OBJECTIVETo establish a calculated panel reactive antibody (CPRA) method to analyze the donor-recipient incompatibility rate in PRA-positive kidney recipients and estimate the probability of these recipients to receive kidney transplantation.

METHODSBased on the database of HLA-A, -B, -DR genes and A-B, A-DR, B-DR, A-B-DR haplotype frequencies collected from 2004 donors from Jan 2000 to Dec 2012, we analyzed CPRA in 202 PRA-positive recipients and evaluated the consistency between PRA and CPRA assessments using a CPRA-Java calculator software, which returned a percentage of CPRA (representing the probability of unacceptable HLA in the donor group) after input of HLA-specific antibodies of a PRA-positive recipient.

RESULTSThe mean PRA intensity of the 202 PRA-positive recipients was (23.12∓17.83)% with a mean CPRA% of (46.07∓23.30)%. A significant difference was found between the mean PRA% and CPRA% in low sensitized recipients (PRA 0-10%) [(6.87∓2.41)% vs (21.63∓11.75)%, P<0.05) and in moderately sensitized recipients (PRA 10%-30%) [(20.15∓5.70)% vs (50.56∓16.86)%, P<0.05), but not in highly sensitized recipients (PRA>30%); The concordance rates between PRA% and CPRA% in the 3 groups were 19.35% (P<0.05), 10.99% (P<0.05), and 100% (P>0.05), respectively.

CONCLUSIONSLowly sensitized kidney recipients might have a lower probability of actually receiving a transplant than PRA% shows. A PRA%>30% is a risk factor for kidney transplantation. PRA reflects the sensitized level of a renal recipient, and reliable detection of HLA antibody specificity is of critical importance. CPRA accurately reflects the probability of a recipient to receive a transplant to assist clinicians in predicting the waiting time and selecting the transplant approach.

Antibodies ; Antibody Specificity ; Graft Rejection ; immunology ; Graft Survival ; immunology ; HLA Antigens ; genetics ; Haploidy ; Histocompatibility Testing ; methods ; Humans ; Kidney Transplantation

Gene chip has been an important way to detect gene changes of organism in different condition. It is a new tendency to use gene chip to research graft rejection at gene level. Ischemia reperfusion injury (IRI) accurs early in organ transplantation. Studying IRI with gene chip is beneficial to understand the mechanism of graft rejection and will provide guidance for surveying and treating graft rejection after organ transplantation.
Animals ; Disease Models, Animal ; Graft Rejection ; genetics ; Mice ; Oligonucleotide Array Sequence Analysis ; Rats ; Reperfusion Injury ; genetics ; Transplants

OBJECTIVETo evaluate the influence of major histocompatibility complex class I chain-related gene A (MICA) antibodies on acute rejection (AR) and renal function in early stage after renal transplantation.

METHODSA total of 197 renal transplant candidates admitted in Nanfang Hospital in 2009-2010 were enrolled in this study. MICA antibodies and their specificity were detected in all the patients, and 139 patients were followed up for early acute rejection (AR) and graft function after transplantation.

RESULTSMICA antibodies were positive before transplantation in 45 candidates (22.84%). Eleven specific MICA antibodies were identified, among which the frequency of MICA019 antibody (65.7%) was significantly higher than that of MICA015 (8.6%) and MICA017 (8.6%) (P<0.01). Eighteen patients with positive MICA antibodies were single-specific and 17 were polyspecific (51.4% vs 48.6% ). Of the 139 patients undergoing renal transplantation, 39 developed early AR (28.1%). Of the 45 candidates positive for MICA antibodies, 38 received renal transplantation and early AR occurred in 14 of them (36.8%); 101 of 152 candidates negative for MICA antibodies underwent renal transplantation, and 25 experienced early AR (24.8%).

CONCLUSIONMICA019 antibody is a frequent MICA antibody possibly due to the high frequency MICA019 gene in Chinese population.

