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

China ; Graft Survival ; HLA Antigens ; genetics ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; methods ; mortality ; Humans ; Transplantation, Homologous

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

OBJECTIVEThe purpose of this study was to determine the effects of local delivery of vascular endothelial growth factor( VEGF) transferred with adenovirus-mediated gene on the survival of ischemic random skin flap in rats.

METHODSThe animals were divided into three groups randomly (n = 10) . A 2 cm x 8 cm dorsal skin flap was designed with the pedicle at the level of the iliac crest. In group A (AdCMV-VEGF), each animal received 10(12) PR replication-deficient recombinant adenovirus (AdCMV-VEGF) in the distal two-thirds of the proposed flap by means of the subdermal injection at ten different locations. In group B (AdCMV-GaI), each received 1012 PR AdCMV-Gal. In Group C (Saline), each received 1 ml saline. Three days after the treatment, the flap was elevated as planed way and re-sutured back to its donor site. All the animals were evaluated 7 days after the operation.

RESULTSThe mean percentage of surviving flap area was (85.91 +/- 2.54)% in group A, (59.56 +/- l.18)% in group B, and (61.48 +/- l.09)% in group C. There was a significant increase in the percentage of the survival area in the flaps of the group A, compared with the group B and group C (Group B vs. Group A, P < 0.01; Group C vs.Group A, P < 0.01, Group B vs. Group C, P >0.05). Hybridization in the situ, the immunohistochemical stain showed that the VEGF was expressed in the survival tissue of the flap treated with the AdCMV-VEGF, but it was not found in the control groups. Histological analysis demonstrated qualitatively greater amount of granulation tissue and angiogenesis was found in the group treated with the AdCMV-VEGF than the controls.

CONCLUSIONSThe results may indicate that Ad vector carrying VEGF cDNA could be useful in enhancing the survival of the skin flap due to the effect of the local delivery of the VEGF.

Adenoviridae ; genetics ; Animals ; DNA, Complementary ; Genetic Therapy ; Genetic Vectors ; Graft Survival ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; genetics ; Surgical Flaps ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo explore the immunosuppressive effect of local transfection of Molluscum contagiosum virus 148 (MC148) gene to allogenous skin graft against rejection.

METHODSMC148 gene was cloned from molluscum contagiosum virus (MCV), and was employed to construct recombinant adenovirus vector (Ad-MC148). The recombinant Ad-MC148 was then locally transfected into a part of the tail skin of eight Lewis rats, which served as skin donors for grafting. The wounds (1 cm x 1 cm) were produced on the tails of 16 Wistar rats, and they were then randomly divided into control (C, n=8, with grafting of skin from donor rats without transfection), and transfection (T, n=8, with grafting of skin from donor rats with transfection of the recombinant Ad-MC148) groups. The expression of MC148 mRNA gene in T group was detected on 6 post operation hour( POH) and 2, 3, 7 and 10 post operation day (POD), and the results were expressed by the ratio of absorption value (A) between MC148 gene and beta-actin. The survival time of skin grafts in both groups was compared. Gross examination of grafted skin was carried out from 7 POD on in both groups, and the pathomorphological changes were examined in both groups on 7 POD.

RESULTSThe MC148 gene expression in rat skin of T group could be identified in 6 POH, and it reached the peak on 3 POD (A(MC148 mRNA) / A(beta_actin) = 0.86), and then subsided thereafter, but it maintained for 10 days. The survival time of the grafts in T group was (15.0 +/- 2.0) days, and it was significantly longer than that in C group (8.5 +/- 3.4) days, (P < 0.01). Gross and microscopic examination showed that the tail skin of T group appeared ruddy on 7 POD, with little leukocytic infiltration in subcutaneous tissue; it began to turn black after 12 to 20 PODs. On the other hand, the tail skin of C group began to turn black and to shed off on 7 POD, with evident leukocytic infiltration in subcutaneous tissue and dermis.

CONCLUSIONLocal transfection of MC148 gene may promote immunosuppression by inhibiting leukocytic infiltration after allogenous skin transplantation.

Adenoviridae ; genetics ; Animals ; Chemokines, CC ; genetics ; Genetic Vectors ; Graft Survival ; Rats ; Rats, Inbred Lew ; Rats, Wistar ; Skin Transplantation ; Transfection ; Transplantation, Homologous ; Viral Proteins ; genetics

BACKGROUNDCostimulatory signals play a vital role in T cell activation. Blockade of costimulatory pathway by CTLA4Ig or CD40LIg have enhanced graft survival in experimental transplantation models yet mechanisms remain undetermined. We investigated the effects of CTLA4Ig and CD40LIg gene transfer on islet xenografts rejection in rats.

METHODSHuman islets were infected with recombinant adenoviruses containing CTLA4Ig and CD40LIg genes and implanted beneath the kidney capsule of diabetic rats. Levels of blood sugar, morphological changes, and survival of grafts were recorded. Expressions of CTLA4Ig, CD40LIg and insulin were detected by immunohistochemical staining and cytokines levels were quantified by enzyme-linked immunosorbent assay (ELISA).

RESULTSBlood glucose levels in transplant rats decreased to normal level on the 2nd day post transplantation. The mean blood glucose in the control group, CTLA4Ig transfected group, CD40LIg transfected group and CTLA4Ig + CD40LIg cotransfected group increased on days 8, 24, 21, 68, post transplantation respectively. The grafts in control group, CTLA4Ig transfected group, CD40LIg transfected group and CTLA4Ig + CD40LIg cotransfected group survived for (8 ± 1), (29 ± 4), (27 ± 3), and (74 ± 10) days, respectively. Survival in CTLA4Ig + CD40LIg cotransfected group was significantly longer. Survivals of CTLA4Ig transfected group and CD40LIg transfected group were significantly longer than control group. In control animals, serum interleukin-2 and tumor necrosis factor α concentration significantly increased within seven days post transplantation. Haematoxylin eosin staining of grafts showed live islets in situ of transplant rats without inflammatory cell infiltration. Immunohistochemical staining confirmed the expression of insulin at islets in all experimental groups.

CONCLUSIONSTransfer of CTLA4Ig and CD40LIg genes, especially the cotransfer of both, inhibits rejection of murine islet xenografts. Downregulated expressions of Th1 cells related cytokines might be related to the beneficial effects.

Abatacept ; Animals ; Enzyme-Linked Immunosorbent Assay ; Graft Rejection ; therapy ; Graft Survival ; genetics ; physiology ; Humans ; Immunoconjugates ; genetics ; metabolism ; Immunohistochemistry ; Insulin ; metabolism ; Islets of Langerhans Transplantation ; immunology ; methods ; Rats ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transplantation, Heterologous ; immunology ; methods

Child ; Graft Survival ; HLA Antigens ; genetics ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Polymerase Chain Reaction ; Tissue Donors ; Transplantation Chimera ; genetics ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; immunology

OBJECTIVETo investigate the effects of rhVEGF on autologous free granular fat grafts in rats.

METHODSForty-eight Sprague-Dawley rats were randomly divided into three groups, sixteen of each. After the autologous free granular fat transplantation, all groups were treated with the plasmid DNA containing cDNA encoding rhVEGF, the blank plasmid DNA and normal saline respectively as the experimental group, the negative group and the saline group. After 3, 7, 15, 30 days, the rats were sacrificed and the grafts were weighted accurately. Histological pathology was evaluated. Micro-vessel count and the expression of vascular endothelial growth factor (VEGF) were examined by immunohistochemical staining.

RESULTSThe weights of the two latter groups were significantly reduced on the 7, 15, 30 day compared with the experimental group. The expression of VEGF and the micro-vessel count in the experimental group were significantly higher than the other two groups during the latter periods.

CONCLUSIONThe cDNA encoding VEGF can induce the expression of VEGF in fat graft, angiogenesis and reduce the free fat graft absorption.

Adipose Tissue ; transplantation ; Animals ; Female ; Gene Transfer Techniques ; Graft Survival ; Male ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; genetics ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo investigate the effect of cytotoxic T lymphocyte-associated antigen 4 immunoglobulin (CTLA4-Ig) gene transfer on rejection of rat islet xenografts.

METHODSHuman islets were infected with the recombinant adenoviruses containing CTLA4-Ig gene, and the transduced islets were transplanted under the left kidney capsule of diabetic rats to establish rat models bearing human-rat islet xenografts. The blood sugar of the rats receiving the transplantation was measured and the xenografts and host survival were observed after transplantation. The morphological changes of grafts were examined, in which the expression of CTLA4-Ig and insulin were also detected by immunohistochemical staining and the cytokines were quantified using enzyme-linked immunosorbent assay (ELISA).

RESULTSThe blood glucose of the rats bearing the grafts decreased to the normal level on day 2 after transplantation. The average blood glucose level of the CTLA4-Ig gene transfected group and the control group increased on day 25 and 8 post-transplantation, respectively. The grafts of the transfected group survived for an average of 28-/+6 days, significantly longer than that in the control group (10-/+2 days, t=10.52, P<0.01), and the host survival were 48-/+8 and 21-/+6 days in the two groups, showing also significant difference between them (t=12.23, P<0.01). In the control rats, serum IL-2 and TNF-alpha concentration drastically increased within 7 days after transplantation to levels significantly higher than that before transplantation (P<0.01), but in the transfected group, the levels were decreased compared with the preoperative levels. In the transfected grafts, positive staining for CTLA4-Ig and insulin were detected.

CONCLUSIONCTLA4-Ig gene transfer may lower the rejection of rat islet xenografts and prolong the survival time of both the grafts and hosts.

Abatacept ; Adenoviridae ; genetics ; Animals ; Blood Glucose ; Graft Rejection ; prevention & control ; Graft Survival ; Humans ; Immunoconjugates ; genetics ; Interleukin-2 ; blood ; Islets of Langerhans Transplantation ; Rats ; Rats, Wistar ; Transduction, Genetic ; Transplantation, Heterologous ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo investigate the effect of recombinant human vascular endothelial growth factor (rhVEGF) on autologous free granular fat grafts in rats.

METHODSForty-eight Sprague Dawley (SD) rats, weighing 190-280 g and regardless sex, were randomly divided into three groups, sixteen in each. After fat transplantation, the rats were treated with plasmid DNA encoding rhVEGF protein (the experimental group), plasmid DNA (the negative group) and normal saline (the blank control group), respectively. At 3, 7, 15 and 30 days after transplantation, the rats were killed and the grafts were weighed, respectively. Histopathological changes were evaluated. Microvessel density and the expression of VEGF were examined by immunohistochemical staining and Western blotting.

RESULTSThe weights of the negative and blank control groups were significantly reduced on the 7th, 15th and 30th days compared with those of the experimental group. The expression of VEGF and the microvessel density in the experimental group were significantly higher than the other two groups during the latter periods.

CONCLUSIONThe plasmid encoding VEGF can induce expression of VEGF and angiogenesis in fat grafts and reduce the absorption of free fat grafts.

Adipose Tissue ; transplantation ; Animals ; Female ; Graft Survival ; physiology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transduction, Genetic ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics