1.Effects of anti-infection treatment on expressions of HLA-DR and CD86 in dendritic cells in rabbit buccal VX2 squamous cell carcinoma tissue with inflammation.
Lin ZHANG ; Nini ZHANG ; Guilin HUANG ; Xiaohua HU ; Jie YI ; Li YAO
West China Journal of Stomatology 2015;33(2):141-144
OBJECTIVETo investigate the effects of anti-infection treatment on the expressions of antigen-presenting-related membrane-surface molecules HLA-DR and CD86 in dendritic cells (DCs) in rabbit buccal VX2 squamous cell carcinoma tissue complicated with local inflammation.
METHODSRabbit buccal VX2 squamous cell carcinoma with local inflammation models that were established by inflammation was induced by inoculation VX2 tumor, mechanical trauma, and drinking of milk with high sugar viscosity. The animals were divided into four groups. Group A (n=12): rabbit buccal VX2 squamous cell carcinoma with local inflammation, procaine penicillin was intramuscularly given, and tinidazole tablets were given by gavage for three consecutive days. Group B (n = 12): rabbit buccal VX2 squamous cell carcinoma with local inflammation, normal saline was intramuscularly given, and aspirin were given by gavage for three consecutive days. Group C (n = 12): rabbit buccal VX2 squamous cell carcinoma with local inflammation, normal saline was given intramuscularly and by gavage for three consecutive days. Group D (n = 10): rabbit buccal VX2 squamous cell carcinoma, normal saline was given intramuscularly and by gavage for three consecutive days. All the rabbits were sacrificed for collection of tumor specimens, and the expression levels of membrane-surface HLA-DR and CD86 in DCs of tumor specimens were detected viaflow cytometry.
RESULTSThe positive expression rate of HLA-DR and the double positive expression rate of HLA-DR and CD86 were group A > group D > group B > group C. The positive expression rate of CD86 were group A > group D > group B and group C (P < 0.05).
CONCLUSIONAnti-infection treatment significantly increased the expressions of HLA-DR and CD86 in DCs of rabbit buccal VX2 squamous cell carcinoma tissue complicated with local inflammation.
Animals ; Carcinoma, Squamous Cell ; immunology ; Dendritic Cells ; HLA-DR Antigens ; metabolism ; Inflammation ; Rabbits
2.Immunohistochemical studies from vitiligo: comparison between active and inactive lesions.
Sung Ku AHN ; Eug Ho CHOI ; Seung Hun LEE ; Jin Hyung WON ; Seung Kyung HANN ; Yoon Kee PARK
Yonsei Medical Journal 1994;35(4):404-410
Vitiligo is an acquired, progressive depigmenting disorder of unknown etiology. In this study, to clarify pathogenesis of vitiligo, the marginal skin of actively spreading and stable vitiligo was examined using ICAM-1, HLA-DR, CD4 and CD8 monoclonal antibodies. In immunohistochemical study, ICAM-1 was expressed in four of five epidermis in active lesions, but not in stable lesion. Dermal ICAM-1 was also expressed in all active and stable lesions. HLA-DR was also expressed in all active epidermis in active lesions, but two of five epidermis in stable lesion. Dermal HLA-DR was also expressed in all active and stable lesion. CD4 lymphocytes were expressed more strongly in active lesion, but CD8 lymphocytes were not different in both lesions. There was no significant difference of degree of positivity with CD4 and CD8 in normal control specimens. In conclusion, we think that ICAM-1 and HLA-DR expression, cytokines released from keratinocytes, melanocytes or lymphocytes and infiltration of activated T-lymphocytes play an important role in disease activity.
Antigens, CD4/metabolism
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Antigens, CD8/metabolism
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Comparative Study
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HLA-DR Antigens/metabolism
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Human
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Immunohistochemistry
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Intercellular Adhesion Molecule-1/metabolism
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Skin/immunology
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Vitiligo/*immunology
3.Immunophenotypic Analysis of Acute Promyelocytic Leukemia.
Fang CHEN ; Yan-Ping HU ; Xiao-Hui WANG ; Shuang FU ; Yu FU ; Xuan LIU ; Min-Yu ZHANG ; Shao-Kun WANG ; Ji-Hong ZHANG
Journal of Experimental Hematology 2016;24(2):321-325
OBJECTIVETo investigate the immunophenotype of leukemia promyelocytes (LP) in bone marrow of patients with acute promyelocytic leukemia (APL) and to explore their characteristics and significance.
