Development strategy and clinical research progress of universal chimeric antigen receptor T-cell drugs / 中国药科大学学报

@#Chimeric antigen receptor T-cell (CAR-T) immunotherapy has made a breakthrough in the clinical treatment of a variety of hematological tumors.However, the CAR-T cell products listed at China and abroad are all autologous CAR-T.Compared with autologous CAR-T treatment, universal CAR-T exhibits significant advantages, which could fulfill the treatment demand of more patients, but also displays high technical barriers.This paper reviews the universal CAR-T, clearly points out the two major challenges faced by the development of universal CAR-T, and then summarizes and analyzes the feasible solutions according to the mechanism causing the two major problems.This paper also summarizes domestic and foreign companies producing universal CAR-T and the latest clinical progress of their superior products, and then discusses the feasibility of the development strategy from another aspect, in order to provide ideas for developing a new generation of universal CAR-T cell therapy products.

Recent advances in preclinical research of renal xenotransplantation and selection of potential recipients in the future / 中华器官移植杂志

After continuous efforts from generations of transplant surgeons, kidney transplantation (KT) has become an optimal treatment for end-stage renal disease.However, an imbalance between supply and demand of organs has always restricted the development of KT.For this clinical dilemma, xenotransplantation is expected to become one practical alternative for alleviating organ shortage.This review summarized recent literature reports of kidney xenotransplantation and the latest cases of pig-to-human kidney and heart transplantations.Also clinical transformations and applications of kidney xenotransplantation were discussed.

Chimeric antigen receptor T cell immunotherapy for refractory and relapsed acute lymphoblastic leukemia: Challenge and Countermeasures / 中华检验医学杂志

Chimeric antigen receptor(CAR)-T cell immunotherapy for refractory and relapsed acute lymphoblastic leukemia (R/R ALL) is one of the breakthroughs in the field of hematological malignant disease treatment. However, several challenges remain, such as immune rejection of allogeneic CAR-T cells, leukemia relapse, as well as could we apply CAR-T cell immunotherapy to ALL patients with positive measurable residual disease and those of newly diagnosed cases. The safety and efficiency of CAR-T therapy will be further improved for patients with ALL only by establishing appropriate strategies to address these challenges.

Effect of immature dendritic cells derived from bone marrow on rejection of orthotopic liver transplantation in rats / 中国组织工程研究

BACKGROUND: Immature dendritic cells have strong antigen uptake and processing ability, but the lack of a variety of costimulatory molecules cannot activate and proliferate initial T cells to produce immune response, which can lead to T cell anergy, thus inducing low immune response or antigen immune specific tolerance. Simultaneously, immature dendritic cells can induce hyporeactivity of allogeneic antigen-specific T cells, thus prolonging the survival time of transplanted organs. OBJECTIVE: To investigate the effect of immature dendritic cells on liver rejection after orthotopic liver transplantation in rats and its mechanism. METHODS: According to the weight of DA and Lewis rats, the rats were randomly divided into three groups, and the liver transplantation model of DA-Lewis rats was established by “two-cuff” method. The rats of control group received no measures. The rats of cyclosporine group were treated with 10 mg/kg cyclosporine from the second day after operation, once a day, for 7 days. The rats of the immature dendritic cell group were injected with 1×106 immature dendritic cells from bone marrow of DA rats one day before operation through dorsal penile vein; the injection was repeated twice with an interval of 10 minutes. The livers of all these rats were removed 7 days after operation. Hematoxylin-eosin staining was used to observe the pathological changes. The mRNA and protein expressions of SHIP, AKT, IKK and IKβ in these three groups were detected by qRT-PCR and western blot assay. RESULTS AND CONCLUSION: (1) Compared with the control group, the survival time of cyclosporine group and immature dendritic cell group was significantly longer (P < 0.05). (2) In cyclosporine group and immature dendritic cell group, the number of infiltrating mononuclear cells and lymphocytes in the portal area of liver tissue was less, the structure of hepatic lobule was not significantly damaged, and the inflammatory cells in hepatic artery, portal vein and bile duct were significantly less than those in control group, which did not reach the level of severe acute rejection. (3) Compared with the control group, the mRNA expression of IKβ in the cyclosporine group and immature dendritic cell group was increased, while the mRNA expression of SHIP, AKT and IKK significantly decreased (P < 0.05). (4) Compared with the control group, the expression of SHIP and IKβ protein significantly increased, IKK and AKT protein significantly decreased in the immature dendritic cell group (P < 0.05). Compared with the control group, the expression of SHIP and IKK protein significantly decreased, AKT and IKβ protein expression significantly increased in the cyclosporine group (P < 0.05). (5) Results confirm that immature dendritic cells can slow down the severe acute rejection, delay the survival time of liver and reduce the T cell immune response ability of allogeneic liver transplantation.

