Allo-cell transplant rejection is associated with class II major histocompatibility complex (MHC II), while its transactivator (namely C II TA) regulates MHC II molecules expression strictly and exclusively. The aim of this study was to investigate the inhibiting effect of C II TA anti-sense RNA on MHC II expression. The cDNA for anti-sense RNA recognizing the 114-523 sequence of C II TA (arC II TA) was obtained from Raji cell by RT-PCR, and then inserted into the pcDNA3.1B plasmid (pcDNA3.1B-arC II TA, pD-arC II TA). Raji cells were transfected stably with pD-arC II TA, classic MHC II antigen (HLA-DR, -DP, -DQ) expression was assayed by flow cytometry (FCM). mRNA abundance of C II TA, invariant chain and classic MHC II were detected by RT-PCR. The results showed that compared with control (sense C II TA), the expression inhibition of HLA-DR, -DP, -DQ on pD-arC II TA positive Raji cell was 65.93%, 54.14%, 68.32% respectively. The mRNA contents of C II TA, invariant chain and classic MHC II also decreased. In conclusion, arC II TA inhibited C II TA and thus the family of MHC II molecules were regulated by it, therefore these results provide a novel approach for the control of graft versus host diseases.
Cell Line, Tumor ; Flow Cytometry ; Graft Rejection ; genetics ; prevention & control ; HLA-DP Antigens ; biosynthesis ; genetics ; HLA-DR Antigens ; biosynthesis ; genetics ; Humans ; Nuclear Proteins ; genetics ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; Transfection

A study of histopathologic changes, ultrastructure, and expression of the HLA-Dr antigen within the giant papillae of patients with giant papillary conjunctivitis was performed to determine whether cell-mediated immune response is related to this condition. Conjunctival giant papillae from ten patients with giant papillary conjunctivitis were examined by light and electron microscopy and by the indirect immunofluorescent staining method with HLA-Dr antibody. The infiltration of eosinophilic neutrophils and granules was most prominent, with the occasional infiltration of mast cells, as shown by light microscopy. The infiltration of activated fibroblasts and Langerhans cells was also observed. Cells expressing HLA-Dr antigen were also markedly increased, as shown by the immunofluorescent method. These findings suggest that delayed hypersensitivity may, along with the processes of antigen presentation by HLA-Dr-expressing (including Langerhans) cells, contribute to the pathogenesis of giant papillary conjunctivitis.
Adolescent ; Adult ; Child ; Conjunctiva/metabolism/*ultrastructure ; Conjunctivitis, Allergic/*metabolism/*pathology ; Fluorescent Antibody Technique, Indirect ; HLA-DR Antigens/*biosynthesis ; Humans ; Immunity, Cellular ; Langerhans Cells/*ultrastructure ; Microscopy, Electron ; Middle Aged

OBJECTIVETo investigate the changes of human leukocyte antigen DR (HLA-DR) expression in the monocytes of elderly patients with sepsis.

METHODSA total of 213 elderly patients were divided into 4 groups according to the criteria defined by the American College of Chest Physicians/ Society of Critical Care Medicine (ACCP/SCCM), namely the non-sepsis group (group A), sepsis group (group B), severe sepsis group (group C) and septic shock group (group D). Another 60 healthy subjects were recruited as the control group. HLA-DR expression in the monocytes from each subject was determined by flow cytometry, and the HLA-DR levels were compared between the groups.

RESULTSCompared with that of the healthy control, HLA-DR expression was significantly increased in group A and lowered in group D. No significant differences were found between the control group and groups B and C. HLA-DR expression was significantly different between groups A, B, C, and D (P<0.01), increasing in the order of group D (32.74-/+13.4)%, group C (61.9-/+14.29)%, group B (69.99-/+12.8)%, and group A (85.06-/+15.37)%.

CONCLUSIONSAlthough with multiple organ diseases and repeated or long-term hospitalization, the non-septic elderly patients do not show lowered expression of HLA-DR. In the elderly patients with sepsis and severe sepsis, HLA-DR expression is not a sensitive index, but obviously lowered HLA-DR expression may serve as a specific indicator in elderly patients with septic shock.