METHODSThe immunophenotypes of leukemia cells in 43 patients with APL were analyzed by means of 4 color immunophenotypes; the cell population in which CD45 strength localized at 10(2) and the SSC strength locatized at 10(2) was defined as R3, the cell population in which CD45 strength localized at 10(3) and the SSC strength localized at 10(2) was defined as R5, moreover the ratio of positive cells >80% was defined as strong positive expression, the ratio of positive cells between 20%-80% was difined as weak positive expression, the ratio of positive cells <20% was difined as negative by gating method of CD45/SSC.
RESULTSThere was a abnormal cell population (R3) in 79.07% cases; the immunophenotypes of R3 was cheracteried by high SSC, weaker expression of CD45, the rate of CD38, CD9 and CD13 all was 100%, moreover their bright expression (>80%) was 86.05%, 90.70% and 86.05%, respectively; the positive expression rate of CD33, CD117 and CD64 was 97.67%, 95.35% and 83.80% respectively, moreover thier bright expression was 84.04%, 69.77% and 30.23% respectively; the CD15 was weakly expressed in 39.53% cases, the CD34 and HLA-DR were weakly expression in 16.28% and 6.98% cases respectively. All the cases did not express CD116. There were 2 cell populations (R3 and R5) in 20.93% cases, the immunophenotypic features of R3 were cosistant with above mentioning, while the immunophenotypes of R5 were lower than those of R3 SSC; the fluorescence intensity of CD45 was higher, but lower than that in normal lymphycytes, the positive rate of CD9, CD13, MPO was 100%, moreover thier fluorescence intensity was high; they did not expressed CD123, CD25, CD22, CD4, CD64 and CD14. Thereby it can be concluded that the typical immunophenotypes is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-) in APL. There was a special immunophenotype in the APL with basophilic granules. Conclusoin: APL has a characteristic immunophenotypic profile, whose typical immunophenotype is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-). The special immunophenotype exists in the APL with basophilic granules. Flow cytometric immunophenotyping may be a useful for rapid recognition of APL and has significant for prognosis.
Antigens, CD ; metabolism ; Cell Count ; Flow Cytometry ; Granulocyte Precursor Cells ; classification ; HLA-DR Antigens ; metabolism ; Humans ; Immunophenotyping ; Leukemia, Promyelocytic, Acute ; classification ; immunology ; Leukocyte Common Antigens ; metabolism ; Prognosis
4.Statistical analysis of 4000 umbilical cord blood units preserved in shandong cord blood bank.
Jie PAN ; Shen-Li ZHOU ; Bai-Jun SHEN ; Wen-Ying YAN ; Ri XU ; Dao-Gang SONG
Journal of Experimental Hematology 2002;10(3):257-260
The current study analyzed the data of 4 000 umbilical cord blood (UCB) units collected in Shandong Cord Blood Bank from the end of 1999 to March 2001. The averages of nucleated cells and CD34(+) cells were more than 1.2 x 10(9) and 3.9 x 10(6) per UCB unit respectively, and more than 1.5 x 10(9) nucleated cells per UCB unit were obtained in 768 UCB units. These UCB units are suitable for transplantation in patients with a body weight greater than 40 kg. The analysis of HLA gene frequency showed that A2, A24, A11, B13, B51, DR15, DR7 and DR9 are the common halotypes in Shandong population and similar to those in the other areas of China. 40% patients could search out at least 1 UCB unit with 1 mismatched HLA locus in Shandong Cord Blood Bank.
Antigens, CD34
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immunology
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Blood Banks
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Blood Preservation
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Cell Count
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China
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Data Interpretation, Statistical
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Fetal Blood
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cytology
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immunology
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metabolism
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Gene Frequency
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HLA-A Antigens
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genetics
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HLA-B Antigens
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genetics
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HLA-DR Antigens
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genetics
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Humans
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Leukocyte Count
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Leukocytes
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cytology
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immunology
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Time Factors
5.Establishment of a method for HLA-DRB genotyping in cord blood by reverse dot-blot hybridization technique.
