Xiong Zou

HLA‐G on peripheral blood CD4+ T lymphocytes: a potential predictor for acute renal rejection

HLA‐G Expression in grafts and serum has been shown to improve graft acceptance. However, its expression on peripheral blood lymphocytes (PBLs) during acute rejection (AR) remains unknown. In this study, we serially monitored HLA‐G expression on CD4+ and CD8+ PBLs of 66 recipients undergoing renal transplantation using flow cytometry at different time points before and after transplantation, as well as during AR episode. In stable recipients, HLA‐G expression on CD4+ PBLs declined during the first week after transplantation and increased continuously with immunosuppressive therapy. Then, expression declined gradually after 1 month and remained at a higher level compared with pretransplantation. When AR occurred, HLA‐G expression decreased significantly compared with the stable level. In three recipients suffering from recurrent rejection, it remained at a low level despite impact immunosuppressive treatment. With mix lymphocyte assay, HLA‐G+ CD4+ T cells showed inhibitory role on proliferation of peripheral blood mononuclear cell. HLA‐G expression on CD8+ PBLs was almost undetectable at different time points in the recipients and healthy controls. Our results suggest that HLA‐G on CD4+ PBLs might provide a potential marker for the early diagnosis of renal AR and for the immunosuppressive status of recipients.

HLA Class I (ABC) Upregulation on Peripheral Blood CD3+/CD8+ T Lymphocyte Surface is a Potential Predictor of Acute Rejection in Renal Transplantation

Background. Renal transplantation is currently the prevalent therapy for most patients with end-stage renal disease. No clinical markers for such rejection have been universally accepted. We aimed to investigate the possibility of use of human leukocyte antigen (HLA) class I (ABC) on peripheral blood CD3+/CD8+ T lymphocytes as a marker of acute rejection. Methods. For recipients undergoing renal transplantation from September 2007 to November 2008, peripheral blood samples were obtained pretransplantation and at days 3 and 7 posttransplantation when the patients were still hospitalized and at weeks 2 and 3 and months 1, 2, 3, and 6 posttransplantation. For patients with fever, lumbodynia, gross hematuria, or oliguria after transplantation, blood samples were collected immediately before and at days 3 and 7 after the administration of anti-inflammatory regents. The level of HLA class I (ABC) on peripheral-blood CD3+/CD8+ T lymphocytes was measured on flow cytometry. Results. For the 79 transplant recipients, the level of HLA class I (ABC) on peripheral-blood CD3+/CD8+ T lymphocytes was consistently elevated during the first 3 weeks after transplantation, declined gradually to pretransplantation levels, then tapered off and remained stable. Patients experiencing acute rejection (AR) or not after transplantation did not differ in level of HLA class I (ABC) up to 6-month follow-up, except at days 14 and 21 after transplantation, when the level was higher for patients experiencing AR (P<0.01). Conclusions. Upregulation of HLA class I (ABC) on peripheral-blood CD3+/CD8+ T lymphocytes could be used as an accurate and reliable predictor of AR after renal transplantation.

Human leukocyte antigen class I on peripheral blood mononuclear cells as a non‐invasive biomarker for esophageal cancer

Esophageal cancer (EC) is a common malignant cancer threatening peoples health. There are no universally accepted parameters for its early diagnosis. The aim of this study was to observe the expression of human leukocyte antigen class I (HLA-I) on peripheral blood mononuclear cells (PBMCs) of EC patients and in individuals of high-incidence area of EC so as to evaluate the feasibility of using this parameter as a potential non-invasive biomarker for the early diagnosis of EC. The present study enrolled 58 pathological confirmed EC patients, 46 patients with benign esophageal disease, and 65 healthy volunteers. Expression levels of HLA-I protein and mRNA on PBMCs were determined by flow cytometry and quantitative reverse transcriptase polymerase chain reaction, respectively. Then, 181 volunteers from Lijiadian, a village with high morbidity of EC, and 153 age- and gender-matched health volunteers were involved in this study to observe HLA-I expressions in individuals of high-incidence area of EC. Compared with benign esophageal disease and health volunteers, the expressions of HLA-I protein and mRNA on PBMCs of EC patients are significantly decreased, especially in patients with stage III and IV EC, but was not influenced by patients age and gender. Furthermore, individuals of high-incidence area of EC also show downregulated HLA-I protein, but not mRNA, expression on PBMCs. Altogether, HLA-I expression on PBMCs of EC patients and individuals from high-incidence area of EC is downregulated, and this parameter might be used as a potential predictor of EC.

Expression of the genes encoding human leucocyte antigens-A, -B, -DP, -DQ and -G in gastric cancer patients.

This study compared the expression of the genes encoding human leucocyte antigens (HLA)-A, -B, -DP, -DR and -G in peripheral blood mononuclear cells (PBMCs) in gastric cancer patients and healthy controls. Using reverse transcription– polymerase chain reaction, levels of classical HLA-A, -B, -DP and -DR and non-classical HLA-G mRNA were studied in 43 gastric cancer patients and 22 controls. In addition, the levels of HLA-A,B,C and -G antigens on the surface of PBMCs were measured in 30 gastric cancer patients and 15 controls using flow cytometry. The mean fluorescence intensity of HLA-A,B,C antigen in the gastric cancer group was significantly lower than in controls. The HLA-G antigen was mainly present on CD4+CD8− T-lymphocytes. The percentage of CD4+CD8− T-lymphocytes positive for HLA-G antigen was significantly lower in the gastric cancer group compared with the healthy controls. Levels of HLA-A, -B and -G mRNA in the gastric cancer group were significantly lower than in controls. The HLA-G mRNA levels were significantly lower in gastric cancer of histological grades III and IV than in grades I and II. These data may provide a novel diagnostic and research tool for gastric cancer.

HLA class I expressions on peripheral blood mononuclear cells in colorectal cancer patients

ObjectiveTo investigate the expression change of human leukocyte antigen (HLA) class I on human peripheral blood mononuclear cells (PBMCs) at both mRNA and protein levels, and to evaluate its roles in the development of colorectal cancer (CRC).MethodsIn the present study, 50 patients with CRC, 35 patients with benign colorectal lesion and 42 healthy volunteers were enrolled. Expression levels of HLA class I mRNA and protein were determined using real-time quantitative reverse transcription PCR (RT-PCR) and flow cytometry analysis, respectively.ResultsThe expression levels of HLA class I mRNA and proteins were not influenced by age and gender. The relative ratios of HLA class I mRNA were 0.99±0.27 in healthy controls, 0.76±0.19 in benign patients, and 0.48±0.21 in CRC patients. Mean fluorescence intensities of HLA class I were 145.58±38.14 in healthy controls, 102.05±35.98 in benign patients and 87.44±34.01 in CRC patients. HLA class I on PBMCs was significantly down-regulated at both mRNA and protein levels in patients with stage III and IV CRC. CRC patients with lymph node metastasis also showed a decreased HLA class I expression at protein level.ConclusionHLA class I expressions on PBMCs are associated with staging of CRC and lymph node metastasis. Monitoring the expression of HLA class I on PBMCs may provide useful information for diagnosis and metastasis judgement of CRC.

Acute Rejection Correlates with Expression of Major Histocompatibility Complex Class I Antigens on Peripheral Blood CD3+CD8+ T-Lymphocytes following Skin Transplantation in Mice

This study investigated major histocompatibility complex class I (MHC-I) antigen expression on peripheral blood T-cells after transplantation to assess its potential as an early marker of acute graft rejection (AGR). Using a mouse model with or without immunosuppressive treatment, the expression of MHC-I antigens on CD3+CD8+ T-lymphocytes was assessed by flow cytometry following syngeneic graft (n = 138) or allograft (n = 138) skin transplantation. The occurrence of AGR was assessed by examining the degree of lymphocyte and monocyte infiltration in transplant biopsies. During AGR, expression of MHC-I antigens increased significantly compared with pre-transplant levels in the allograft group, even with immunosuppressive treatment. The highest expression of MHC-I antigens occurred 5 – 6 days before macroscopic rejection. These results suggest that expression of MHC-I antigens on peripheral blood CD3+CD8+ T-lymphocytes could be used as an early marker for predicting AGR.

Imbalance of Interleukin-18 and Interleukin-18 Binding Protein in Children with Henoch-Schönlein Purpura

The balance between interleukin-18 (IL-18) and its endogenous antagonist, IL-18 binding protein (IL-18BP), was evaluated in children with Henoch-Schönlein purpura (HSP). Plasma IL-18 and IL-18BP levels and peripheral blood mononuclear cell IL-18 mRNA expression were significantly higher in patients with active HSP (n = 30) than in healthy controls (n = 20); IL-18BP mRNA expression was similar in active HSP and controls. Plasma levels and mRNA expression of IL-18 and IL-18BP in patients in remission (n = 19) were similar to those in controls. The ratios of IL-18 / IL-18BP plasma levels and IL-18 / IL-18BP mRNA levels in active HSP were significantly higher than in patients in remission and healthy controls. Thus, adequate IL-18BP to block the proinflammatory activity of IL-18 may not be present in active HSP and regulation of the IL-18 / IL-18BP balance might provide a potential therapeutic strategy.

The amount of surface HLA-I on T lymphocytes decreases in breast infiltrating ductal carcinoma patients.

Human leucocyte antigen class I (HLA-I), which includes HLA-A, -B and -C, is an essential immune factor participating in the antitumour immune response. The changes in HLA-I expression in peripheral blood T lymphocytes in cancer patients have yet to be defined. This study examined the expression of HLA-I on CD4+ and CD8+ T lymphocytes in female patients with stage I — IV breast infiltrating ductal carcinoma, benign breast tumour diseases (mammary intraductal papilloma or breast fibroadenoma), and in healthy controls. HLA-I was down-regulated on CD4+ T lymphocytes from patients with stage III and IV cancer, and on CD8+ T lymphocytes in patients with stage I — IV cancer compared with healthy controls. HLA-I expression in T lymphocytes may contribute towards immune-balance disorders in tumour patients.

Lymphocytic HLA-A mRNA is a reliable indicator of acute rejection in renal transplantation.

BACKGROUND Rejection in renal transplantation is the most frequent event causing transplant failure. It is important to identify parameters to predict rejection, which are helpful in a timely fashion. METHODS Fifty-nine renal transplant recipients were divided into two groups: group 1 (stable renal function) and group 2 (acute rejection episodes). The levels of HLA-A mRNA in peripheral blood lymphocytes (both pre- and posttransplantation) were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) with glucose-6-phosphate dehydrogenase (G6PDH) as an internal reference. The TEST software was used to analyze the relative expressions of HLA-A mRNA. RESULTS There was no statistical significance between features of the two groups pretransplant versus normal controls. Posttransplant, the HLA-A mRNA levels decreased significantly compared to those of pretransplant and normal control individuals. The levels of HLA-A mRNA among the 10 patients with acute rejection episodes were significantly increased. There was no significant change in the lymphocyte populations in the early stage of an acute rejection episode compared with the prerejection value. CONCLUSION HLA-A mRNA expression was strongly correlated with immune status. The HLA-A mRNA levels may provide an effective and reliable indicator to predict acute rejection episodes in renal transplantation.