Aged ; Aged, 80 and over ; Biomarkers ; analysis ; Female ; Flow Cytometry ; HLA-DR Antigens ; analysis ; biosynthesis ; Humans ; Leukocytes, Mononuclear ; immunology ; Male ; Sepsis ; immunology ; Shock, Septic ; immunology

OBJECTIVEThe purpose of this study was to investigate the transcription and expression levels of human leukocyte antigen-DR at different stages of oral squamous cell carcinomas.

METHODSThe specific monoclonal antibody and beta-locus specific oligonucleotide probes of human leukocyte antigen-DR were employed in this study. A total of 32 primary oral squamous cell carcinomas, 15 metastatic focuses and 26 histologically normal oral epithelia, were detected for the presence of the human leukocyte antigen-DR molecule by using immunohistochemistry and in situ hybridization.

RESULTSThe human leukocyte antigen-DR expression of the primary focuses was significantly higher than that of the normal epithelia (P < 0.05), but their expression did not show statistically difference between the metastatic focuses and the normal epithelia. The immunohistochemistric results were identical with those of in situ hybridization.

CONCLUSIONThe abnormal higher expression of the HLA-DR is a common character of primary oral squamous cell carcinomas, but it may be not relevant to the metastasis of tumors.

Carcinoma, Squamous Cell ; genetics ; immunology ; HLA-DR Antigens ; biosynthesis ; genetics ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Mouth Mucosa ; immunology ; Mouth Neoplasms ; genetics ; immunology

OBJECTIVETo investigate the effect of a tight control of blood glucose by intensive insulin therapy on human sepsis, and to explore the potential mechanism of the intensive insulin therapy.

METHODSEligible patients were randomized by a blinded pharmacist to receive tight control of blood glucose by intensive insulin therapy (maintenance of blood glucose at a level between 4.4 and 6.1 mmol/L) or to receive conventional treatment (maintenance of glucose at a level between 10.0 and 11.1 mmol/L). The expression of HLA-DR on peripheral monocytes was measured in 54 patients by flow cytometry on 24 h, 3 d, 5 d, 7 d, 10 d and 14 d of intensive care in parallel with serum c-reactive protein (CRP), severity of the disease (APACHE II score, SOFA score) and clinical data collection.

RESULTSPatients receiving intensive insulin therapy were less likely to require prolonged mechanical ventilation. Tight control of blood glucose significantly reduced the number of days during which leukopenia or leukocytosis and the days with hypo- or hyperthermia (P < 0.05). Hypoglycemia occurred in 3 patients (10.7%) in the tight control of blood glucose group. There were no instance of hemodynamic deterioration or convulsions. Compared with the conventional treatment, tight control of blood glucose also increased the HLA-DR expression of peripheral monocytes, and there were significantly difference on 3 d, 5 d and 7 d (P < 0.05). Whereas it suppressed the elevated serum CRP concentrations, there was significantly difference on 7 d (P < 0.05).

CONCLUSIONSTight control of blood glucose by intensive insulin therapy expedited healing of human sepsis, and increased the HLA-DR expression of peripheral and suppressed the elevated serum CRP. So, it is necessary to use insulin to strict control the glucose levels in human sepsis.

Blood Glucose ; metabolism ; C-Reactive Protein ; metabolism ; HLA-DR Antigens ; biosynthesis ; Humans ; Hyperglycemia ; drug therapy ; etiology ; metabolism ; Hypoglycemic Agents ; therapeutic use ; Insulin ; therapeutic use ; Sepsis ; complications