Individualized induction of immune tolerance and regulatory T cell / 器官移植

The induction of immune tolerance is an essential component and the utmost goal in the field of organ transplantation immunity, which depends upon the recognition and presentation of transplantation antigens, the activation and response of the immune system and other immune essence. However, before successfully inducing immune tolerance, how to carry out individualized induction of immune tolerance in organ transplant recipients to optimize the combination of immunosuppressive agents and individualized treatment and achieve the ideal state of optimal prevention and treatment of immune rejection and minimal adverse reactions, remains to be further resolved by the organ transplantation practitioners. Based on the reports of international core journals, the individualized induction strategy of immune tolerance and the future prospects were reviewed in this article from the following aspects including the mechanism underlying induction of immune tolerance, realization of operational immune tolerance, novel strategy of individualized induction of immune tolerance and application of regulatory T cell in individualized immune tolerance in combination with clinical and laboratory research results of regulatory T cell in our center.

Preparation and In Vitro and In Vivo Evaluation of Sirolimus Ophthalmic Micelles / 中国药学杂志

OBJECTIVE: To prepare sirolimus micelles for the purpose of improving the solubility and applicability in the eye. METHODS: Sirolimus micelles were prepared by thin-film dispersion with Tween-80 and polyoxyethylene stearate (P40S) as carriers. Size distribution, X-ray diffraction spectra, IR spectra and encapsulation efficiency were used to evaluated the character of micelles. The in vitro release and scleral permeability were investigated, and the pharmacokinetics of micelles formulation in rabbit aqueous humor was preliminarily studied. RESULTS: The average particle size was (12.36±0.21) nm, the mean potential was (-6.08±0.98) mV, and the entrapment efficiency was (99.6±0.20)%. Both X-ray diffraction and IR spectra showed that there was almost no free drug. The in vitro release and scleral permeation profiles were in accordance with Higuchi equation. The pharmacokinetic RESULTS: of aqueous humor showed that sirolimus micelles could reach and maintain the therapeutic concentration in the eye for a long time. CONCLUSION: The prepared sirolimus ophthalmic micelles is suitable for ocular administration and is expected to become a promising formulation in the treatment of immune rejection for corneal transplantation.