Yi-Ning HUANG ; Can LIAO ; Xue-Wei TANG ; Yan LI ; Xing-Mei XIE ; Rui-Ping ZENG
Journal of Experimental Hematology 2002;10(2):148-152
The HLA system was discovered by virtue of the fact that it was polymorphic. The impetus for its discovery was the search for polymorphic antigens to match for transplantation, by analogy with the human red cell blood groups. The most usually DNA method of HLA typing is sequence specific oligonucleotides (SSO) and PCR sequence specific primers (SSP). SSO technique is perfectly suited for analyzing large number of samples, it is not suitable for individual or small numbers. The SSP method is ideal for typing individual samples, but it is costly and requires high capacity thermal cycles for larger numbers of samples. To set up a simple, quick, cheap and high resolution DNA method, were collected sixty-three cord blood samples from Guangzhou Cord Blood Bank, got DNA from blood by the traditional guanidine hydrochloride distillation method. Each sample was simultaneously typed by SSOP, PCR-SSP and reverse dot-blot hybridization (RDB) methods. All of typed is success. The results of three DNA methods are consistent each other. 60 HLA-DRB1 alleles could be accurately distinguished with the RDB method. Our results show that RDB method is a simple, quick, cheap and high resolution method for HLA-DRB types. It can be used in any HLA typing.
Fetal Blood
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immunology
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metabolism
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Genotype
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HLA-DR Antigens
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genetics
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Histocompatibility Testing
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methods
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Humans
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Nucleic Acid Hybridization
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methods
6.Interleukin-1 gene polymorphism disease activity and bone mineral metabolism in rheumatoid arthritis.
Xiao ZHANG ; L LLAMADO ; I PILLAY ; P PRICE ; R WILL
Chinese Medical Journal 2002;115(1):46-49
OBJECTIVETo determine whether interleukin-1 alpha and 1 beta gene polymorphism is associated with rheumatoid arthritis disease activity and bone mineral metabolism, and whether there is any relationship between IL-1 beta and rheumatoid arthritis (RA) motif gene.
METHODSIL-1 gene polymorphisms were analyzed in 65 RA patients who met American College of Radiology (ACR) criteria and 60 controls. From genomic DNA, 2 polymorphisms in each gene for IL1 alpha-889 and IL-1 beta + 3953 were typed by PCR-RFLP and HLA-DRB1 allele typing was also undertaken by PCR-SSOP. Some clinical and laboratory parameters were collected. The allelic frequencies and carriage rates were compared between RA patients and controls and between patients with active and quiescent disease. Comparison was also made between IL-1 polymorphism and parameters of bone mineral metabolism and between patients with the HLA-DRB1 RA motif plus IL-1 beta 2 and patients without the two alleles. Fisher test and the analysis of variance was used to analyze the data.
RESULTSThere was no significant difference in the frequency and carriage rate of IL-1 alpha polymorphisms between RA patients and the controls. The beta 2/2 genotype of IL-1 beta was more common in female RA patients compared with controls (P = 0.001). A lower carriage rate of IL-1 beta 2 occurred in male RA patients (P = 0.001). A higher carriage rate of IL-1 alpha 2 is associated with a higher ESR (P = 0.008), HAQ score (P = 0.03), and vit-D3 (P < 0.001), but conversely a lower SJC (p = 0.002), a lower RF (P = 0.002) and a lower BMD at the lumbar spine (P = 0.001). A higher frequency of IL-1 alpha 1 is associated with a lower CRP value (P = 0.009). An increased IL-1 beta 2 carriage is associated with active rheumatoid disease as indicated by a higher CRP (P < 0.001), ESR (P < 0.001) and pain score (P = 0.001) and a higher BMD at the lumbar spine (P = 0.007), lower vit-D3 and. Udpd/Crea level The presence of the HLA DRB1 RA motif and IL-1 beta allele 2 at same time did not contribute to disease activity.
CONCLUSIONPolymorphisms of the IL-beta gene may affect the RA occurrence. Carriage of IL-1 beta 2 polymorphisms is associated with more active disease in RA and the presence of both the IL-1 alpha 2 and the IL-1 beta 1 allele in RA influences bone resorption.
Alleles ; Arthritis, Rheumatoid ; genetics ; metabolism ; Bone Density ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Interleukin-1 ; genetics ; Male ; Polymorphism, Genetic
7.Study on the relationship between the human leucocyte antigen-DR expression on CD14+ monocytes and sepsis.
Hong-ming YANG ; Yan YU ; Jia-ke CHAI ; Sen HU ; Shu-hong LEI ; Dong-qing ZHANG ; Bai-jiang TAO ; Guo-an ZHANG ; Li-xin HE ; Yong-ming YAO
Chinese Journal of Burns 2007;23(4):272-275
OBJECTIVETo investigate the changes in the expression of HLA-DR on CD14+ monocytes of burn patients with delayed resuscitation, and to analyze the relationship between it and sepsis.
