Animals ; CD4 Antigens ; blood ; CD4-Positive T-Lymphocytes ; metabolism ; radiation effects ; CD8 Antigens ; blood ; CD8-Positive T-Lymphocytes ; metabolism ; radiation effects ; Male ; Mice ; Ultraviolet Rays

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 ; Antigens, CD8/metabolism ; Comparative Study ; HLA-DR Antigens/metabolism ; Human ; Immunohistochemistry ; Intercellular Adhesion Molecule-1/metabolism ; Skin/immunology ; Vitiligo/*immunology

Objective To identify the potential long non-coding RNA (lncRNA) expressed in rheumatoid arthritis (RA) synovium key to RA onset and investigate its association with immune cell infiltration. Methods RA synovium data were downloaded from the GEO database and normalized. The lncRNAs key to RA onset were identified using multiple machine learning methods. Infiltration of 22 immune cell populations in RA synovium was measured by cell-type identification by estimating relative subsets of RNA transcripts (CIBER-SORT). The relationship between the key lncRNA and infiltrating immune cells was analyzed. Finally, real-time quantitative PCR was applied to validate the expression of the key lncRNA in RA synovial cells. Results lncRNA human leukocyte antigen complex P5(HCP5) was identified as the key lncRNA associated with RA onset. Infiltration analysis revealed increased abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages while decreased abundance of M2 macrophages in RA synovial tissue. Correlation analysis demonstrated that the lncRNA HCP5 expression was positively associated with the infiltration abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages in RA synovial tissue. Furthermore,the expression of lncRNA HCP5 in RA synovial cells was up-regulated. Conclusion lncRNA HCP5 expression is up-regulated in RA synovial tissue and potentially associated with immune cells infiltration.
Humans ; Arthritis, Rheumatoid ; CD8-Positive T-Lymphocytes ; HLA Antigens/metabolism* ; RNA, Long Noncoding/metabolism* ; Synovial Membrane/metabolism*

OBJECTIVETo determine the changes in levels of D-dimer, prothrombin time (PT), fibrinogen (Fib), CD4 and CD8 in relation to hepatopulmonary syndrome (HPS) by using a rat model system and to assess the association with pathologic changes in lung.

METHODSForty male Sprague-Dawley rats were divided into equal groups for modeling of cirrhosis and HPS. The two groups were assessed by blood gas analysis, standard biochemical tests to measure D-dimer, PT, Fib, CD4 and CD8, and pathological examination of lung tissues.

RESULTSThe HPS rats showed significantly lower PaO2 than the cirrhosis rats (58.20+/-3.19 mmHg vs. 85.00+/-2.53 mmHg, P = 0.000). The HPS rats showed significantly higher levels of D-dimer, Fib and CD8 than the cirrhosis rats (0.39+/-0.09 mg/ml vs. 0.25+/-0.05 mg/ml, P = 0.000; 1.77+/-0.10 g/L vs. and 1.49+/-0.09 g/L, P = 0.010; 32.32+/-4.45/mm3 vs. 20.13+/-6.09/mm3, P = 0.014). The HPS rats showed significantly lower levels of PT, CD4 and CD4/CD8 than the cirrhosis rats (14.86+/-1.04 s vs. 16.23+/-0.75 s, P = 0.036; 20.45+/-3.86/mm3 vs. 26.75+/-5.32/mm3, P = 0.000; 0.64+/-0.09 vs. 1.32+/-0.13, P = 0.000). The lung tissues of the HPS rats showed microthrombosis in pulmonary vessels, which were not observed in lung tissues of the cirrhosis rats.

CONCLUSIONHPS-related differential levels of D-dimer, PT, Fib, CD4, CD8 and CD4/CD8 may represent a biomarker profile suggestive of incidence of thromboembolism in lung.