Effect of electroacupuncture combined with Schwann cell transplantation on limb locomotor ability, regional remyelination and expression of spinal CD4 and CD8 proteins in compressive spinal injury rats / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) combined with transplantation of Schwann cells (SCs) on limb locomotor, myelin sheath repair and expression of CD4 and CD8 in compressed spinal cord injury (CSCI) rats, so as to explore its mechanisms underlying improvement of CSCI. METHODS: A total of 45 female SD rats were randomly divided into normal control, model, EA, Schwann cell (SC) transplantation, and EA＋SC transplantation groups (n＝9 rats in each group). The CSCI model was established by laminectomy at T12-L2 and clip compression. Rats of the SC transplantation group accepted injection of the cultured SC suspension (2×106/6 µL) into the central, upper and lower sites of the injured spinal cord (5 mm in depth) 7-8 days after CSCI modeling. EA (2 Hz) was applied to bilateral "Zusanli" (ST36) and "Sanyinjiao" (SP6) for 10 min, once daily and 6 days a week for 3 weeks. The Basso, Beattie and Bresnahan locomotor rating scale (BBB scale) was used to evaluate the function state of CSCI. Morphological changes of the regional injured tissue were observed under light microscope after H．E. staining. The myelin sheath repair state and survival of SCs were detected by Luxol fast blue (LFB) staining and immunofluorescence histochemistry, and the expression of CD4, CD8 and P0 of the injured spinal cord was detected by Western blot. RESULTS: Compared with the normal control group, the BBB scores at the time-points of 0 d, and 1, 2, and 3 weeks were significantly decreased in the model group (P0.05). LFB staining showed a disordered arrangement of the nerve fibers in the white matter, myelinociasis and obvious decrease of the medullated fibers in the model group, and these situations were relatively milder in both EA and SC transplantation groups and obviously milder in the EA＋SC transplantation group. H．E. staining displayed that the structure of the injured region of the spinal cord was incomplete, accompanied with a large number of defect cavities and neuronal karyopyknosis in the model group, while the structure was relatively clear, with an increase of the normal neurons and fewer neuronal karyopyknosis in the EA＋SC transplantation group. Compared with the normal control group, MBP in the model group was significantly decreased (P0.05), while after the intervention and in comparison with the model group, the expression levels of P0 protein were significantly increased in the EA, SC transplantation and EA＋SC transplantation groups (P<0.05), and was significantly higher in the EA＋SC transplantation group than in both EA and SC transplantation groups (P<0.05). The expression levels of CD4 and CD8 proteins were significantly lower in the EA＋SC transplantation group than in the SC transplantation group (P<0.05)．. CONCLUSION: EA＋SCs transplantation can improve the locomotor function in CSCI rats, which may be related to its effects in increasing the survival of transplanted SCs to promote the remyelination and in reducing the immune rejecting reaction.

hCD47 induces immune tolerance of human macrophages to porcine endothelial cells / 器官移植

Objective To investigate the effect of human CD47 (hCD47) in inducing the immune tolerance of human macrophages to porcine endothelial cells. Methods The porcine iliac endothelial cell (PIEC) transfected with pCDH-hCD47-FLAG plasmid was assigned into the pCDH-hCD47 group, PIEC transfected with pCDH-FLAG empty vector plasmid was assigned into the pCDH group, PIEC transfected with hCD47-dN was assigned into the pCDH-hCD47-dN group and human umbilical vein endothelial cell (HUVEC) was assigned into the positive control group. The cells were co-cultured with human macrophages to detect and analyze the phosphorylation of signal regulatory protein α (SIRPα) and the killing effect of human macrophages on PIEC. Furthermore, porcine arteriae endothelial cell (PAEC) was isolated from GT－/－ and GT－/－/ hCD 47 gene editing pigs to analyze the phosphorylation of SIRPα and the killing effect of human macrophages on PAEC. Results The pCDH group cells could not induce the phosphorylation of SIRPα, whereas the pCDH-hCD47 group cells could activate the phosphorylation of SIRPα after 10 min co-culture with human macrophages, and the degree of phosphorylation of SIRPα was increased with the prolongation of the co-culture time. The pCDH-hCD47-dN group cells failed to activate the phosphorylation of SIRPα. Human macrophages exerted significant effect on killing the pCDH group cells. The pCDH-hCD47 group cells could evidently inhibit the killing effect of human macrophages (P < 0.05), whereas the pCDH-hCD47-dN cells failed to suppress the killing effect of human macrophages. GT－/－-PAEC could not activate the phosphorylation of SIRPα after co-culture with human macrophages. However, GT－/－/hCD47-PAEC significantly activated the phosphorylation of SIRPα after co-culture with human macrophages. Human macrophages exerted significant killing effect on GT－/－-PAEC, and GT－/－/hCD47-PAEC could obviously inhibit the killing effect of human macrophages (P < 0.05). Conclusions The expression of hCD47 in the porcine endothelial cells can inhibit the killing effect of human macrophages on endothelial cells by activating the phosphorylation of SIRPα.