METHODSTwenty-five patients with total burn surface area over 30% TBSA and delayed resuscitation were enrolled in the study, among which 7 were complicated by sepsis during hospitalization. Peripheral blood was collected on 1, 3, 7, 14 and 28 post-burn days (PBD), and the blood of the patients with sepsis were also collected on the 1 and 2 days after the occurrence of sepsis. Twenty healthy volunteers were enrolled as controls. Expression rate of HLA-DR on CD14+ monocytes was determined by flow cytometry. The level of TNF-alpha and IL-10 were measured by ELISA.
RESULTSExpression rate of HLA-DR antigen on CD14+ monocytes in burn patients without sepsis on 1, 3, 7, 14, 28 PBD were (15 +/- 6)%, (7 +/- 5)%, (26 +/- 17)%, (28 +/- 16)% and (47 +/- 16)%, respectively, which were obviously lower than that of healthy people [(92 +/- 10)%, P < 0.01], and it was also markedly lower on 1 and 2 days after the occurrence of sepsis than that of controls and those of patients without sepsis on 1, 7, 14, 28 PBD (P < 0.01). The positive rate and concentration of TNF-alpha in patients with sepsis were obviously higher than that of healthy people and patients without sepsis (P < 0.05 or P < 0.01). There was a negative correlation between the expression rate of HLA-DR on CD14+ monocytes and IL-10 levels, and it showed significant difference on 1, 7, and 28 PBD (r = -0.9963, -0.7459, -0.8474, respectively, P < 0.01).
CONCLUSIONImmune function is suppressed and proinflammatory mediators are excessively released in severely burn patients after delayed resuscitation, especially when complicated with sepsis. Expression of HLA-DR on CD14+ monocytes may be an useful parameter for monitoring the immune function of burn patients.
Burns ; immunology ; metabolism ; HLA-DR Antigens ; metabolism ; Humans ; Interleukin-10 ; blood ; Lipopolysaccharide Receptors ; metabolism ; Monocytes ; immunology ; metabolism ; Sepsis ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism
8.A Case of Natural Killer Cell Leukemia Misdiagnosed as Tuberculous Lymphadenopathy.
A jin LEE ; Sang Gyung KIM ; Chang Ho JEON ; Hun Suk SUH ; Ghil Suk YOON ; An Na SEO
The Korean Journal of Laboratory Medicine 2009;29(3):194-198
Natural killer (NK) cell neoplasms are a group of rare but highly malignant tumors. We report here one case of NK cell leukemia. A 54-yr-old woman presented with a 2-month history of progressive left neck mass. Based on the positive result of tissue PCR for Mycobacterium tuberculosis, she was at first diagnosed with tuberculous lymphadenopathy. After two weeks, she developed generalized lymphadenopathy, hepatosplenomegaly, fever and anemia. Subsequent evaluation was performed including bone marrow aspiration and biopsy. Peripheral blood smear showed leukoerythroblastic features with 31% blasts. Bone marrow was packed with agranular blastoid cells, which were periodic acid-Schiff (PAS) positive and myeloperoxidase (MPO) negative. Immunophenotyping showed that these cells were positive for CD45 and HLA-DR, whereas negative for CD3, CD5, CD7, CD10, CD13, CD14, CD19, CD20, CD22, CD33, CD34, and CD61. Because of the absence of the markers of T-cell, B-cell, and myeloid lineage-specific antigens, we added CD16/56 for the immunophenotyping and the blasts were positive (94%). The tumor cells of biopsied lymph node were only positive for CD56, consistent with NK cell lymphoma. Epstein-Barr virus (EBV) was not detected by RNA in situ hybridization. Culture for M. tuberculosis was negative. Thus this patient was diagnosed with blastic NK cell lymphoma/leukemia involving bone marrow and lymph node.
Antigens, CD45/metabolism
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Bone Marrow/pathology
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Female
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HLA-DR Antigens/metabolism
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Humans
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Killer Cells, Natural/immunology/*pathology
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Leukemia/*diagnosis/immunology/pathology
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Middle Aged
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Tuberculosis, Lymph Node/diagnosis
9.Expression and analysis of HLA-A, B and DRB1 genes in patients with chronic myelogenous leukemia in Guangdong area.