Adult ; Antibodies ; immunology ; Antibody Specificity ; Female ; Graft Rejection ; immunology ; Graft Survival ; immunology ; Histocompatibility Antigens Class I ; genetics ; immunology ; Humans ; Kidney Transplantation ; Male ; Middle Aged

OBJECTIVETo study the role of interleukin (IL)-10 in acute-graft-versus-host disease (aGVHD) and graft rejection.

METHODSSerum concentrations of IL-10 in 28 patients who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) were measured by enzymed-linked immunosorbent assay (ELISA). IL-10 gene expression in peripheral mononuclear cells was measured by reverse-transcriptase polymerase chain reaction (RT-PCR) after transplantation.

RESULTSSeven patients developed grade I GVHD, 7 patients developed grade II-IV GVHD, 4 patients had graft rejection. Before transplantation, the concentrations of IL-10 were higher in patients who later did not developed aGVHD. After transplantation, IL-10 levels increased in patients without aGVHD, but decreased in patients with aGVHD or graft rejection. And IL-10mRNA was more frequent in patients without aGVHD compared to those with aGVHD.

CONCLUSIONSIL-10 plays a negative role in the development of aGVHD and graft rejection.

Acute Disease ; Adult ; Child ; Graft Rejection ; etiology ; Graft vs Host Disease ; etiology ; Humans ; Interleukin-10 ; blood ; genetics ; physiology ; Middle Aged ; RNA, Messenger ; analysis

To investigate the gene expression profiles in acute allograft rejection of renal transplantation, and identify the markers for the early diagnosis of acute rejection, heterotopic kidney transplantation was performed by using F344 or Lewis donors and Lewis recipients. No immunosuppressant was used. Renal grafts were harvested on days 3, 7, and 14. A commercial microarray was used to measure gene expression levels in day-7 grafts. The expression levels of 48 genes were up-regulated in the allograft in comparison with the isograft control, and interferon-gamma-induced GTPase gene was most significantly up-regulated in allografts. It is concluded that a variety of pathways are involved in organ transplant rejection which is dynamic and non-balanced. IFN-inducible genes, such as IGTP, may play an important role in the rejection. A lot of important factors involved in acute rejection are unnecessary but sufficient conditions for the rejection. We are led to conclude that it is virtually impossible to make an early diagnosis based on a single gene marker, but it could be achieved on the basis of a set of markers.
Gene Expression Profiling ; Gene Expression Regulation ; Graft Rejection/*genetics ; Graft Rejection/metabolism ; Kidney/*metabolism ; Kidney Transplantation ; MAP Kinase Signaling System ; Oligonucleotide Array Sequence Analysis ; Rats, Inbred F344 ; Rats, Inbred Lew ; Signal Transduction ; Species Specificity ; Time Factors ; Transplantation, Homologous

OBJECTIVETo investigate the correlation between major histocompatibility complex (MHC) class I chain-related gene A (MICA) gene alleles matching rates and graft rejection in small intestine, liver and kidney transplantation.

METHODSGenome DNA were extracted from blood samples or pathological sections collected from donors and recipients of living-related transplantation, included 4 cases of small bowel transplantation, 5 cases of liver transplantation and 6 cases of kidney transplantation. The correlation between MICA alleles matching rates and acute graft rejection was analyzed following 13 MICA alleles determination by polymerase chain reaction based on sequence-specific primers (PCR-SSP).

RESULTSHLA zygosity of all donors and recipients was confirmed to be half-matching. The recipients displaying higher matching rates of MICA alleles with donors showed lighter clinical and pathological rejection and longer survival time. On the contrary, recipients with lower matching rates of MICA alleles with donors showed severer clinical and pathological rejection and shorter survival time relatively.

CONCLUSIONMatching rates of MICA alleles has negative relevance to acute rejection, and positive relevance to survival time of recipients in small bowel, liver, and kidney transplantation.