Next generation sequencing screening for human parvovirus B19 infection after liver transplantation and the analysis of related risk factors / 器官移植

Objective To summarize the screening Methods for human parvovirus (HPV) B19 infection after liver transplantation and analyze the related risk factors. Methods Clinical data of 86 recipients were retrospectively analyzed. According to the Results of next generation sequencing (NGS), all recipients were divided into the HPV B19 infection group and control group. Clinical characteristics, treatment regime and clinical prognosis of patients infected with HPV B19 were analyzed. The risk factors of HPV B19 infection were analyzed using univariate and multivariate Logistic regression model by forward LR step method. Results Nine of the 86 recipients developed fever and progressive anemia with unexplained reasons at approximately 2 weeks after liver transplantation. NGS detection demonstrated that HPV B19 was positive and they were diagnosed with pure red cell aplasia (PRCA) caused by HPV B19 infection. After intravenous immunoglobulins (IVIG) was given and the immunosuppressant therapy was adjusted, the hemoglobin levels in all patients were significantly increased. The Results of multivariate analysis revealed that low serum globulin level in peripheral blood at postoperative 7 d [odds ratio (OR) =0.749, P=0.040] and young age (OR=0.937, P=0.038) were the independent risk factors of HPV B19 infection after liver transplantation. Conclusions HPV B19 infection should be considered in relatively young patients with unexplained hemoglobin decline early after liver transplantation. NGS screening is an effective method for early diagnosis of HPV B19 infection. Low serum globulin level in peripheral blood at postoperative 7 d and young age may be independent risk factors of the incidence of HPV B19 infection.

Mechanism of IL-34 in rat corneal allograft rejection / 眼科新进展

Objective To investigate the expression of interleukin-34 (IL-34) after corneal transplantation in rats and its mechanisms of action in immune rejection.Methods SD rats were used as donors and Wistar rats as the recipients to establish corneal transplantation experimental models.Together 60 Wistar rats were randomly divided into 3 groups according to the random number table methods,and rats in B group was transplantated with autologous cornea,while C and D groups received allogeneic corneal transplantation.Another 10 rats were collected as normal controls (A group).After B,C group was treated with postoperative Tarivid eye drops,D group was treated with TobraDex eye drops postoperatively.The survival rate of corneal grafts and survival analysis were evaluated in the 10 rats from the 4 groups,and then the corneal grafts were taken from the rest of rats at the fourteenth day after the operation for hstopathological,immunohistochemical and RT-PCR examinations.Results Survival analysis showed that immune rejection did not occur in A and B group,and the mean survival time (MST) was (26.00 ± 0.97) days in D group,which was much higher than that in group C[(10 ± 1.55) days] (P ＜0.001).HE staining showed that there was obvious inflammatory cell infiltration and neovascularization in the corneal tissue of C group,and there were only a few inflammatory cells and neovascularization in D group.Immunohistochemistry showed that IL-34 protein expression in C group (0.089 4 ± 0.005 6) was significantly higher than that of A group (0.037 7 ± 0.002 3),B group (0.068 4 ±0.004 4) and group D (0.044 5 ± 0.004 5) (F =145.21,P ＜ 0.01),and its expression was mainly located in the epithelial and stromal layer.RT-PCR showed that the expression levels of IL-34,IL-1β,IL-17A,TNF-α mRNA in corneal tissue of C group were significantly higher than those in A,B and D group (all P ＜ 0.05).Conclusion IL-34 may involve in the rejection after corneal transplantation in rats,and TobraDex eye drops can delay the rejection by inhibiting the expression of IL-34 and related signaling pathways.

Immunotherapy of tacrolimus in small volume liver transplantation of rats / 中华肝胆外科杂志

