCL10) as described previously (26). Samples were standardized for HPRT and quantification of the gene of interest is given by 2…”

Section: Real-time Pcrmentioning

confidence: 99%

Location and Time-Dependent Control of Rejection by Regulatory T Cells Culminates in a Failure to Generate Memory T Cells

Carvalho-Gaspar

Jones

Luo

et al. 2008

The Journal of Immunology

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Adaptive CD25+CD4+ regulatory T cells (Treg) can be induced following exposure to alloantigen and may function alongside naturally occurring Treg to suppress allograft rejection when present in sufficient numbers. However, the location of the Treg as they function in vivo and the mechanisms used to control donor-reactive T cells remains ill-defined. In this study, we used a CD8+ TCR transgenic model of skin allograft rejection to characterize in vivo activity of donor-reactive Treg cells during induction of transplantation tolerance. We demonstrate that, initially after skin transplantation, Treg attenuate the priming of donor-reactive naive CD8+ T cells in the lymphoid tissue draining the graft site. However, with time, peripheral suppression is overcome despite the continued presence of Treg, resulting in the priming of donor-reactive CD8+ T cells and graft infiltration by the resultant effector T cells and induction of a “Tc1-like” intragraft gene expression profile. These intragraft effector CD8+ T cells are then prevented from eliciting rejection by Treg that simultaneously infiltrate the skin allografts, resulting in a failure to generate donor-reactive memory CD8+ T cells. Overall, these data demonstrate for the first time that donor-reactive Treg can suppress allograft rejection using distinct mechanisms at different sites in vivo with the overall outcome of preventing the generation of donor-reactive memory T cells.

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“…Chemokine receptors are also divided into four corresponding groups. 3 One or three amino acids separate the first and second cysteines in the CXC and CX3C chemokines, respectively, the two cysteines are adjoining in the CC subfamily, and the C subfamily lacks the first and pairing third conserved cystein residues. The fifth receptor subfamily, CX, reported only in zebrafish lacks the two N-terminal residues, but retains the third and fourth residues.…”

Section: Introductionmentioning

confidence: 99%

The integrative roles of chemokines at the maternal–fetal interface in early pregnancy

2014

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Embryos express paternal antigens that are foreign to the mother, but the mother provides a special immune milieu at the fetal-maternal interface to permit rather than reject the embryo growth in the uterus until parturition by establishing precise crosstalk between the mother and the fetus. There are unanswered questions in the maintenance of pregnancy, including the poorly understood phenomenon of maternal tolerance to the allogeneic conceptus, and the remarkable biological roles of placental trophoblasts that invade the uterine wall. Chemokines are multifunctional molecules initially described as having a role in leukocyte trafficking and later found to participate in developmental processes such as differentiation and directed migration. It is increasingly evident that the gestational uterine microenvironment is characterized, at least in part, by the differential expression and secretion of chemokines that induce selective trafficking of leukocyte subsets to the maternal-fetal interface and regulate multiple events that are closely associated with normal pregnancy. Here, we review the expression and function of chemokines and their receptors at the maternal-fetal interface, with a special focus on chemokine as a key component in trophoblast invasiveness and placental angiogenesis, recruitment and instruction of immune cells so as to form a fetus-supporting milieu during pregnancy. The chemokine network is also involved in pregnancy complications.

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Chemokine gene expression during allograft rejection: Comparison of two quantitative PCR techniques

Cited by 42 publications

References 40 publications

Allograft rejection mediated by memory T cells is resistant to regulation

Allograft rejection mediated by memory T cells is resistant to regulation

Location and Time-Dependent Control of Rejection by Regulatory T Cells Culminates in a Failure to Generate Memory T Cells

The integrative roles of chemokines at the maternal–fetal interface in early pregnancy

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