Animals ; CD4 Antigens ; metabolism ; CD4-CD8 Ratio ; CD8 Antigens ; metabolism ; Disease Models, Animal ; Fibrin Fibrinogen Degradation Products ; metabolism ; Fibrinogen ; metabolism ; Hepatopulmonary Syndrome ; blood ; Liver Cirrhosis ; blood ; Lung ; pathology ; Male ; Prothrombin Time ; Rats ; Rats, Sprague-Dawley

BACKGROUND: We developed a single-color multitarget flow cytometry (SM-FC) assay, a single-tube assay with graded mean fluorescence intensities (MFIs). We evaluated the repeatability of SM-FC, and its correlation with multicolor flow cytometry (MFC), to assess its application as a routine FC assay. METHODS: We selected CD19, CD3, CD4, and CD8 as antigen targets to analyze a lymphocyte subset. MFIs were graded by adjusting monoclonal antibody (mAb) volumes to detect several cell populations. Dimly labeled mAb was prepared by decreasing mAb volume and the optimum diluted volume was determined by serial dilution. SM-FC repeatability was analyzed 10 times in 2 normal controls. The correlation between SM-FC and MFC was evaluated in 20 normal and 23 patient samples. RESULTS: CV values (0.8-5.0% and 1.3-4.1% in samples 1 and 2, respectively) acquired by SM-FC with CD3-fluorescein alpha-isothyocyanate (FITC)dim+CD4-FITCbright and with CD19-FITCdim+CD3-FITCbright showed good repeatability, comparable to that acquired by MFC (1.6-3.7% and 1.0-4.8% in samples 1 and 2, respectively). Excellent correlation was observed between the 2 methods in the 20 normal samples (B cells, T cells, non-Thelper cells, and Thelper cells; r2=0.87, 0.97, 0.97, and 0.98, respectively; P<0.05). There were also linear relationships between SM-FC with CD19-FITCdim+CD3-FITCbright and CD8-PEdim+CD4-PEbright, and MFC, in the 23 patient samples (B cells, T cells, Tcytotoxic cells, and Thelper cells; r2> or =0.98, 0.99, 0.99, and 0.99, respectively; P<0.05). CONCLUSIONS: The multicolor, single-tube SM-FC technique is a potential alternative tool for identifying a lymphocyte subset.
Antibodies, Monoclonal/chemistry/*immunology ; Antigens, CD19/chemistry/metabolism ; Antigens, CD3/chemistry/metabolism ; Antigens, CD4/chemistry/metabolism ; Antigens, CD8/chemistry/metabolism ; B-Lymphocyte Subsets/immunology/metabolism ; Color ; Flow Cytometry/*methods ; Fluorescein-5-isothiocyanate/*chemistry ; Humans ; T-Lymphocyte Subsets/immunology/metabolism

OBJECTIVETo characterize the phenotypic and biological properties of CD56(+) natural killer cells from human peripheral blood mononuclear cells (PBMCs).

METHODSSurface markers and intracellular cytotoxic molecules were stained with multi-color-labeled monoclonal antibodies and analyzed at the single cell level the relation between NK subsets and biological characteristics by flow cytometry.

RESULTSNK cells in PBMCs could be divided into two major populations, CD56(bright) and CD56(dim), based upon the expression of CD56 molecules. Both CD56(bright) and CD56(dim) expressed CD95 (Fas) with CD95(bright) and CD95(dim) subsets. CD56(dim) subsets had higher percentage of CD8, granzyme B and perforin expression compared to those of CD56(bright) subsets. In CD56(bright) and CD56(dim) subpopulations, CD95(bright) and CD8(+) subsets had higher percentage of granzyme B and perforin expression.

CONCLUSIONCD56(+) NK cells in PBMCs are composed of distinct subpopulations, CD56(dim) and CD56(dim) CD8(+) NK subsets have higher percentage of granzyme B and perforin and may play an important role in the killing of target cells.

CD56 Antigen ; metabolism ; CD8 Antigens ; metabolism ; Granzymes ; metabolism ; Humans ; Killer Cells, Natural ; classification ; immunology ; metabolism ; Lymphocyte Subsets ; immunology ; Perforin ; metabolism ; Phenotype ; fas Receptor ; metabolism

OBJECTIVETo examine the expressions of HLA class I antigen and CD8 in various cervical diseases and investigate their association with cervical cancer.

METHODSThe expressions of HLA class I antigen and CD8 in cervical tissues sampled from patients with cervical cancer, cervical intraepithelial neoplasia (CIN), and chronic cervicitis were detected using SP immunohistochemistry. The association of the expressions of HLA class I antigen and CD8 with the clinicopathologic indices of the patients was analyzed.