Alleles ; Graft Rejection ; genetics ; immunology ; Histocompatibility Antigens Class I ; genetics ; Humans ; Intestine, Small ; transplantation ; Kidney Transplantation ; immunology ; Liver Transplantation ; immunology ; Living Donors ; Organ Transplantation

BACKGROUNDDonor organ rejection continues to be a significant problem for patients receiving transplants. We therefore tested whether transferring a donor's major histocompatibility complex (MHC) gene to the recipient would mitigate the rejection of transplanted hearts in mice.

METHODSH-2K(k) gene from donor mice was amplified using nested polymerase chain reaction (PCR) and ligated into a mammalian expression vector, which was then transfected into thymus ground mass cells collected from the recipients. Clones stably expressing the transgene were then injected into the recipients' thymus visualized using ultrasound. Control mice were administered cells previously transfected with empty vector. Following heart transplantation, cardiac activity was monitored electrocardiographically. Recipient thymus cells were tested for MHC antigenicity using flow cytometry and spleen cells were subjected to mixed lymphocyte culture tests. Finally, the transplanted hearts were sectioned, stained and examined under light microscopy.

RESULTSSouthern analysis following nested PCR revealed clear expression of H-2K(k) gene. Following transplantation, electrocardiosignals were detectable highly significantly longer in recipients administered thymal cells expressing donor H-2K(k) than in those receiving control cells. Flow cytometric analysis using an anti-H-2K(k) antibody confirmed its expression in H-2K(k) treated recipients but not in control mice. Mixed lymphocyte cultures containing H-2K(k) treated cells showed significantly less proliferation than those containing control cells. Hearts from control mice showed substantially greater lymphocyte infiltration than those from H-2K(k) treated mice and large areas of necrosis.

CONCLUSIONRejection of transplanted hearts can be mitigated substantially by introducing the donor's MHC into the recipient.

Animals ; Blotting, Southern ; Electrocardiography ; Female ; Flow Cytometry ; Graft Rejection ; genetics ; immunology ; Heart Transplantation ; immunology ; methods ; Major Histocompatibility Complex ; genetics ; immunology ; Male ; Mice ; Polymerase Chain Reaction

BACKGROUNDWe investigated the role of 1,25-dihydroxyvitamin D3(1,25-(OH)2D3) in preventing allograft from acute rejection following orthotopic liver transplantation.

METHODSA rat orthotopic liver transplantation model was used in this study. SD-Wistar rats served as a high responder strain combination. Recipients were subjected to administration of 1,25-(OH)2D3 at dosages ranging from 0.25 microg.kg(-1).d(-1) to 2.5 microg.kg(-1).d(-1). Survival after transplantation as well as pathological rejection grades and IFN-gamma mRNA, IL-10 mRNA transcription intragraft on day 7, and day 30 post-transplantation were observed.

RESULTSAfter recipients were treated with 1,25(OH)2D3 at dosages of 0.5 microg.kg(-1).d(-1) or 1.0 microg.kg(-1).d(-1), survivals of recipients were prolonged. Ninety-five percent confidence intervals of survival were 46 - 87 days and 69 - 102 days (both P = 0.0005 vs control group), respectively. On day seven post-transplantation, relative levels of IFN-gamma mRNA transcription were 0.59 +/- 0.12 and 0.49 +/- 0.16, which was higher than the control group (P = 0.005, P = 0.003, respectively). Relative levels of IL-10 mRNA transcription were 0.83 +/- 0.09 and 0.76 +/- 0.09, which was lower than the control group (P = 0.002, P = 0.003, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and on day thirty post-transplantation were 1.5 and 2.0 in comparison with the CsA-treated group (P = 0.178, P = 0.171, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and day thirty post-transplantation were 1.5 and 1.5 in comparison with CsA-treated group (P = 0.350, P = 0.693, respectively).