Li WEI ; Lu-Lu XIAO ; Xiang-Yuan WU ; Qu LIN ; Ming DONG ; Jing-Yun WEN ; Xiao-Kun MA ; Fei CHONG
Journal of Experimental Hematology 2008;16(4):915-918
To study the gene polymorphism of HLA-A, B, DRB1 alleles in patients with chronic myelogenous leukemia and to explore the correlation of HLA with chronic myelogenous leukemia, the polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-RSSO) was used to analyze the polymorphism of HLA-A, B, DRB1 alleles of 293 CML Patients and 406 randomized and synchronous blood donors (healthy and unrelated with patients) from Guangdong Han population. The results indicate that the gene frequency of HLA-A*24 in CML group was 15.53% lower than that of control group (22.09%, RR = 0.63, p = 0.005); the gene frequency of HLA-B*13 in CML group was 10.41% higher than that of control group (6.74%, RR = 1.68, p = 0.016). The gene frequency of HLA- DRB1*14 in CML group was 7.51% lower than that of control group (11.89%, RR = 0.58, p = 0.008). The differences were all statistically significant. It is concluded that the gene frequency of HLA-A*24, HLA- DRB1*14 in CML patients is significantly lower than normal people in Guangdong. The gene frequency of HLA-B*13 in CML patients is significantly higher than normal people in Guangdong. Further study is needed to make sure whether HLA-A*24 and HLA- DRB1*14 are protective gene markers for CML acquisition on Guangdong Chinese Han population and whether HLA-B*13 is a gene marker for CML susceptibility on this population.
Adolescent
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Adult
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Aged
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Blood Donors
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Child
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Child, Preschool
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China
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Female
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HLA-A Antigens
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genetics
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metabolism
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HLA-A24 Antigen
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HLA-B Antigens
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genetics
;
metabolism
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HLA-B13 Antigen
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HLA-DR Antigens
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genetics
;
metabolism
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HLA-DRB1 Chains
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Humans
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
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genetics
;
immunology
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Male
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Middle Aged
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Young Adult
10.Human leucocyte antigen-DR expression on CD(14)(+) monocytes and its relationships with multiple organ dysfunction syndrome in severe sepsis.
Yi-Nan LI ; Li-Xin ZHOU ; Bing FANG ; Ke-Jiang MAO ; Wei-Biao WEN ; Tie-Ou YU ; Yi-Cheng ZOU ; Wen-Yan LI ; Chang LI
Journal of Southern Medical University 2009;29(7):1372-1374
OBJECTIVETo explore the changes of CD(14)(+) monocyte human leucocyte antigen DR (HLA-DR) and their relationship with multiple organ dysfunction syndrome (MODS) in severe sepsis.
METHODSNinety-one patients with a definite diagnosis of severe sepsis in the intensive care unit (ICU) were included. CD(14)(+) monocyte HLA-DR levels were detected by flow cytometry on the first, 4th and 7th days of the study, and Marshall scores and prognosis on day 28 were evaluated.
RESULTSThirty-four patients died within 28 days following the onset with a mortality rate of 37.4%. Persistently lowered levels of HLA-DR were detected and significantly increased Marshall scores were found in the fatal cases at all the time points (P<0.001). In the surviving patients, the levels of HLA-DR were significantly increased (P<0.01) and Marshall scores were gradually decreased (P<0.001). During the observation period, the levels of HLA-DR decreased significantly as the number of dysfunctional organs and Marshall scores increased (P<0.001). The levels of HLA-DR were significantly increased in severe sepsis patients with 2-4 dysfunctional organs and Marshall score of 5-12 (P<0.05 or P<0.001). No changes in HLA-DR levels in severe sepsis patients with 5-6 dysfunctional organs and Marshall scores of 13-22. The levels of HLA-DR showed a significant inverse correlation to Marshall scores (r=-0.368, P<0.001).
CONCLUSIONIn patients with severe sepsis, persistent low CD(14)(+) monocyte HLA-DR levels predicts high mortality. The levels of HLA-DR are significantly correlated to the severity of organ dysfunction.
Adult ; Aged ; Aged, 80 and over ; Female ; HLA-DR Antigens ; metabolism ; Humans ; Lipopolysaccharide Receptors ; Male ; Middle Aged ; Monocytes ; immunology ; metabolism ; Multiple Organ Failure ; pathology ; Sepsis ; immunology ; metabolism