RESULTSThe positive expression rates of HLA class I antigen in cervical cancer, CIN, and chronic cervicitis were 22.6%, 100.0%, and 100.0%, and the positive expression rates of CD8 were 22.6%, 95.5%, and 100.0%, respectively. The positive rates of HLA class I antigen and CD8 were significantly lower in patients with cervical cancer (P<0.01). Patients with stage I cervical cancer had significantly higher positive rates of HLA class I antigen and CD8 than those with stage II cervical cancer (46.7% vs 0.0%, 46.7% vs 0.0%, both P<0.01). The expressions of HLA class I antigen and CD8 decreased with the progression of the clinicopathological stages, and may even become undetectable. The expressions of HLA class I antigen and CD8 were not related to the differentiation degree of the tumor or lymph node metastasis (P>0.05). A positive correlation was found between HLA class I antigen expression and CD8 expression.

CONCLUSIONThe expressions of HLA class I antigen and CD8 are down-regulated or deleted in CIN and cervical cancer, and they may play important roles in the development and progression of CIN and cervical cancer.

Adult ; Aged ; CD8 Antigens ; immunology ; metabolism ; Cervical Intraepithelial Neoplasia ; immunology ; pathology ; Down-Regulation ; Female ; Histocompatibility Antigens Class I ; immunology ; metabolism ; Humans ; Middle Aged ; Uterine Cervical Neoplasms ; immunology ; pathology

Wolynes argued that the track of a protein's folding was directed by the tendency of lowering its energy, and thus when a local minimum of its energy was reached, a relatively stable conformation was formed. However not all of the local minimums will lead the protein to a biologically useful conformation, for those otherwise are called energy traps. Wolynes energy landscape theory and natural selection have well explained the high efficiency of protein folding in vivo, instead of being stuck in energy traps. As to whether a protein can assume different conformations with the same bioactivity, there is no clear answer yet. In this paper, two conformational states of a pHLA-A*2402 are discovered after refolding, and by studying their interactions with TCR and CD8alphaalpha, two conformations of pHLA-A*2402 are confirmed of having escaped from natural selection.
CD8 Antigens ; chemistry ; Energy Metabolism ; HLA-A24 Antigen ; chemistry ; Humans ; Protein Conformation ; Protein Folding ; Receptors, Antigen, T-Cell ; chemistry

This article reviews that as a functionally and phenotypically distict immunoregulatory T cell subpopulation, CD4(+)CD25(+) regulatory T cells can suppress the activation and proliferation of CD4(+)CD25(-) T cells and CD8(+) T cells and the production of IL-2 and IFN-gamma. These regulatory cells play an important role in allograft tolerance, although the mechanisms are not completely understood to date. CD4(+)CD25(+) regulatory T cells can be isolated, activated and expanded in vitro without loss of their immunoregulatory function. The suppressive function of activated CD4(+)CD25(+) cells is antigen non-specific. Ex vivo activated and expanded regulatory T cells have a perspective for practical use.
CD4 Antigens ; blood ; CD8 Antigens ; blood ; Cell Division ; immunology ; Humans ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Receptors, Interleukin-2 ; blood ; T-Lymphocytes ; cytology ; immunology ; metabolism ; Transplantation Tolerance ; immunology

To obtain large quantity of human leukocyte antigen F (HLA-F) and cluster of differentiation 8alpha homodimers (CD8alphaalpha) proteins and to study their relationship, HLA-F and CD8alpha genes with rare codon in Escherichia coli were cloned using an N-terminal synonymous mutation method. High-efficiency expression protein inclusion bodies were acquired. The proteins were refolded using the dilution method and purified with gel-filtration and anion exchange chromatography. The results of gel-filtration and native-PAGE indicate that HLA-F interacts with CD8alphaalpha. This interaction may affect the binding between CD8alphaalpha and other MHC molecules to regulate immune responses. These results provide a basis for further research of HLA-F.
CD8 Antigens ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Histocompatibility Antigens Class I ; biosynthesis ; genetics ; Humans ; Mutation ; Protein Interaction Domains and Motifs ; Protein Multimerization ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification