Ultraestructura del compartimiento clase II en macrófagos humanos en rosetas macrófago-linfocitarias autólogas / Ultrastructure of the class II compartment in human macrophages in autologous macrophage-lymphocyte rosettes

Introducción: El procesamiento y presentación de antígenos está involucrado en el fenómeno de múltiples sinapsis inmunológicas de la Roseta Macrófago-Linfocitaria humana (RML) entre macrófagos derivados de monocitos y linfocitos T CD4+ de cultivos autólogos leucocitarios totales extraídos de la sangre; aquí los antígenos autólogos de los neutrófilos apoptóticos son presentados por la vía endocítica o vía Clase II. El Compartimiento Clase II (CCMII) ha sido caracterizado en células B y células dendríticas en modelos murinos. Objetivo: estudiar la evolución ultraestructural de la organización espacial CCMII en macrófagos en el fenómeno de RML humana. Métodos: Se utilizaron muestras de sangre humana sana, anticoagulada con heparina (n=10) donadas por Banco de Sangre, UNC, en anonimato. Cultivos leucocitarios autólogos en medio TC199 (SIGMA, St. Louis, MO). Se tomaron muestras de cultivo celular a: 1, 2, 3, 20, 48 y 96 horas. Se aplicó la técnica de RML. Las citopreparaciones se sometieron a técnicas de procesamiento para su estudio ultraestructural con el MET: Zeiss LEO-906E. Resultados: Observamos cuerpos multivesiculares, multilaminares y tubulares en la organización espacial del CCMII a lo largo del tiempo de cultivo. Estructuras tubulares aparecieron a las 48 horas de cultivo. Se concluye que organización espacial del CCMII toma diversos aspectos en coincidencia con la ocurrencia de transformación macrofágica en cultivo y su rol como célula presentadora de antígenos (CPA) en RMLs. Dado el origen autólogo de los antígenos presentados postulamos que el perfil de los macrófagos en RMLs podría corresponder al alternativo o M2

Introduction: Processing and presentation of antigens is involved in the phenomenon of multiple immunological synapses of human Macrophage-Lymphocyte Rosette (MLR) between monocyte-derived macrophages and CD4 + T cells of total leukocyte cultures autologous extracted from blood; here autologous apoptotic neutrophil antigens are presented by the endocytic pathway or via Class II. Class II Compartment (MIIC) has been characterized by B cells and dendritic cells in murine models. Objective: To study the ultrastructural evolution in the spatial organization MIIC in macrophages in the phenomenon of human MLR. Methods: healthy human blood samples were used, anticoagulated with heparin (n = 10) donated by Blood Bank, UNC, anonymous. Autologous leukocyte cultures in TC199 medium (SIGMA, St. Louis, MO). Cell culture samples were taken at 1, 2, 3, 20, 48 and 96 h. MLR technique was applied. The citopreparations underwent processing techniques for ultrastructural study with MET: Zeiss LEO-906E. Results: We observed multivesicular, multilamellar, and tubular bodies in the spatial organization of MIIC throughout the culture time. Tubular structures appeared at 48 h of culture. We conclude that spatial organization of MIIC takes various aspects coinciding with the occurrence of macrophage transformation in culture and its role as antigen presenting cell (APC) in MLRs. Given the autologous antigens presented we postulate that the profile of macrophages in MLRs could correspond to alternative or M2 (AU) .

Association of CD58 polymorphism with multiple sclerosis and response to interferon beta therapy in a subset of Iranian population

Multiple sclerosis [MS] is one of the leading neurodegenerative causes of physical disability world-wide. Genetic aberrations of autoimmunity pathway components have been demonstrated to significantly influence MS development. Cluster of Differentiation 58 [CD58] is pertained to a group of genes which had been assayed in several recent association studies. Given the significance of CD58 in modulation of T regulatory cells that control autoimmune responses, the present study was conducted to investigate the frequency of rs12044852 polymorphism and its effect on the outcome of interferon beta [IFN- beta] therapy in a subset of Iranian MS patients. Two hundred MS patients and equal number of healthy controls were recruited to be genotyped in an experimental case-control based study through polymerase chain reaction using specific sequence primers [PCR-SSP]. Relapsing remitting multiple sclerosis [RRMS] patients administered IFN- beta therapy were followed up with clinical visits every three months up to two years. The mean of multiple sclerosis severity score [MSSS] and expanded disability status scale [EDSS] were measured to monitor the change in severity of MS in response to IFN- beta therapy. Pearson's Chi-square and analysis of variance [ANOVA] tests were the main statistical methods used in this study. Strong association was found between the CC genotype and onset of MS [p=0.001, OR=2.22]. However, there was no association between rs12044852 and various classifications and severity of MS. Pharmacogenetics-based analysis indicated that carriers of CC genotype had the highest MSSS score compared to others, implying a negative impact of rs12044852 on response to IFN- beta t herapy. Taken together, our findings revealed the critical effect of rs12044852 polymorphism of CD58 on the progression of MS disease. This indicates that genotyping of MS patients may expedite achieving personalized medical management of MS patients