CONCLUSIONAfter each recipient was treated with 1,25-(OH)2D3 at a dosage of (0.5 - 1.0) microg.kg(-1).d(-1), transcription of cytokine intragraft was accommodated effectively and deviated to Th2 type, resulting in alleviation of acute rejection. 1,25-(OH)2D3 can prolong survival of recipient after orthotopic liver transplantation.

Animals ; Calcitriol ; pharmacology ; physiology ; Graft Rejection ; prevention & control ; Interferon-gamma ; genetics ; Interleukin-10 ; genetics ; Liver Transplantation ; mortality ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Transcription, Genetic

OBJECTIVETo gain insight into the role of Toll-like receptor 2 (TLR2) in graft rejection following penetrating keratoplasty, and investigate the expression of TLR2 mRNA in the corneal graft.

METHODSPenetrating keratoplasty was performed in 3 groups of rats for orthotopic autologous corneal transplantation (group A), allograft corneal transplantation (group B), or allograft corneal transplantation with hormone treatment (C). The transparency and neovascularization of the cornea were observed using a slit-lamp microscope and scored according to the rejection index, with normal cornea serving as the control. The corneal tissues were sampled at 5, 7, and 9 days after the transplantation for histopathological examination and detection of TLR2 mRNA expression using RT-PCR.

RESULTSWith the passage of time, edema, opacities and neovascularization of the corneal graft occurred after the operation in all the groups. Seven days after the operation, the rejection index of group B, but not that of groups A and C, met the diagnostic criteria for graft rejection with also support by histopathological evidence. The expression of TLR2 mRNA was detected in normal corneas and augmented in the corneal grafts in the 3 transplantation groups. TLR2 mRNA expression in group B was significantly higher than that of group A, and the expression in group C decreased significantly in comparison with that in group B (P<0.05).

CONCLUSIONAs the recognition receptors of native immune system, TLR2 in the rejected corneal grafts may recognize the allograft antigen and play a role in acute graft rejection after penetrating keratoplasty.

Animals ; Cornea ; metabolism ; Female ; Graft Rejection ; immunology ; Keratoplasty, Penetrating ; Postoperative Complications ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism

OBJECTIVEThis paper studied the effect of RNaseP against CIITA on repressing class II MHC (MHCII) expression.

METHODSIt was constructed that M1-RNA with guide sequences (GS), recognizing the 629 site of CIITA (M1-629-GS), by PCR from pTK117 plasmid, then was cloned into psNAV (psNAV-M1-629-GS). CIITA target gene was obtained from Raji cell by RT-PCR, and then inserted into pGEM-7zf (+) (pGEM-800). psNAV-M1-629-GS and pGEM-800 were transcribed and then mixed up and incubated in vitro. Stable transfectants of hepatocyte with psNAV-M1-629-GS by nanometer were tested for MHCII induction by recombinant human interferon-gamma (IFN-gamma). mRNA abundance of CIITA was measured by RT-PCR.

RESULTSIt showed that M1-629-GS could exclusively cleave pGEM-800 that formed a base pair with the GS. When induced with IFN-gamma, the expression of HLA-DR, -DP, -DQ on psNAV-M1-629-GS+ hepatocyte was (1.01+/-0.51)%, (4.37+/-1.28)%, (1.98+/-0.42)% respectively, was down-modulated 90.65%, 89.11% and 65.32% compared with control, while the mRNA content of CIITA reduced significantly (P<0.01).

CONCLUSIONM1-629-GS could effectively repress MHCII expressing through cleaving CIITA mRNA. These results provided insight into the future application of it as a new nucleic acid drug against the rejection of hepatic transplantation.

Graft Rejection ; prevention & control ; Histocompatibility Antigens Class II ; analysis ; Humans ; Liver Transplantation ; immunology ; Nuclear Proteins ; genetics ; RNA, Messenger ; analysis ; Ribonuclease P ; pharmacology ; Trans-Activators ; genetics