The present study was purposed to investigate the relation and difference of expression phase between class II transactivator (CIITA) and HLA-DR antigens after IFN-gamma induction, and the inhibition of CIITA and HLA-DR by STAT1-alpha antisense oligonucleotides (STAT1-alpha AS); and to explore the potential effect and significance of CIITA and STAT1-alpha AS in transplantation immunity. T lymphocytes from peripheral blood of healthy subjects were incubated with IFN-gamma at different doses. RT-PCR was used to detect CIITA mRNA and Western blot was used to analyze HLA-DR antigen. Then the optimum dose of IFN-gamma was chosen for the experiment. CIITA mRNA and HLA-DR antigen were detected at various time points. Different doses of STAT1-alpha AS and sense oligonucleotides (STAT1-alpha S) were added to T lymphocytes followed by IFN-gamma. After incubation with IFN-gamma, the expression of CIITA mRNA and HLA-DR was detected once again. The results showed that CIITA mRNA was detectable at 5 hours after IFN-gamma incubation and reached the peak at 14 hours, then declined, but the CIITA mRNA was still found at 23 hours. HLA-DR antigen was detectable at 28 hours after IFN-gamma incubation and reached a peak at 52 hours, then declined. CIITA mRNA expression was positively correlated to HLA-DR expression, and was earlier than the latter. The expression of CIITA mRNA in the AS groups was significantly lower than that in the control group after 5 micromol/L, 10 micromol/L and 20 micromol/L STAT1-alpha AS treatment (P < 0.01). The expression of CIITA mRNA in the S groups was higher than that in the AS groups (P < 0.01), but there was no significant difference between the S group and the control group. The expression of HLA-DR antigen was significantly inhibited by STAT1-alpha AS, and the expression level of HLA-DR protein in the AS group was about 64.3% of that in the control group (P < 0.01), while there was no significant difference in HLA-DR expression between the S group and the control group. The changes in HLA-DR expression were similar to those in CIITA expression after STAT1-alpha AS treatment. It is concluded that CIITA expression is positively correlated with HLA-DR expression, and was detectable earlier than that of latter after IFN-gamma incubation. Stat1-alpha antisense oligonucleotides may have a sequence-specific inhibiting effect on the expression of CIITA and HLA-DR antigen after IFN-gamma incubation in vitro culture, and can prevent T lymphocyte activation. CIITA may play an important role in pathogenesis of transplantation immunity.
Cells, Cultured ; HLA-DR Antigens ; biosynthesis ; genetics ; Humans ; Interferon-gamma ; pharmacology ; Nuclear Proteins ; biosynthesis ; genetics ; Oligonucleotides, Antisense ; antagonists & inhibitors ; RNA, Messenger ; biosynthesis ; genetics ; STAT1 Transcription Factor ; antagonists & inhibitors ; T-Lymphocytes ; cytology ; Trans-Activators ; biosynthesis ; genetics ; Transplantation Immunology ; immunology

To investigate the influence and mechanisms of CD47 engagement by its soluble mAb B6H12 on the maturation and function of cultured dendritic cells (DCs), monocyte-derived DCs were propagated in granulocyte-macrophage colony stimulating factor (GM-CSF) combined with lipopolysaccharide (LPS) and interleukin (IL)-4, in the presence or absence of soluble anti-CD47 monoclonal antibodies (anti-CD47 mAbs, B6H12). The characteristic morphology of DCs was identified by using the transmission electron microscopy. Flow cytometry was used to detect the cell surface phenotypes. The concentration of IL-12 P70 in supernatant was measured by ELISA. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by BrdU-ELISA. Electrophoretic mobility shift assay (EMSA) was applied to examine the activity of NF-kappaB. The results indicated that the anti-CD47 mAbs markedly suppressed the expression of CD80, CD86, CD83, CD1a and HLA-DR on the surface of DCs (P < 0.05). The data of the mixed leukocyte reaction and IL-12 P70 production were consistent with the results by flow cytometry (P < 0.01). Pre-exposure to B6H12 mAb during the development of DCs resulted in a dramatic depletion of the DNA binding activity of NF-kappaB toward nucleus protein. Moreover, such an inhibition effect seemed to be dose dependent. In conclusion, the soluble mAb B6H12 inhibits the expression of the costimulatory molecules and MHCII molecules on the DCs. The antigen-presenting function of DCs was also impaired by B6H12. And these modulations are closely related with the depletive DNA binding activity of NF-kappaB. It is suggested that the soluble B6H12 exerts a negative effect on the maturation and function of in vitro cultured DCs due to inhibition of NF-kappaB binding activity.
Antibodies, Monoclonal ; immunology ; pharmacology ; Antigens, CD ; biosynthesis ; Antigens, CD1 ; biosynthesis ; B7-1 Antigen ; biosynthesis ; B7-2 Antigen ; biosynthesis ; CD47 Antigen ; immunology ; Cell Size ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; metabolism ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; HLA-DR Antigens ; biosynthesis ; Humans ; Immunoglobulins ; biosynthesis ; Interleukin-12 ; metabolism ; Membrane Glycoproteins ; biosynthesis ; NF-kappa B ; metabolism ; Oligonucleotides ; metabolism ; Protein Binding

Staphylococcus aureus may perform an crucial function in atopic dermatitis (AD), via the secretion of superantigens, including staphylococcal enterotoxins (SE) A or B, and toxic shock syndrome toxin-1 (TSST-1). Dysregulated cytokine production by keratinocytes (KCs) upon exposure to staphylococcal superantigens (SsAgs) may be principally involved in the pathophysiology of AD. We hypothesized that lesional KCs from AD may react differently to SsAgs compared to nonlesional skin or normal skin from nonatopics. We conducted a comparison of HLA-DR or CD1a expression in lesional skin as opposed to that in nonlesional or normal skin by immunohistochemistry (IHC). We also compared, using ELISA, the levels of IL-1alpha, IL-1beta, and TNF-alpha secreted by cultured KCs from lesional, nonlesional, and normal skin, after the addition of SEA, SEB and TSST-1. IHC revealed that both HLA-DR and CD1a expression increased significantly in the epidermis of lesional skin versus nonlesional or normal skin in quite a similar manner. IL-1alpha, IL-1beta, and TNF-alpha secretion was also significantly elevated in the cultured KCs from lesional skin after the addition of SsAgs. Our results indicated that KCs from lesional skin appear to react differently to SsAgs and increased proinflammatory cytokine production in response to SsAgs may contribute to the pathogenesis of AD.
Tumor Necrosis Factor-alpha/biosynthesis/genetics ; *Superantigens/administration & dosage/immunology ; Staphylococcus aureus/*immunology/pathogenicity ; Male ; Keratinocytes/immunology/*microbiology ; Interleukin-1/biosynthesis/genetics ; Inflammation Mediators/metabolism ; Humans ; HLA-DR Antigens/metabolism ; Enterotoxins/administration & dosage/immunology ; Dermatitis, Atopic/etiology/immunology/*microbiology ; DNA, Complementary/genetics ; Case-Control Studies ; Base Sequence ; Bacterial Toxins/administration & dosage/immunology ; Antigens, CD1/metabolism ; Adult

OBJECTIVETo probe the effect and mechanism of Compound Glycyrrhizin in treating AIDS.

METHODSForty AIDS patients were randomly divided into a treatment group and a control group, both treated with HAART. In addition, the former was given Compound Glycyrrhizin for 6 months, and the CD4+ T count and the expressions of CD8+ and HLA-DR on the surface of peripheral blood lymphocytes (PBL) were studied before and after the treatment.

RESULTSAfter 6 months of treatment, the expressions of CD8+ and CD38+ of PBL in the treatment group [(6.6 +/- 2.1)%] were found lower than in the control [(11.4 +/- 3.8)%] (t = 5.043, P < 0.01) and CD4+ T count [(243.6 +/- 91.2) x 10(6)/L vs (170.8 +/- 55.7) x 10(6)/L] rose more significantly (t = 3.045, P < 0.01).

CONCLUSIONCompound Glycyrrhizin can lower the expression of active T-lymphocyte subset, inhibit HIV and help immune reconstitution.

ADP-ribosyl Cyclase 1 ; biosynthesis ; Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adolescent ; Adult ; Anti-HIV Agents ; therapeutic use ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Female ; Glycyrrhizic Acid ; therapeutic use ; HLA-DR Antigens ; biosynthesis ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; drug effects ; immunology