Recombinant expression and characterization of CD2-binding domain of Macaca mulatta lymphocyte function-associated antigen 3 in Pichia pastoris / 生物医学工程学杂志

Human lymphocyte function-associated antigen 3 (hLFA3) has been identified as an important T cell accessory molecule. Rhesus monkeys (Macaca mulatta) have been widely used as animal models for human immune disorders. Due to the species-specificity of immune system, it is necessary to study M. mulatta LFA3 (mmLFA3). In this study, the gene encoding mmLFA3 CD2-binding domain (mmLFA3Sh) was amplified by polymerase chain reaction (PCR) and genetically fused to human IgG1 Fc fragment in pPIC9K to construct the expression plasmid pPIC9K-mmLFA3Sh-Ig. Approximately 3-4 mg mmLFA3Sh-Ig protein was recovered from 1 L of inductive media, and mmLFA3Sh-Ig produced by the P. pastoris can bind to the CD2 positive cells, and suppress the monkey and human lymphocytes proliferation induced by Con A and alloantigen in a dose-dependent manner. These results suggested that mmLFA3Sh-Ig might be used as a novel tool for pathogenesis and experimental immunotherapy of Rhesus monkey immune disorders.

Expression of CD58 in childhood B-lineage acute lymphoblastic leukemia and its feasibility in minimal residual disease detection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To measure the expression of lymphocyte function-associated antigen-3 (CD58) in childhood B-lineage acute lymphoblastic leukemia (B-ALL) and to explore the feasibility of CD58 as an indicator for minimal residual disease (MRD) detection in childhood B-ALL.</p><p><b>METHODS</b>Eighty-seven children diagnosed with B-ALL between January 2014 and September 2014 were enrolled, and 20 hospitalized children who had no tumor or blood disease and had normal bone marrow cell morphology served as the control group. The expression features of CD58 in bone marrow samples from the two groups (at diagnosis, on day 15 of induction chemotherapy) were analyzed by four-color flow cytometry (FCM). Quantitative real-time polymerase chain reaction (qRT-PCR) and FCM were used to detect MRD in B-ALL patients on day 33 of induction chemotherapy.</p><p><b>RESULTS</b>The mean fluorescence intensity of CD58 expression in the 87 B-ALL cases (91±33) was significantly higher than that in the 20 controls (14±6) (P<0.01); CD58 was over-expressed in 44 of the B-ALL cases. In the B-ALL children, the expression of CD58 on day 15 of induction chemotherapy (105±22) was not significantly different from that at diagnosis (107±26) (P>0.05). In the 44 B-ALL patients with CD58 over-expression, FCM showed 9 MRD(+) cases and 35 MRD(-) cases, while qRT-PCR showed 11 MRD(+) cases and 33 MRD(-) cases; 42 cases (95%) showed consistent results of the two tests, so there was no significant difference between the two methods in detecting MRD (P>0.05).</p><p><b>CONCLUSIONS</b>CD58 is over-expressed and stable in children with B-ALL, and it can be considered as an indicator for MRD detection in childhood B-ALL.</p>

Effects of herbal cake-separated moxibustion on the expression of erythrocyte CD58 in different ages of healthy people / 中国针灸

<p><b>OBJECTIVE</b>To observe the efficacy of herbal cake-separated moxibustion on the expression of erythrocyte CD58 in different ages of healthy people and explore the differences of the therapeutic effect in different ages and its mechanism.</p><p><b>METHODS</b>A total of 82 health participants were divided into a young age group and a middle-old age group according to the ages. They were treated with herbal cake-separated moxibustion on Shenque (CV 8), Guanyuan (CV 4), Zusanli (ST 36), Pishu (BL 20), Shenshu (BL 23) with cake made by Shudihuang (Radiz Re hmanniae Preparata), Shanyao (Rhizoma Dioscoreae), Shanzhuyu (Fructus Corni ), etc. The treatment was given for 10 sessions once other day and each acupoint for 3 successive dosages. The mean fluorescence intensities of erythrocyte CD58 were measured by flow cytometry before and after moxibustion.</p><p><b>RESULTS</b>After moxibustion, erythrocyte CD58 expression were significantly higher than that before moxibustion in two groups (both P < 0.01), particularly in young age group, which was significantly higher than that in middle-old age group (P < 0.01).</p><p><b>CONCLUSION</b>The effect of moxibustion in youth is evidently superior to that in middle-old age. Its mechanism is connected with that moxibustion can enhance the expression of erythrocyte CD58.</p>