Objective To detect the plasma concentration of tacrolimus and the survival time of rats after small-volume liver transplantation,and to investigate the criteria for immunological rejection after small-volume liver transplantation.Methods Lewis rats and Brown Norway rats were used to establish a small volume and normal liver volume transplantation model,which were divided into 7 groups:whole liver transplantation group (WI),small volume allogeneic liver transplantation group (SI),and whole liver allograft group (WA),small volume allogeneic liver transplantation group (SA),whole liver allograft immunotherapy group (WAT),small volume allogeneic liver transplantation immunotherapy group (SAT),small volume allogeneic liver transplantation immunotherapy modulation group (SATa).Morphological and functional changes of liver tissue were studied postoperatively,AST and tacrolimus plasma concentrations were detected,and survival was recorded.Results Compared with the WA group,the inflammatory cells infiltrated in the portal area of the SA group,the inflammatory changes of the sinusoidal endothelial cells,and the proportion of TCRpositive lymphocytes increased.Four days after transplantation,peripheral blood tests showed that CD4+CD25+ double positive lymphocytes were significantly lower in the allograft group than in the allograft group,and the positive expression rate in the SA group (0.6％) was significantly lower than that in the WA group (1.8％).The differences were statistically significant (P＜0.05).In the SAT group,the blood concentration of tacrolimus was significantly higher than that in the WAT group at each time point (P＜0.05).The blood concentration of tacrolimus in the SATa group was relatively stable,and the plasma concentration of the SATa group was stable.And AST was significantly lower than the SAT group,the differences were statistically significant (P＜0.05).Compared with WAT group,the proliferation and apoptosis rate of hepatocytes in SAT group and SATa group were significantly increased.The proliferation of hepatocytes in SATa group was significantly higher than that in SAT group (P＜0.05).Survival analysis showed that the cumulative survival rate of the WA group was 85.7％,which was significantly higher than that of the SAT group (28.6％).The difference was statistically significant (P＜0.05).The cumulative survival rate of the SATa group was 51.7％.The survival time of WAt group was (57.4±25.0) days,SAT group was (28.0±29.10) days,SATa group was (39.7± 29.0) days,which were longer than untreated groups.The ratio of proliferation to apoptosis (PRA) increased with increasing time of tacrolimus.Regardless of blood concentration,tacrolimus plasma concentration was positively correlated with AST (R =0.758,P＜0.05),indicating RPA was inversely correlated with AST (R=-0.962,P＜0.05).Conclusion The use of tacrolimus significantly prolonged the survival time of small-volume allogeneic liver transplantation rats.Adjusting the amount of tacrolimus under the guidance of tacrolimus plasma concentration and AST serum value equation TD =-0.494TC-0.0035AST+ 260.487 to make the blood concentration relatively stable,it can further extend the allogeneic liver transplantation rat time to live.

Study of high-risk corneal transplantation rejection and the expression of VEGF-C/D / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the expression and the significance of VEGF-C/D in rat cornea after alkali burning as well as the role of lymphangiogenesis in the high-risk corneal transplantation rejection. ●METHODS:The model of alkali burn corneal was made. Different times corneas were taken to electron microscope for vascularization, and examined the expression of VEGF-C/D and VEGFR-3 in l, 3, 5, 7, 14, 28d. The other rat cornea after alkali burn were divided into four parts to penetrate keratoplasty, containing only blood vessels in the cornea ( group A ) , angiogenesis and lymphangiogenesis ( group B ) , lymphangiogenesis degenerating period ( group C ) , angiogenesis degenerating period ( group D ) . ln addition, there are also normal groups ( group N ) to compare the Rl values and survival time of corneal graft. ●RESULTS: Electron microscopy showed that, when the first 7d rat cornea appeared neovascularization after alkali burn, but not lymphangiogenesis. The occurrence of new blood vessels and lymphatic in 2wk. There were no obvious lymphangiogenesis in 5wk and the angiogenesis gradually subside in 8 wk. The expression of VEGF-C/D and VEGFR-3 in the corneas of rats were up-regulated in the third days after the injury, and reached its peaks at 5d. The average survival time of group N, A, B, C, D were (14.25±0.62)d, (9.35±1.02)d, (5.06±1.13)d, (8.71±0.83) d, (9. 44±1. 05)d after transplant cornea. Compared to the rest of the group, group B plant average survival time significantly shortened (P ● CONCLUSION: VEGF - C/D and VEGFR - 3 are expressed significantly after corneal alkali burn. New lymphatic vessels can accelerate high - risk corneal transplantation immune rejection.

Characterization of swine leukocyte antigen class I genes and homology analysis of the similarity to HLA in GGTA1 -/- Wuzhishan minipigs / 中国实验动物学报

Objective This study was aimed to characterize the swine leukocyte antigen( SLA) class I genes of GGTA1 -/ - Wuzhishan minipigs and compare their similarity to human leukocyte antigen( HLA) . It has important implica?tions for understanding the cellular rejection in xenotransplantation. Methods Specimens of ear tissue from six founding GGTA1 -/ - Wuzhishan minipigs were collected, and the SLA class I genes (SLA?1, SLA?3, SLA?2) were amplified by RT?PCR. Purified products were cloned into pEASY?T1 vectors and sequenced, followed by BLAST alignment and using bioin? formatc analysis to characterize the SLA class I genes and compare with the similarity to HLA. Results A total of six al?leles were detected, among them alleles were previously reported (SLA?1?0703,SLA?2?1102, SLA?3?0401, SLA?3?0403), and the other were novel (SLA?1?0401wz01, SLA?2?11wz01). The homology between alleles of SLA class I genes in Wuzhishan minipigs and HLA was from 70?5% to 72?1%. The homology analysis of critical amino acid residues on HLA binding with human CD8 + molecules showed that SLA?1?0401wz01, SLA?1?0703, SLA?2?11wz01, SLA?2?1102 and SLA?3?0401 occurred mutant at amino acid positions 225 and 228 ( T→S,T→M) , whereas the other loci were highly conserved. There was a high homology at amino acid level between SLA?2?11wz01, SLA?2?1102 and HLA class I genes which are NK cell KIRs binding sites. Conclusions The amino acid sequences of SLA class I genes of GGTA1 -/ -Wuzhishan minipigs have a high homology to HLA. From the point of view of cell?mediated xenograft rejection, the amino acid sequences of SLA class I genes of GGTA1 -/ - Wuzhishan minipigs have a high homology to HLA, therefore, Wzhishan minipigs may become a good potential donor for pig?human xenotransplantation.

Functional disruption of human leukocyte antigen II in human embryonic stem cell

BACKGROUND: Theoretically human embryonic stem cells (hESCs) have the capacity to self-renew and differentiate into all human cell types. Therefore, the greatest promise of hESCs-based therapy is to replace the damaged tissues of patients suffering from traumatic or degenerative diseases by the exact same type of cells derived from hESCs. Allo-graft immune rejection is one of the obstacles for hESCs-based clinical applications. Human leukocyte antigen (HLA) II leads to CD4+ T cells-mediated allograft rejection. Hence, we focus on optimizing hESCs for clinic application through gene modification. RESULTS: Transcription activator-like effector nucleases (TALENs) were used to target MHC class II transactivator (CIITA) in hESCs efficiently. CIITA(-/-)hESCs did not show any difference in the differentiation potential and self-renewal capacity. Dendritic cells (DCs) derived from CIITA(-/-)hESCs expressed CD83 and CD86 but without the constitutive HLA II. Fibroblasts derived from CIITA(-/-)hESCs were powerless in IFN-γ inducible expression of HLA II. CONCLUSION: We generated HLA II defected hESCs via deleting CIITA, a master regulator of constitutive and IFN-γ inducible expression of HLA II genes. CIITA(-/-)hESCs can differentiate into tissue cells with non-HLA II expression. It's promising that CIITA(-/-)hESCs-derived cells could be used in cell therapy (e.g., T cells and DCs) and escape the attack of receptors' CD4+ T cells, which are the main effector cells of cellular immunity in allograft.