CD58 expression of liver tissue in patients with chronic hepatitis B virus infection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Several kinds of intercellular adhesion molecules closely relate to hepatitis B. The complex of CD(2) and CD58 plays an important role in enhancing the adhesion of T lymphocytes to target cells, hyperplasia and activation of T lymphocytes. In this study, we explored the relationship between the expression of CD58 in liver tissue and chronic hepatitis B infection.</p><p><b>METHODS</b>We determined the expression of the CD58 molecule on the surface of hepatocytes by using immunohistochemistry and the levels of serum HBV DNA from patients with HBV infection and from normal controls. The biochemical parameters of hepatic function were analyzed as well.</p><p><b>RESULTS</b>CD58 expression in hepatocytes significantly increased with the severity progression of chronic HBV infection. The IOD levels (log10) of CD58 in the control, mild, moderate, and severe chronic HBV infection groups were 0, (7.20+/-4.64) x 10(3), (25.63+/-7.41) x 10(3) and (37.47+/-11.17) x 10(3) respectively (P<0.05 compared with the control group, respectively).</p><p><b>CONCLUSION</b>CD58 probably increases cell mediated immunity to eliminate hepatitis B virus and leads to damage of hepatocytes.</p>

Quality control methods and requirements for recombinant human lymphocyte function associated antigen 3 IgG1 fusion protein (rhLFA3-IgG1) / 药学学报

To establish methods and requirements for quality control of rhLFA3-IgG1, biological potency of rhLFA3-IgG1 was determined by CD2 molecule competitive binding assay on Jurkat cell surface. Purity of rhLFA3-IgG1 was analyzed by SEC-HPLC and IEC-HPLC. Peptide mapping was preformed by tryptic digestion and RP-HPLC after sample reduced and carboxymethylation by DTT and indoacetic acid, respectively. CHO host cell protein and Protein A residual were detected by ELISA separately. The quality control methods and requirements, such as biological potency, the physical-chemical characteristic of rhLFA3-IgG1 had been established. The methods and requirements for quality control of rhLFA3-IgG1 showed advantages of assuring the products safety and efficacy, which can be used for routine quality control of rhLFA3-IgG1.

Prognostic significance of lymphocyte function associated anti-gen-3 (CD58) in childhood B cell-acute lymphocytic leukemia / 中国实验血液学杂志

This study was aimed to investigate the value of CD58 in evaluation of early therapeutic effect on childhood B-ALL. The expression features of CD58 in 135 cases of childhood B-ALL were analyzed by four-color flow cytometry; MRD detection protocol for B-ALL using CD58/CD10/CD34/CD19 combination was established; the correlation between the expression features of CD58 and MRD detection was analyzed for the early therapeutic response in childhood B-ALL. The results showed that the mean value of CD58 MFI in 135 cases of B-ALL was 113.08 +/- 63.33, which was significantly higher than that in 15 cases of normal bone marrow controls (14.68 +/- 5.26, P < 0.01). In addition, CD58 was over expressed in 51.9% (70/135) of B-ALL patients, indicating that CD58 could be an effective marker in MRD detection. The CD58/CD10/CD34/CD19 was the second most effective combination next to TdT/CD10/CD34/CD19 in B-ALL MRD detection with flow cytometry. Meanwhile, the positive rate of MRD detection by flow cytometry was significantly lower in CD58 over expression group (P < 0.05). It is concluded that CD58 may be used as an indicator for detection of MRD in B-ALL patients, which would enrich the combination of MRD detection. The CD58 over expression may be considered as a marker of a favorable prognosis in childhood B-ALL.

Study on the relationship between level of CD58 expression in peripheral blood mononuclear cell and severity of HBV infection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>As one of the intercellular adhesion molecules, CD58 plays important roles in promotion of the adhesion between T cells and target cells, hyperplasia, activation of T cells and natural killer cells, and balance between Th1 and Th2. We studied the relationship between the levels of CD58 expression in peripheral blood mononuclear cells (PBMCs) and severity of HBV infection.</p><p><b>METHODS</b>The levels of CD58 mRNA in PBMCs were detected using quantitative reverse transcription PCR. The percentage of CD58 positive cells was detected by flow cytometry in patients and healthy controls.</p><p><b>RESULTS</b>The levels of CD58 mRNA and the percentage of CD58 positive cells in patients infected with HBV were significantly higher than that in the control. Based on severity of HBV infection, the patients were classified into four groups. The expression of CD58 increased significantly in an order from mild chronic, moderate chronic, severe chronic to severe hepatitis groups. The levels of CD58 mRNA and the percentage of CD58 positive cells in PBMCs from patients with HBV infection were both positively correlated with serum levels of ALT and AST.</p><p><b>CONCLUSION</b>The level of CD58 expression is related with the severity of HBV infection and the degree of liver damage.</p>

Expression of Antigen Presenting Function-Associated Surface Molecules on Interferon gamma-Treated Gingival Fibroblasts and Periodontal Ligament Fibroblasts / 대한치주과학회지

It is becoming increasingly clear that human gingival fibroblasts(HGF) may play a role in regulating immune responsiveness in inflammatory periodontal lesions. Stimulation of HGF with locally-secreted T cell cytokine IFNgamma induces human leukocyte antigen class II(HLA II) expression on HGF, which is one of the characteristic feature of professional antigen presenting cells(pAPC). However, IFNgamma-treated HGF and other nonprofessional antigen presenting cells(npAPC) are known to be ineffective or less effective antigen presenter to resting T cells. This study, therefore, was undertaken in an effort to elucidate the differences in expression of cell surface molecules between npAPC in periodontal tissues, such as HGF and periodontal ligament fibroblasts(PDLF), and pAPC such as monocytes/macrophages. Using flow cytometry, the levels of cell surface expression of HLA-D, ICAM-1, LFA-3, and B7-1, which are involved in antigen presentation, were determined in HGF, PDLF and human myelomonocytic cell line THP-1. IFNgamma clearly induced HLA-D expression on both of fibroblasts and monocytes dose dependently. However, expression level on monocytes were 4 to 5 times higher than that on fibroblasts, and induction rate was faster in monocytes than in fibroblasts. The levels of ICAM-1 expression on fibroblasts and monocytes were enhanced by IFNgamma in a dose dependent manner. On the other hand, the expression of LFA-3 molecule, which could be detected in fibroblasts and monocytes without cytokine stimulation, was no more enhanced by addition of IFNgamma. B7-1, important costimulatory molecule in T cell activation and proliferation, was not detected on both of fibroblasts and monocytes even when stimulated with IFNgamma, except on monocytes fully differentiated by pretreatment of PMA and treated by IFNgamma. These results suggest that delayed expression of HLA-D and absence of B7-1 on IFNgamma -treated fibroblasts may at least in part be involved in the ineffectiveness of fibroblasts as primary APC. And it is postulated that although periodontal fibroblasts may not serve as primary APC in normal periodontium, sustained expression of HLA II on ubiquitous fibroblasts in inflammatory lesions may perpetuate immune responses and produce chronic inflammation and tissue injury.

Expression of LFA-3(CD58) After Intracorneal Infection of Candida albicans in Rabbits

In this immunohistochemical study we applied a monoclonal antibody(mAb) to evaluate the expression pattern of lymphocyte functionassociated antigen 3(LFA-3) in rabbit`s corneas before and after intracorneal injection of Candida albicans. Ten right eyes were induced to get immunocompromized cornea with subconjunctival injection of 2mg of dexamethasone once a day for 3 days(group I), and 10 left eyes had normal cornea without subconjunctival injection of dexamethazone(group II). Each 2 corneas in both group I and II were resected at 3, 12, 24 and 72 hours after intracorneal injection of C. albicans. Each 2 corneas without intracorneal injection of C. albicans in both groups were used as a control. The results were as follows: LFA-3 was expressed weakly on corneal epithlium in control of group I and group II. Expression of LFA-3 on vascular endothelium of group II was somewhat stronger than that of group I, LFA-3 was expressed moderately on vascular endothelium, and was detected on corneal stroma at 3 hors after intracorneal injection in both groups. Expression of LFA-3 on corneal stroma was slightly increased in both group II, and markedly increased in group I at 12 hours after intracorneal injection. Group II showed slightly increased LFA-3 expression on corneal and II to be expressed on corneal endothelium and inflammatory cells at 24 hours after injection. Its expression on corneal epithelium, stroma and endothelium was more increased in group II than in group I at that time. Group I showed moderate LFA-3 expression on corneal epithelium, corneal endothelium and inflammatory cells, and strong expression on corneal stroma and vascular endothelium at 72 hours after infection. Otherwise, LFA-3 expression in group II was weak to moderate n corneal epithelium, corneal endothelium and inflammatory cell, and moderate on corneal stroma and vascular endothelium. In this study, it was found that expression of LFA-3 in group I was weaker than that in group II in control and at 3 hours after intracorneal injection of C. albicans, but group I showed more strong LFA-3 expression than group II after 12 hours of intracorneal injection.