Prevention of cyclosporine A in different methods of administrations on the rejection after penetrating keratoplasty in rabbit eyes / 中华实验眼科杂志

Background Cyclosporine A (CsA) is an effective drug to prevent rejection response after penetrating keratoplasty (PKP).Appropriate dosage forms and right administrating route is very important for improving the bioavailability of CsA.Objective This study was to investigate the preventing effect of CsA microspheres via subconjunctive and anterior chamber injection and CsA eye drops on immune rejection after PKP.Methods Sixty eyes of 60 clean adult New Zealand white rabbits served as receipts,and 60 eyes of 30 clean adult pigmented rabbits were used as the donors.The receipts were randomized into the blank control group (only PKP group),subconjunctival CsA injection group,subconjunctival vector injection group,anterior chamber CsA injection group,anterior chamber vector injection group,and 1％ CsA eye drops group.PKP was performed on the all rabbits,and then the CsA microsphere (0.1 ml,12 g/L) or blank microsphere (0.1 ml) were respectively administered in the corresponding groups.The corneal grafts were examined by slit lamp microscope regularly,and rejection index (RI)was calculated based on the corneal opacity,edema and neovascularization.The intraocular pressure (IOP) of operative eyes was measured by Tono-pen tonometer before operation,3 days,1 week,2 weeks,3 weeks,1 month,2 months and 3 months after operation,respectively.Histopathological examination on corneal grafts was performed 1month and 3 months after operation.Results The IOP of all the rabbits lowed after operation,but there was no statistical difference in different time points and various groups (Ftime =29.210,P =0.000; Fgroup =0.254,P =0.938).The grafts of the blank control group,subconjunctival vector injection group and the anterior chamber vector injection group showed varied degrees of corneal opacity and neovascularization 2-3 weeks after operation,and the degree of graft opacity was aggravated obviously in the fourth week,with the RI 8.60±1.52,8.60±0.55 and 8.80±0.84 individually.Neovascularization of the grafts in the subconjunctival CsA injection group,anterior chamber CsA injection group,and 1％ CsA eye drops group was found 3 weeks after operation,and the RI were 4.40±0.89,3.20±0.84 and 3.00±0.71,showing a significantly lower than that of the control groups (P＜0.05).Mild inflammatory response of the grafts was seen in the anterior chamber CsA injection group,but it was lightened over time.The histopathological examination revealed obvious thickening of corneal grafts,neovascularization and infiltration of lots of inflammatory cells in the blank control group,subconjunctival vector injection group and the anterior chamber vector injection group.However,only slight new blood vessels and inflammatory response were seen on the subconjunctival CsA injection group,anterior chamber CsA injection group,and 1％ CsA eye drops group.Conclusions Administration of CsA in different methods can prevent the immune rejection after PKP in rabbit eyes,the effect of giving drugs via anterior chamber injection is better than that via subconjunctive pathyway.

Inhibitory effect of neutralizing interleukin-17 antibody on corneal allograft rejection / 中华实验眼科杂志

Background Interleukin-17 (IL-17)is a potent pro-inflammatory cytokine and plays a pathogenic role in autoimmune disease.It was confirmed that IL-17 is implicated in allograft rejection of many transplanted organs.Recent studies have foensed on the effect of IL-17 antagonists on allograft rejection.Objective This study aimed to investigate the inhibitory effect of anti-mouse IL-17 monoclonal antibody (mAb) on corneal allograft rejection.Methods Twenty-five 8 to 10-week-old C57BL/6 mice and 50 BALB/c mice were collected.Donor cornea grafts with 2 mm diameter from 25 C57BL/6 mice was transplanted to 50 eye of BALB/c mice to establish a model of corneal transplantation.The recipients were randomized into 2 groups,and neutralizing mouse IL-17antibody or isotype control antibody was intraperitoneally injected immediately after transplantation for experimental treatment,respectively.Allografts were scored clinically at appropriate time points after treatment based on Plskova criteria,and ≥5 was confirmed as rejection.Infiltrating cells in corneal graft were detected qualitatively and quantitatively by immunohistochemistry and reverse transcription-PCR separately.The cytokine levels of T helper type 1 (Th1),Th2,and Th17 in recipients' spleen wer(c) analyzcd by ELISA.The use of the animals followed the Statement of ARVO.Results Compared with the isotype control antibody group,the survival of grafts was improved in the IL-17mAb group(P＜0.05).The levels of neutrophile granulocyte mRNA,CD4+ and CD8+ T lymphotes mRNA were 2.22±0.10,1.64±0.04 and 1.32±0.10 in the IL-17 mAb group,showing a significant decline in comparison with those of the isotype control antibody group(3.61 ±0.08,2.69±0.06 and 2.17±0.04) (P=0.000,0.000,0.000).Interferon-γ(IFN-γ),IL-12 p40 and IL-17 concentrations in recipients ' splenocytes were (529.80 ± 13.83) ng/L,(539.58 ±10.74) ng/L and(173.70±8.11)ng/L in the IL-17 mAb group,and thosc in the isotype control antibody group were (741.48± 10.51) ng/L,(1156.90 ± 69.93) ng/L and (366.13± 7.93) ng/L,with significant differences between them (P=0.000,0.001,0.000).Conclusions Neutralization IL-17 bioactivity inhibits mouse corneal allograft rejection to a certain extent.

Preliminary research on 16 peripheral blood monocyte immune rejection relative genes expression after heart transplantation with fluorescent quantitation RT-PCR techniques / 中国综合临床

Objective To construct the micro-invasive immune rejection monitoring methods with peripher-al blood mononuclear cell gene expression detection and evaluate the clinic rejection estimation value. Methods The SYBR Green Ⅰ was used as fluorescent dye and the GAPDH as house keeping gene control in the quantitatiun RT-PCR technique to observe the 16 immune rejection relative genes expression features after heart transplantation. results were also compared with that of the normal people. Results The 16 immune rejection relative genes expres-sion were no different between normal people and the transplantation recipients before surgery (P>0.05). After heart transplantation the expression of ITGA4, FKB, ILI R-2 up regulated and the level of PF4、ITGAM、TGFβ1、 RHOU down regulated. The results were similar with the clinic observation that the immune rejection often occurs in the first 3 months after heart transplantation. It implied that these 7 genes may play an important role in the acute im-mune rejection after transplantation. Conclusion The real time quantitation RT-PCR methods were constructed suc-cessfully to detect the multiple immune relative genes expression and is of chnic applicable.

Research of immunoreaction of heterogeneic articular cartilage extracellular matrix derived scaffold / 中国矫形外科杂志

0.05).[Conclusion]Porcine articular cartilage extracellular matrix derived scaffold used in heterogenic transplantation or allograft have no immunologic rejection.This research provides a feasibility for application of xenogenic acellular cartilage to clinic.

Immunosuppression of tripterygium wilfordii hoof-F on acute rejection of bone xenograft in mice / 吉林大学学报(医学版)

Objective To investigate the effect of tripterygium wilfordii hoffF on acute immune rejection of bone xenograft in mice.Methods Male Neijiang swine and BALB/c mice were used as donor and receipts,respectively.Forty mice were divided into experimental group and control group at random.The mice in experimental group were administered intragastrially with tripterygium wilfordii hoofF before bone xenograft.At two weeks postoperatively,CD4+,CD8+,CD4+/CD8+ T lymphocytes in spleen were detected by flow cytometry,and the levels of IL-2,IFN-? and IL-4 in blood were analyzed by quantitative ELISA assay.And the infiltration of lymphocytes and macrophages surrounding soft tissues of bone graft was also observed in order to evaluate the immunosuppression of tripterygium wilfordii hoof-F.Results At two weeks postoperatively,compared with control group,CD4+,CD8+,CD4+/CD8+ T lymphocytes in spleen,the levels of IL-2,IFN-? and IL-4 in blood of mice in experimental group were significantly decreased(P

Heterologous corneal endothelial cell transplantation: human corneal endothelial cell transplantation in Lewis rats

A heterologous corneal endothelial transplantation was attempted using human endothelial cells and a Lewis rat penetrating keratoplasty model. Cultured human endothelial cells were seeded to a Lewis rat cornea, which was denuded of its endothelium. When grafted into the syngeneic Lewis rat, the graft remained clear for at least five days, and then became opaque and edematous because of immune rejection reaction. In contrast, corneas denuded of their endothelium became opaque and edematous immediately after transplantation. These results demonstrate that transplanted endothelial cells have enough antigens to induce rejection reaction even though they have the functional capacity to deturge the cornea.